Screening of Linseed (Linum Usitatissimum) Accessions For Resistance To Fusarium Oxysporum F. Sp. Lini

Results of
Plant Protection
Research
2018
Editors
Birhanu Bekele
Takele Negewo
Shashitu Bedada
Getachew Ayana
Gashawbeza Ayalew
Mohammed Yesuf
የኢትዮጵያ የግብርና ምርምር ኢንስቲትዩት

Ethiopian Institute of Agricultural Research
Results of
Plant Protection
Research
2018
Proceedings of the Completed Plant

Protection Research Activities
Addis Ababa, Ethiopia
Copy Editing and Design: Elizabeth Baslyos

©EIAR, 2020
Website: http://www.eiar.gov.et
Tel: +251-11-6462633
+251-11-6-676926
P.O.Box: 2003
Addis Ababa, Ethiopia
ISBN: 978-99944-66-56-6
Citation:
Birhanu Bekele et al., (ed.). 2020. Results of Plant Protection Research. Proceedings of the Completed
Plant Protection Research Activities, 2018. Addis Abeba, Ethiopia.
Contents
Foreword i
PART I: Plant Pathology

Survey of Wheat Rust Diseases in Ethiopia
Netsanet Bacha, Tsegaab Tesfaye, Kitesa Gutu, Getaneh Woldeab, Tamrat Negash, Daniel Kassa, Fikirte Yirga,
Megersa Debela, Gudeta Bedada, Tesfaye Dessale, Ashenafi Gemechu, Tilahun Bayisa, Zerihun Eshetu, Zerihun
Tomas, Girma Teshome, Mitiku Kebede, Bilal Temam, Misgana Mitiku, Bekele Kassa, Yitagesu Tadesse, Mequnanint
Andualem, Tsehaye Brhane, and Wulita Wondwosen 3
Physiological Races of Puccinia graminis f. sp. tritici in Ethiopia in 2018 Cropping Season
Netsanet Bacha, Tsegaab Tesfaye, Kitesa Gutu, Getaneh Woldeab, Tamrat Negash, Daniel Kassa, Fikirte Yirga, Megersa
Debela, Gudeta Bedada, Tesfaye Dessale, Ashenafi Gemechu, Tilahun Bayisa, Zerihun Eshetu, Zerihun Tomas, Girma
Teshome, Mitiku Kebede, Bilal Temama, Misgana Mitiku, Asfaw Azanaw, Bekele Kassa, Yitagesu Tadesse, Mequnanint
Andualem, Tsehaye Brhane, Wulita Wondwosen, and Kalkidan Yallew
15
Survey and Identification of Major Diseases of Black Cumin (Nigella sativa L.) in Ethiopia
Merga Jibat, Wakjira Getachew, Habetewold Kifelew, and Abukiya Getu 25
Major Diseases of Coriander (Coriandrum sativum L.) in Different Growing Areas of Ethiopia
Merga Jibat, Wakjira Getachew and Abukiya Getu 31
Evaluation of Trichoderma Species as Biocontrol Agents against Fusarium Wilt of

Rosemary and Sage
Mihiret Mekonnen and Begashaw Manahlie 35
Collection, Isolation and Identification of Entomopathogenic Fungi from

Different Agro Ecological Zones of Ethiopia
Solomon Yilma, Denberu Kebede and Tsegaye Mihrete 47
Chitinolytic Activity of Entomopathogenic Fungi Collections from Ethiopia

Solomon Yilma and Denberu Kebede 55
Evaluation of Faba Bean Genotypes for Faba bean Gall Resistance

Musa Jarso, Asnakech Tekalign, Nigussie Kefelegn, Gemechu Keneni, Simegnew Anley
Abebe Hailu and Fantanesh Sendek 65
Evaluation of Sweetpotato Varieties against Major Sweetpotato Viruses in Ethiopia

Shiferaw Mekonen and Fekadu Gurmu 69
Screening of Rice Germplasms for Resistance against Sheath Rot

(Sarocladium oryzae) at Fogera, Ethiopia
Desalegn Yalew Fetene, Muluadam Birhan and Tekalign Zeleke 79
Screening of Quality Protein Maize Inbred Lines for Turcicum Leaf Blight and Gray Leaf Spot
Midekssa Dida, Temesgen Deressa and Debela Diro 93
Screening of Maize Inbred Lines for Gray Leaf Spot and Turcicum Leaf Blight Resistance
Midekssa Dida, Temesgen Deressa and Debela Diro 105
Evaluation of Soybean Genotypes (Glycine max (L.) Merr.) for Asian Soybean Rust
(Phakopsora pachyrhizi ) in South Western Ethiopia
Yechalew Sileshi, Mesfin Hailemariam, Abush Tesfaye, Kifile Belachew and Negassa Dechasa 117
Screening of Linseed (Linum usitatissimum) Accessions for Resistance to
Fusarium oxysporum f. sp. Lini., at Holetta, Ethiopia
Habtewold Kifelew, Misteru Tesfaye and Birhanu Mengistu 129
Development of Integrated Management of Faba bean Gall

(Olpidium viciae Kusano) in West Shewa Highland
Tajudin Aliyi, Alemayehu Hailu, Bayoush Birke and Gudisa Hailu 135
Screening of Fungicides for the Control of Aloe vera Rust under Field Condition
Mihiret Mekonnen and Begashaw Manahlie 147
Evaluation of Fungicides Efficacy against Rice sheath Rot (Sarocladium oryzae)

in Rain-fed Lowland Rice (Oryzae sativa L.) in Fogera Hub
Muluadam Berhan, Desalegn Yalew, Tekalgn Zeleke 153
PART II: Weed Science

Survey and Identification of Major Weeds of Seed Spices in Ethiopia
Merga Jibat, Wakjira Getachew, Abukiya Getu, Habetewold Kifelew 167
Critical Weed Competition Period on Roselle (Hibiscus sabdariffa L.)

at Wondo Genet, Southern Ethiopia
Ano Wariyo and Negasu Guteta 175
Critical Weed Competition Period on Basil (Ocimum basilicum L.)

at Wondo Genet and Alage, Southern Ethiopia
Critical Weed Competition Period on Chamomile (Matricaria chamomilla L.)

Critical Weed Competition Period on Roselle (Hibiscus sabdariffa L.)

at Arba Minch, Southern Ethiopia
Getachew Gudero and Ano Wariyo 215
Herbicidal Control of Chrysanthemum segetum in Wheat Field at Cheliya District

Shashitu Bedada, Nigusie Hundesa, Gectachewu Bekele, Tesfaye Abdissa and Tadele Gudeta
231
Herbicides Efficacy on Weeds of Upland Rice (Oryza sativa L.) in Fogera, Ethiopia
Muluadam Berhan, Desalegn Yalew, Tekalgn Zeleke and Esuyawukal Demis 239
Herbicidal Control of Weeds in Turmeric at Tepi, South western Ethiopia

Wakjira Getachew, Merga Jibat and Habetewold Kifelew 253
Management of Striga (Striga hermonthica) in Sorghum (Sorghum bicolor) at Jeldu District

Shashitu Bedada, Dessalegne Gella, Niguse Hundessa, Getachew Bekele, Tesfaye Abdisa,
Dereje Geremew and Tadele Gudeta 259
Integrated Management of Wild Radish (Raphanus raphanistrum)

in Faba Bean in South West Shewa Zone, Ethiopia
Shashitu Bedada, Niguse Hundessa, Shugute Addisu, Dessalegne Gella, Getachew Bekele,
Tesfaye Abdissa and Tadele Gudeta 265
Integrated Management of Cotton Weeds at Middle Awash Rift Valley of Ethiopia

Workishet Taye, Zemedkun Alemu, Sharew Abate and Bedane Gudeta 279
PART III: Agricultural Entomology
Studies on Geographic Distribution, Seasonal Abundance and Insecticidal
Control of White Mango Scale (Aulacaspis tubercularis) (Hemiptera: Diaspididae)
Birhanu Sisay, Abiy Fekadu, and Gashawbeza Ayalew 289
Screening of Common bean Genotypes for their Resistance to the

Mexican Bean Weevil (Zabrotes subfasciatus)
Mulatwa Wondimu and Berhanu Amsalu 299
Evaluation of Botanical and Synthetic Insecticides against the Cabbage Aphid

Brevicoryne brassicae (Linnaeus) on Ethiopian Kale at Debre Zeit, Ethiopia
Ermias Shonga, Fasil Tadesse and Getachew Tabor 309
Foreword
Agriculture is the backbone of the Ethiopian economy. The sector determines the
growth of all other sectors and consequently the whole economy. It constitutes
over 50% of the gross domestic product (GDP), accounts for over 85% of the
labor force and earns over 90% of the foreign exchange. On average, crop
production makes up 60% of the sector’s outputs. The sector is dominated by
small-scale farmers who practice rain-fed mixed farming based on traditional
technology. Land area under smallholding accounts for 95% of the total
agricultural land, and the smallholder system is responsible for more than 90% of
the total agricultural output.
The productivity of crops is severely threatened by various factors in which pests

(diseases, insect pests, weeds and migratory pests) are taking the lion share. Crop
losses due to pests are a serious concern where in most cases small holder farmers
have limitation both in technical know-how and financial resources to apply
effective and sustainable pest control tactics and reduce crop damage. Apart from
the yield loss, pests also affect the quality of agricultural products, where the
concern of food safety, environmental pollution and market competence are
becoming issues for various stakeholders in the product value chain that includes
growers, traders, consumers and policy makers.
Considering the current plant protection problems, the plant protection directorate
has been re-structured in 2019. During the restructuring, national programs were
identified, and research strategy developed. Accordingly, Plant Pathology,
Agricultural Entomology, Weed Science, Quarantine and Pesticide research are
the identified national programs under the directorate. Both basic and applied
research has been conducted during the last three years to generate information
and technology towards effective and sustainable pest management. Several
information and technologies that can be used by the various beneficiaries have
been generated. Apart from generation of technologies and information, the
various national programs provide services associated with identification of
diseases, insects and weeds. Moreover, quarantine and pesticide efficacy services
are also provided by the directorate for the various national and international
stakeholders.
The information presented in this proceeding is aimed at availing information and

technology generated in plant protection research directorate for use by the
various users engaged in agricultural research and development.
Diriba Geleti (PhD)

Deputy Director General
[i]
PART I
Plant Pathology
[1]
[2]
Survey of Wheat Rust Diseases in Ethiopia
Netsanet Bacha1, Tsegaab Tesfaye1, Kitesa Gutu1, Getaneh Woldeab1, Tamrat Negash2, Daniel
Kassa2, Fikirte Yirga2, Megersa Debela3, Gudeta Bedada 3, Tesfaye Dessale4, Ashenafi
Gemechu5, Tilahun Bayisa6, Zerihun Eshetu 6, Zerihun Tomas7, Girma Teshome8, Mitiku
Kebede9, Bilal Temam9 , Misgana Mitiku9, Bekele Kassa10, Yitagesu Tadesse10, Mequnanint
Andualem11, Tsehaye Brhane12, and Wulita Wondwosen 13
1
Ethiopian Institute of Agricultural Research (EIAR), Ambo Agricultural Research Center P.O. Box 37,
Ambo, Ethiopia; 2EIAR, Kulumsa Agricultural Research Center, Assela; 3Oromia Agricultural Research
Institute (OARI), Bako Agricultural Research Center, Bako, Ethiopia; 4Amhara Region Agricultural Research
Institute (ARARI), Sirinka Agricultural Research Center; 5EIAR, Debre Zeit Agricultural Research Center;
6
OARI, Sinana Agricultural Research Center, Bale Robe; 7Southern Agricultural Research Institute (SARI),
Areka Agricultural Research Center; 8OARI, Bore Agricultural Research Center, Bore, Ethiopia; 9SARI,
Werabe Agricultural Research Center, Werabe; 10EIAR, Holetta Agricultural Research Center; 11ARARI, Adet
Agricultural Research Center; 12Tigray Agricultural Research Institute, Alamata Agricultural Research
Center, Alamata; 13ARARI, Debrebirhan Agricultural Research Center.
Abstract
Wheat is one of the most important crops in the world. Ethiopia is the largest wheat
producer in Africa. Wheat rusts are among the most important constraints in wheat
production in Ethiopia. The aim of this study was to study the geographical distribution
of rusts in major wheat growing regions of Ethiopia and determine the reaction of
wheat cultivars to wheat rusts in the country during 2018 cropping season. A total of
727 wheat fields were surveyed in major wheat growing areas of Oromia, Amhara,
Tigray and Southern Nations, Nationalities Peoples Region (SNNP). Yellow rust was the
most widely distributed disease in the surveyed fields with a prevalence percentage of
61.3. The overall mean incidence values for yellow rust, stem rust and leaf rust were
36.3, 20.9 and 4%, respectively. Likewise, the overall mean severities were 15, 9 and
1.7%, respectively in the same order. The incidences and severities of wheat rusts varied
among wheat varieties in the season. Most varieties showed moderately susceptible to
susceptible responses against yellow rust and stem rust populations. The most
commonly grown cultivars such as Danda’a, Digelu, Hidasse and Ogolcho were
affected by yellow rust and stem rust disease in the season and showed MSS-S reaction
types in most of the study areas. The predominance of susceptible varieties can be an
important recipe for the development of rust epidemics in Ethiopia. Hence, continuous
supply of resistance varieties is needed to avoid wheat rust epidemics in the country.
Introduction
Wheat (Triticum spp.) is the most widely grown crop worldwide and is the crucial
component for the global food security. Wheat provides one-fifth of the total
caloric intake for the world population and is produced on an area of over 200
million hectares worldwide (Wang et al., 2015). It is cultivated on 15.4% of the
arable land, accounting for around 30% of global grain production and 44% of
cereals used as food (FAO, 2017).
Ethiopia is the largest wheat producer in Sub-Saharan Africa (FAO, 2017). The
crop ranked fourth in land coverage and total production after tef, maize and
[3]
sorghum (CSA, 2017) in the country. Wheat is widely grown by subsistent
farmers under rain-fed conditions and over one-third of cereal farm households are
dependent on wheat farming (Shiferaw et al., 2013). According to CSA (2017), in
Ethiopia wheat is produced on over 1.7 million hectares of land with annual
average productivity of about 2.67 metric tons/ha, which is by far below the
world’s average yield of about 3.3 tones/ha. This relatively low mean national
yield may be partially attributed to diverse biotic and abiotic stresses among which
rust diseases are the most important.
Wheat rust pathogens are known for their various races. New races of rust
pathogens can occur during a single growing season due to mutation,
recombination, and selection for virulence against rust resistance genes in wheat.
Hence, each season it is crucial to carry out surveys to detect new and highly
virulent pathogen phenotypes as they appear. According to Huerta-Spino (1994)
and Park et al. (2011), surveys also provide essential information to determine the
gene combinations to be considered by breeding programs using major gene
resistance. The improved wheat varieties and local landraces which are under
production has to be evaluated their status of resistance to wheat diseases.
Accordingly, the present study was conducted with the objectives of determining
the distribution and intensity of wheat rust diseases in the country and assessing
the diversity and of wheat varieties deployed to farmers.
Materials and Methods

Description of the study area
The surveys of wheat rust were carried out in the major wheat growing regions of
Ethiopia: Oromia, Amhara, Tigray and Southern Nations, Nationalities Peoples
(SNNP) in 2018 cropping season. The surveys were carried out at 5-10 km
interval following main and feeder roads on preselected routes in areas where
wheat is important and diseases are known to be present.
Disease assessment
Disease assessments were made at five points along the two diagonals (in an ‘’X’’
pattern) of the field using quadrant and used to calculate average values. The
incidence was calculated by using the number of infected plants and expressed as
a percentage of the total number of plants assessed.
Disease incidence: DI (%) = Number of diseased plants in a quadrant X 100

Total No. of plants in a quadrant
[4]
The disease severity was examined visually on the whole plants within the
quadrants as the percentage of plant tissue affected and recorded according to
modified Cobb's scale (Peterson et al., 1948).
Disease severity: DS (%) = Area of a plant tissue affected X 100

Total area of a plant part assessed
The host plant response to infection was also scored using the description of
Roelfs et al. (1992). The prevalence of disease was measured by using the number
of fields affected divided by total number of fields assessed and expressed in
percentage.
Prevalence (%) = No of fields affected X 100

Total fields assessed
In addition, data on geographical information (latitude, longitude and elevation) of

each field were recorded using GPS.
Data analysis
The data were analyzed by using the descriptive statistics (Gomez and Gomez
1984). The SPSS IBM20 and Arc GIS version 10.5.1 computer software programs
were used to compute the mean and map the surveyed areas, respectively.
Results and Discussion

Wheat rusts status in the wheat growing zones
A total of 727 wheat fields with 313 in Oromia; 46 in Tigray, 94 in SNNP and 274
fields in Amhara regions were evaluated for prevalence, incidence and severity of
rusts from September to December, 2018 (Table 1). In the season, major wheat
growing highlands of 19 zones found in the four regions were assessed for the
three rust diseases.
The mean number of yellow rust affected fields was 61.3%. Yellow rust was
widely (79.6%) distributed in Amhara followed by 76.6% in SNNP region. In
Oromia region the distribution was 49.5%, while it was 2.2% in Tigray. The peak
yellow rust prevalence (100%) was recorded in Guji zone of Oromia and Gedeo
zone of SNNPN region. The incidence of yellow rust ranged from 0 to 100% with
overall mean of 36.3%. The maximum mean of 56.0% was recorded in SNNP
region followed by mean of 53.7% in Amhara region. The least mean incidence of
0.4% was noted from Tigray region. Among the different zones, the highest mean
incidences of yellow rust were recorded in Silte (73.5%); Hadiya (72.9%) and
Gedeo (60.7%) zones. Severity of yellow rust varied from 0-100% with overall
[5]
mean severity of 15%. The mean severities of 20.6%, 19.6%, 11.2% and 0.04%
were recorded from SNNP, Amhara, Oromia, and Tigray regions, respectively.
The highest yellow rust severity (28.1%) was registered in Silte, while the lowest
mean (0.04%) was noted in S. Tigray.
The overall mean prevalence of stem rust in the growing season was 39.1%. Stem
rust affected fields reach 66%, 49.8%, 21.2, 17.4% in SNNP, Oromia, Amhara and
Tigray regions, respectively. This disease was absent in Sidamo zone of SNNP,
while all farms assessed were affected in E. Shewa, Halaba, and Gurage zones.
Stem rust prevalence in S. Wellega and Guji zones were 92.3 and 96.9%,
respectively. Stem rust incidences ranged from 0-100%, while the mean for the
regions were below 21.5% with the exception of 4.5% for Tigray region. Stem
rust incidence was high (> 60%) in four zones of SNNP region and moderate (>
37.0%) in W. Shewa, W. Arsi and E. Wellega zones of Oromia region. However,
there was no stem rust recorded in Sidamo zone of SNNP region. Stem rust
severities ranged from 0-95% with overall mean of 9.0%. The means over the
regions varied from 1.6% in Tigray to 21.5% in SNNP region. The lowest stem
rust severity of 1.6% was recorded in S. Tigray zone. Stem rust infection was not
observed in Sidama zone of the SNNP region.
Prevalence of leaf rust in the season was 19.1%. The distribution of leaf rust was
less than 20% in three of the regions while it was 38.3% in SNNP region. No leaf
rust was observed in five zones of the regions (Table 1). The incidence of leaf rust
ranged from 0- 100% with means of less than 11.6% of each region. Maximum
incidence of the disease (32.9%) was noted in Hadiya zone and the least (0%) in
E. Wellega, E. Shewa, N. Shewa, Awi and Sidama zones. leaf rust severity ranged
from 0-70% with overall severity of 1.7%. A maximum severity of 6.7% was
noted in Hadiya, while the lowest zero was recorded in many zones of the four
regions (Table 2).
The field assessment showed that the wide distribution of yellow rust across the
regions. According to Wellings (2011), yellow rust is causing significant damage
in the major wheat growing regions of the world including China, the USA, India,
Pakistan, and Nepal, Yemen and East England, Australia, New Zealand, Ethiopia
and Kenya. Carver (2009) also indicated that the importance of yellow rust has
been reported from over 60 countries in the world and has caused significant yield
losses. In Ethiopia, frequent epidemics of yellow rust have been a major challenge
to wheat production in the country. The breakdown of resistance conferred by Yr9
due to the emergence of virulent races of P. striiformis f. sp. tritici, yellow rust
causing pathogen, in 1989 resulted in major epidemics of yellow/stripe rust that
challenged wheat production in Ethiopia and the rest of the world (Chen et al.
2014; Wellings, 2011). In 2010, Ethiopia experienced a devastating stripe rust
[6]
epidemic that affected more than 600,000 ha of wheat and significant widespread
losses were realized (Abeyo et al., 2014).
Likewise, Mesifin (2019) reported that stem rust is a major concern in East
African highlands while it has only minor importance in countries of North
America and Europe. According to Periyannan et al. (2013), Ethiopia is
considered as a hot spot for the development and spread of new stem rust
diversity. These new races were virulent on most of varieties grown in the country
and are among the most virulent in the world. The high prevalence of the rust
diseases in wheat growing areas of Ethiopia might be the result of cultivation of
susceptible wheat cultivars and favorable climatic conditions (Hei et al., 2017).
On the other hand, wheat leaf rust is more frequent in almost all global regions
than stem rust or stripe rusts of wheat (Bolton et al., 2008). However, it has minor
importance in wheat production in Ethiopia.
Growing resistant cultivars is a mostly preferred approach for control of rusts

because it is an economic, effective, and environmentally friendly strategy (Chen
2005; 2013). However, resistant cultivars can become susceptible due to changes
in the pathogen population dynamics. Virulence characterization of the pathogen
population on a regular basis is required to devise effective disease management
by deploying effective resistance genes.
[7]
Table 1. Prevalence of wheat rusts in 2018 main cropping season
No of Yellow rust Stem rust Leaf rust

No of No of No of
Region fields
Zone infected Prevalence infected Prevalence infected Prevalence
assessed
fields fields fields
Oromia W. Shewa 51 23 45.1 12 23.5 5 9.8
SW Shewa 25 14 56.0 2 8.0 4 16.0
West Arsi 40 9 22.5 5 12.5 4 10.0
S. Wellega 13 12 92.3 12 92.3 0 0.0
Horoguduru 51 33 64.7 32 62.7 2 3.9
E. Shewa 7 1 14.3 7 100.0 0 0.0
N. Shewa 17 13 76.5 1 5.9 0 0.0
Bale 77 31 40.3 54 70.1 3 4.0
Guji 32 32 100 31 96.9 32 100
Mean 313 155 49.5 156 49.8 50 16.0
Amhara S. Gonder 35 28 80.0 5 14.3 1 2.9
Awi 4 3 75.0 1 25.0 0 0.0
W. Gojam 35 14 40.0 10 28.6 3 8.6
E. Gojam 55 36 65.5 22 40.0 2 3.6
N. Shewa 145 137 94.5 20 13.8 45 31.0
Mean 274 218 79.6 58 21.2 51 18.6
Tigray S. Tigray 46 1 2.2 8 17.4 2 4.3
SNNP Sidamo 20 17 85.0 0 0.0 0 0.0

Gedeo 7 7 100 2 28.6 5 71.4
Halaba 14 6 42.9 14 100.0 14 100.0
Silte 34 31 91.2 30 88.2 7 20.6
Hadia 12 10 83.3 9 75.0 8 66.7
Gurage 7 1 14.3 7 100.0 2 28.6
Mean 94 72 76.6 62 66.0 36 38.3
Grand total 727 446 61.3 284 39.1 139 19.1
[8]
Table 2. Incidence and severity ranges and mean of rust diseases of wheat in 2018 main season
Region Zone Yellow rust Stem rust Leaf rust

Incidence Severity Incidence Severity Incidence Severity
Range Mean Range Mean Range Mean Range Mean Range Mean Range Mean
Oromia W/ Shewa 0-100 17-5 0-80S 7.8 0-100 37.0 0-95S 14.2 0-20 1.1 0-15MS 0-7
W/Arsi 0-100 14.9 0-60MS 5.4 0-100 42.6 0-80S 11.8 0-100 6-5 0-50MS 2-5
E/Wellega 0-90 54.6 0-40S 21.2 0-100 54.6 5MS-60S 19.7 0 0 0 0
Horoguduru 0-95 34.0 0-80S 17.5 0-90 22.4 0-60S 13.2 0-80 2.4 0-70S 1.9
E/Shewa 0-100 14.3 0-60S 5.6 5-70 40.7 TR-50MSS 11.2 0 0 0 0
N/Shewa 0-100 28.8 0-80S 18 0-30 2.1 0-25S 1.6 0 0 0 0
Bale 0-100 13.5 0-30S 2.83 0-100 23.9 0-80S 8.6 0-100 2.0 0-20S 0.46
Guji 0-50 18.1 0-60S 20.0 0-30 18.8 0-40MS 20.5 5-25 11.7 10MR- 13.3
35MS
Mean 0-100 20.6 0-80S 11.2 0-100 27.6 0-95S 12.1 0-100 3.1 0-70S 2.2
Amhara S/Gonder 0-100 54.1 0-100S 23.1 0-100 10 0-60MS 6.7 0-100 2.9 0-40MS 1
Awi 0-100 35 0-20S 0.4 0-5 1.25 0-TR 0.25 0 0 0 0
W/Gojam 0-100 12.9 0-40S 3.4 0-100 4.3 0-60S 5.5 0-60 1 0-60S 1.9
E/Gojam 0-100 21.7 0-80S 5.6 0-60 7.8 0-60S 4.3 0-5 0.1 0-10MS 0.16
N/Shewa 0-100 176.1 0-90S 28.4 0-100 3.7 0-40MS 0.9 0-100 3.6 0-40S 0.9
Mean 0-100 53.7 0-100S 19.6 0-100 6.3 0-60S 2.4 0-100 2.4 0-60S 0.9
Tigray S/Tigray 0-20 0.4 0-5MR 0.04 0-80 4.5 0-60S 1.6 0-80 2.1 0-30|MS 0.6
SNNP Sidamo 0-100 52.8 0-60S 17.2 0 0 0 0 0 0 0 0
Gedeo 0-100 60.7 0-60S 24.1 0-25 5.7 0-10MS 2.3 0-35 10 0-25MS 3.7
Halaba 0-100 20.7 0-45MS 5.9 0-100 60.4 10MR-90S 24.2 10-35 19.6 5R-25MS 5.8
Silte 0-100 73.5 0-90S 28.1 0-100 66.3 0-90MS 27.9 0-100 9.1 0-40MSS 1.9
Hadiya 0-100 72.9 0-60MS 2.9 0-100 64.6 0-85S 27.4 0-100 32.9 0-40MSS 6.7
Gurage 0-100 14.3 0-50MS 5.7 95-100 99.3 35MSS- 55.1 0-20 5.7 0-10MR 1.1
80S
Mean 0-100 56.0 0-90S 20.6 0-100 49.0 0-90S 21.5 0-100 11.6 0-40MSS 2.8
Grand mean 0-100 36.3 0-100 15.0 0-100 20.9 0-95S 9.0 0-100 4.0 0-70S 1.7
MR: Moderately Resistance, MS: Moderately Resistance, S: Susceptible, tr: trace
[9]
Prevalence and intensity of wheat stem and yellow rusts by
wheat variety
In 2018 main cropping season Danda’a (175 fields), Ogolcho (52), Hidasse (51),
Kakaba (41), Digelu (23) and Local (21) were the dominant wheat varieties grown
across the regions. Of these, Danda’a, Ogolcho, Digelu and Huluka were the most
popular and widely grown variety in Oromia region. Kingbird in Oromia, Kakaba,
Hidase and Kingbird in Tigray were not affected by yellow rust. While Liben in
Oromia, Kingbird in Tigray and Alidoro, Degelu, Kakaba, Gundile and Shamet in
Amhara region were free from stem rust (Table 3).
The incidences and severities of wheat rusts varied among wheat varieties. Most
varieties showed moderately susceptible to susceptible response against yellow
rust and stem rust populations. However, the local cultivar showed susceptible
reaction to yellow rust and resistance response to stem rust diseases in most of the
regions. The most dominant variety Danda’a showed susceptible reaction against
yellow rust in the regions. The yellow rust incidence and severity on Danda’a
varied between 17.7-80.3% and 7.9-25.5%, respectively. The highest yellow rust
incidence of 100% was recoded on variety Menze. In Oromia, the highest yellow
rust incidences were recorded from varieties Digelu and Triticale with 54.8 and
37.5%, respectively. The maximum yellow rust severity of 80% was observed on
variety Menze in Amhara region (Table 3).
Stem rust mean severity up to 36.7% was noted on Hidase in Oromia. Similarly,
2.5, 3.6 and 4.3% mean severities were recorded on varieties Triticale, Hidasse
and some unknown improved varieties in Amhara region. The highest stem rust
severity of 36.7 % was recorded on variety Hidasse followed by Sanate with
severity of 26.0%. Liben in Oromia, Kingbird in Tigray were not affected by
yellow rust. Similarly, Alidoro, Digelu, Kakaba, Gundile, Shamet in Amhara and
some unknown varieties in SNNP region were free from stem rust.
[10]
Table 3. Reaction of dominant wheat cultivars to yellow and stem rusts in 2018 cropping season
Yellow rust Stem rust

Region Variety TNF Severity Severity
Incidence Incidence
Range Mean Range Mean
Danda’a 79 17.7 0-60S 7.9 28.7 0-60S 10.8
Ogolcho 52 21.2 0-60S 6.6 32.5 0-40S 7.6
Digelu 19 54.8 0-80S 35.8 40 0-40S 16.1
Huluka 18 15 0-40S 16.3 15.9 0-35MS 17.2
Hidasse 12 24.1 0-60S 11.6 69.25 0-80S 36.7
Kakaba 13 12.8 0-40S 7.6 26 0-70S 16.2
Kubsa 10 31.6 0-60S 20.7 29 0-95S 15.35
Alidoro 6 12.5 0-25MS 9.8 22.5 0-40S 12.6
Oromia Kingbird 5 0.2 0-Tmr 0.08 0.4 0-Tms 0.32
Liben 4 8.8 0-25MSS 8.3 0 0 0
Local 4 29 0-70S 19 3 0-5R 0.25
Triticale 4 37.5 0-40S 17.5 8.8 0-10MSS 3.5
Senate 4 7.5 10MR-20MSS 9 22.5 30MS-40MS 26.0
Lemu 3 27 0-30S 10.1 0.7 0-5MSS 2.25
Simba 3 35 0-60MSS 19.3 33.3 0-40MSS 12
Unknown 50 19.4 0-80S 10.9 27.4 0-60S 9.9
Kakaba 25 0 0 0 1.4 0-5MR 0.144
Local 12 1.7 0-5MR 0.2 0.8 0-2MR 0.1
Tigray
Hidasse 4 0 0 0 40 3MR-60S 17.8
Kingbird 3 0 0 0 0 0 0
Alidoro 2 70 15S-45S 1.5 0 0 0
Danda’a 75 80.3 0-80S 25.5 2.4 0-40MS 0.6
Hidasse 23 84.6 5MR-81MS 30.6 12.7 0-30S 3.6
ET-13 18 52.3 0-70MS 9.9 0.1 0-1R 0.01
Triticale 11 28.6 0-100S 20.4 11.8 0-20S 2.5
Menzie 6 100 60S-90S 80 0.2 0-1R 0.03
Local 5 14 0-40S 14.4 10 0-10MS 1.6
Amhara Digelu 4 75 15S-80S 51.3 0 0 0
Tsehay 4 52.5 0-40MS 15 15 0-10MR 1
Kakaba 3 60 10MS-60S 29.3 0 0 0
Unknown 3 66.7 0-80S 53.3 0.3 1-1MR 0.1
Gundile 2 80 10S-50S 30 0 0 0
Shamet 2 80 20S-80MS 42 0 0 0
Improved 112 28.3 0-90S 8.5 8.9 0-60S 4.3
Danda’a 21 61 0-60S 22.5 45.5 0-40MS 14.5
Kakaba 18 64.7 0-60S 25.8 80.3 0-70MS 32.8
Shorima 13 48.8 0-35MS 12.5 13.8 0-40MSS 5.1
Hidasse 12 73.8 0-60MS 24.3 58.3 0-90S 32.9
SNNP
Ogolcho 5 63 0-60S 26 37 0-45MS 16.2
Local 4 50 0-50MSS 20.3 50 0-65MSS 23.6
Unknown 4 56.3 0-45MS 15 0 0 0
Wane 2 20 0-5MR 1 47.5 10R-20MSS 10
TNF, Total Number of Fields; MR, Moderately Resistance; MS, Moderately Resistance;
S, Susceptible; tr, trace.
Generally, most recently released varieties in the country were succumbed to

either yellow rust or stem rust diseases shortly after their introduction. According
[11]
to this study, the dominant wheat cultivars Danda’a, Kakaba, Hidase and Ogolcho
were susceptible to rust diseases and covered 50% of the wheat fields inspected. In
most cases, the failures have been due to the virulence present in the pathogens
population and deployment of qualitative type of resistance in wide array of wheat
cultivars (Hei et al., 2017). The predominance of susceptible varieties can be an
important recipe for the development of rust epidemics in Ethiopia. Another
problem with the continued use of susceptible varieties is that it increases the
chances of a new mutant race developing to attack presently resistant varieties
(CIMMYT, 1989). Hence, continuous supply of resistance varieties is needed to
avoid wheat rust epidemics in the country.
Several efforts were made towards resistant cultivars development in Ethiopia and
several bread and durum wheat cultivars with various levels of rust resistance
were released for production. However, the resistance to stem rust of most of the
released bread wheat and durum wheat varieties has been broken soon after their
release. This experience in the country emphasizes the need for genes with
broader resistance or for combinations of resistance genes that can confer a
broader and more durable resistance (Zhang et al., 2017). Combining seedling
resistance with adult plant resistance in the field will provide valuable indications
to select resistant varieties.
Conclusion and Recommendation

The present results indicated that the distribution of wheat rusts especially of
yellow and stem rusts is remarkably on increasing trend. The increased intensity
of these diseases might be associated with the evolution of new pathogen races,
long distance dispersal of the pathogen, and extensive cultivation of the
susceptible wheat varieties. Building strong surveillance network, which
accompanies surveys, intensive sampling and surveillance is highly
recommendable in partnership within pathologists, breeders, modelers,
epidemiologists, molecular geneticists and policy makers.
The current study reported that most wheat varieties cultivated in farmers’ fields
were susceptible to rusts. The most principal and excellent management strategy
that provides adequate protection without the need for chemicals to control wheat
stem rust has been host resistance, which is effective and affordable to small-scale
farmers. However, the narrow genetic base of stem rust resistance in the Ethiopian
wheat cultivars aggravates their ineffectiveness against rusts. Therefore, efforts
need to be directed to widen the genetic bases of wheat varieties for stem rust
resistance to control the diseases in the country. Thus, further research on adult
resistance stage should be conducted to come up with a full package of
recommendation on those varieties against selected dominant races.
[12]
Acknowledgements
The authors greatly acknowledge the financial support from Biotechnology and
Biological Sciences Research Council (BBSRC, UK Research and Innovation) and
the Delivering Genetic Gains in Wheat (DGGW) Project.
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[14]
Physiological Races of Puccinia graminis f. sp.
tritici in Ethiopia in 2018 Cropping Season
Netsanet Bacha1, Tsegaab Tesfaye1, Kitesa Gutu1, Getaneh Woldeab1, Tamrat Negash2,
Daniel Kassa2, Fikirte Yirga2, Megersa Debela3, Gudeta Bedada 3, Tesfaye Dessale4,
Ashenafi Gemechu5, Tilahun Bayisa6, Zerihun Eshetu6, Zerihun Tomas7, Girma
Teshome8, Mitiku Kebede9, Bilal Temama9, Misgana Mitiku9, Asfaw Azanaw10, Bekele
Kassa11, Yitagesu Tadesse11, Mequnanint Andualem12, Tsehaye Brhane13, Wulita
Wondwosen14, and Kalkidan Yallew14
1
Ethiopian Institute of Agricultural Research (EIAR), Ambo Agricultural Research Center P.O. Box 37,
Ambo, Ethiopia; 2EIAR, Kulumsa Agricultural Research Center, Assela; 3Oromia Agricultural Research
Institute (OARI), Bako Agricultural Research Center, Bako; 4Amhara Region Agricultural Research Institute
(ARARI), Sirinka Agricultural Research Center; 5EIAR, Debrezeit Agricultural Research Center; 6OARI,
Sinana Agricultural Research Center, Bale Robe; 7Southern Agricultural Research Institute (SARI), Areka
Agricultural Research Center; 8OARI, Bore Agricultural Research Center, Bore; 9SARI, Werabe Agricultural
Research Center, Werabe; 10ARARI, Gonder Agricultural Research Center; 11EIAR, Holeta Agricultural
Research Center; 12ARARI, Adet Agricultural Research Center; 13Tigray Agricultural Research Institute,
Alamata Agricultural Research Center, Alamata; 14ARARI, Debrebirhan Agricultural Research Center
Abstract
Stem rust caused by Puccinia graminis f.sp. tritici (Pgt) is one of the most important
diseases of wheat (Triticum aestivum) across the globe. Because of the sudden changes
in stem rust race patterns, commercial varieties are becoming vulnerable globally at
large and particularly in Ethiopia. In the virulence survey, seven stem rust races
namely TKTTF, TTTTF, TKKTF, TTKTT, TKPTF, TTKTF and TTRTF identified.
Northern American Nomenclature system was used to pathotype the races. Most genes
possessed by the differentials were ineffective against one or more of the tested
isolates. The Digelu race, TKTTF and TKKTF were identified from 93 (28%) stem rust
isolates each, while TTTTF was detected from 90 (27%) samples. The new race for the
country, TTKTT was isolated from samples collected in W. Arsi, Hararge, Wollega and
Guji zones of Oromia region and E. Gojam and S. Wollo zones of Amhara region.
Varieties such as Huluka, Sanate, Ogolcho, Shorima, Hidase and Danda’a were highly
affected by the race TTKTT. This race has been confirmed to belong to TTKSK (Ug99)
race group. This first confirmation of TTKTT in Ethiopia is important because of its
virulence to the Sr24 gene, indicating the increase in virulence and variability in the
Ethiopian P. graminis f. sp. tritici population. The gene Sr24 was a valuable source of
resistance and had been effective so far against most stem rust races. The virulence
observed on Sr24 might force majority of Ethiopian varieties to be out of the
production and indicates the need for developing cultivars with combinations of
effective resistance genes or cultivars with multiple minor genes to achieve durable
resistance.
Introduction
The mobility of stem rust coupled with their inherent ability to change through
mutation, genetic recombination and new introductions from other countries
makes continual monitoring important (Park, 2007). Furthermore, some studies
[15]
that were carried out in Ethiopia showed that most previously identified races
were virulent on most of the varieties grown in the country and they are among the
most virulent in the world (Admassu et al., 2009). Keeping this in mind,
continuous and exhaustive surveys require to be carried out to give an
understandable picture of the virulence pattern of Pgt in Ethiopia. The survey of
races helps to generate information regarding virulence of races, their frequency
and distribution patterns and to identify the evolution of new races and forecasting
the virulence shifts in a population. This is the greatest significance in developing
wheat cultivars with durable stem rust resistance (Admassu et al., 2009).
According to Belayneh and Emebet (2005), rusts should be surveyed regularly to
determine its current status and to take action before the pathogen becomes a risk
to wheat production. Major wheat growing areas of Ethiopia have been hammered
several times by rust diseases such as yellow, stem and leaf rust (AARC, 2017).
Moreover, due to rapid changes in stem rust race patterns, commercial cultivars
are inclined to become at risk. Therefore, detail information on wheat stem rust
physiological races in Ethiopia is crucial to detect new races as they appear and
respond accordingly. Consequently, the present study was conducted to identify
stem rust races prevalent in major wheat growing areas

Collection of rust samples
Stem rust samples were collected and received from Oromia, Amhara, SNNP and
Tigray regions during 2018 main season from wheat fields and trial plots every 5-
10 km or at the first field thereafter. Stems and/or leaf sheath of wheat plants
infected with stem rust were cut into small pieces of 5 to 10 cm in length using
scissors and placed in paper bags after the leaf sheath was separated from the stem
in order to keep stem and/or leaf sheath dry. The samples were then labeled with
the name of the zone, district, and variety, GPS (altitude, latitude and longitude)
data and date of collection and transported to Laboratory for analysis. Race
analysis was undertaken independently at Ambo Agricultural Research Center
(AmARC), Ethiopia and Regional Cereal Rust Research Center (RCRRC), Turkey
following the International Nomenclature System of P. graminis f. sp. tritici
(Roelfs and Martens, 1988).
Isolation and multiplication of single-pustule isolates

Bulked urediniospores from each field were suspended in lightweight mineral oil,
Soltrol 170 (Chevron Phillips Chemical Company, the woodlands, Texsas, United
States) and sprayed onto 7-day-old seedlings of variety McNair, which does not
carry known stem rust resistance genes (Roelfs et al., 1992). After seven to ten
days of inoculation (when the flecks/symptoms were clearly visible) leaves
containing a single fleck that produce single pustule was selected from the base of
[16]
the leaves and the remaining seedlings within the pots were removed using
scissors. Only leaves containing single pustules from each location were
separately covered with cellophane bags and tied up at the base with a rubber band
to avoid cross contamination (Fetch and Dunsmore, 2004).
Two weeks later (when the pustule was well developed), spores from each pustule
were collected and a suspension, prepared by mixing urediospores with Soltrol
170, was inoculated on seven-day-old seedlings of the susceptible variety McNair
for multiplication purpose for each of the single pustules on separate pots. The
urediniospores descending from one pustule made up a single pustule isolate. One
isolate was developed from each wheat field and used for the final race analysis.
Inoculation of wheat stem rust differential lines

The seedlings of 20 wheat differential hosts with known stem rust resistance genes
and a susceptible variety McNair were grown in 10 cm diameter pots. The
differential wheat lines possessed resistance genes Sr5, Sr6, Sr7b, Sr8a, Sr9a,
Sr9b, Sr9d, Sr9e, Sr9g, Sr10, Sr11, Sr17, Sr21, Sr24, Sr30 Sr31, Sr36, Sr38,
SrTmp and SrMcN. Each rust isolate was suspended in Soltrol 170. The
suspension adjusted to 4 X 106 spores ml-1 was sprayed onto seedlings of the
differentials using spore inoculators. Inoculated seedlings were then placed in a
dew chamber in darkness for 18 hours at 18-22oC and 98-100% relative humidity.
Upon removal from chamber, plants were exposed to 4 h of fluorescent light to
provide condition for infection and allowed to dry their dew for about 1-2 hours.
Inoculated plants were then transferred to greenhouse benches where the
temperatures were kept between 18 and 25oC and the relative humidity at 60-70%
(Stubbs et al., 1986).
Determination of races
Seedling infection types were scored 14 days after inoculation using a 0 to 4 scale
(Stakman et al., 1962). The IT readings of 3 (medium-size uredia with/without
chlorosis) and 4 (large uredia without chlorosis or necrosis) were regarded as
susceptible. Other readings, i.e. 0 (immune or fleck), 1 (small uredia with
necrosis), and 2 (small to medium uredia with chlorosis or necrosis) were
resistant. The variations were refined by modifying characters as follows: -,
uredinia somewhat smaller than normal for the infection type; +, uredinia
somewhat larger than normal for the infection type.
Race designation was done by grouping the differential lines into five subsets as
indicated in Table 1. Each isolate was assigned using a five-letter designation
based on its reaction on the differential lines (Roelfs and Martens, 1988; Jin et al.,
2008).
[17]
Table 1. Nomenclature of Puccinia graminis f. sp. tritici based on 20 differential wheat hosts
Infection types produced on near-isogenic Sr lines

Set 1 5 21 9e 7b
Pgt - code Set 2 11 6 8a 9g
Set 3 36 9b 30 17
Set 4 9a 9d 10 Tmp
Set 5 24 31 38 McN
B Lowa Low Low Low
C Low Low Low Highb
D Low Low High Low
F Low Low High High
G Low High Low Low
H Low High Low High
J Low High High Low
K Low High High High
L High Low Low Low
M High Low Low High
N High Low High Low
P High Low High High
Q High High Low Low
R High High Low High
S High High High Low
T High High High High
Source: Roelfs and Martens (1988); Jin et al. (2008)
aLow = Infection types 0, 1, and 2 and combinations of these values
bHigh = Infection types 3 and 4 and a combination of these values
Data analysis
Data on physiological races were analyzed by using the descriptive statistics
(Gomez and Gomez 1984).

A total of 623 stem rust samples were collected from all surveyed regions during
2018 cropping season. Of these, 333 samples, 228 from Oromia, 60 from Amhara,
40 from South, 4 from Afar and 1 from Tigray were analyzed. The analysis
identified seven stem rust races namely TKTTF, TTTTF, TKKTF, TTKTT,
TKPTF, TTKTF and TTRTF (Table 2). TKTTF and TKKTF were identified from
93 (28%) stem rust isolates each, while TTTTF was detected from 90 (27%)
samples. In addition, TTKTT was isolated from 20 samples; however, TKPTF,
TTKTF and TTRTF were recorded from 12, 10, and 5 samples, respectively. Race
TKTTF is virulent to all of differential lines except, Sr11, Sr24 and Sr31 (Table
4). Race TTTTF has wide virulence spectrum. It is virulent to all resistant genes,
except two genes Sr24 and Sr31. Race TTTTF thus poses a serious threat to the
country’s wheat production. The wheat cultivars, Hitosa, Kakaba, Danda’a,
Emmer, Ogolcho, Kingbird, Kubsa, Digelu, Alidoro, Limu, Mangudo, Huluka,
Senate, Shorima, Dashen, Hidase, Messeba and Durum wheats were infected by
one or more of these races (Table 2).
[18]
The identification of the 7 races (Table 3) from samples analyzed in the
season confirmed the presence of wider range of virulence within the
Puccinia graminis f. sp. tritici population in Ethiopia and is in line with
previous studies conducted in the country (Belayneh and Emebet, 2005;
Admassu et al., 2009; Hailu et al., 2015; Hei et al., 2018). The continual
wheat production, favorable microclimates and the presence of the alternate
host, Berberris holistii in proximity to wheat production areas of Ethiopia
could be the main reasons for the rapid evolution and high virulence diversity
in the country (Hei et al, 2018). According to Hei et al. (2018), most of the
races in Ethiopia varied from one another by single-gene changes. Such
single-step changes in virulence were reported to be the main process of
evolutionary change in Puccinia graminis f. sp. tritici populations.
Of the races identified, TTKTT was detected for the first time in the country
from repeated experiments. Race TTKTT was virulent to wheat differential
lines containing Sr5, Sr6, Sr7b, Sr8a, Sr9a, Sr9b, Sr9d, Sr9e, Sr9g, Sr10,
Sr11, Sr17, Sr21, Sr24, Sr30, Sr31, Sr38, SrTmp, SrMcN and avirulent on
lines with Sr36. This race was detected from 18 samples collected in W. Arsi
(Central Ethiopia), Hararge (Eastern Ethiopia), Wollega (Western Ethiopia)
and Guji (Southern Ethiopia) zones of Oromia region (Table 2) and 2 isolates
from East Gojam and South Wollo zones of Amhara region. It was identified
from samples collected from Huluka, Sanate, Ogolcho, Shorima, Hidase and
Danda’a varieties. This race was reported in Kenya for the first time in 2014,
and belongs to the TTKSK (Ug99) race group (Patpour et al. 2016). TTKTT
has the most complex virulence combination of all known Ug99 races, and
range expansion out of Kenya is highly significant.
This first confirmation of TTKTT in Ethiopia is important because of its

virulence combinations for SrTmp, Sr24, and Sr31, indicating the increase in
virulence and variability in the Ethiopian Pgt population. The gene Sr24 is a
valuable source of resistance, effective against most stem rust races, including
race TTKSK (Ug99) worldwide (Jin et al., 2008). The detection of TTKTT in
Ethiopia is great concern since a high frequency of widely grown cultivars
and advanced breeding lines in Ethiopia are known to possess Sr24 resistance
gene (Hundie et al. 2019). According to Jin et al. (2008), commercial cultivars
with resistance based on Sr24, particularly in the East African region, must be
viewed with caution in terms of potential stem rust epidemics (Table 4).
[19]
Table 2. Stem rust races detected in zones surveyed and varieties from which the races were detected in 2018 cropping
season
Region Zone Races Detected Varieties from which races detected from
TTTTF(13) Unknown (10) Hitosa (2) Durum (1)
TKTTF(6) Unknown (1)
Bale TKKTF (20) Unknown (14) Hitosa (1) Kakaba (1), Danda’a (10 Hidase (1) Emmer (1)
TTKTF (3) Unknown (3)
TTRTF (1) Unknown (1)
Nursery lines (10) Ogolcho (4) Kingbird (1), LMPG (2), Hidase (1)
TTTTF (20) Unknown (2)
TTRTF (4) nursery lines (4)
TKTTF (6) Ogolcho (1), Danda’a (1), Kubsa (1), Hidase (1), Unknown (2)
TKKTF (11) Ogolcho (3), Hidase (3) LMPG (5)
TTKTT (2) Ogolcho (2)
Arsi TTKTF (1) LMPG (1)
TTTTF (6) Danda’a (4), Unknown (1), Digelu (1),
TKTTF (8) Danda’a (6) Digelu (1) Unknown (1)
S.W. TKKTF (3) Kubsa (1) Danda’a (2)
Shewa TKPTF (1) Danda’a (1)
Oromia
Kakaba (12), Kubsa (5) Kingbird (1) Unknown (4) MacNair (1) Digelu
TKTTF (41) (10) Danda’a (7), Alidoro (1)
Kakaba (5) Unknown (4) Kubsa (20 Kingbird (1) Danda’a (2) Hidase (1)
TTTTF (17) Digelu (1) Ogolcho (1)
TKKTF (5) Danda’a (1) Mixed (1) Limu (1), Unknown (1) Alidoro (1)
TKPTF (6) Kakaba (3) Kubsa (1) Digelu (1) Unknown (1)
W. Shewa TTKTF (1) Hidase (1)
TKTTF(2) Unknown (1), Kubsa (1)
TKPTF (1) Unknown (1)
E. Shewa TKKTF(4) Unknown (3), Mangudo (1)
TKTTF (3) Unknown (3)
TKKTF (9) Digelu (2) Unknown (7)
TKPTF (2) Unknown (1) Hidase (1)
Hararge TTKTT (1) Hidase (1)
TKKTF(2) Hidase (2)
TTKTT(14) Huluka (8), Senate (3), Shorima (3)
TTTTF(1) Kubsa(1)
Guji TTKTF(1) Shorima (1)
TKTTF(3) Unknown(3)
TTTTF(4) Danda’a (1),Digelu (1), Unknown (2),
TTKTT(1) Unknown
TKPTF(2) Danda’a (1), Unknown (1)
Wellega TKKTF(1) Triticale(1)
Unknown (12), Danda’a (1),Ownless (1), Hidase (4), Kakaba(2),
TKKTF(22) Mangudo (1), Kenya (1)
TKTTF(8) Unknown (5), Digelu(2), Alidoro (1)
TTTTF(7) Unknown (4), Danda’a (2), Kubsa (1)
N. Shewa TTKTF(2) Ownless (1), Unknown (1)
Amhara TKTTF(5) Unknown (5)
TTTTF(2) Unknown (2)
TKKTF (2) Unknown (1)
E. Gojjam TTKTT (1) Unknown (1)
W. TKKTF(1) Unknown (1)
[20]
Gojjam
TKTTF(1) Unknown (1)
S. Gonder TKKTF(1) Unknown (1)
TKKTF(1) local (1)
TKTTF (4) Unknown (3), Kakaba (1)
S. Wello TTKTT (1) Danda’a (1)
TKTTF(14) Unknown (3), Wane (1), Kakaba (6), Lemu (1), Ogolcho (1), Danda’a (2)
Silte, TTTTF(15) Ogolcho (1), Kakaba (5), Danda’a (5), Dashen (1), Messebo(1), Hidase (2)
SNNP TKKTF(10) Hidase (5), Kingbird (1), Kakaba (3), Danda’a (1)
Werabe &
Hadiya TTKTF(1) Unknown( 1)
TTTTF(1) Unknown (1)
TKTTF(1) Kakaba (1)
Afar
TKKTF(1) Kakaba (1)
Werer TTKTF(1) Kakaba (1)
Tigray S. Tigray TKKTF(1) Kakaba (1)
Table 3. Stem rust races identified in the surveyed regions in 2018 cropping season
Samples Races
analyzed
Regions TTTTF TKTTF TKKTF TTKTF TTRTF TTKTT TKPTF
Oromia 228 63 69 55 6 5 18 12
Amhara 60 11 19 26 2 - 2 -
South 40 15 14 10 1 - - -
Afar 4 1 1 1 1 - - -
Tigray 1 - - 1 - - - -
Table 4. Virulence spectrum of the Pgt races identified in Ethiopia in 2018 main cropping season
Races Virulence Avirulence

TTTTF 5, 21, 9e, 7b, 11, 6, 8a, 9g, 36, 9b, 30, 17, 9a, 9d, 10, Tmp, 38, McN 24, 31,
TKTTF 5, 21, 9e, 7b, 6, 8a, 9g, 36, 9b, 30, 17, 9a, 9d, 10, Tmp, 38, McN 11, 24, 31
TKKTF 5, 21, 7b, 6, 8a, 9g, 9b, 30, 17, 9a, 9d, 10, Tmp, 9e, 38, McN 11, 36, 24, 31
TTRTF 5, 21, 9e, 7b, 11, 6, 8a, 9g, 36, 9b, 17, 9a, 9d, 10, Tmp, 38, McN 30, 24, 31
TKPTF 5, 21, 9e, 7b, 6, 8a, 9g, 36, 30, 17, 9a, 9d, 10, Tmp, 38, McN 11, 9b, 24, 31
TTKTF 5, 21, 9e, 7b, 11, 6, 8a, 9g,9b, 30, 17, 9a, 9d, 10, Tmp, 38, McN 36, 24, 31
TTKTT 5, 21, 9e, 7b, 11, 6, 8a, 9g, 9b, 30, 17, 9a, 9d, 10, Tmp, 24, 31, 38, McN 36

Seven races were identified from 333 samples analyzed in the country. Of these
races, TKTTF and TKKTF were dominant races with frequencies of 28% each
followed by the highly virulent race TTTTF (27%). The present study was the first
confirmation of Pgt race TTKTT in Ethiopia. This race is Ug99 related race and
virulent on Sr24, the valuable source of resistance that has remained effective in
Ethiopia for a long time. Moreover, the result highlighted further spread of Ug99
race groups and the need for close monitoring.
[21]
In the present study, all the Sr genes in the differential lines were ineffective
against one or more races detected in the country indicating the need for
developing cultivars with combinations of effective resistance genes to enhance
their longevity. Alternatively, cultivars with multiple minor genes to achieve
durable resistance could be developed. The current result also emphasizes the
importance of regular monitoring to timely identify new races, and utilize this
information in screening and identification of effective sources of resistance.
Moreover, race analysis work with additional Sr genes need to be done to confirm
the correlation between phenotypic and genotypic variations.
Acknowledgements
The authors are greatly indebted to the Delivering Gentic Gain in Wheat (DGGW)
and Real-time and seasonal forecasting of wheat rust epidemics to inform
surveillance and control: Ethiopia as a LMIC test case (BBSRC) projects for
financial support of the study. The Authors thank technical staff of wheat research
section of Ambo Agricultural Research Center for their valuable technical support
during the whole period of the study.
References
AARC, 2017 APPRC. 2017. Ambo Plant Protection Research center. Plant Pathology Department
Progress Report for the 2016-2017. Pp.24- 60, Ambo.
Admassu B, Lind V, Friedt W, Ordon F, 2009. Virulence analysis of Puccinia graminis f.sp. tritici
populations in Ethiopia with special consideration of Ug99. Plant Pathology 58, 362-69.
Belayneh A and Emebet F. 2005. Physiological races and virulence diversity of Puccinia graminis
f.sp. tritici on wheat in Ethiopia. Phytopathologia Mediterranea 44:313-318.
Fetch Jr TG and Dunsmore KM. 2004. Physiological specialization of Puccinia graminis on wheat,
barley, and oat in Canada in 2001. Canadian Journal of Plant Pathology 26: 148-155.
Gomez KA and Gomez AA. 1984. Statistical procedures for agricultural research. 2nd ed. John
wiley and sons. New York. 680pp.
Hailu E, Woldeab G, Denbel W, Alemu W, Abebe A, Mekonnen A. 2015. Distribution of stem rust
(Puccinia graminis f. sp. tritici) races in Ethiopia. Plant 3, 15-9.
Hei NB, Tsegaab T, Getaneh W, Endale H, Bekele H, Daniel K, Fikirte Y, Fufa A, Wubishet A,
Teklay A, Miruts L, Alemar S, Tesfaye G. 2018. Distribution and frequency of wheat stem
rust races (Puccinia graminis f. sp. tritici) in Ethiopia. Journal of Agricultural and Crop
Research 6(5)፡ 88-96.
Jin Y, Szabo L, J Pretorius Z, A Singh, RP Ward R and Fetch T. 2008. Detection of virulence to
resistance gene Sr 24 within race TTKS of Puccinia graminis f sp. tritici. Plant Disease 92:
923-926.
Park RF. 2007. Stem rust of wheat in Australia. Australian Journal of Agricultural Research
58:558-566.
Patpour M, Hovmoller M, Justesen AF, Newcomb M, Olivera Firpo PD, Jin Y. 2016. Emergence
of virulence to SrTmp in the Ug99 race group of wheat stem rust, Puccinia graminis f. sp.
tritici, in Africa. Plant Disease 100: 522
[22]
Roelfs AP and Martens JW. 1988. An international system of nomenclature of Puccinia garaminis
f.sp. tritici. Phytopathology 78: 526-533.
Roelfs AP, Singh RP and Saari EE.1992. Disease of wheat: Concepts and method of disease
management. CIMMYT, Mexico City. 150pp.
Stackman EC, Stewart DM, Loegering WQ. 1962. Identification of physiologic races of Puccinia
graminis var. tritici.’ USDA ARS, E716. United States Government Printing Office:
Washington, DC. pp 5-50.
Stubbs RW, Prescott JM, Sarrri EE, Dubin HJ. 1986. Cereal Disease Methodology Manual.
CIMMYT, El Batan, Mexico.
[23]
[24]
Survey and Identification of Major Diseases of
Black Cumin (Nigella sativa L.) in Ethiopia
Merga Jibat1, Wakjira Getachew2, Habetewold Kifelew3, Abukiya Getu1
1
Ethiopian Institute of Agricultural Research (EIAR), Tepi Agricultural Research Centre, P.O.Box 34, Tepi,
Ethiopia; 2EIAR, Jimma Agricultural Research Centre, P.O.Box 192, Jimma; 3EIAR,Holeta Agricultural
Research Centre, P.O.Box 2003, Addis Ababa.
Abstract
Black cumin plays an important role in food flavors, perfumes and preparation of
medicine. However, diseases are major limiting factors for the cultivation and
production of this crop in many agro-ecologies of Ethiopia. This study was aimed to
identify major diseases of black cumin in different agro-ecologies of Ethiopia. A
survey was conducted in major black cumin growing areas of Ethiopia during 2016
and 2018 main cropping season. Disease assessments were made by using 0.5m x
0.5m quadrate laid along diagonal of the fields. Moreover, plants with in the
quadrants were thoroughly examined from base to the apex for disease incidence and
severity. The major cumin diseases observed wilt (Fusarium oxysporum) (0-72%),
blight (Alternaria burnsii) (0-88%), and powdery mildew (Erysiphe polygoni) (0-
60%) at moderate to severe level. Plant parasitic Orobanche was observed as the
emerging problem in black cumin.
Introduction
Black Cumin (Nigella sativa L.) is a member of Apiaceae (Umbelliferae) family.
This species is originated in Egypt and East Mediterranean, but is widely
cultivated in Iran, Japan, China, Turkey and Ethiopia (Shewaye, 2011). Black
Cumin has a long history of uses for food flavors, perfumes and medicinal values.
Its oil has been used for good smell to some medicines, sterilizing of surgical
operation fiber, production of some veterinary and agricultural medicines and
plastic components (Aminpour and Karimi, 2004).
Seeds of black cumin have an aromatic odor and bitter taste. They are used as an
essential ingredient in soup component, sausages, cheese, cakes and candies. The
different variety of black cumin seed accumulate up to 50% thymol, a monocyclic
phenolic compound. The presence of this compound makes black cumin valuable
source for health care Industry (Black et al., 2005) and medicinal purposes
(Ashraf and Orooj, 2006). In Ethiopia, it is commonly used in the preparation of
local spice called in Amharic "Berbere", in which it tends to reduce its hotness
(Hedberg et al., 2003), for preparation of curries, bread, traditional beverage such
as “katikala or Areke”, traditional Ethiopian stews "Wot" and preservation of
butter (Jansen, 1981). However, diseases are major limiting factors for the
cultivation and production of black cumin in many agro-ecologies of Ethiopia.
[25]
In agricultural production, pest control is one of the most challenges for the
farmers throughout the world. It causes a serious damage to crops by competing
for resource, damaging plant and plant parts via other chemical effects. In
addition, the pest increases cost of production, reduces quality and yields of crops.
Survey of the status of diseases in crops is essential to determine general levels of
crop health, or the presence of particular diseases of quarantine significance,
prioritization of problems to enable proper allocation of resources, and to assess
the crop losses due to particular diseases.
Yet, there was no research based information on the major disease of black cumin
in Ethiopia. Therefore, this study was carried out with the objectives to provide
baseline information on diseases associated with black cumin in Ethiopia and
prioritize economically important ones for further research.

A survey was conducted in major black cumin growing areas of Ethiopia during
2016 and 2018 main cropping season in East Showa (Ada'a, Chafe donsa, and
akaki), Arsi (Hela-Zambaba, Hela-Tareta, Zambaba-Hela, Birbof -Chole) and Bale
zones (Goro, Ginnir and Gololcha districts) of Oromia regional state. Disease
samples were collected for isolation and identification of pathogens, and fields
were assessed for incidence of major diseases of black cumin. Within selected
fields a quadrant of 0.5m x 0.5m was thrown and disease incidence was recorded
for every quadrant diagonally across the fields. Data were recorded for wilt,
blight, powdery mildew and other diseases.
Disease incidence was recorded as percentage of infected plants in a field for wilt,
while foliar diseases such as blight and powdery mildew were scored on 0-4 scale,
where 0= completely free from disease, and 4 >75% of the area infected (Uppal et
al., 1938). Other information like crop stage, cultivar, cropping sequence and
pesticides applied were also taken into consideration in each district. Detailed of
study area for black cumin diseases are given in table 1.

The major disease of black cumin observed on farmers' field were wilt (Fusarium
oxysporum) (0-72%), blight (Alternaria burnsii) (0-88%), and powdery mildew
(Erysiphe polygoni) (0-60%) in moderate to severe level. Prevalence of Orobanche
(parasitic plant) was also observed in few fields, these can be considered as the
emerging problems in black cumin. During the survey, the crop was at different
growth stages of ranging from flowering to grain maturity. Wilt, powdery mildew
and blight diseases were observed at the status of moderate to severe in one or
[26]
the other surveyed areas. The district based prevalence of different diseases is
presented in table 1.
Table 1. Distribution of black cumin diseases and plant parasitic weed ‘orobanche’in different production areas during
2016 and 2018 main cropping seasons
.
Variety
Zone Districts grown Disease recorded and pathogen identified
Chafe Donsa Dershaye Wilt(Fusarium oxysporum), Blight(Alternaria burnsii),
East Showa Powdery mildew(Erysiphe polygoni)
Akaki Eden Wilt(Fusarium oxysporum), Blight(Alternaria burnsii),
Powdery mildew(Erysiphe polygoni)
HelaZambaba Darbera Wilt(Fusarium oxysporum)
Arsi HelaTareta Local Wilt(Fusarium oxysporum), Blight(Alternaria burnsii)
Birbof Cole Local Wilt(Fusarium oxysporum), Blight(Alternaria burnsii)
ZambabaHela Local Wilt(Fusarium oxysporum), Blight(Alternaria burnsii)
Ginnir Eden Wilt(Fusarium oxysporum), Blight(Alternaria burnsii),
Bale Powdery mildew(Erysiphe polygoni)
Goro Local Wilt(Fusarium oxysporum)
Gololcha Local Wilt(Fusarium oxysporum)
Incidence of wilt caused by Fusarium oxysporum was recorded in almost all fields
of each rural community surveyed and varied from 0 to 72% with a mean
incidence of 2.9 to 32.6% in different districts of surveyed area (Table 2). The
second highest disease incidence was recorded for blight caused by Alternaria
burnsii on foliage where the percent disease incidence varied from 0 to 88% with
a mean score of 2.1 to 35.8%. Powdery mildew caused by Erysiphe polygoni was
observed in some areas, and percent disease incidence ranged from 0 to 60% with
mean score of 13.9-42%. This result was in agreement with Gaur (2005), who
reported the existence of wilt disease in black cumin caused by Furasium species.
On the basis of the specificity of the wilt pathogen to its host, Patel et al.,(1957)
identified the pathogen as Fusarium oxysporum f. sp. cumini. During the survey,
it was noted that early sown black cumin matures early and escapes powdery
mildew. The incidence of blight was severe in most of fields surveyed during 2018
as compared to 2016, perhaps due to variation in the current environmental
conditions. Alternaria blight of black cumin was first reported from Bombay in
1930 and the causal organism was identified as Alternaria burnsii (Uppal et
al.,1938). In India Erysiphe polygoni is the most important causative of powdery
mildew diseases in seed spices including black cumin (Uppal and Desai, 1933).
[27]
Table 2. Prevalence of major cumin diseases in different production areas of Ethiopia during the 2016 and 2018 main
cropping seasons
Zone District Willt Blight Powdery mildew

Mean Range Mean Range Mean Range
East Showa Chafe- Donsa 7.6 1.7-21.8 29.3 0.75 16.6 3.5-50
Akaki 9 0-10 20.1 0-65 - 4-25
Hela-Zambaba 32.6 0-18 18.6 0-54 13.9 0-29.5
HelaTareta 7.7 0-39 27.5 0-88 - -
Arsi Birbof- Chole 4.9 0-42 35.8 0-71 - -
Zambaba-Hela 12.9 5-30 30.2 0-52 - -
Ginnir 22.6 0-72 2.1 0-16 - -
Bale Goro- - - 24.8 14-38 42 20-60
Gololcha 2.9 0-17 4.2 0-12 - -
The collected information showed wide spread occurrence of wilt and blight in
Akaki and Chafe-Donsa districts of East Showa; Hela-Zambaba, Hela-Tareta,
Zambaba-Hela, and Birbof Chole districts of Arsi zone; and Goro, Ginnir and
Gololcha districts of Bale Zone in moderate to severe level. Whereas, powdery
mildew appeared only in few districts surveyed. Few fields were also infested with
parasitic weed 'Orobanche' in Hela-Zambaba and Hela-Tareta districts of Arsi
zone. The two air-borne diseases, powdery mildew and blight, are highly weather
dependent, hence integrated disease management strategy should be developed to
combat them. The emerging problem of Orobanche also needs proper attention to
understand the host-pathogen interaction and development of integrated
management strategy to control its spread in wide areas with time.

An extensive survey was conducted to identify diseases associated with black
cumin in Ethiopia. It is important to give priority to develop new varieties resistant
to single or multiple diseases identified as important with desired qualities. The
production technology should give attention on cultural practices which may
suppress the pathogens and diseases caused by them. Eco-friendly disease
management system needs to be developed primary to the integrated approach.
Acknowledgments
The authors thank the Ethiopian Institute of Agricultural Research (EIAR) and
Tepi Agricultural Research Centre for financing and providing all the necessary
facilities and support. Sincere thanks also extended to Debrezeit, Kulumsa and
Sinana Agricultural Research Centers, spice crops research team for their kind
support during data collection.
[28]
References
Aminpour and Karimi. 2004. Underutilized medicinal spices. Spice India. 17: 5-7.
Ashraf M, Orooj A. 2006. Salt stress effects on growth, ion accumulation and seed oil
concentration in an arid zone traditional medicinal plant ajwain (Trachyspermum ammi L.
Sprague). J. Arid Environ., 64:209-220.
Black M, Bewley D, Halmer. 2005. The Encyclopedia of seed science, technology and
uses.wallinoford. CAB P 7.
Gaur M M. 2005. Plant protection work in Ajmer Merwara in 1948 Plant Diseases. Plant
Protection Bulletin Govt. of India1:20-21.
Hedberge I, Edwards S, SileshiNemomissa. 2003. Flora of Ethiopia and Eriteria.4: 2, Apiaceae to
Dipsaceae. The Natural Herbarium.Addis Ababa University, Addis Ababa and Uppsala.P. 21.
Mogessie A and Tetemke M. 1995. Some microbiological and nutritional properties of Borde and
Shamita. Traditional Ethiopian fermented beverages. Ethiop. J. Health Dev. 9:105-110.
Patel PN, Prasad N, Mathur RL and Mathur BL. 1957. Fusarium wilt of cumin. Current Science26:
181-182.
Shewaye L. 2011. Antifungal Substances from Essential Oils. M.Sc. Thesis. Addis Ababa,
University. 8.
Uppal BN. 1930.Macrosporium blight of Cuminum cyminumin the Bombay Presidency. Internal
Inst Agric Internal Bull Plant Protect4.
Uppal BN and Desai MK. 1933.Cumin powdery mildew in Bombay, Bombay Dept.Agric.Bull169:
1-16.
Uppal BN, Patel MK, Kamat M N. 1938.Alternaria blight of cumin.Indian Journal ofAgricultural
Sciences8: 49-62.
[29]
[30]
Major Diseases of Coriander (Coriandrum
sativum L.) in Different Growing Areas of Ethiopia
Merga Jibat1, Wakjira Getachew2 and Abukiya Getu1
1
Ethiopian Institute of Agricultural Research (EIAR), Tepi Agricultural Research Centre, P.O.Box 34, Tepi,
Ethiopia; 2EIAR, Jimma Agricultural Research Centre, P.O.Box 192, Jimma.
Abstract
Coriander (Coriandrum sativum L.) is an important annual herb used extensively all
over the world. In Ethiopia it is intensively cultivated in various districts of East
Showa, Arsi, and Bale administrative zones. A survey to determine the identity, status
and economic importance of diseases associated with coriander was carried out in
major growing districts of the three zones covered. The result showed that root rot and
wilt are important diseases prevalently occurring and causing yield loss in most of
coriander growing areas assessed in Ethiopia. Highest incidences of 49.5% and 14%
were recorded, respectively, for root rot and wilt in Arsi zone. Fusarium solani and
Fusarium oxysporum were isolated from the diseased samples and found to have
associated with root rot and wilt, respectively, based on their cultural and
morphological characteristics. These diseases are causing a direct loss in crop
productivity with an incidence of root rot ranged from 24-49% and wilt ranged from 3-
14%.
Introduction
Coriander (Coriandrum sativum L.) is an annual herb prominently used as a leaf
and seed spice. Since from the ancient period, it was used as an important
ingredient of different food. Coriander is popularly used in soups, salads,
seasoning and chutney all over the world. It is cultivated in India on area of 552.7
thousand hectares and productivity of 0.8 metric tons per hectare in 2014/15
(Spice Board, 2015). In Ethiopia, the production and exports of coriander have
been increased significantly in the past few years due to strong demand from the
local and export markets. But, the crop is highly affected with soil borne diseases
which causes heavy yield loss every year. These pathogens are microscopic,
hidden and unevenly distributedin the soil or in infected plant material. These
pathogens enter through roots and become systemic causinga broad range of
diseases on various hostplants; such as vascular wilts, pre- and post-emergence
blights as well as root and stem rots (Pascale et al., 2002; Schollenberger et al.,
2006). The disease symptoms were observed in scattered patches in the cultivated
fields. Soil borne pathogens cause a major damage to crop and cause 10 per cent
yield loss on Coriander (Muthulakshmi et al., 2002). The present investigation
was undertaken to detect pathogen (s) associated with Coriander, understand the
status and economic importance of the disease (s) caused by them in the major
growing areas of Ethiopia.
[31]
A survey was conducted during 2016 and 2018 main cropping season to recordthe
diseases incidence in coriander growing areas of Ethiopia (East Showa, Arsi and
Bale zones). Fields were assessed to determine the incidence of major diseases of
coriander and identify the causal pathogen(s). Within selected fields, a quadrant of
0.5m x 0.5m was thrown and disease incidences were taken for every quadrant
diagonally across the fields and thenumber of plants showing typical root rot and
wilt symptoms and the total number of plants were recorded. Percent disease
incidence was calculated using the following formula.
Disease Incidence (%)
Isolation and identificationof the pathogen

The infected portions of plants showing symptoms of root rot, vascular
discoloration and premature dried plants were selected for tissue isolation and cut
into 3mm small pieces, washed with sterile distilled water, surface sterilized with
0.1 percent sodium hypochlorite solution for 60 seconds. The samples were
subsequently washed three times in sterilized distilled water, blot dried and then
transferred to sterilized Petri-dishes containing potato dextrose agar (PDA) media.
The plates were then incubated at roomtemperature (28±10C) and observed
periodically for fungal mycelial growth. Hyphae from thesebits were again
purified and pure cultures weremaintained on PDA slants. The identities of these
cultureswere confirmed through microscopic examinations and by referring
toliteratures.

Incidence of root rot and wilt diseases were recorded during survey in most of the
coriander growing areas of Ethiopia (Table 1). The percent disease incidence in
the surveyed locations ranged from 24 to 49%. Commonly encountered symptoms
include yellowing of leaves followed by vascular discoloration and drying of tap
root. Initially, the tap root of infected plants showed a reddish brown discoloration
which later become larger and darker. In severe stage of infection, the death of
plants was observed.
During survey, majorly two fungal pathogens viz. Fusarium solani and Fusarium
oxysporum were isolated from the diseased samples. Pure cultures were obtained
with hyphal tip isolation technique. Pure culture of Fusarium spp. in Petriplate
looked off white in color and under compound microscope, hyphae looked white
to creamy aerial mycelia with abundant chlamydospores in single or in pairs on
[32]
terminal either solitary or sometimes in chain (Figure 1). The pathogens were
identified based on their morphological and cultural characters (Bhaliya and
Jadeja, 2014).
Table 1. Incidences of root rot (Fusarium solani) and wilt (Fusarium oxysporum) diseases of coriander at major growing
areas of Ethiopia.
S/no Zone Districts Disease incidence (%)

Fusarium solani Fusarium oxysporum
1 East showa Chafedonsa 25.16 5
Akaki 32.28 7.2
Debrezeit 28.2 5.8
Mean 26.1 3.25
2 Arsi Hela-Zambaba 49.5 14
Hela-Tareta 24.23 11.3
Zambaba-Hela 43.5 5.2
Mean 39.36 11.56
3 Bale Goro 34.56 6.8
Ginnir 44.52 7.8
Gololcha 28.72 8.9
Mean 34.54 8.21
Morphological view of the cultures
Figure 1. Morphological and Microscopic characteristics of Fusarium isolates from coriander, M= Macroconidia;
m=Microconidia; C=Chlamydospore
Highest disease incidence of root rot (49.5%) and wilt (14%) was observed in Hela-
Zambaba district. High disease incidence and severity in this district may be
[33]
attributed to the use of infected planting materials or growing in contaminated
soils or both along with conducive environmental conditions favoring pathogen
multiplication. Least disease incidence of root rot (24.23%) and wilt (5%) was
observed in Hela-Tareta and Chafe-donsa districts, probably because farmers in
this district may use pesticides and the area may be free of soil borne dormant
chlamydo spores.

In general, root rot and wilt disease caused by Fusarium solani and Fusarium
oxysporum, respectively, were identified in coriander and are prevalent in some
districts surveyed causing greater yield loss. This is the first survey report on
incidences of root rot and wilt diseases in coriander growing areas of Ethiopia. It
is therefore imperative to suggest that integrated disease management options
should be sought in the future research before the diseases getting a significant
coriander production threat all over the growing areas in the country.
Acknowledgements
The authors thank the Ethiopian Institute of Agricultural Research (EIAR) for
financing this study. Thanks also go to all Tepi, Kulumsa, Sinana and Debrezeit
Agricultural Research Center Spice Crops Research Section for their assistance
during survey and data collection. Authors are also very grateful to peoples and
daily laborers who closely assisted during the laboratory analysis.
References
Bhaliya CM and Jadeja KB. 2014. Efficacy of different fungicides against Fusariumsolani causing
coriander root rot. The Bioscan, 9(3), pp.1225-1227.
Muthulakshmi P, Chezhiyan N, Muthukrishnan K and Doraisamy S. 2002. Management of
coriander wilt using biocontrol agents. Journal of Spices and Aromatic Crops, 11(2), pp.138-
140.
Pascale M, Visconti A and Chelkowski J. 2002. Ear rot susceptibility and mycotoxin
contamination of maize hybrids inoculated with Fusarium species under field conditions.
In Mycotoxins in Plant Disease (pp. 645-651).Springer, Dordrecht.
Schollenberger M, Müller HM, Rüfle M, Suchy S, Plank S and Drochner W. 2006. Natural
occurrence of 16 Fusarium toxins in grains and feedstuffs of plant origin from
Germany. Mycopathologia, 161(1), pp.43-52.
Spice Board. 2015. Spices Board, India Ministry of Agriculture and Farmers Welfare, Govt. of
India.
[34]
Evaluation of Trichoderma Species as
Biocontrol Agents against Fusarium Wilt of
Rosemary and Sage
Mihiret Mekonnen1 and Begashaw Manahlie2
1
Wondo Genet Agricultural Research Center (WGARC), Ethiopian institute of Agriculture Research,
P.O.Box.198, Shashemene, Ethiopia; 2Debre Berhan University, College of Natural and Computational
Science, Department of Biotechnology, Debre Berhan, Ethiopia
Abstract
Rosemary and sage, aromatic and medicinal plants in Lamiaceae family, are
economically important crops all over the world. Fresh and dried leaves of rosemary
and sage have been used in cosmetics, perfumery and pharmaceutical industries and
for food flavoring and herbal tea throughout the world. However, these economically
important crops suffer from Fusarium wilt (F. oxysporium), one of the most
economically important diseases of both plants. The aim of this study was to examine
the efficacy of Trichoderma species against Fusarium wilt of rosemary and sage in
vitro and in vivo. In this study, six Trichoderma species were examined for their
efficacy of controlling Fusarium wilt of rosemary and sage. Trichoderma species
namely T. harzianum, T. hamatum, T. viride, T. asperillium, T. artroviride and T.
longibrachiatum inhibited the mycelial growth of F. oxysporium, isolated from
rosemary, by 23.90, 17.57, 19.70, 17.33, 16.50 and 19.07%, respectively. Similarly,
the above antagonists inhibited the growth of the pathogen isolated from sage by
24.57, 19.90, 22.00, 16.67, 17.90 and 21.07%, respectively in a separate in vitro
experiment. On the other hand, application of T. harzianium, T. longibrachiatum and
T. viride in vivo significantly reduced (p < 0.05) Fusariums wilt of both rosemary and
sage. Maximum disease control of 79.17 and 65% were achieved by T.
longibrachiatum on rosemary and sage, respectively. Generally, T. harzianium
revealed the highest pathogen inhibition effect in vitro followed by T. viride and T.
longibrachiatum. In vivo application of T. viride, T. harzianum and T.
longibrachiatum on rosemary and sage caused significant difference (p < 0.05%) in
plant height, fresh leave weight, fresh stem weight and essential oil yield compared to
the control. Hence, these Trichoderma species can be used as potential biocontrol
agents against the fusarium wilts in combination with other management options.
Introduction
Rosemary (Rosmarinus officinalis L.), a Lamiaceae family, an ornamental and
medicinal plant, is a very significant crop all over the world. Both fresh and dried
leaves of rosemary have been used for its characteristic aroma in food cooking or
consumed in small amount as herbal tea, while rosemary extracts are routinely
employed as natural antioxidant to improve the shelf life of perishable foods. The
other major use of rosemary is in the perfumery and cosmetic industries where the
essential oils are employed as natural ingredients of fragrances (Oji-Ardabily et.
al., 2006). The essential oil can be distilled from the flowering tops; however,
large amount of commercial oil is distilled from the stem and leaves of the wild
[35]
plants before they get to the flower. Chemical products of Rosemary can also be
used for managing many human disorders such as headache, hair and skin
problems (Hosseinzadeh et al., 2006; Tironi et al., 2009).
Rosemary is native to Mediterranean and Asia. However, it is still reasonably

found around the world due to its many uses (Verhoeven et al., 2008; Türe et al.,
2009). In some places, it is cultivated to make drugs for controlling spasm and
emphysema disease, which reduces human peace (Hosseinzadeh et al., 2006).
Sage (Salvia officinalis) is an aromatic, rather woody perennial shrub in the mint
family (Lamiaceae). It is native to the shores of the northern Mediterranean and
eventually spreads to different countries (Lakusic et al., 2013). The plant grows
best in a well-draining, rich, clay loams and requires high intensity of sunlight.
The planting site should be warm, dry and protected from wind. Sage plants
should be pruned in early spring to promote new growth, or after flowering in the
summer.
S. officinalis is economically most important species being widely used in
medicine, cosmetic and food industry (Al-Tawaha et al., 2013; Grzegorcazyk and
Wysokinska, 2008). It is one of the most economically important herbs for its rich
essential oil and its plethora of biologically active compounds extensively used in
folk medicine (Aziz et al., 2013). It is used as herbal tea, for food flavoring, in
cosmetics, perfumery and pharmaceutical industries throughout world (Chalchat et
al., 1998). It is generally known for its multiple pharmacological effects including
its antibacterial (Delamare et al., 2007; Kamatou et al., 2008), antiviral (Loizzo et
al., 2008), antioxidative (Kelen and Tepe, 2008), antimalarial (Kamatou et al.,
2008), anti-inflammatory (Baricevic et al., 2001), antidiabetic (Eidi et al., 2009)
activities.
However, these economically important aromatic and medicinal plants suffer from
Fusarium wilt caused by F. oxysporium. This is one of the most economically
important diseases on both plants at different experimental fields of Wondo Gent
Agricultural Research Center. For example, this disease causes remarkable yield
reduction (50-60%) in sage plant at Wondo Gent Agricultural Research Center
experimental field. The disease is characterized by dropping of leaves, yellowing
and drying of leaves from the base upwards, improper branching, browning of
vascular bundles, and wilting and premature death of infected plants. Fusarium
wilt is a soil born disease that can be introduced into crop fields by contaminated
seeds or infected seedlings. Once introduced, the fungus can survive for many
years in the soil and invade the plants through the fibrous root system and disrupt
water and mineral uptake within the plant. Infection and disease development are
favored by warm soil temperature and low soil moisture.
[36]
Trichoderma species, one of the common fungal biocontrol agents, is being used
worldwide for proper management of various foliar and soil borne plant
pathogens. Members of the genus Trichoderma are imperfect fungi, fast growing
in culture and produce numerous green spores. These fungi species occur
worldwide and are commonly found associated with plant roots, soil and plant
debris (Howell et al., 2003). These species have long been recognized as
biological agents to control various plant diseases. Since the first application in
1930s, Trichoderma species became popular biological agents to protect crops
against plant pathogens all over the world. The use of biocontrol agents, like
Trichoderma spp. are acclaimed as effective, ecofriendly and cheap, nullifying the
ill effects of chemicals (Harman et al., 2004). Trichoderma species have superior
capacity to mobilize and take up soil nutrients compared to other organisms
(Schirmbock et al., 1994). This study was aimed to examine the efficacy of
existing Trichoderma biocontrol agents against Rosemary and Sage Fusarium wilt
under laboratory and greenhouse conditions

Preparation of Trichoderma species
In this study, six species of Trichoderma species namely T. harzianium, T. viride,
T. hamatum, T. artroviride, T. longibrachiatum and T. asperillium were tested
against F. oxysporium which causes Fusarium wilt on Rosemary and Sage. The
species were selected because of their antagonistic activity against other pathogens
in other studies. The Trichoderma species were obtained from mycology research
Laboratory of Ambo Agricultural Research Center, Ethiopia. Each Trichoderma
species was multiplied on Potato Dextrose Agar (PDA) media separately and
stored in refrigerator at 4oC for further experimental use.
Isolation and preparation of F. oxysporium
Infected roots and leaves of Rosemary and Sage were cut into pieces (2-3 mm)
and surface sterilized with 10% sodium hypochlorite for 5-10 seconds. Then, the
sterilized plant pieces were washed three times with sterilized distilled water and
transferred aseptically on to PDA. The inoculated plates were incubated at 27±2oC
and observations were made daily for emergence of culture. After the
development of the fungal spores, stock cultures were prepared using PDA in test
tubes and stored in refrigerator at 4°C for further use.
In vitro antagonistic activity of Trichoderma species against the pathogen
Dual culture technique was used to test the antagonistic activity of Trichoderma
species against F. oxysporium. Nine-millimeter disc of ten days old cultures of
Trichoderma species and F. oxysporium were inoculated on PDA medium in
opposite sides one cm away from the edges of the Petri plate. The six
Trichoderma species were cultured with F. oxysporium separately each with three
[37]
replications and incubated at 25±30C. Percent inhibition over control was
calculated using formula developed by Vincent (1927) as follows:
PI = C-T x 100
C
Where, PI = Percent inhibition over contro; C = Growth of test pathogen with
absence of antagonist (mm); T = Growth of test pathogen with antagonist (mm)
Antagonistic activity of Trichoderma species against

the pathogen in green house
Antagonistic activities of three in vitro effective Trichoderma species against F.
oxysporium were evaluated on rosemary and sage in green house. Forest soil, sand
and FYM potting soil mixture in 1:1:1 ratio (w/w/w) was prepared, autoclaved for
2hrs and filled in earthen pots of 5kg capacity. Rosemary and sage seedlings were
planted separately in the pots. After 3 months, the seedlings were pulled out from
the pots and dipped in respective Trichoderma species formulation containing 108
spores/ml for 2hrs, ensuring that the roots alone were immersed in the inoculums.
Trichoderma treated seedlings were then transplanted in to new earthen pots filled
with sterilized potting soil mixture. Spore suspension of the pathogen containing
108 spores/ml was incorporated in the potting soil mixture, one day before
transplanting. Then, two inoculated seedlings were planted per pot with three
replications. Soil drenching with formulation of the pathogen was done at 15 th and
30th days after transplantation. The pots were maintained in greenhouse at 26 ± 2
°C and 50–60% relative humidity. F. oxysporium inoculated plants which were
not inoculated with Trichoderma species were used as control. The treatments
were arranged in completely randomized design (CRD).
Percent disease control and disease severity were recorded at fifteen days interval
from the first appearance of symptom till harvest. Disease severity was assessed
visually using 0 - 4 scale, where 0, 1, 2, 3 and 4 represent 0 to 24%, 25 to 49%, 50
to 74%, 75 to 99% and 100% (dead plant), respectively of wilted leaves. Plant
height, branch number, fresh leaf weight, fresh stem weight and essential oil yield
were recorded at the time of harvest. Essential oil was extracted by hydro
distillation as illustrated by Guenther (1972). Disease control was calculated using
the following formula:
% DC = C - T/C x 100; where, % DC = percent disease control; C = control; T = treated
Data were statistically analyzed using analysis of variance (ANOVA) and
difference between means was assessed using Duncan’s Multiple Range Test at
5% probability level using SAS version 9.3.
[38]
In vitro antagonistic activities of Trichoderma species against F. oxysporium
The results of dual culture tests revealed that all Trichoderma species significantly
(p < 0.05) inhibited the mycelial growth of F. oxysporium. Clear zones of
inhibition were observed between the colonies of the pathogen and Trichoderma
species. The antagonists T. harzianium, T. hamatum, T. viride, T. asperillium, T.
artroviride and T. Longibrachatum, respectively, inhibited the mycelial growth of
F. oxysporuim isolated from rosemary by 23.90, 17.57, 19.70, 17.33, 16.50 and
19.07%. Whereas, the above antagonists, respectively, inhibited the growth of F.
oxysporuim isolated from sage by 24.57, 19.90, 22.00, 16.67, 17.90 and 21.07% in
a separate experiment (Table 1, Figure 1). The inhibition zone could be due to the
effect of diffusible inhibitory volatile and non-volatile compounds such as
tubercidin, candicidin, phosphlactomycin, phenasin and 4-diacetylphloroglucinol
produced by the Trichoderma species (Hwang et al., 1994; Ajith et al., 2010). In a
report by Ajith et al. (2010), volatile and non-volatile compounds produced by
Trichoderma species, T. saturnisporum, T. harzianum, T. viride and T. reesei were
studied against Colletotrichum capsici. The volatile compounds produced by these
selected Trichoderma species were found to show 30 to 67% inhibition of C.
capsici. Mohiddin et al. (2013) also reported that T. viride inhibited sclerotia
production of S. rolfsii by 48% and S. sclerotium by 78% and development of
mycelia by 40%. Other possible mechanisms of antagonism employed by
Trichoderma species includes nutrient and niche competitions, mycoparasitism,
hyphal interactions, and secretion of cell wall degrading enzymes such as lytic and
proteolytic enzymes (Ponnusamykonar et al., 2011; Vinale et al., 2008).
The respective Trichoderma species showed nearly comparable inhibition
percentage in rosemary and sage experiments. T. harzianium revealed the highest
inhibition effect followed by T. viride and T. longibrachiatum on F. oxysporium
isolated from rosemary and sage plants (Table 1). Similar in vitro experiments
conducted by Barari and Foroutan (2013) and Shukla et al. (2016) showed that T.
harzianum inhibited the radial mycelial growth of Fusarium solani by 68.22% and
S. rolfsii by 76.3%. In the present study, T. hamatum, T. asperillium and T.
artroviride achieved comparatively less inhibitory effects on mycelia growth of
the pathogen. On the other hand, full mycelia growth of the pathogen was
observed on the control after 10 days of culture (Figure 1). Some of the
antagonists tested in this study were not found to be very effective against the test
pathogen under the in vitro observations but they may show better result under
their natural field condition as their activities depend on the environment.
[39]
Figure 1. Inhibition effects of Trichoderma species on mycelia growth of F. oxysporium cultured on PDA from both
rosemary and sage leaves discs. The six Trichoderma species show varying degrees of effectiveness on
inhibition of mycelial growth of the pathogen. Full mycelial growth is demonstrated from untreated plants.
Table 1: In vitro growth inhibition effects of Trichoderma species against F. oxysporum on PDA
Experiment
Treatments (cultures) One (F. oxysporium from rosemary) Two (F. oxysporium from sage)
Inhibition Zone (%) Inhibition Zone (%)
T. harzianium +F. oxysporium 23.90 + 2.80a 24.57 + 2.60a
T. hamatum+F. oxysporium 17.57 + 1.77c 19.90 + 2.80b
T. viride +F. oxysporium 19.70 + 1.30b 22.00 + 1.20b
T. asperillium+F. oxysporium 17.33 + 1.33 c 16.67 + 1.66c
T. artroviride+F. oxysporium 16.50 + 1.60c 17.90 + 1.80c
T. longibrachiatum+F. oxysporium 19.07 + 1.20b 21.07 + 2.20b
Control (F. oxysporiumonly) 0.00 + 0.00d 0.00 + 0.00d
LSD 1.46 1.63
CV 2.178 2.179
Means with the same letter within the same column are not statistically different (P < 0.05).
[40]
Effectiveness of Trichoderma species against Fusarium
wilt of rosemary and sage in vivo
Application of the three in vitro best effective Trichoderma species, namely T.
harzianium, T. longibrachiatum and T. viride significantly reduced (p < 0.05)
Fusarium wilt of both rosemary and sage in greenhouse condition. Maximum
disease controls of 79.17 and 65% were achieved by T. longibrachiatum on
rosemary and sage, respectively (Table 2 and3, Figure 2). Despite the fact that T.
viride and T. harzianium were not quite as effective as T. longibrachiatum, both
antagonists significantly reduced disease severity on sage and rosemary compared
to the control. T. viride and T. harzianium demonstrated 48% and 45% disease
control efficacy, respectively, on sage while both species revealed comparable
disease control efficacy (58.33%) on rosemary. Disease severity level was high on
untreated plants with corresponding decrease in essential oil yield (Table 2 and 3,
Figure 2).
Successful control of Fusarium wilt in many crops by application of different

species of Trichoderma has been reported by many researchers (Shahriari and
Barari 2008a, b; Rinu et al., 2013; Ramezani, 2009). A study by Barari (2016)
indicated that application of Trichoderma antagonists was effective in suppressing
Fusarium wilt incidence of tomato from 14.75 to 20.15%. However, it is widely
known that environmental parameters such as abiotic and biotic factors as well as
method and timing of applications may have influence on the biological control
efficacy of Trichoderma species (Behzad et al., 2008). According to Vinale et al.
(2008) and Harman et al. (2004) the role of Trichoderma species is not only to
control the growth of pathogenic microbes, but also enhance plant defense
responses, stimulates colonization of rhizosphere and plant growth.
Significant differences (P < 0.05) were also observed between treated and
untreated rosemary plants for plant height and essential oil yield measurements
while there were significant differences (P < 0.05) between treated and untreated
sage plants for all measured parameters except plant height (Table 2 and 3).The
highest essential oil yield (52.62 g/p) was obtained from T. longibrachiatum
treated rosemary plants. Significantly much higher essential oil yields were also
recorded from T. viride (32.64 g/p) and T. harzianum (29.45 g/p) treated rosemary
plants compared to untreated (11.09g/p) plants. Similarly, maximum plant height
(79.75cm), branch number (30.25) and fresh leave weight (48.26g/p) were
recorded from T. longibrachiatum treated rosemary plants. Conversely, nearly
comparable fresh stem weights (32.87 and 31.26g/p) were recorded from T.
longibrachiatum and T. viride treated rosemary plants, respectively. No significant
difference was observed for branch number (27.50 and 27.00), fresh leave weight
(36.93 and 34.26g/p) and fresh stem weight (22.77 and 20.21g/p) between T.
harzianum treated and untreated rosemary plants. T. viride treated rosemary plants
[41]
showed significant yield performance compared to T. harzianum treated plants
except for branch number (Table 2).
Table 2: Effectiveness of Trichoderma species on Fusarium wilt control and yield parameters of rosemary under
greenhouse conditions
Treatment PH BN FLW FSW DS (%) DC (%) EOY
T. viride 69.33b 29.75ab 42.09b 31.26a 10b 58.33b 32.64b

T. harzianum 64.88c 27.50b 36.93c 22.77b 10b 58.33b 29.45c
T. 79.75a 30.25a 48.26a 32.87a 5c 79.17 a 52.61 a
Control 62.38d 27.00b 34.26c 20.21b 24a 0.00c 11.09d

LSD 2.0223 3.01 3.028 2.447 4.7559 9.1737 2.3332
Cv 2.45 2.446 2.446 2.14 2.44 2.44 2.446
Means with the same letter within the same column are not statistically different (P < 0.05). Where, PH = plant height; BN
= branch number; FLW = fresh leave weight; FSW = fresh stem weight; DS = disease severity; DC = disease control and
EOY = essential oil yield.
Similarly, application of T. viride and T. longibrachiatum on sage caused

significant difference in plant height (70.33 and 75.00 cm), fresh leave weight
(76.51 and 92.12 g/p), fresh stem weight (42.87 and 45.39 g/p) and essential oil
yield (40.34 and 41.82 g/p) compared to T. harzianum treated (63.33 cm, 57.23
g/p, 36.65 g/p and 25.23 g/p) plants, respectively. No significant difference was
observed in plant height, branch number, fresh stem weight and essential oil yield
among T. viride and T. longibrachiatum treated plants except for fresh leave
weight (57.23 g/p) (Table 3). However, higher branch number (34.37), fresh leave
weight (57.23 g/p) and fresh stem weight (36.65 g/p) were recorded from T.
harzianum treated plants compared to untreated (28.33, 40.29 g/p and 30.52 g/p,
respectively) plants. Whereas no significant difference was observed in plant
height and essential oil yield (63.33 cm and 25.23 g/p) between T. harzianum
treated and untreated (61.80 cm and 20.40 g/p) sage plants, respectively.
Table 3: Effectiveness of Trichoderma species on Fusarium wilt control and yield parameters of sage under greenhouse
conditions
Treatment PH BN FLW FSW DS (%) DC (%) EOY
T. viride 70.33a 38.50a 76.51b 42.87a 24.38b 48b 40.34a

T. harzianum 63.33b 34.37a 57.23c 36.65b 25.00b 45b 25.23b
T. longibrachiatum 75.00a 35.00a 92.12a 45.39a 18.33c 65a 41.82a
Control 61.80b 28.33b 40.29d 30.52c 38.50a 0.00c 23.40b
LSD 5.2725 4.8975 4.7457 4.9111 1.8829 6.629 5.7708
CV 2.447 2.44 2.446 2.447 2.446 2.44 2.446
Means with the same letter within the same column are not statistically different (P < 0.05). Where, PH = plant height; BN
= branch number; FLW = fresh leave weight; FSW = fresh stem weight; DS = disease severity; DC = disease control and
EOY = essential oil yield.
[42]
Sage T. longi Rosemary T. longi SageT.viridae
Rosemary T.viridae Sage T. harzianum Rosemary T. harzianum
Sage Control Rosemary control
Figure 2: Effect of Trichoderma species against Fusarium wilt of rosemary and sage in greenhouse condition. High
disease severity occurs in the pots planted with untreated seedlings, while seedlings treated with
Trichoderma species show better growth.
In the present study, plant height, branch number, fresh leave weight, fresh stem
weight and essential oil yield were increased in T. viride, T. harzianum and T.
longibrachiatum treated plants. Similar findings were reported on plant growth of
cereal and legume crops due to application of Trichoderma gamsii (Rinu et al.,
2013). The effectiveness of inoculation of tomato plants with Trichoderma species
in increasing of yield was reported by Shahriari and Barari (2008a, b). The
increase in plant growth and biomatter production may be due to the production of
plant growth promoters or through indirect stimulation of nutrient uptake and by
producing siderophore or antibiotics to protect plants from deleterious rhizosphere
organisms (Hwang et al., 1994).

From the present study, it can be concluded that the six in vitro tested
Trichoderma species inhibited the mycelia growth of F. oxysporium at different
level. The three in vitro selected Trichoderma species namely: T. harzianium, T.
longibrachiatum and T. viride significantly reduced Fusarium wilt of rosemary
and sage in greenhouse condition. Significant differences were also observed in
plant height, branch number, fresh leaf weight, fresh stem weight and essential oil
[43]
yield between Trichoderma species treated and untreated plants. Hence, all the
three Trichoderma species can certainly be used as potential biocontrol agents
against the fusarium wilt in integrated pest management. Additional efforts should
also be done for isolation of new Trichoderma species with high antagonistic
abilities against the soil borne pathogens as an alternative to synthetic pesticides.
Moreover, further actions are needed to prepare and apply formulations of highly
antagonistic Trichoderma species for the control of the disease.
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[46]
Collection, Isolation and Identification of
Entomopathogenic Fungi from Different Agro
Ecological Zones of Ethiopia
Solomon Yilma, Denberu Kebede and Tsegaye Mihrete
Ethiopian Institute of Agricultural Research, Ambo Agricultural Research Center,
P.O.box 37, Ambo, Ethiopia
Abstract
Entomopathogenic fungi have a high potential for biological control of insect pests.
Natural habitats are the most important resources for these micro-organisms. The aim
of this study was to isolate, identify and assess occurrence of entomopathogenic fungi
in the soil from different part of the country. Soil samples was taken from Wolega,
Horo Gudru, Jimma zone, Gamo Gofa, Dawuro, Kembata, Wolayta Sodo, Gurage, and
Silte, Wonji- Showa, Metahara and Finchaa between October and November of each
study year of 2016-2018. Entomopathogenic fungi were isolated from soil using
Galleria mellonella as bait trap (insect bait trap method). From the total 251 soil
samples collected, 89 fungal isolates were identified of which 50 of them were B.
bassiana and 39 were M. anisopliae. 61.9% of the identified Entomopathogenic fungi
were obtained from forest followed by farm land (28.08%). The abundance of
Metarihizium anisopliae and Beauveria bassinia in the forest and farm land are vice
versa. The availability of Metarihizium anisopliae in the forest are higher whereas
Beauveria bassiana in the farmland are more abundant. Evaluation of biodiversity and
pathogenicity of these isolates can help to develop effective biological agents to control
insect pest.
Introduction
Use of entomopathogenic fungi as biological control agents for insect species has
increased the global attention during the last few decades. The myco-insecticide
based on Beauveria bassiana Metarizihium anisopliae, Vaillemin (Babu et al.,
2001; Sharma, 2004), Paecilomyces fumosoroseus (Wize) Brown and Smith (Alter
and Vandenberg, 2000; Avery et al., 2004) and Verticillium lecanii (Zimm.)
Viegas (Butt et al., 2001) have been used to control various insect pests.
There is an increasing interest in the exploitation of Hyphomycetous fungi for the

control of invertebrate pests and diseases. Fungal biological control is an exciting
and rapidly developing research area with implications for plant productivity,
human health and food production. Entomopathogenic fungi (EPF) and several
taxa of the other fungi have demonstrated excellent suppression of insect pests in
green house conditions (McCoy et al., 1988; Ferron et al., 1991; Tanada and
Kaya, 1993; Inglis et al., 2001). Beauveria bassiana has recently been registered
against an array of greenhouse pests, including stem borers, aphids, thrips, white
flies and spider mites (Shah and Goettel, 1999).
[47]
In Ethiopia, research on entomopathogenic fungi that control of insect pests at
different commodity has been carried out during the past several years. According
to Tesfaye and Emiru (2010); Seneshaw et al. (2003); Tesfaye et al. (2012), the
Metarihizium fungus was isolated and identified from parts of different insects and
soil from various parts of Ethiopia including Alamata, Arba Minch, Metahara,
Wonji-Shoa and Finchaa. The ubiquitous fungus, Beauveria bassiana, was also
isolated and identified from parts of different insect specimens collected in
different parts of Ethiopia including Fura, Sekota, Wikro, Erer, Gusquam,
Debremarkos, Ashengie, Tikurinchini, Metahara, Wonji-Shoa and Finchaa
(Tesfaye and Emiru, 2010; Seneshaw et al., 2003; Tesfaye et al., 2012). However,
the potential of these bio-agents is not exploited well.
Various studies in Ethiopia involving laboratory experiments and small-scale

applications have demonstrated the effectiveness of fungal pathogens for the
control of spotted stem borer (Chilo partellus) and pink stem borer (Sesamia
calamistis Hampson) in Maize (Tadele & Pringle, 2004), Acanthoscelides
Obtectus (Julia & Lina, 2010), the greater wax moth (Galleria mellonella)
(Namusana & Emiru, 2010), Desert Locust (Seneshaw, Emiru and Dawit, 2003),
cotton/melon aphid (Tesfaye and Emiru, 2010) and pink stem borer (Tesfaye,
Selvaraj, Leul and Mulugeta, 2012). Almost all of these researches have been
undertaken with B. bassiana and Metarhizium anisopliae isolates of Ambo
Agricultural Research Center collections. Not only isolates of these collections are
small in number, but also weak in virulence due to poor preservation that limited
their mass production, formulation, shelf life determination and delivery for field
application. In Ethiopia, the locally isolated entomopathogenic fungi are not
extensively being in use as biopesticide, although the demand for
entomopathogenic fungi is increasing among research centers and universities.
This study was, therefore, undertaken to isolate and identify potential EPF from
different agro-ecological zones of Ethiopia for future use.

Collection of soil samples
Soil sample collection was carried out in an areas where the presence of
entomopathogenic fungi are earlier reported in Ethiopia, namely Wolega, Horo
Gudru, Jimma zone, Gamo Gofa, Dawuro, Kembata, Wolayta Sodo, Gurage Silte,
Wonji- Showa, Metahara and Finchaa between October and November of each
study year. From each surveyed area, 600g soil samples were taken at a depth of
5-10 cm by removing the most top surface soil (Tsay et al., 2006). A total of 251
soil samples were collected in ‘W’ shape pattern in every 1km intervals. The
collected samples were packed in plastic bags and transported to Ambo
Agricultural Research Center Bio-control Laboratory.
[48]
Isolation of EPF
Isolation of EPF was done using adult greater wax moth (Galleria mellonella)
obtained from Ambo Agricultural Research Centre, Ethiopia. Rearing was
performed in plastic boxes incubated in dark inside incubator at 20oC. In a flask,
adult moths (in 1:1 female to male ratio) were kept by providing honey and water.
In addition, folded tissue papers were placed for oviposition. The papers were
removed with the eggs attached and placed in new plastic jar with feed ingredient
(Meyling, 2007). The feed composition was 180 g honey, 180 g glycerine and 50
g wheat bran. First honey and glycerine were melted in a cooking pot and mixed
thoroughly with wheat bran after 15 minutes. The feed and eggs attached on tissue
papers were transferred in to a bigger jar for rearing inside incubator at 20°C. The
feed was changed periodically based on the larval growth stage of G. mellonella.
Larvae of approximately 2.5-3 cm in length (4 weeks after hatching) were used for
baiting soil samples (Meyling, 2007).
Before inoculating in the soil, larvae were immersed in boiled water (at 56oC) for
10 seconds and transferred for cooling in a running bath water for 30 seconds in
order to prevent webbing. Third instar 10 G. mellonela larvae were placed into
small glass jars of about 500 ml (on average 300 g soil). Soils were placed on top
of the larvae until approximately 2/3 of the containers are filled and incubated at
22°c. The containers were inverted every day, so that the larvae continually have a
chance to move through the soil and repeatedly exposed to infective conidia. In
the course of the experiment, data on mortality were recorded daily for ten days.
The dead larvae were collected and submerged into 1% sodium hypochlorite, 70%
ethanol for one minute, respectively and washed in sterile distilled water for three
minutes to remove saprophytes and all conidia found on the outer surface of the
larvae body (Odindo, 1994). The disinfected cadavers (dead larvae) were allowed
on filter paper to dry for three minutes. This step were added to ensure that,
mycosis observed on the surface of the cadavers would not be attributed to spores
used during the treatment but rather to growth from the interior to the exterior of
the insect after colonization of internal organs. Cadavers were held under high
humidity on Petri dishes containing damp filter paper to provide sufficient humid
conditions to promote fungal outgrowth. Petri dishes were sealed with Para film to
maintain greater than 95% RH and incubated in the dark at 27°C. Larvae
considered mycosis when growth of the fungus is visible on the external surface
and those which showed hyphal growth characteristics of the EPF were recorded
as infected.
Purification of isolated EPF

Mortality of G. mellonela larvae were checked continuously for 10 successive
days after inoculating in the soil sample. Fungi samples outgrowing and
sporulating on the cadaver were cultured on artificial media (Sabouraud’s
[49]
Dextrose Yeast Agar, SDYA) and pure cultures were obtained through successive
transfer for identification. Tentative identification was done based on macro and
micro growth and morphological characters of fungi (Subramanian, 1971; Lacey,
1997; Sinishaw, 2002). Pure cultures of EPF were sub-cultured on to Sabouraud’s
dextrose agar with yeast extract (SDAY) and incubated at 27°C, 75% RH and
photoperiod of 12:12h light and dark for ten successive days till they were
developed to full culture and good sporulation.
Confirmatory test against G. melonella

The surface of ten-day old cultures were scrapped with a sterile scalpel and
suspended in aqueous solution of 0.01% Tween 80. The fungal suspension were
vortexed for one minute to break up the conidial chains or clumps and filtered
through several layers of sterile cheesecloth to remove mycelia. The dose of
conidia in the filtrate was estimated using haemocytometer under a light
microscope (40 x magnifications). Conidial suspension (1x107, 1x108 and 1x109
conidia per ml) were prepared for each isolate and applied on 2.5-3cm larvae of
Galleria melonella. Ten larvae inoculated with 1x108 ml-1 conidia were introduced
in each Petridish with filter paper. Sterile distilled water with 0.01% Tween 80
was used as free control treatment. The treated larval insects were incubated at 27
°C and 70 ± 5% RH and maintained for 10 days. The dead larvae were collected
and submerged into 70% ethanol for three seconds and 5% sodium hypochlorite
for two minutes (Odindo, 1994) and washed in sterile distilled water for three
minutes to remove saprophytes and all conidia found on the outer surface of
cadavers. The disinfected cadavers (dead larvae) were allowed to dry for ten
minutes on Watmann No.1 filter paper. Cadavers were held under high humidity
on Petridishes containing damp filter paper to provide sufficient humid conditions
to promote fungal outgrowth and sealed with Para film to maintain greater than
95% RH and incubated in the dark at 27°C. Larvae were considered mycosis when
growths of the fungus are visible on the external surface and those which showed
hyphal growth characteristics of the EPF were recorded as infected.

Entomo-pathogenic fungi isolates obtained from different parts of the country are
presented in Table 1. Two hundred fifty one soil samples were collected from
Wolega, Horo Gudru, Jimma zone, Gamo Gofa, Dawuro, Kembata, Wolayta
Sodo, Gurage, Silte, Wonji- Showa, Metahara and Finchaa between October and
November of each study year of 2016-2018. A total of 89 entomopathogenic fungi
were isolated from the soil collected from 10 zones using G. mellonella larvae as a
baiting trap. From the different agro-ecology of the country, forest found
relatively richer in entomopathogenic fungi composition followed by grazing land.
Similarly, study in Syria confirmed that EPF isolates were obtained from most
[50]
locations, the highest percentage of EPF identified was reported in soil collected
from forest (Almanoufi et al., 2012). In addition, apart from typical insect-
pathogenic fungi, some accompanying fungal species with unproved
entomopathogenic activities were isolated from investigated soils by means of
Galleria bait method.
Table 1. Sampling site and ecology of Entomopathogenic fungi
Site Ecology No of No of dead Galleria No of EPF isolates

Sample /positive sample/ recorded
West Showa Semi-forest 2 2 1
Sugarcane farm 18 16 2
Semi forest 10 9 3
Acacia 9 6 2
Maize farm 1 1 1
Semi forest 3 3 1
Acacia 6 5 2
Grazing land 1 1 1
Jimma zone Forest 19 13 11
Farm land 38 21 17
Grazing land 3 1 1
Horo Gudru Rod side 8 2 1
Grazing land 1 1 1
Farmland 2 1 1
Forest 6 2 2
Coffee farm 3 3 2
Banana farm 1 1 1
Mango farm 1 1 1
Shrub 1 1 1
Wooden land 4 2 1
West Wolega Rod side 6 2 1
Grazing land 4 2 1
Farmland 5 3 2
Forest 1 1 1
Chat farm 1 1 1
Niger farm 1 1 1
Wood land 7 3 2
Gamo Gofa Enset 19 9 7
Wolayta Enset 22 13 10
Dawuro Enset 12 7 3
Hadya Enset 10 4 1
Kembata Enset 6 2 1
Gurage Enset 5 3 1
Silte Enset 4 1 1
Total 251 149 89
Based on this study, EPF were dominantly identified from forest. Similarly, the
high proportion of infected cadavers of Galleria melonella by fungi obtained in
soil samples collected from forest habitats. This result is in parallel with findings
reported by Maryam et al. (2014) in Iran who claimed that EPF is high in soil
samples collected under Oak trees.
[51]
Following isolation of EPF using the Galleria bait method, identification was done
using morphological and cultural characteristics such as mycelial growth pattern,
color and spore shape aided by microscopic examination. As shown in figure 1,
the geographical distribution of EPF was high in in Jimma, East Showa, Horo
Gudru and Wolayta Sodo zones compared to other areas assessed. The abundance
of Beauveria bassinia in Jimma zone was higher than in the other sites covered,
while its availability is highly reduced in Gamo Gofa, wolayta, Dawuro, Hadya,
Kembata, Gurage and Silte.
Beauveria bassiania
Metarhizium anisopliae
Figure 1. Distribution of identified Entomopathogenic fungi from different locations studied in Ethiopia
The highest proportion (61.79%) of the isolated entomopathogenic fungi were from forest
followed by farm land (28.08%). Soils from the agricultural field (farm land) most
frequently harbored Beauveria bassiana (21.35%), while soil from the forest most often
contained Metarhizium anisopliae (33.7%) (Table 2). This result is in line with findings
reported by Meyling and Eilenberg (2005) who stated that soils from agricultural
field most frequently harboured B. bassiana while soil from the hedgerow most
often contained Paecilomyces fumosoroseus. However, B. bassiana was also common
in forest soil (28.08%). In the soils from farm land, Metarhizium anisopliae was less
frequently isolated than Beauveria bassiana. M. anisopliae was not found in soil samples
collected from the road side. The species detected in the present study were within
the expected range based on previous studies performed at similar ecology using
insect bait (Meyling and Eilenberg, 2005; Steenberg, 1995; and Chandler et al.,
1997. Grazing land and road sides had relatively higher proportion of Beauveria bassiana
than Metarhizium anisopliae. Generally, the proportion of EPF in soil samples from
uncultivated land is very low. This finding is substantiated by similar studies from Syria
that reported no EPF isolates were obtained from uncultivated lands assessed
(Almanoufi et al., 2012).
[52]
Table 2. Distribution of entomopathogenic fungi isolated from major ecological habitats
EPF isolates Habitat

Forest Grazing land Wood land Farm land Road side
Beauveria bassiana 25(28.08%) 3(3.4%) 1(1.1%) 19(21.35%) 2(2.25%)
Metarhizium anisopliae 30(33.7%) 1(1.1%) 2(2.25%) 6(6.74%) 0
Total 55(61.79%) 4(4.5%) 3(3.35%) 25(28.08%) 2(2.25%)
*Numbers in parenthesis indicates the proportion occurrence of EPF from the respective isolates
Conclusion and Recommendations

The study showed that the soils from forest and farm lands are characterized by
having higher composition of entomopathogenic fungi than of adjacent grazing
and road sides. Natural habitats are rich in biodiversity of entomopathogenic
fungi. Based on findings of this study, it is recommended that future research
work should focus in optimizing techniques for mass production and appropriate
formulation to keep the quality and guarantee large scale production and
application of EPF. Furthermore, as the biological control is important
components of integrated pest management, genetic diversity of indigenous
populations of B. bassiana and M. anisopliae should be studied in detail using
molecular markers.
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environment. Laboratory manual, January 2007. (http:// orgprints.org/11200)
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(ed). Techniques of insect rearing for the development of integrated pest and vector
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Seneshaw Aysheshim, Emiru Seyoum and Dawit Abate. 2003. Evaluation of Ethiopian Isolates of
Entomopathogenic Fungi as Potential Biological Control Agents of the Desert Locust,
Schistocerca gregaria. Pest Management Journal of Ethiopia. 7:1-9.
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Indian council of Agricultural. Research, New Delhi, India
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infectivity to Sitona species and other insects in Lucerne. Ph.D. Thesis. The Royal Veterinary
and Agricultural University, Denmark.
Tadele T and Pringle KL. 2003. Food consumption by Chilo partellus (Lepidoptera: Pyralidae)
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[54]
Chitinolytic Activity of Entomopathogenic
Fungi Collections from Ethiopia
Solomon Yilma and Denberu Kebede
Ethiopian Institute of Agricultural Research, Ambo Agricultural Research Center,
P.O. Box 37, Ambo, Ethiopia
Abstract
Chitin, a -1,4-linked polymer of N-acetyl-D-glucosamine, is one of the most
abundant polysaccharides in nature next to cellulose. It is a major cell wall
constituent of higher fungi belonging to chitridiomycetes, ascomycetes,
basidiomycetes and deuteromycets, insect exoskeletons and crustacean shells. The
present study aims to investigate the production of chitinase enzyme by
Entomopathogenic fungi of pervious collections of Ambo agricultural research
center and Desert Locust Control Organization (DELCO). Further, to elucidate
the effects of process parameters on chitinase production. EPF strains were
evaluated for chitinase production by carrying out enrichment in minimal salt
medium containing colloidal chitin as sole carbon source. The chitinolytic activity
of the strains was investigated by the ability of the isolate to produce zone of
clearance on colloidal chitin agar. A total of 16 Entomopathogenic fungi were
obtained from Ambo agricultural research center, biological control laboratory
and DELCO. The production of chitinase by Entomopathogenic fungi was
optimized using different growth media, substrate concentrations, pH and
temperature. Among them, a strain designated as DD produced 23.65 mm radial
clearance zone and the highest chitinolytic activity in Chitin amended synthetic
media was 2.5 % colloidal chitin at pH 5.6 and 25°C. Generally, except strain
9604 the other strains of EPF produced chitinase enzyme at various range and the
virulence of the strains could increase by incorporating chitin as carbon source.
Introduction
Chitin is considered to be the second most abundant polymer after cellulose. This
polymer of N- Acetylglucosamine (GlcNAc) is widely distributed in nature in the
form of exoskeletal material of crustaceans and insects (Wang et al., 2012). Insect
cuticles are composite structural materials with mechanical properties that are
optimal for their biological function. The cuticle consists of a thin outer epicuticle
containing lipids and proteins and a thick procuticle consisting mainly of chitin
and protein (Kang et al., 1999). Entomopathogenic fungi enter their hosts by
direct penetration of the cuticle by their chitinase enzyme, which is a barrier
against most microbes. Consequently, fungal pathogens have a potential as a
biological means of controlling sap-sucking insects that have not been controlled
with chemical pesticides (Aida and Taghreed, 2014).
Chitinases are a group of enzymes which are responsible for degradation of chitin.
They play a pivotal role in recycling chitin in nature. Chitinases are known to
[55]
perform many biological functions and they occur in organisms such as bacteria,
fungi, actinomycetes, insects and higher plants (Jolapara et al., 2013).
Microorganisms produce chitinase in order to utilize chitin as energy source
whereas fungi and insect produce chitinases as they are involved in
morphogenesis. In plant, they are involved in defense against pathogens (Aida and
Taghreed, 2014).
Thus presence of chitin degrading organisms in nature presents an economical and

environmental friendly alternative in order to obtain chitinases. Over the decade,
the exploration of microbes as the source of chitinases has increased due to its
various applications. Chitinases obtained from microbial sources have been
employed in various fields of medicine, biotechnology, food, wastewater and
agricultural industries (Jolapara et al., 2013). High production costs of chitinases
necessitate the need for scientists to explore and understand the properties of
microbial chitinases in order to formulate inexpensive and reliable chitinases
mixture. This serve two purposes: reducing the environmental hazard and
generating various value added compounds of industrial interest.
Beauveria bassiania and Metarihzium anisopilae are one of the most studied
entomopathogenic fungi with a wide host range. Cuticle degrading enzymes from
entomopathogenic fungi can be attributed to the comparison of isolates differing
in pathogenicity. The degree of pathogenicity is related to the production of the
chitinolytic enzymes in entomopathogenic fungi (Dhar and Kaur, 2010; Leger et
al., 1991; Havukkala et al., 1993 and Sayed et al., 1989). In the last many years
anattempts have been made on isolation and identification of entomopatogenic
fungi from different source, namely soil and dead insects at Ambo Agricultural
Research Center and over 53 isolates of Beauveria and Metarihzium spp. are
available in the Biological Control Laboratory. However, their chitinolytic activity
not yet studied. Therefore, the present study aims to investigate and identify
chitinolytic entomopathogenic fungi isolates collections found in Ambo
Agricultural Research Center and Desert Locust Control Organization (DLCO),
and optimize process parameters in order to maximize microbial production of
extracellular chitinase.

Chemicals used
Chitin powder , potato dextrose agar (PDA), Saboaraud dextrose yeast extract agar
(SDYA) Agar Powder, KH2PO4, K2HPO4, MgSO4, FeSO4.7H2O, MnCl2.4H2,
ZnSO4.7H2 were purchased from commercial suppliers.
[56]
Inoculums reinitiating
All 16 strains of Beauveria and Metharhzium spp. used in this study were obtained
from Ambo agricultural research center (AARC) biological control laboratory and
Deseret Locust Control Organization (DLCO). To reactivate the strains, they
were inoculated in Petri dishes containing potato dextrose agar plus
chloramphenicol (0.05% v/v), supplemented with 0.5% of yeast extract (PDAY)
and incubated at 26°C for 12 days for conidiation. Following incubation,
harvested conidia were prepared in 0.01% v/v Tween 80 in sterilized distilled
water and sprayed by micro-atomizer on Galleria mellonella. Newly emerged
conidia from the insect were subcultured not more than four times, at ten days
intervals in PDAY. To confirm viability, the conidia were spread on PDAY and
incubated for 16 h at 26°C. Germination rates were scored for the presence of
germ tube on 100 conidia at random using compound microscope at 400 ×
magnification.
Preparation of colloidal chitin

Colloidal chitin was prepared from commercial chitin powder by the modification
of the published method (Hsu and Lockwood, 1975). Twenty gram chitin was
dissolved in 200 ml of concentrated HCl stirring for 3 min at 40oC. The chitin was
precipitated as colloidal suspension by slowly adding water (2 L) adjusted to 5oC.
Colloidal suspension was collected by filtering through coarse filter paper; the
filtered colloidal suspension was washed with tap water until the PH of the
suspension reaches about 7.0 (Narayanan et al., 2013).
Disc preparation
Spore suspension of 100 μl @ 1 × 106 spores/ml of four days old cultures of chitin
embedded were spread plated on Sabaured Dextrose Agar and the plates were
incubated for 3 days at 28 °C. At the end of 3 days, 5 mm mycelial disc with agar
was retrieved with the help of cork borer and placed in the middle of fresh test
substrate containing chitin agar plates and incubated at 28 °C for 10 days.
Determination of EPF strains for their chitinolytic enzyme activities

The effect of chitin agar media on fungal growth was studied under in vitro
condition. Fungal growth were determined by inoculation of SDYA and CA
(chitin Agar) plates with an agar disc (5 mm) diameter of actively growing young
mycelium placed in the center of the medium. Four plates were used as replicates
for each treatment (Patil et al., 2000). For determination of chintinase producing
fungi, the agar medium amended with colloidal chitin were used (Aida and
Taghreed, 2014). The mineral synthetic medium consists of (g/l) (NH 4)2SO4,1:
K2HPO4, 1: MgSO4, 0.5: KCl, 0.5: NaCl, 0.5: CaCl2, 0.2: FeSO4.7H2O, 2g:
colloidal chitin, 20: agar, 15 and distilled water 1 L ( Bansode and Bajekal, 2006).
The PH of the medium was adjusted to 5.6 with KOH and HCl. The medium were
[57]
sterilized by autoclaving at 121oC for 15 min (Aida and Taghreed, 2014). A 3 day
5mm mycelium disc with agar were retrieved with the help of corkborer and
placed in four places of fresh test substrate and incubated at 28 °C for 8 days.
Chitinase producing fungi strains were determined on the basis of larger
hydrolysis zone after 8 day of incubation. Enzyme activities were measured using
digital caliper and calculated as an index of the total diameter of the colony plus
halo divided by the diameter of the colony (Sanivada and Challa, 2014).
Optimization of cultural conditions

Effect of pH, temperature & chitin concentration
The effects of PH, temperature and chitin on the chitinolytic enzyme production by
entomopathogenic fungi were studied by growing cultures at temperatures between
23-30 degrees centigrade, PH 5.5-7.5 and chitin concentration 1.25-5g/l .
Experimental design and statistical analysis

The experiment was laid out in CRD with five replications on freshly prepared
fungal conidial suspensions. Chitinase activities of the 16 different isolates were
analyzed by Minitab version 20 and treatment means were compared at 5% level
of significance.

Determination of EPF strains for their chitinolytic enzyme activities
Chitinase enzyme production potential of Entomopathogenic fungi (EPF) are
presented in Table1 and Fig.1. They were tested for the production of chitinase
enzyme at the temperature, pH and coloidal chitin of, respectively, 26oC, 5.6 and
2g/l as done by Shaukat (2010). Among the 16 EPF tested, only one isolates
(9604) showed none chitinase activity, while the other 15 isolates produced zone
of clearance over 9 mm. The highest measurement of chitinase activity (23.8 mm)
was observed for isolate DD. The influence of environmental factors such as pH
and temperature has an impact over biological processes such as enzyme
production by controlling the availability of metabolic ions. Previous reports have
shown that species of entomopathogenic fungi are known to produce chitinolytic
enzyme (Sayed et al., 1989; Farag et al., 2016). It should be noted, however, that
not all entomopathogenic fungi are considered as a potential chitinase producer, as
witnessed in this study. The present result is in agreement with the findings of
Farag et al. (2016) who reported that the majority of entomopathogenic fungi
tested were produced maximum chitinase at slightly acidic pH.
[58]
Figure 1. Hydrolysis of chitin produced by the isolate ‘DLCO-73’
Tabe1: Individual and mean chitinase enzyme activity measurement in mm at 26.2o C, PH 5.6 and 2% coloidal chitin
Isolate code Growth PH Media used Replication Mean

temp. R1 R2 R3 R4
DLCO-141 26.2 5.6 Chitin media 14.2 13.7 15.4 14.8 14.5
S1 26.2 5.6 Chitin media 13.8 14.0 13.5 14.0 13.8
ZG1 26.2 5.6 Chitin media 16.4 15.3 18.8 14.4 16.2
P-18 26.2 5.6 Chitin media 16.9 17.5 19.9 18.0 18.1
B4M 26.2 5.6 Chitin media 16.8 16.2 16.6 18.5 17.0
PPRC-51 26.2 5.6 Chitin media 17.6 12.7 16.4 17.2 16.0
9609 26.2 5.6 Chitin media 11.0 16.3 11.0 10.1 12.1
DD 26.2 5.6 Chitin media 23.6 23.6 24.0 24.0 23.8
ZG3 26.2 5.6 Chitin media 10.6 8.1 10.0 9.8 9.6
PPRC-2 26.2 5.6 Chitin media 9.4 9.4 8.9 8.7 9.1
9604 26.2 5.6 Chitin media 0 0 0 0 0
PPRC-1a 26.2 5.6 Chitin media 10.8 9.6 8.9 8.8 9.5
9615 26.2 5.6 Chitin media 9.7 10.0 10.2 10.5 10.1
F111B 26.2 5.6 Chitin 9.6 9.2 9.8 9.7 9.6
Optimization of cultural conditions

Effect of pH, temperature & chitin concentration:
The effects of the pH, temperature and chitin on the chitinolytic enzyme
production by entomopathogenic fungi were studied by growing cultures at
temperatures between 23-30 degrees centigrade, pH 5.5-7.5 and chitin
concentration 1.25-5g/l
Optimum pH for enzyme production:

The influences of initial pH on the culture medium of tested strains are shown in
figure 2. Maximum chitinase production was observed over a range of pH 5-5.6.
At pH higher and lower than the optimum pH range, enzyme production was
[59]
repressed. Chitinases of Beauveria bassinia and Metarihzium anisopliae pH
optima is between 5.5 to 5.7. However, its optima are not stable over broad pH
range (Brurberg et al., 1996). According to this study the activity of chitinase
dramatically drops when the pH of the media changed to 7. However, it was fairly
stable from PH 7 to 8.5
Optimum temperature for enzyme production:

The influence of temperature on chitinase production by EPF is depicted on Figure
3. Maximum chitinase production was observed at a temperature ranging from 24-
25oC. Increase in the temperature beyond this range resulted in decrease in
enzyme production. These results show the organism’s mesophilic preference for
enzyme production. Optimum temperature is crucial in production of chitinase
since it influences cell growth. The effect of temperature on chitinase activity was
also determined using temperature range between 23-30oC. In this study, the
temperature optima for chitinases ranged from 24.5 - 25.5°C and then it showed a
gradual decrease until the temperature reached about 26oC and from which it was
slightly stable up to 30o C. The observations made related to optimum
temperature is in complete agreement with the Shaukat et al. (2010) who reported
that 25oC as the optimum temperature for chitinase production by entomo-
pathogenic fungus Isaria fumosorosea.
Optimization of substrate concentration

The optimization of substrate concentration is presented in Figure 4. Chitinase
production was observed with respect to substrate concentration, the optimum
level being observed was in the range of 1.5-2.5g/l (Fig 4). The optimum
concentration of chitin as the substrate in the production medium is crucial for
chitinase induction in order to produce maximum chitinase. Five different
colloidal chitin concentrations (1.25% - 5%) were used for chitinase production
and the result coincided with what reported by Taechowisan et al. (2003). Results
of this study revealed that the activity of chitinase is not efficient below 1% and
above 2.5% chitin concentration. Its production was maximum at 2% chitin
concentration (Table 2). The study also indicates the enzyme activity slightly
decreases above 3% chitin concentration.
[60]
Table 2. Upper and lower limit of mean chitinase activity measured in mm
Isolate code Replication Mean SDev 95% CI

DLCO-141 5 14.108 1.060 (13.008, 15.208)
ZG1 5 16.040 0.034 (14.940, 17.140)
PPRC-1a 5 8.594 0.741 (7.494, 9.694)
PPRC-51 5 16.139 1.963 (15.039, 17.239)
PPRC-2 5 8.132 1.664 ( 7.032, 9.232)
9609 5 11.69 2.60 ( 10.59, 12.79)
DLCO-73 5 14.6450 0.1406 (13.5455, 15.7445)
DD 5 23.65 0.00 (22.55, 24.74)
ZG3 5 7.458 0.839 (6.358, 8.558)
DLCO-39 5 7.6550 0.0909 (6.5555, 8.7545)
9604 5 0.000000 0.0000 (-1.099508, 1.099508)
9615 5 9.994 0.406 (8.894, 11.094)
F111B 5 9.476 0.309 (8.376, 10.576)
B4M 5 16.767 1.055 (5.667, 17.867)
P-18 5 17.770 1.338 (16.670, 18.870)
S1 5 14.108 1.060 (13.008, 15.208)
halo zone /mm/
Figure 2. Influence of temperature on chitinase production

halo zone /mm/
Figure 3. Influence of initial pH on the culture medium
[61]
Halo zone /mm/
Figure 4. Effect of chitin concentration on chitinase production

The present study confirmed that soil and insect derived entomopathogenic fungi
produce chitinase enzyme used as a catalyst for the degradation of chitin.
Maximum chitinolytic enzyme production is achieved by controlling the
temperature, substrate concentration and pH values. 95.57 % of the tested
entomopathogenic fungi isolates produced chitinase enzyme in the presence of
chitin media. Production of chitinase enzyme by EPF isolates are fairly stable over
PH range, temperature and chitin concentrations used.
The production of microbial chitinases attracted great attention in the last few
decades and microorganisms which produce a complex of insectlytic enzymes are
considered to be possible biological control agents. Many microbes showed the
ability to produced chitinases, among them entomopathogenic fungi species are
the most famous. Indeed, the fungal strains of Beauveria bassiana and
Metarhizium anisopliae used throughout this work were proved to possess a
relatively high chitinase activity. Therefore, while using entomopathogenic fungi
for the management of insect pests, researchers could grow them on chitin
amended culture medium for the increment of its virulence. These properties
indicate that the chitinase enzyme obtained from entompathogenic fungi may be a
good candidate for biocontrol of pathogenic fungi and some pathogenic bacteria.
Crude chitinase enzyme could also used as an alternative pest management option
for fungal pathogen. In addition, this enzyme can be used commercially for the
preparation of Nacetylglucosamine which has a pharmacological activity. A
[62]
detailed study is necessitated in order to harness the ability of the isolate to
produce commercial value added products.
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of chitinase produced by Aspergillus terrus. African Journal of Biotechnology,13(14):1567-
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Chul kang S, Park S and Lee G. 1998. Purification and characterization of a novel chitinase from
the entomopathogenic fungi, Metarihzium anisopliae . Journal of Invertabrate Pathology.73,
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Chitinase Production by Isolated Bacillus Spp. Int J Pharm Pharm Sci,5 ( Suppl): 412-418
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[64]
Evaluation of Faba Bean Genotypes for
Faba bean Gall Resistance
Musa Jarso1, Asnakech Tekalign1, Nigussie Kefelegn2, Gemechu Keneni1,
Simegnew Anley,2 Abebe Hailu3 and Fantanesh Sendek3
1
Ethiopian Institute of Agricultural Research, Holetta Research Centre, P.O.Box36, Ethiopia
2 Amhara Regional Agricultural Research Institute (ARARI), Debrebirhan Research Center, Ethiopia
3
ARARI, Gonder Research Center, Ethiopia
Abstract
Faba bean (Vicia faba L.) is one of among the most important food legumes and
widely considered for its high nutritional value a good resource of protein. Faba Bean
Gall (Olpidium viciae) is one of the major faba bean challenge; a new disease
devastating the crop and threatens its production in major faba bean growing
highland areas of the Ethiopia. Efforts were made on evaluation of many genotypes
to develop resistant varieties; however, none of them were resistant. Therefore, the
present study was initiated to find faba bean varieties resistant/tolerant for faba bean
gall, stable high yielding and stress tolerant for different agro-ecologies in Ethiopia.
A total of 12 faba bean materials selected from previous preliminary variety trials
were evaluated using RCBD with three replications as national variety trial over
three hot spot locations for faba bean gall for two years in 2017 and 2018 main
cropping seasons. Significant differences (p<0.001) were observed among the
genotypes for most of the yield and yield component traits. Genotype EH06070-1
showed tolerance for the faba bean gall disease consistently over the three locations
and years with better yield. However the data was not adequate to conclude this
genotype as tolerant. Thus, this genotype will be further evaluated to be used as a
parent material.
Introduction
Faba bean (Vicia faba L.) is one among the most important food legumes and
widely considered for its high nutritional value as a good resource of protein,
starch, cellulose and minerals (Haciseferogullari et al., 2003). It constitutes the
main dish particularly for the low income groups in the Middle East and most part
of Mediterranean, china and Ethiopia (Ferris and Kaganzi, 2008). Faba bean is
also good break crop in cereal production system for its potential of fixing
considerable amount of atmospheric nitrogen hence reduces the need of farmers to
use artificial fertilisers (Lindemann and Glover, 2003).
Despite its importance the higher productivity of faba bean is below its potential
due to different biotic and abiotic factors. Faba bean gall (FBG) designated as
caused by Olpidium viciae is a newly emerged and being considered one of the
major faba bean production challenge in the country. It is already devastating the
crop and threatening its production in major highland growing areas of Ethiopia
since 2012.
[65]
Little is known about the disease and managing it is a priority of the faba bean
improvement program of Ethiopia. Faba Bean Gall (FBG) was found to spread
rapidly through farmers’ fields in Northern part of the country and the central
highlands where it is now widespread and increasing in incidence and severity
(Hailu et al., 2014). Typical symptoms include green and sunken lesions on the
upper side of the leaf and bulge to the back side of the leaf, and finally develop
light brownish color lesions, chlorotic galls, and progressively broaden to become
circular or elliptical uneven spots. When there are numerous galls on leaves and
stems these coalesce later in the season and cause necrosis of plant tissue.
Affected plants become stunted and do not develop seed. Efforts have been made
to find host resistant by evaluating germplasms under natural infestation but none
of the materials were tolerant for this disease. Therefor the present study was
initiated to evaluate materials for their resistance to faba bean gall and to develop
stable, high yielding faba bean variety for faba bean growers in different agro-
ecologies in Ethiopia.

A Total of 12 faba bean selected materials from previous preliminary variety trials
based on their tolerance for the faba bean gall disease were evaluated as national
variety trial over three hot spot locations (North Shewa DebreBirehan /Mush and
Gonder / Debark) for two years from 2017 to 2018 main cropping season. The
design was RCBD with three replications. Each germplasm planted in 4 rows with
0.4 m between rows, 0.1 m between plants in 4 m lengths of 0.6 m between
adjacent plots. NPS at the rate of 121 kg h-1 was applied at planting. Weeding and
other management practices were done as per recommendation. Data for the
following agronomic traits were recorded on a plot basis at all locations; days to
50% flowering (DTF) and days to physiological maturity (DTM) for the entire
plot. Other agronomic data viz., Number of pods per plant (PPP), number of seeds
per pod (SPP) and plant height (PH) were also collected. Disease scoring was
made from the entire plot using percentage scoring method. Data on thousand seed
weight (TSW) from randomly selected 1000 seeds from each plot and grain yield
(GY) was taken from the entire plot. Grain yield adjustments were made by
weighing the oven dried and adjusting to constant moisture level of 10%. The
grain yield recorded on plots basis were converted to kg ha-1 for statistical
analysis. The analysis of variance (ANOVA) was performed using the PROC
GLM in SAS (SAS Institute, 2012).
Result and Discussion

The sporadic nature of the disease across location and year to evaluate the
materials was the major challenge. Thus, the data used for the analysis were only
three; these were two years data of 2017 and 2018 from Mush and only one year
[66]
data of 2018 from Gonder. The analysis of variance (ANOVA) revealed that
significant (p < 0.001) variation among the twelve genotypes for faba bean gall
disease reaction, TSW, grain yield (GY) and other agronomic traits at Mush (in
2017 and 2018) and at Debark (in 2018) (Tables 1, 2 & 3). Out of the 12 faba bean
materials, genotype EH06070-1 revealed moderately tolerance to the faba bean
gall disease consistently across location and year with better yield than other
genotypes and the standard checks. The data used for this result was not sufficient
to develop confidence on the tolerance level of this genotype. Therefore, this
genotype (EH06070-1) has to be further evaluated to be used as a parental
material for faba bean gall resistance breeding in faba bean improvement program.
Table 1: Mean yield and yield related traits of faba bean genotypes and their response to the disease at Mush in 2017
Entry PH PPP SPP Gall (%) BM (t ha-1) GY (kg ha -1) TSW g

Walki 56.0b 10.4a 2.1bcd 77.8bc 2026cd 264g 281d
EH09019-4 60.2b 7.6ab 2.0cd 50de 3095bc 1634a 681ab
Gora 74.3a 8.8ab 2.4a 66.7cde 3000bcd 1751a 597b
EH010008-3 61.6b 5.9b 2.2abcd 61.1cde 1861d 540defg 645b
EH010008-5 64.3b 6.2b 2.4ab 75bc 2697bcd 746cdef 623b
Dosha 57.7b 7.2ab 2.0cd 88.9b 2027cd 321efg 446c
EH010050-1 55.2b 6.6b 2.2abcd 77.8bc 2223cd 799cd 604b
EH010054-1 54.2b 6.6b 2.0bcd 63.9cde 2344cd 1134bc 731a
EH010058-1 60.2b 6.0b 2.3abc 69.4cd 2436cd 779cde 631b
CS20DK 57.0b 5.9b 2.1abcd 91.7a 2840bcd 280fg 381c
EH09017-5 61.9b 6.9b 1.9d 72.2c 3633b 1098bc 600b
EH06070-1 75.3a 10.6a 2.4ab 33.3e 6003a 1329ab 637b
Mean 61.5 7.4 2.2 69 2848.7 889.6 57.1
CV 10.2 29.3 10.4 16.5 24.4 33.6 9.8
Where: PH: plant height; PPP: Pod per plant; SPP: Seed per pod; Gall: faba bean gall disease score; BM: Biomass; GY:
Grain yield;
Table 2: Mean yield and yield related traits of faba bean genotypes and their response to the disease at Mush in 2018
Entry DF DM PH PPP TSW Gall (%) BM (t ha-1) GY (kg ha -1)

Walki 33d 110d 55bc 4 490d 28ab 1c 487c
EH09019-4P 34ab 116a 57bc 5 850a 22b 3b 1371b
Gora 33abcd 115ab 64ab 5 810ab 44ab 3b 1609ab
EH010008-3 34ab 117a 61abc 4 860a 33ab 3b 1503ab
EH010008-5 33cd 116ab 61abc 5 910a 22b 3b 1620ab
Dosha 32d 113bc 53c 4 680bc 56a 2b 1282b
EH010050-1 35a 117a 57bc 4 820ab 50ab 3b 1557ab
EH010054-1 34ab 112cd 62abc 4 920a 56a 3b 1514ab
EH010058-1 34ab 117a 68A 4 850a 39ab 4a 2059a
Cs20DK 33bcd 113bc 57bc 5 600cd 50ab 2b 1310b
EH09017-5 33bcd 113bc 66ab 5 840a 28ab 3ab 1791ab
EH06070-1 34ab 116a 59abc 5 680bc 28ab 3b 1518ab
Mean 33 115 59.84 4.4 77.7 38 2.77 1468.46
CV 2.7 1.5 10.6 26.5 11.7 49 24.6 26.4
Where: PH: plant height; PPP: Pod per plant; SPP: Seed per pod; Gall: faba bean gall disease score; BM: Biomass;
GY: Grain yield;
[67]
Table 3: Mean yield and yield related traits of faba bean genotypes and their response to the disease at Debark, Gondar
in 2018
Entry DF DM Gall% PH PPP SPP TSW BM (t ha-1) Gy(kg ha -1)

Walki 60 148 63 92 6 2.3 660 2.89 1111
EH09019-4 63 152 46 84 5.1 2.4 980 3.99 1486
Gora 60 150 42 94 6 2.7 910 3.99 1779
EH010008-3 61 152 34 91 5.1 2.7 1030 3.8 1586
EH010008-5 60 146 37 89 4.3 2.6 920 3.56 1425
Dosha 59 149 74 93 5.9 2.5 730 3.52 1443
EH010050-1 60 146 44 82 4.5 2.6 850 3.36 1309
EH010054-1 64 147 12 79 4.1 2.6 1030 3.28 1470
EH010058-1 61 148 47 87 4.9 2.7 970 4.53 1960
CS20DK 60 148 50 90 5.4 2.3 650 3.36 1262
EH09017-5 60 144 47 83 4.6 2.5 880 3.52 1345
EH06070-1 61 148 23 82 4.5 2.6 880 3.91 1606
Mean 61 148 43 87 5 2.5 870 3.6 1482
CV (%) 3 2.9 44.9 7.6 27 11.8 9.1 15.46 23.7
LSD (5%) 2.3 28 9 12 0.8 506
sig (entry) ** Ns ** * ns Ns *** * *
Where: DF: Days to flowering; DM: Days to maturity; PH: plant height; PPP: Pod per plant; SPP: Seed per pod; Gall: faba
bean gall disease score; BM: Biomass; GY: Grain yield.
References
Bouhassan A, Sadiki M and Tivoli B. 2004. Evaluation of a collection of faba bean (Vicia faba L.)
genotypes originating from the Maghreb for resistance to chocolate spot (Botrytis fabae) by
assessment in the field and laboratory. Euphytica 135:55–62.
CSA. 2018. Report on area and production of crops. Central Statistics Agency agricultural sample
survey for 2017 / 2018 Addis Ababa, Ethiopia
Ferris S and Kaganzi E. 2008. Evaluating marketing opportunities for haricot beans in Ethiopia.
IPMS (Improving Productivity and Market Success) of Ethiopian Farmers Project Working
Paper 7. ILRI (International Livestock Research Institute), Nairobi, Kenya. p. 68.
López-Bellido L, López-Bellido R, Redondo R and Benítez J. 2006. Faba bean nitrogen fixation in
a wheat-based rotation under rainfed Mediterranean conditions: Effect of tillage system.
Field Crops Research 98:253-260.
Haciseferogullari H, Gezer I, Bahtiyarca Y and Menges HO. 2003. Determination of some
chemical and physical properties of Sakız faba bean (Vicia faba L. Var. major). Journal of
Food Engineering 66:475-479.
Hailu E, Getaneh G, Sefera T, Tadesse N, Bitew B, Boydom A, Kassa D, Temesgen T. 2014. Faba
bean gall: a new threat for faba bean (Viciafaba) production in Ethiopia. Advances in Crop
Science and Technology 2:4 http://dx.doi.org/10.4172/2329-8863.1000144
Lindemann WC and Glover CR. 2003. Nitrogen fixation by legumes. In: Cooperative Extension
Service Guide A-129, editor New Mexico State University and the USDA.
SAS Institute. 2012. SAS proprietary software. Release 9.3 SAS Inst., Cary, NC, USA.
Torres AM, Rom B, Avila CM, Satovic Z, Rubiales D, Sillero JC. 2006. Faba bean breeding for
resistance against biotic stresses: Towards application of marker technology. Euphytica
147:67-80.
[68]
Evaluation of Sweetpotato Varieties against Major
Sweetpotato Viruses in Ethiopia
Shiferaw Mekonen and Fekadu Gurmu
South Agricultural Research Institute, Hawassa Research Centre, P.O. Box 6, Hawassa, Ethiopia.
Abstract
Sweetpotato virus disease (SPVD) is one of the most destructive diseases in East
Africa including Ethiopia. Since 2004, the disease becomes an important threat in
Ethiopia, particularly in Southern region. Use of virus resistant varieties is the most
effective means of reducing sweetpotato yield losses due to virus infection. Six
sweetpotato varieties introduced from abroad were evaluated in Hawassa and Dilla
research sites in 2017 and 2018 cropping season to determine the reaction of the
varieties to major viruses in the region. The treatments were arranged in a
Randomized Complete Block Design (RCBD). Standard and susceptible checks were
included in the treatment. Virus symptom identification and severity score were
executed in the field and in the net tunnel using biological property related to virus
interaction with its host such as leaf and plant color (purple, mosaic, and vein
clearing) and size deviation from normal appearance such as deformation of leaf
size and shape. In addition, to confirm the presence of virus and to identify the type
of virus, diagnosis was made using serological test. The result revealed that there
was a significant difference (P<.0.05) among varieties in responding to sweetpotato
virus disease. The two introduced varieties, NASPOT 13 O and NASPOT 12 O,
showed consistent resistance reaction to the virus across locations and years. The
minimum disease severity (1.3 and 1.5) score and the maximum root yield (28.98 and
25.19 t ha-1) were recorded for the two varieties, respectively. Whereas, the
maximum disease severity (3.54 and 3.66) score and the lowest yield (14.57 and
16.16 t ha-1) were obtained from variety Guntute and New Kawogo, respectively.
Therefore, the two high yielding varieties, NASPOT 13 O and NASPOT 12 O, which
showed resistance reaction to SPVD, can be used for breeding purpose as sources of
resistance and to encourage production in areas where sweetpotato virus disease is
a serious problem for sweetpotato production.
Introduction
Sweetpotato virus disease (SPVD) is the most important biotic constraint of
sweetpotato [Ipomoea batatas (L.) Lam.] in the world as well as in Africa next to
sweetpotato weevil (Lenné, 1991; Qaim, 1999). More than 20 viruses are known
to infect cultivated sweetpotato worldwide (Fuglie, 2007). In Ethiopia, since 2004,
SPVD has been recognized as an important disease that greatly reduces the yield
of the crop in the country. Subsequent studies and assessments in southern
Ethiopia have indicated the incidence and occurrence of the virus to reach 35-
100% and 20-80%, respectively. Sweetpotato Feathery Mottle Virus (SPFMV),
Sweetpotato virus G (SPVG), Sweetpotato chlorotic stunt virus (SPCSV) and
Sweetpotato virus 2 (SPV2) were the major viruses identified in Southern
[69]
Ethiopia and known infecting the crop either singly or in combination (Tamru,
2004; Abraham, 2010; Teddy et al., 2011).
The yield loss incurred due to virus infection reported to be varied depending on
the agroecology of the country. For instance, in Nigeria and Uganda, viruses
account for 50% yield loss; whereas, in East Africa, over 90% of yield reductions
have been associated with viruses (Gibson et al., 1998). Shiferaw et al. (2013)
reported that the yield loss due to virus infection can be varying depending on the
quality of the vine used at planting and the level of reaction of varieties to the
disease. In Ethiopia, the root yield reduction due to the synergistic infection of
SPFMV and SPCSV has been reported to be 37% (Tesfaye et al., 2013).
Although control of viral diseases remains difficult in subsistence cropping
systems (Rukarwa et al., 2010), disease management strategies such as cultural
practices, phytosanitary measures, control of vectors and deployment of genetic
resistance to prevent or limit the extent of damage have been recommended
(Maule et al., 2007). Among all management options, use of disease resistant
plants is ideal in terms of effectiveness and sustainability for managing any plant
disease in general and for SPVD in particular (Maule et al., 2007; Valverde et al.,
2007). However, the incidence and severity of the pathogen is strongly influenced
by the interaction of temperature, vectors, hosts and pathogen genetics (Adiki et
al., 2016). The outbreak of the disease in resistant variety reported to be
fluctuating depending on the environment due to climatic aspects and emergence
of new virus strains. This implies the varieties showed resistance reaction in one
environment might be susceptible when introduced to another new environment.
Therefore, the aim of this study was to evaluate the introduced tolerant
sweetpotato varieties against SPVD under Ethiopian agro-ecology.

Description of the site
The study was conducted at Hawassa and Dilla in Southern Nation, Nationalities
and People Regional State in 2017 and 2018 main rainy seasons. Hawassa is
located at an altitude of 1700 masl and 07°03′54″N, 38°28′59″E coordinates,
receiving annual rainfall of 1046.3 mm, whereas Dilla is found at an altitude of
1519 masl, 06°22′49″N, 38°18′25″E coordinates with annual rainfall of 1354.6
mm. The locations are the known hot spot areas for sweetpotato viruses, where the
two important viruses infecting sweetpotato, namely SPFMV and SPCSV is
equally prevalent (Abraham, 2010).
Planting materials
Six introduced tolerant orange fleshed sweetpotato (OFSP) varieties, namely
Tomurabuka, Naspot-8, Wagabolige, New Kawago, NASPOT 12 O and NASPOT
[70]
13 O were used along with locally released tolerant (Kulfo) and susceptible
(Gunute) OFSP varieties which were used as checks.
Experimental design and spacing
The experiment was laid out in a randomized complete block design with three
replications. Vines were planted in four rows with a plot size of 3 m x 2.4 m and
were spaced at 60 and 30 cm between rows and plants, respectively. The known
susceptible variety Guntute (Shiferaw et al., 2014) was planted around the
experimental plot to serve as a source of inocula in the trial. The transmission
efficiency of SPVD by vectors may vary with environmental conditions and
vector population size which fluctuates over seasons and years. To avoid such
suspicion, the varieties evaluated at field condition also planted in a net tunnel
using pots. Then virus infected sweet variety (Guntute) having white flies and
aphids introduced in the net tunnel for artificial inoculation.
Assessment of disease incidence and severity scoring
Assessment and disease scoring in the field was started two months after planting
and data collected at one month interval until the crop gets matured. Severity and
incidence of disease was determined using the symptom on the host, for instance
leaf and plant color deviation from normal appearance (purple, mosaic, vein
clearing) and deformation of leaf size and shape (Gutierrez et al., 2003).
Severity was recorded on scale of 1-5, where 1 = no visible symptoms, 2 = mild
symptoms (a few local lesions on a few leaves), 3 = moderate symptoms (mosaic
symptoms on leaves), 4 = severe symptoms (mosaic symptoms with plant
stunting) and 5 = very severe symptoms of purpling/yellowing or mosaic on
leaves, severe leaf distortion, reduced leaf size and severe stunting (Mwanga et al.,
2013; Njeru et al., 2004). Incidence (I) of virus for each plot was determined by
counting the number of diseased plants out of a total in a plot of 3m x 1.2 m area
and expressed in percent (%). The resistant reaction was determined by severity
score and purple color intensity of NCM-ELISA test. 1= immune; 1.1-2= resistant;
2.1-3 = moderately susceptible; 3.1-4 = susceptible; 4. 1-5= highly susceptible.
Virus detection: Three months after planting, leaf samples were collected using
plastic bag from the top, middle and basal parts of the plant of each experimental
field and from artificially inoculated sweetpotato plant in the net tunnel. The
serological test was conducted at Areka Agricultural Research Center
biotechnology laboratory using Nitrocellulose membrane enzyme-linked immune
sorbent assay (NCM-ELISA) (Aritua et al., 1998). Accordingly, each sample was
tested against antibodies of 10 economically important sweetpotato viruses.
Yield and yield component
Data related to marketable yield (MY) and unmarketable yield (UMY) was
determined from two central rows in each plot. Storage roots (t ha-1) were sorted
[71]
as marketable and unmarketable (blighted, rotten and deformed) roots. Storage
root diameter and length were determined from 10 storage roots randomly taken
from each plot by using a measuring caliper.
Data analysis
The data on disease incidence and severity, storage root yield (marketable,
unmarketable and total yield), root diameter and length were subjected to analysis
of variance (ANOVA). Means were separated following Fisher’s protected least
significant difference test at 5% significance level (LSD 5%). All analyses were
done with the Statistical Analysis System (SAS) version 9.1 (SAS Institute Inc.,
2003.).

Virus detection and reaction of varieties
Natural infection of virus disease was appeared during the study periods. The
SPVD inoculum pressure was high and devastated the susceptible clones under
natural condition. In all locations, the susceptible variety Gunute showed the
typical symptom of virus infection. The six introduced varieties had showed
different level of resistance to sweetpotato viruses under the main rainy season at
field condition.
Different level of virus severity were recorded and observed depending on the
genetic makeup of the varieties against sweetpotato virus. Variety NASPOT 13 O
and NASPOT 12 O showed resistant reactions in the field with mean severity
score of 1.3 and 1.5, respectively and negative color intensity for NCM-ELISA
test. Variety Tomurabuka, NASPOT-8, Wagabolige and Kulfo showed mild
symptom with severity score of 2.1 - 2.5 (moderately susceptible). The susceptible
check variety (Guntute) and the introduced variety New Kawogo showed
susceptible reaction to SPVD with severity score of 3.5 and 3.66, respectively
(Table 1).
Also the serological test result revealed different color intensity for the tested
varieties. Accordingly, the varieties NASPOT 13 O and NASPOT 12 O samples
showed negative color intensity for virus infection indicating that the two varieties
were resistant to SPVD. The remaining varieties Tomurabuka, NASPOT-8,
Wagabolige, New Kawogo, Kulfo and Guntute showed positive color intensity to
Sweetpotato Virus, which shows that these varieties were susceptible to SPVD.
However, the intensity of the virus concentration varied from weak (+) to strong
color intensity (+++). Three viruses namely SPFMV, SPCSV and Cucumber
Mosaic Virus (CMV) were detected during the serological test (Table 1), which
implies that the three virus types were prevalent in the study areas.
[72]
SPFMV and SPCSV were detected in all varieties except NASPOT 12 O and
NASPOT 13 O as revealed by the serological test (Table 1). The result is in
agreement with Tewodros et al., (2011) and Abraham (2010) who reported the
prevalence of the two (SPFMV and SPCSV) viruses in Ethiopia. CMV detected
only for variety New Kawogo and it was also previously reported in Ethiopia by
Shiferaw et al., (2016).
Table 1. Response of sweetpotato varieties to major sweetpotato viruses using serological test and Mean severity score
at Hawassa and Dilla research sites
Type of virus
Treatment Severity (1-5) FMV SPVG CSV C-6 CALV CFV LV MSV MMV CMV
Tomurabuka 2.2 (MS) + - + - - - - - - -
Naspot-8 2.3 (MS) + - + - - - - - - -
Wagabolige 2.1 (MS) + - ++ - - - - - - -
New Kawogo 3.66 (S) ++ - ++ - - - - - - +
NASPOT 12 O 1.5 (R) - - - - - - - - - -
NASPOT 13 O 1.3 (R) - - - - - - - - - -
Kulfo 2.5 (MS) + - + - - - - - - -
Guntute 3.5 (S) ++ ++ - - - - - - -
*- = negative; + = low color intensity/weak reaction/; ++ = Medium color intensity/medium reaction/; +++ = strong color
intensity/strong reaction/. MS = moderately susceptible, S = Susceptible, R = Resistant
FMV = Sweetpotato feathery mottle virus; PVG= Sweetpotato virus G; CSV= Sweetpotato chlorotic stunt virus; C-6 virus;
CaLV = Sweetpotato caulimo-like virus; CFV= Sweetpotato chlorotic flecks virus; LV= Sweetpotato latent virus; MSV=
Sweetpotato mild speckling virus; MMV= Sweetpotato mild mottle virus; CMV = Cucumber mosaic virus.
seveity (1-5 ) scale
Dilla 2017
Dilla 2018
mean
Hawassa 2017
Hawassa 2018
mean
Figure 1.The response of sweetpotato varieties to virus disease at hot spot areas as expressed by severity scale.
Response of varieties to virus severity, incidence and marketable yield

The combined analysis of variance showed that treatment, years and year x
treatment interaction effects were significant (p < .01) in controlling the disease
and increasing storage root yield (Table 2). The treatment and year effects on
[73]
storage root yield had accounted for the large portion of variation as expressed by
mean squares. The mean square of treatment and year were 2035.6 and 787.47,
respectively. The interactions of year x treatment effect on yield were accounted
for small portion with mean square of 67.26 (Table 2).
Similarly for virus incidence and severity, the influence of treatment and year
were the highest as expressed by the mean square. The mean square of virus
severity for treatment and year was also significant and 8.54 and 3.56, respectively
(Table 2). Similarly the virus incidence for treatment and year effect were
significant with mean square of 2714.39 and 622.25, respectively. In general, the
study revealed that, the presence of variability within the sweetpotato varieties to
sweetpotato viruses. Next to varieties, year effect showed higher significant
difference and contributed for the variation in disease incidence and the obtained
yield. This might be due to climate change across the experimental years, notably
variability in rain fall, humidity and temperature over the growing seasons.
Table 2. Analysis of variance for sweetpotato storage root yield, and virus disease measurement
Source of variation Mean square

DF Yield Virus severity Virus incidence
Year 1 787.47** 3.56* 622.25*
Location 1 1.11ns 0.75ns 115.21ns
Treatment 7 2035.6** 8.54 * 2714.39*
Year*location 1 14.46ns 2. 5ns 1067.80*
Year x treatment 7 67.26* 0.29ns 127.44ns
location x treatment 7 29.67ns 0.38ns 134.50ns
Year x location x treatment 7 9.25ns 0.21ns 67.24 ns
Effect of varieties on virus incidence

There was significant difference between tested sweetpotato varieties in reducing
the incidence of SPVD (Table 5). The three varieties NASPOT 13 O, NASPOT 12
O and Tomurabuka were significantly reduced the disease incidence as compared
to the checks and the remaining tested varieties. The virus disease pressure was
very high in 2018 than in 2017 cropping season. The mean disease incidence for
NASPOT 12 O (8.75 and 20) and NASPOT 13 O (11.38 and 16.5) were recorded
in 2017 and 2018, respectively, which is very low as compared to the standard
check Kulfo (33.62 and 35.82) and the susceptible check Guntute (51.49 and 60)
in 2017 and 2018 cropping seasons (Table 3). Among the introduced varieties,
NewKawago showed susceptible reaction and high disease incidence.
[74]
Table 3. The performance of varieties against sweetpotato virus as expressed by disease incidence across years and
locations
Incidence (%)
Varieties 2017 2018 Over all mean
Dilla Hawassa Mean Dilla Hawassa Mean
Tomurabuka 14.36d 13.33de 13.845 24.17cde 29.17cde 26.67 20.28
NASPOT-8 23.56dc 20.74cd 22.15 25cde 30cde 27.5 24.83
Wagabolige 33.2bc 26.23bc 29.71 30.83bcd 35.8bcd 33.315 31.51
New Kawogo 37.66bc 33.43ab 35.545 40.83b 55.83a 48.83 42.19
NASPOT 12 O 10.18d 7.32e 8.75 17.5de 22.5de 20 14.38
NASPOT 13 O 12.77d 10de 11.38 15e 18e 16.5 13.94
Kulfo 32.52bc 34.72ab 33.62 33.33bc 38.33bc 35.83 34.72
Guntute 58.73a 44.25a 51.49 57.5a 62.5a 60 55.75
Mean 27.87 23.75 25.81 30.52 36.52 33.02 30.63
LSD 18.83 12.59 - 14.52 13.5 - -
CV% 32.96 30.28 - 27.17 23.34 - -
Performance of the Varieties for Marketable Root Yield

There was no significant difference among the test locations; however there were
significant differences between the experimental years. The mean root yield
performance of each location in 2017 was higher than 2018 cropping season
There was significant difference (p< 0.01) between genotypes for marketable root
yield. High mean marketable root yields of 28.01, and 24.5 t ha-1 were obtained
from NASPOT 13 O and NASPOT 12 O, respectively. These varieties provided
high yield advantage of 93.17% and 68.97% and 48.67% and 30.04% over local
and standard checks, respectively (Table 4). The two high yielding varieties were
resistant to SPVD as indicated in Tables 1 and 3.
On the other hand, low marketable root yields were obtained from susceptible
variety Guntute (14.5 t ha-1). The significant yield difference observed in yield
between resistant and susceptible varieties might be mainly due to the effect of
virus on the susceptible ones. The current result is in line with the previous works
reported by Ngailo et al. (2013) who reported that variation among sweetpotato
varieties in terms of virus resistance is expressed by increased yield and decreased
quality loss of tested varieties.
[75]
Table 4. Performance of sweetpotato varieties evaluated for root yield at hot spot areas in 2017 and 2018 cropping
seasons.
Yield ( t ha-1 )
Variety 2017 2018 Overall Yield advantage
Dilla Hawass Mean Dilla Hawasa Mean mean YAOSC YAOLC
Tomurabuka 25.93b 24.07ab 25 12.96c 12e 12.48 18.74 -0.53 29.24
NASPOT-8 20.92c 23.6abc 22.26 16.48b 15.5dc 15.99 19.12 1.49 31.86
Wagabolige 19.28cd 23.24ab 21.26 12.78c 12.79de 12.78 17.02 -9.66 17.38
New Kawogo. 17de 16.57bc 16.78 17.54b 14.8cde 16.17 16.47 -12.58 13.59
NASPOT 12 28.15ab 27.7a 27.92 21.39b 20.77b 21.08 24.5 30.04 68.97
NASPOT 13 29.83a 28.28a 29.05 30.11a 23.83a 26.97 28.01 48.67 93.17
Kulfo 20.37cd 24.35ab 22.36 14.35c 16.3c 15.32 18.84 0.00 29.93
Guntute 14.12e 15.74 14.92 12.78c 15.46dc 14.08 14.5 -23.04 0.00
LSD 3.79* 8.51 7 2.95
CV% 9.87 20.28 23 10.26
YAOSC = Yield advantage over standard check (%), YAOLC= Yield advantage over local check
Table 5. The combined mean response of sweetpotato varieties for virus severity and incidence
Treatment name Yield (t ha-1) Severity( 1-5 ) Incidence (%) ⃰

Tomurabuka 18.28cd 2.2cd 19.21cd
NASPOT-8 17.93cd 2.3bc 26.49c
Wagabolige 16.75cde 2.1 bc 38.14b
New Kawogo 16.16de 3.66a 49.35a
NASPOT 12 O 25.192b 1.5de 14.16d
NASPOT 13 O 28.98a 1.3e 13.61d
Kulfo 18.81c 2.58b 35.1b
Guntute 14.57e 3.54a 56.78a
LSD 2.34 0.53 7.66
CV% 14.68 27.34 30.56
Values followed by the same letter were not significantly different (p< 0.05)

There was significant difference among the tested sweetpotato varieties against
sweetpotato viruses at the test locations. Two varieties, namely NASPOT 13 O
and NASPOT 12 O showed better performance in terms of resistance to SPVD
and high storage root yield. Therefore, the two varieties can be used as a major
component of integrated disease management to control SPVD and to promote
production in areas where sweetpotato virus disease is highly prevalent in the
country. In addition, since the identified genotypes possess high levels of
resistance, they can be used in breeding program as parental lines.
Acknowledgements
The financial support of the South Agricultural Research Institute (SARI) and the
Ethiopian Institute of Agricultural Research (EIAR) is duly acknowledged. The
[76]
Authors also extend their acknowledgement to Areka Agricultural Research
Center for their technical and material support during serological virus detection
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(Ipomea batatas) From Ethiopia. Doctorial dissertation. University of Bonn, pp. 126.
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[77]
[78]
Screening of Rice Germplasms for Resistance
against Sheath Rot (Sarocladium oryzae)
at Fogera, Ethiopia
Desalegn Yalew1*, Muluadam Birhan1, Tekalign Zeleke2
1
Ethiopian Institute of Agricultural Research (EIAR), Fogera National Rice Research and Training Center
(FNRRTC), P. O. Box 1937, Bahir Dar, Ethiopia , 2EIAR, Ambo Agricultural Research Center,
P.O.Box. 37 Ambo, Ethiopia
Abstract
Sheath rot of rice, Sarocladium oryzae [(Sawada) W. Gams & D. Hawksw], is
currently regarded as one of the most serious rice diseases in Fogera plains. Varietal
resistance is the most cost-effective and eco-friendly management strategy for the
small-scale farmers. An investigation was conducted to identify resistant sources
among rice germplasm introductions that could be employed in varietal development
program. Eighty germplasms along four checks were evaluated in field trials for two
years at two locations arranged in an augmented design. Results indicated that three
immune germplasms (scrid014-1-1-1-1, Scrid037-4-2-2-5-2 and YUNLU
N0.33) were found to be suitable for resistant variety development. The remaining,
germplasms were found to exhibit different levels of reactions, of which 27 resistant,
35 moderately resistant, 13 moderately susceptible and two susceptible, with PSI
ranging between 1.48–56.17%. The immune germplasms perform better than the
standard check varieties in all agronomic and phenological traits considered. Based
on these results, the high yielder immune germplasms could be used to develop
resistant varieties so as to meet the farmers’ requirement, as far as rice production is
concerned.
Introduction
Sheath rot, caused by Sarocladium oryzae (Sawada) is one of the major diseases
of rice. The pathogen mainly infects the upper most flag leaf sheaths that enclose
the emerging young panicle during the boot stage. The lesions are oblong or
irregular oval spot and usually expressed as a reddish-brown discoloration of the
flag-leaf sheath. Early or severe infection affects the panicle so that it only
partially emerges. The unemerged portion of the panicle rots, turning florets red-
brown to dark brown. Grains from damaged panicles are discolored reddish-brown
to dark brown and may not fill the affected grains, are known as chaffy grains and
the disease is appropriately known as “empty head” and is familiar as “rice
abortion” (Kindo, 2012). Moreover, the pathogen is mostly observed on the entire
seed (about 46%) and on the lemma and/or palea (about 31%) (Mew and Gozales,
2002).
Sheath rot is one of the most serious and devastating rice diseases in wetland rice
growing regions (Lanoiselet et al., 2012). The pathogen attacks flag leaf sheaths
[79]
and grains and yield losses result mainly from poor panicle formation and
exertion, spikelet sterility (80-100%), reduced grain filling, and losses in milling
(Simon, 2016). Quality is also affected as severe attacks lead to chaffy, discolored
grains and affect viability and nutritional value of the grains followed by a
decrease in the protein and starch contents of infected seeds (Reddy et al., 2000).
Seeds from infected panicles become discolored and sterile, thereby reducing
grain yield and quality significantly. Since the pathogen attacks the crop at
maturity starting from panicle initiation stages; its impact is direct to minimize the
crop yields. There was a yield loss report ranging from 20 to 85% in Taiwan and
30 to 80% in Vietnam, the Philippines and India (Anonynomous., 2012).
Variability in yield loss depends upon prevailing favorable conditions under which
rice is grown and the level of susceptibility of the grown cultivar (Ayyadurai et al.,
2005).
In Ethiopia, diseases of rice in general, and sheath rot in particular is not well
studied. This is because rice cultivation in the country is at an infant stage, and
that associated production constraints are not well known along with the fact that
importance of diseases of newly introduced crops are expanding and manifesting
gradually with the time. However, now a day sheath rot becomes major rice
disease especially in Fogera plains with prevalence, incidence and severity of 100,
47 and 44%, respectively (unpublished). Therefore, unless effective management
measure is taken, the disease will cause high yield loss with the consequence that
leads the rice crop to be out of production in the area. Thus, there is a need to
establish appropriate management method to tackle the above mentioned problem.
Most sheath rot management practices in rice fields rely on integration of
chemical control with cultural practices. However, according to Ayyadurai et al.
(2005) fungicide treatments are most of the time unsuccessful under farmers’
conditions or are very expensive as well as harmful to the environment. In the
same context, biological control has been of limited effect due to inconsistency of
antagonists under field conditions (Gnanamanickam, 2009). Therefore, among the
options, use of resistant varieties would offer a better management compared to
other control strategies, as it is inexpensive and eco-friendly strategy to the
environment (Sharanappa et al., 2003; Swami and Alane, 2013). Thus, the most
sustainable solution is the development and deployment of resistant varieties. The
resistant varieties also could be developed either through selection/screening or
crossing (Agrios, 2005). A number of resistant varieties have been developed in
different countries (Pearce et al., 2001), but none of them has been developed and
available in Ethiopia. Hence, screening of introduced and available rice
germplasms for their resistance against rice sheath rot is important to develop
resistant varieties in Ethiopia particularly in Fogera plains. Therefore the objective
of the study was to evaluate the reaction of rice germplasms against sheath rot and
identify resistant sources for future breeding purposes.
[80]
Area description
The experiment was conducted at Fogera National Rice Research and Training
Center during years 2017 – 2018 main cropping seasons in lowland ecosystem.
Geographically the research center is located at latitude of 11° 58′ N and longitude
of 37° 41′ E with an altitude of 1819 meter above sea level. The area receives
average annual rainfall of 1230 mm with mean maximum and minimum
temperatures of 12 and 28°C, respectively.
Experimental materials and design

In this experiment 80 rice germplasms, introduced from different countries to
Ethiopia at different times, along four checks (three recently released rice varieties
and one local cultivar) were screened against rice sheath rot in naturally infested
fields. The experiment was arranged in augmented design with four blocks of non-
replicated plots each containing 24 genotypes. Each genotype was sown in three
rows of 2m long and 0.6m wide plot. All agronomic practices were applied
uniformly for all plots.
Data collection and analysis

Information on agronomic data, disease data and all other necessary parameters
were collected during the study period. Phenological and agronomic data were
collected in plot bases while diseases data were collected from 10 pre tagged
plants in each plot.
Disease incidence was assessed starting from the onset of the disease. It was
recorded by counting the number of plants showing the symptom and dividing by
the total number of plants assessed; then the results were expressed in percentage
of disease incidence using the following formula.
---------------------------------------(1)
where: DI = disease incidence
---------------------------------------(2)
Where: DS = disease severity

The severity was scored four times at weekly interval starting from the onset of
the disease. It was collected by estimating the proportion of infected tissue areas
out of a total area assessed, and rated using 0-9 scale developed by IRR (2013) as
detailed in Table 1.
[81]
Table 1. IRRI standard evaluation system for rice sheath rot severity rating scales (0 – 9) and descriptions
Scale/ grade Description

0 No lesion/spot on flag leaf sheath.
1 Spots visible on the tillers upon very careful examination (<1% flag leaf sheath area covered).
3 Spots visible on the tillers upon careful examination (1-5% flag leaf sheath area covered).
5 Spots easily visible on the tillers (6-25% flag leaf sheath area covered).
7 Spots present on almost whole the tillers parts (26-50% flag leaf sheath area covered) damage
conspicuous.
9 Spots very common on whole the tillers parts (51-100% flag leaf sheath area covered), death of plants
common, damage directly reduce severe yield loss.
Source: IRR (2013)
The numerical values of the severity were further used for the calculation of the
mean percent severity index (PSI) using the following formula as indicated by
Wheeler (1969).
-----(3)
Based on their PSI values of reaction to the disease, the tested germplasms were
classified as resistant, moderately resistant, moderately susceptible, susceptible
and highly susceptible using the Lalan Sharma et al. (2013) standard as indicated
in table 2.
Table 2. Percent severity index (PSI) and Host reaction (HR) to rice sheath rot disease
Percent severity index (PSI) Host reaction (HR)

0% Immune
1-10% Resistant
11-25% Moderately resistant
25-50% Moderately susceptible
50-75% Susceptible
76-100% Highly susceptible
Since our objective was to identify sheath rot resistant germplasms through
screening, statistical analysis and mean separation were performed for all
germplasms and the result is presented here only for immune and resistant
germplasms found in all blocks along checks. Statistical analysis was performed
using Statistical Packages for Augmented Design (SPAD) and excel (Microsoft
office version 2016) following standard and specific procedures applied during
data analysis in augmented design experiments. Mean separation was also
computed for traits which have significant differences among germplasms using
LSD at 5% significant level as described in Gomez and Gomez (1984).
LSD 5% = t 0.025(error df.) x√ -----------------(4)

where C = number of checks
[82]
LSD 5% = t 0.025 (error df.) x √ --------(5)
where b = number of blocks and C= number of checks

Results from the combined analysis of variance for selected important agronomic,
phenological and disease traits are presented in Table 3. Mean squares estimates
from analysis of variance revealed that there were significant differences among
genotypes for some traits and no significant interaction for some other traits
(Table 3).
Table 3. Mean square estimates from analysis of variance (ANOVA) for selected traits across blocks
Sources of DF PH (cm) PL (cm) NFGPP NETPP DM PSI % TGW (g) GY (kg/ha)

variation
Unadjusted 3 611.65 8.46 361.41 4.00 469.45 234.76 55.49 2580646.00
Block(b-1)
Adjusted entries 83 114.61 1.50 131.47 1.04 28.50 137.70 13.17 948483.95
(c+g-1)
Unadjusted 83 136.39 1.72 143.09 1.14 45.07 145.69 15.00 1030999.00
entries(c+g-1)
Among 3 221.47** 1.49 ns 102.96 ns 3.06 * 12.77 ns 622.68 ** 37.99 ** 4013079.14**
controls/Check
(c-1)
Among Test 79 107.64** 1.50 ns 133.95 * 0.97 ns 27.93 ns 120.83 ** 11.93 ns 843148.88 ns
genotypes (g-1)
Test genotype * 1 344.21** 1.05 ns 21.53 ns 0.42 ns 120.97 * 15.81 * 37.12 * 76169.78 ns
check
Error (c-1) (b-1) 9 13.68 0.61 44.57 0.61 13.14 3.36 5.47 326574.81
Total (N-1) 95
CV% 4.92 4.42 9.62 13.27 2.56 11.35 7.50 12.99
DF = degree of freedom, CV = coefficient of variation in percent, PH = plant height in centimeter, PL = panicle length in centimeter,
NFGPP = number of filled grains per panicle, NFTPP = number of fertile tillers per plant, DM = days to maturity, PSI = percent severity
index, HR = host reaction, R = resistant, I = immune, TWG = thousand grains weight in gram, GY = grain yield in kg/ha, ** = significantly
different at 1 % level of significance, * = significantly different at 5% level of significance, ns = no significant difference.
Reactions of genotypes to rice sheath rot disease

Classification of the host reaction based on their PSI value according to Lalan
Sharma et al. (2013) rating scale revealed that the tested genotypes had different
reaction to the disease. Among 80 germplasms, three genotypes were immune, 27
resistant, 35 moderately resistant, 13 moderately susceptible and two germplasms
were susceptible. There were no any sheath rot symptoms observed in the immune
germplasms. While, among the 27 resistant genotypes the lowest PSI value
(1.48%) was scored on the Hangamchal followed by WAS 161-B-6-B-B-1-B
(NERICA-L-38) with PSI value of 2.22%. On the other hand, three genotypes viz.
CHOMRONG, IR 83222-F11-200 and Saegyejinmi scored the highest PSI value
(10% each), though they are within the resistant group. Moreover, the susceptible
germplasms such as Trakya and SCRID091-15-2-2-1-1 scored the highest PIS
[83]
value of 50.15 and 54.56%, respectively. While all the rest germplasms were
either moderately resistant or moderately susceptible with the PSI value ranging
from 11.48 to 39.17% (Table 4).
Similar study was conducted by Jakkuva (2012) using 44 genotypes and got
different reaction levels among the tested genotypes. The results of this study
revealed that out of the 44 genotypes, none was found immune. Whereas, two,
sixteen, fourteen, seven and five genotypes showed highly resistant, resistant,
moderately resistant, moderately susceptible and susceptible reactions,
respectively.
Table 4. Mean Percent Severity Index value and host reaction of rice germplasms against rice sheath rot disease
S.N Name of genotypes PSI % Reaction S/n Name of genotypes PSI % Reaction
1 Aromatic-1 9.85 R 41 IR74052-184-3-3 21.57 MR
2 Edirne 4.44 R 42 YUNJING 23 15.19 MR
3 Halilbey 23.56 MR 43 WAB502-8-5-1 16.30 MR
4 Osmancik-97 16.48 MR 44 PSBRC44 15.56 MR
5 Trakya 50.15 S 45 WAB376-B-10-H3 24.07 MR
6 Tunca 20.74 MR 46 IR 83222-F11-167 24.81 MR
Suitou Chuukanbohon
7 3.33 R 47 IR 83222-F11-18 25.74 MS
Nou 11
8 Condai 12.13 MR 48 IR 83222-F11-200 10.00 R
9 Pepita 11.48 MR 49 IR 83222-F11-209 21.11 MR
10 Saegyejinmi 10.00 R 50 IR 83222-F11-66 5.56 R
11 Lunyuki 7.04 R 51 IR76999-52-1-3-2 5.56 R
12 Hangamchal 1.48 R 52 IR 83249-F9-29 8.52 R
13 Hawaghaelo-2 15.26 MR 53 STEJAREE 45 22.70 MR
14 Namcheobyeo 18.04 MR 54 CHOMRONG 10.00 R
15 Samgangbyeo 34.07 MS 55 WAB880-1-38-20-17-P1-HB 20.19 MR
16 SCRID091-10-1-3-2-5 21.78 MR 56 IRAT112 26.48 MS
WAS 161-B-6-B-B-1-B
17 SCRID091-15-2-2-1-1 54.56 S 57 2.22 R
(NERICA-L-38)
18 SCRID091-18-1-5-4-4 12.22 MR 58 IR 83372-B-B-115-4 8.52 R
19 SCRID091-20-2-2-4-4 16.85 MR 59 IR 83377-B-B-93-3 12.59 MR
20 SCRID091-24-3-2-2-3 36.30 MS 60 IR 83383-B-B-141-2 25.19 MS
21 SCRID090-60-1-1-2-4 38.78 MS 61 IR 83372-B-B-115-3 31.11 MS
22 SCRID090-72-3-1-3-5 25.65 MS 62 IR80420-B-22-2 19.26 MR
23 SCRID090-164-2-1-2-1 6.78 R 63 IR80463-B-39-3 15.19 MR
24 SCRID090-177-2-4-3-4 20.93 MR 64 IR 72768-8-1-1 6.67 R
25 SCRID090-18-1-2-2-1 9.89 R 65 IR 75518-18-1-2-B 19.26 MR
26 SCRID091-20-3-1-3-4 39.17 MS 66 IR 75518-84-1-1-B 8.52 R
27 SCRID122-5-2-1-1-3 17.78 MR 67 YUNLU N0.33 0.00 I
28 SCRID122-13-1-1-4-3 28.52 MS 68 IR 81047-B-106-2-4 27.78 MS
WAS 161-B-6-B-1
29 SCRID186-72-1-1-2 34.44 MS 69 5.19 R
(NERICA-L-36)
30 SCRID198-73-5-1-3 10.74 MR 70 ARCCU16Bar-13-2-16-2-1-1 3.33 R
31 scrid079-1-5-4-2 20.74 MR 71 Yungeng 44 7.78 R
32 exp304 14.81 MR 72 Yungeng 45 8.15 R
33 FOFIFA 171 8.89 R 73 Yungeng 38 5.63 R
34 FOFIFA 172 17.22 MR 74 Fengdao 23 6.30 R
35 FOFIFA 167 3.33 R 75 KB-2 9.26 R
[84]
36 HR 17512-11-2-3-1-4-2-3 15.93 MR 76 Songgeng9 20.74 MR
37 scrid014-1-1-1-1 0.00 I 77 P-28 14.00 MR
38 scrid019-1-1-1-1-2 4.44 R 78 P-38 4.44 R
39 Scrid037-4-2-2-5-2 0.00 I 79 Li Jing 9 15.56 MR
40 Scrid113-3-5-3-5-4 5.56 R 80 Li jing 11 16.67 MR
40
35 35
Number of germplasms
30
27
25
20
15 13
10
5 3 2
0
0 (I) 1-10% R 11-25% (MR)25-50 % (MS) 50-75% (S)
Percent severity index value and host reaction

Figure 1. Reactions of 80 rice germplasms against sheath rot disease
Similarly, classification of host reaction based on their PSI value revealed that the
check varieties had variable response to the disease. Among the four check
varieties, two of them (Erib and Wanzaye) were resistant while Idget was
moderately resistant and X-jigna was moderately susceptible (Table 5).
Table 5. Mean Percent Severity Index value and host reaction of the check
rice varieties against rice sheath rot at Fogera
S.N Check rice varieties PSI % Reaction

1 Erib 5.83 R
2 Idget 20.90 MR
3 Wanzaye 4.074 R
4 X-jigna 30.13 MS
Agro morphological characteristics of immune and resistant germplasms

The mean performance of different germplasms occurring across different blocks
is given in Table 5. The following characteristics were based on the data generated
in sheath rot resistance screening experiment. Most of the immune and resistant
germplasm had better agronomic and morphological performance as compared
with the checks in all aspects, while some of them had equal or less agronomic
and better resistance performance with checks. This is evidence that resistance and
yield response are sometimes having inverse relation. Therefore, the resistant
[85]
genotypes can be used as source of resistant gene(s) for crossing purpose with
high yielding but susceptible varieties.
Among the 27 resistant germplasms, the highest yield was scored on Fengdao 23
(6462.50 kg h-1), followed by Yungeng38 (6301.02 kg h-1), Yungeng 45 (6249.05
kg h-1) and scrid019-1-1-1-1-2 (6102.02 kg h-1) as compared with other resistant
and immune germplasms. In addition, these genotypes also had moderate
performance in other agronomic, phenological and morphological traits (Table 6).
Similarly, of the three immune germplasms, namely scrid014-1-1-1-1, Scrid037-4-

2-2-5-2 and YUNLU N0.33, the highest yield (5974.11 kg h-1) was scored on
scrid014-1-1-1-1 as compared with other immune germplasms and almost nearly
equal with high yielder resistant germplasms. Moreover, scrid014-1-1-1-1 was
superior over the check varieties and other immune germplasms in all other traits.
While Scrid037-4-2-2-5-2 was performed almost as equal as Wanzaye variety and
superior than other check varieties and the immune genotype YUNLU N0.33,
which performs better than Idget and X-jigna varieties (Table 7).
[86]
Table 6. Mean values of agro morphological traits of selected immune and resistant germplasms occurring at different blocks
S.N Genotype PH (cm) PL (cm) NFGPP NFTPP DM PSI % HR TGW (g) GY (kg/ha)
1 Aromatic-1 64.73 16.73 64.29 6.87 140.33 9.85 R 34.73 3211.94
2 Edirne 69.27 16.67 63.71 8.00 128.00 4.44 R 31.98 4239.27
3 Suitou Chuukanbohon Nou 11 80.20 18.13 74.47 4.67 137.33 3.33 R 32.97 4272.01
4 Saegyejinmi 62.93 16.73 74.44 5.27 144.00 10.00 R 26.24 3189.59
5 Lunyuki 79.27 16.33 71.40 5.07 144.33 7.04 R 30.47 2395.36
6 Hangamchal 75.60 16.87 73.07 4.27 137.00 1.48 R 34.79 3840.01
7 SCRID090-164-2-1-2-1 91.33 17.87 72.29 5.73 137.00 6.78 R 33.11 4748.68
8 SCRID090-18-1-2-2-1 79.80 17.60 60.93 4.33 137.00 9.89 R 36.20 4439.29
9 FOFIFA 171 79.00 17.20 61.64 5.07 141.00 8.89 R 33.87 4774.28
10 FOFIFA 167 88.20 15.53 54.62 5.67 133.67 3.33 R 31.18 5501.32
11 scrid014-1-1-1-1 92.00 17.33 76.56 7.00 139.67 0.00 I 27.13 5974.11
12 scrid019-1-1-1-1-2 90.13 17.13 69.53 7.00 140.00 4.44 R 28.40 6102.02
13 Scrid037-4-2-2-5-2 87.73 16.93 82.24 7.13 140.00 0.00 I 28.63 5528.59
14 Scrid113-3-5-3-5-4 84.93 17.53 62.22 6.53 137.33 5.56 R 37.54 4813.07
15 IR 83222-F11-200 63.53 16.93 58.27 6.87 141.67 10.00 R 28.29 4644.31
16 IR 83222-F11-66 84.40 18.40 81.51 5.33 147.00 5.56 R 30.10 4547.39
17 IR76999-52-1-3-2 59.93 17.73 60.00 7.60 150.67 5.56 R 26.08 4966.75
18 CHOMRONG 83.80 17.60 51.04 6.75 133.67 10.00 R 34.28 4138.48
19 WAS 161-B-6-B-B-1-B (NERICA-L-38) 59.33 18.53 58.58 5.47 150.67 2.22 R 25.84 4076.84
20 IR 72768-8-1-1 59.13 18.73 71.24 6.13 152.00 6.67 R 27.85 4575.27
21 IR 75518-84-1-1-B 75.13 18.13 53.47 5.73 155.67 8.52 R 31.19 1917.42
22 YUNLU N0.33 85.47 19.20 86.22 5.27 146.67 0.00 I 32.77 5024.36
23 WAS 161-B-6-B-1 (NERICA-L-36) 54.40 17.87 60.40 5.93 149.67 5.19 R 29.35 3024.07
24 Yungeng 44 85.73 20.60 94.24 5.07 144.00 7.78 R 30.24 5977.10
25 Yungeng 45 83.40 21.20 109.98 4.73 145.67 8.15 R 32.37 6249.05
26 Yungeng 38 72.27 17.27 77.98 8.13 147.67 5.63 R 30.74 6301.02
27 Fengdao 23 78.00 18.60 85.73 5.27 146.00 6.30 R 29.41 6462.50
28 KB-2 68.80 18.13 76.51 5.80 142.67 9.26 R 27.22 3942.43
29 Songgeng9 74.13 17.70 57.78 5.80 147.67 2.59 R 32.13 5987.47
30 P-38 69.93 18.20 66.09 6.47 143.67 8.52 R 28.02 3894.43
Adjusted grand mean 74.58 17.67 69.52 5.91 141.93 16.27 30.96 4388.60
LSD 5% 12.45 2.60 23.25 2.65 11.76 6.49 32.84 2043.57
CV% 4.92 4.42 9.62 13.27 2.56 11.35 7.50 12.99
* PH = plant height in centimeter, PL = panicle length in centimeter, NFGPP = number of filled grains per panicle, NFTPP = number of fertile tillers per plant, DM = days to maturity, PSI =
percent severity index, HR = host reaction, R = resistant, I = immune, TWG = thousand grains weight in gram, GY = grain yield in kg/ha
[87]
Table 7. Mean values of agro morphological traits of selected immune germplasms along checks
S/N Genotype PH (cm) PL (cm) NFGPP NFTPP DM PSI % HR TGW (g) GY (kg/ha)
1 Erib 69.82 17.45 69.34 5.18 138.67 5.83 R 30.44 5027.10
2 Idget 77.28 17.87 66.60 4.82 141.08 20.90 MR 35.09 3825.47
3 X-jigna 85.70 18.82 74.70 6.60 139.67 30.13 MS 29.38 3426.44
4 Wanzaye 84.88 17.62 62.62 6.37 136.83 4.07 R 35.29 5563.33
5 scrid014-1-1-1-1 92.00 17.33 76.56 7.00 139.67 0.00 I 27.13 5974.11
6 Scrid037-4-2-2-5-2 87.73 16.93 82.24 7.13 140.00 0.00 I 28.63 5528.59
7 YUNLU N0.33 85.47 19.20 86.22 5.27 146.67 0.00 I 32.77 5024.36
Adjusted grand mean 74.58 17.67 69.52 5.91 141.93 16.27 30.96 4388.60
LSD 5% 5.91 1.23 11.03 1.26 5.58 3.08 15.58 969.35
CV% 4.92 4.42 9.62 13.27 2.56 11.35 7.50 12.99
* PH = plant height in centimeter, PL = panicle length in centimeter, NFGPP = number of filled grains per panicle, NFTPP = number of fertile tillers per plant,
DM = days to maturity, PSI = percent severity index, HR = host reaction, R = resistant, I = immune, TWG = thousand grains weight in gram, GY = grain yield in kg/ha.
[88]
It is clear from table 7 that the immune genotypes gave high yield ranging from
5024.36 to 5974.11 kg h-1, and all are apparently free of disease as compared with
the checks. In addition, they had better plant height (ranging 85.47 – 92 cm),
panicle length (16.93 – 19.20 cm), field grains per panicle (76.62–86.22) and
fertile tillers per plant (5.27– .13) (Table 7), which all have direct contribution for
yield increment. On the contrary, the moderately resistant and moderately
susceptible checks (Idget and X-jigna, respectively) had less performance in the
above-mentioned traits compared with the immune genotypes. In fact, when a
genotype is susceptible and attacked by sheath rot disease, it tends to give short
plant height, short panicle length (un-emerged panicle) and chaffy grains, leading
to yield reduction (Kindo, 2012). Lalan Sharma et al. (2013) also reported that, the
dwarf varieties appeared to be more prone to sheath rot because of their shortened
internodes and poor exertion of the panicle from the flag leaf sheath. Genotypes
resistant to diseases, high yielder and good with other agronomic traits are of great
interest for researchers as well as rice producers. In cognizant of this, the present
study resulted in promising genotypes possessing traits of good agricultural
importance (high yielder and disease resistance) like scrid014-1-1-1-1 (Table 7),
which the existing varieties under cultivation do not have. This result is in line
with the findings of Simon (2016), who screened 64 rice genotypes and observed
different level of resistance among the genotypes, of which six genotypes were
found to be resistant to the disease. Such genotypes can serve as sources of
qualified variety development so as to ensure the satisfaction of rice producers to
get high yield and high net return.
High net return is achieved by growing varieties having trait of disease resistance,
high yielder and with high market value or high consumer satisfaction.
Unfortunately, the available varieties in Ethiopia lack either one or two of these
important traits. Moreover, farmers in Fogera plain repeatedly reported that the
market demanded cultivar ‘X-jigna’ is severely attacked by sheath rot. It is,
therefore, hopped that the result of the current study will alleviate this problem if
the identified good traits possessed by genotype ‘scrid014-1-1-1-1’ is properly
used as a resistant donor parent to cross with the white color and market
demanded ‘X- jigna’ cultivar.

The aim of this study was to screen rice germplasms for their resistance against
sheath rot, one of the most important rice diseases that seriously threaten rice
productivity in Fogera plains. Because of the fact that rice cultivation is relatively
recent to Ethiopia, more research had not so far been done in the country towards
the control of this disease. Genetic improvement of locally adapted cultivars
through breeding for resistance to this economic disease would be the most
[89]
sustainable and cost-effective strategy to tackle the threat caused by the disease.
To this end, identifying sources of resistance among introduced germplasm was
the first major step forward and results from this study revealed that 30
germplasms had different levels of resistance to sheath rot (3 immunes and 27
resistant).
Moreover, among the three genotypes found to be immune, genotype ‘scrid014-1-

1-1-1’ is more yielder than the newly released high yielder and resistant check,
Wanzaye. Thus, it is possible to conclude that this genotype can be considered as
both resistant and high yielder candidate variety for release having passed through
verification and demonstration stages. Therefore, cultivar ‘scrid014-1-1-1-1’ will
be promoted for national variety verification trial along standard checks for the
next cropping season.
Generally, all the three immune germplasms will be considered the best sources of
resistant genes for sheath rot as far as varietal improvement is concerned,
especially since they are already adapted to most of Fogera’s lowland rice growing
ecosystems. Furthermore, the 27 genotypes found resistant to sheath rot are
additional resource increasing the chance of broadening the genetic bases of rice
as far as developing sustainable, better resistant and high yielder varieties is
concerned.
References
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Microbiology 50: 319-323.
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and Hawksworth. MSc Thesis, Dharwad University of Agricultural Sciences, Dharwad. Pp
87.
Kindo D. 2012. Studies on management of sheath rot disease of rice. MSc. Thesis, Indira Gandhi
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potential and incidence of sheath rot of rice caused by Sarocladium oryzae (Sawada). Journal
of Applied and Natural Science, 5: 24-29.
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of Sheath Rot in Rice and Bamboo Blight. In: Sreenivasaprasad, S. and Johnson R. (eds). Major
fungal diseases of rice: Recent advances. Springer Netherlands, Dordrecht. p. 285-292.
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Accessions for Multiple Disease Resistance. Plant Pathology and Microbiology Publications,
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Simon MM. 2016. Increasing the resilience of elite rice cultivars to sheath rot (Sarocladium oryzae
[(Sawada) W. Gams & D. Hawksw]) in Rwanda through breeding for resistance . PhD
dissertation, University of KwaZulu-Natal Republic of South Africa. Pp 157.
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gram (Phaseolus aureus Roxb.). Bioscience Discovery, 4(1): 107-110.
Wheeler BJ. 1969. An Introduction to Plant Diseases. John Wiley and Sons, Ltd.
[91]
[92]
Screening of Quality Protein Maize Inbred Lines
for Turcicum Leaf Blight and Gray Leaf Spot
Midekssa Dida, Temesgen Deressa and Debela Diro
Ethiopian Institute of Agricultural Research, Bako National Maize Research Centre,
P.O.Box 3, Bako, Ethiopia
Abstract
Maize (Zea mays) is one of the most widely cultivated crops in the world as well in
Ethiopia. But maize production in Ethiopia is constantly threatened by the potential
outbreak of major foliar diseases such as Turcicum leaf blight (TLB) and Gray leaf
spot (GLS). These diseases are characterized by relatively rapid leaf necrosis and
premature death of foliage which eventually reduces grain yield. The diseases become
a major problem in all major maize growing areas. Development of host resistance to
these diseases can provide an important component of integrated disease
management; which is the most effective and practical method of managing maize
diseases. The study was conducted to evaluate the reaction of quality Protein maize
inbred lines to TLB and GLS in the main cropping season during 2018. The inbred
lines were obtained from Bako National Maize Research Center, breeding program,
and it was arranged using alpha-lattice design with two replications. The inbred lines
were evaluated in TLB and GLS screening field under artificial inoculation at Bako
Agricultural Research Center, West Shewa, Ethiopia. Disease severity (1-5 scale) was
used to assess at ten days intervals from disease onset until the maize attained
physiological maturity. All the inbred lines showed symptoms of both diseases during
the season, but the intensity of the diseases differed significantly (P<0.05) among the
inbred lines. Accordingly, out of 42 genotypes screened for both TLB and GLS, lines
MBRC5BcF108-2-3-1-B-5-2-B-B-#,Obatanpa-5-4-1-1-1-#-#-#,([[[CML159/[CML159/
[MSRXPOOL9]C1F2-205-1,(OSU23i)-5-3-X-X-1-BB]F2-3sx]-8-1-BB/CML509]F2-
1/[GQL5/[GQL5/[MSRXPOOL9]C1F2-205-1(OSU23i)-5-3-X-X-1-BB]F2-4sx]-11-3-1-1B*4]-
12/CML373IR)-B-8-B-7-2-1-B-#-#-#,(CML176xKULENI(F2)-4-3-1-1-1-#-#,(CUBA/
GUADC1F27-4-3-3-B-1-BX[KIiIMAiTA94A]-30MSV-03-2-10-B-2-B-B)-306-1-B-2-B-B-B-
#, were selected as resistant based on disease severity measured on 1-5 rating scale.
Introduction
Maize (Zea mays L.) is one of the most important cereal crops in Ethiopia, ranking
second in area coverage and first in total production. Currently, about 2.13 million
ha of land is covered by maize with an average production of 8.39 million tons,
and the average national yield of maize under small-scale farmers reaches 3.94
t/ha (CSA, 2018). However, it is very low as compared to the potential of maize in
the high rainfall and irrigated areas, which is (8 –11 t ha-1) and also low as
compared to world average productivity of 5.78 ton ha-1 (USDA-FAS, 2017). An
important portion of this low yielding in maize is attributable to abiotic and biotic
stresses, and poor management practice as well as climate change. Among biotic
factors, foliar diseases are reported to be widespread and destructive disease of
maize mainly in warm and humid growing regions of Ethiopia (Tewabech et al.,
2001). Out of various maize foliar diseases Turcicum Leaf Blight (TLB) and Gray
[93]
Leaf Spot (GLS) which are caused by Excerohillum turciccum and Cercosporra
zeae maydis, respectively, are the most economically important diseases of maize
crop production (Tewabech et al., 2012). Due to the presence of conducive
environmental factors such as relatively higher humidity, moderate to high
temperature and/ or warm areas during the maize cropping season at mid-altitude
sub-humid agro-ecology of Ethiopia, Turcicum leaf blight and grey leaf spot
diseases are considered as the most economically important, which are identified
to cause significant yield lose. If infection occurs in early developmental stage of
the plant, Turciccum leaf blight can cause leaf necrosis and premature death of
foliage, which reduces grain yield of the crop (Raymundo and Hooker, 1981). On
the other hand, grey leaf spot shows necrotic lesion tend to be long and individual
lesion may merge, lead to leaf senescence and greatly reducing the photo-synthetic
areas leading to poor grain filling (Darera et al., 2008). A yield loss due to GLS
was estimated at about 37 % in Ethiopia (Wegary et al., 2004) and 60% in South
Africa (Ward et al., 1997). Similarly, according to Assefa and Tilahun (1992),
TLB caused the highest mean grain yield loss of 50% and thousand kernel weight
loss of 16.4% on susceptible cultivars. Methods to manage TLB and GLS diseases
include cultural practices, chemical and host plant resistance (Pratt et al., 2003).
Due to its inconsistency with environment and expensiveness, uses of chemical
fungicides are not effective. The most effective and cost-efficient means of
managing these diseases is the use of host plant resistance. It is, therefore,
desirable to identify resistant inbred lines from diverse sources in maize pre-
breeding program in order to improve genetic resistance to these foliar diseases.
Though early research efforts made to identify maize germplasm resistant to these
diseases and utilizing them for maize breeding program, subsequent study for
additional source of maize germplasm should be screened under artificial
inoculation to obtain new and stable resistance. The objective of this study was to
select quality protein maize inbred lines that are resistant/tolerant to GLS and/or
TLB through evaluation from locally developed and adapted maize inbred lines
for use in maize improvement program.

The experiments were conducted at Bako national maize research center
(BNMRC) of maize disease nursery field during main season of 2018. Bako is
located at 9°06’ N and 37°09' E and receives the annual rain fall of about 1237
mm and situated at an altitude of 1650 m above sea level, which represent mid
altitude sub-humid agro-ecology zone of Ethiopia. It has minimum and maximum
average temperature of 15.60C and 30.70C, respectively. Both experiments were
conducted in the field under artificial epiphytotic conditions for evaluation against
TLB and GLS.
[94]
Description of experimental materials and design
A total of forty-two QPM inbred lines were used and arranged in 6x8 alpha lattice
design with three replications. Some of the genotypes were developed by BNMRC
and the rest are obtained from CIMMYT. Initially, inbred lines were developed for
their Quality Protein and yield traits, but their resistance level also very important.
Each inbred line was planted in a plot consisting of two rows of 3.6m long spaced
at 25 and 75cm between plants with in rows and rows, respectively. The non-
quality protein maize inbred lines of CML-197 and SC-22 were used as
susceptible check for TLB and GLS, respectively. Nitrogen (N2) and di-
ammonium phosphate (P2O5) fertilizers were applied at the recommended rates of
92 kg/ha and 69 kg/ha, respectively. All agronomic management practices for the
area were applied as per the recommendations.
Inoculum preparation and inoculation

Inoculum of E. turcicum and C. zeae maydis was prepared a year before
experimentation by collecting from heavily infected maize fields showing distinct
TLB and GLS symptoms, respectively. The infected leaves were dried under
shade and crushed/grounded in to mill about the coarseness of wheat bran and
stored in paper bags at a temperature of 4oC until inoculation date. The pulverized
leaves then dusted in the whorls of the plants according to Dagne et al. (2008) by
placing a pinch of leaf mill when plant attains 6-8 leaf stage during moisten
environments in order to retain long enough to permit spore germination.
Inoculation and data collections were done for both diseases separately in different
experimental fields. Some of the genotypes were developed by BNMRC and the
rest are adopted from CIMMYT. A second inoculation was made ten days later
after the first inoculation to ensure adequate infection.
Assessment of disease reaction

TLB and GLS were visually assessed in the field two weeks after artificial
inoculation on a plot basis from the two rows. Data collected included date first
disease appeared, disease incidence, disease severity, and other agronomic traits
including plant height (cm) and grain yield (t/ha). The progress of severity of
foliar diseases on each inbred lines were quantified at ten days intervals starting
from onset of disease until dent stages and the highest or final severity value of
each inbred lines was used for statistical analysis. Disease incidence was measured
as percent of infected plants per total plant per plot. Disease severity was rated
based on 1-5 scoring scale (CIMMYT, 1985); where 1=no disease symptoms,
2=moderate lesion below the leaf subtending the ear, 3=heavy infestation on and
below the leaf subtending the ear with few lesions above it, 4=severe lesion on all
but the uppermost leaves which may have a few lesions, 5=all leaves dead. The
categorization on each disease reactions was made on the basis of disease severity
ratings using a 1-5 scale (Roane et al., 1974) with some modifications, where;1.0–
[95]
2.0=Resistant (R); 2.1-2.5=Moderately Resistant (MR); 2.6-3.0= Susceptible (S),
and >3.0 Highly susceptible (HS).
Statistical data analysis

Data were analyzed using PROC GLM of SAS version 9.2 (SAS Institute, 2004).
Mean separation was performed to compare treatment means using LSD-test at
5% level of significance.

A total of forty-two QPM inbred lines were screened for resistance to TLB and
GLS. The mean disease severity and yield results indicated significant (P < 0.05)
variation among the inbred lines for TLB and GLS resistance (Tables 1 and 2).
Disease severity ranged from 1.6 to 4.1 for TLB and 1.3 to 4.5 for GLS. Inbred
lines with mean severity values of < 2 were categorized under resistant/tolerant to
TLB and GLS. Whereas inbred lines with mean severity values ranging from 2.1-
2.5 categorized as moderately resistant, from 2.6-3.0 as susceptible and those with
severity value > 3 were considered as highly susceptible to TLB and GLS (Tables
1 and 2). Accordingly, 32 inbred lines were resistant/tolerant, 8 inbred lines
moderately resistant and 2 were Susceptible to TLB (Table 1). Similarly, 7, 6, 3
and 26 inbred lines were categorized, respectively, under resistant/tolerant,
moderately resistant, susceptible and highly susceptible to GLS (Table 2). Those
inbred lines, which have showed resistance/tolerance were compared to the
susceptible checks CML 197 and SC 22 for TLB and GLS, respectively. Best
selected inbred lines could be used as TLB and GLS source of resistance for use in
maize resistance breeding programs.
[96]
Table 1. Mean Turciccm leaf blight severity and yield of 42 QPM inbred lines evaluated under artificial inoculation during 2018 main cropping season at Bako.
Entry TLB (Sev1-5 Reaction Grain Yield

Pedigree scale)
No t/ha
1 BQ00RC3-#-28-2-1-1-1-1-1-#-#-# 1.5 R 3.29
2 MBRC5BcF108-2-3-1-B-5-2-B-B-# 1.5 R 2.00
3 Obatanpa-180-2-1-1-2-2-1-#-#-# 1.5 R 4.28
([[[CML159/[CML159/[MSRXPOOL9]C1F2-205-1(OSU23i)-5-3-X-X-1-BB]F2-3sx]-8-1-BB/CML509]F2- MR
4 2.5 3.51
1/[CML182/[CML182/CML395]F2-3sx]-4-1-B]-3/CML390IR)-BBB-4-1-1-B-#-#-#
5 (CML142/144-7-B) (F 2)- 9-2-1-2-2-1-#-#-# 2.3 MR 2.15
6 (CML197-B/[CML144/[CML144/CML395] F2-8sx]-1-2-3-2-B*5) F2-B-46-1-3-B-#-#-# 2.0 R 2.49
7 BQ00RC3-#-356-1-1-2-1-1-1-#-#-# 1.5 R 1.16
8 BQ00RC3-#-216-2-2-1-1-1-#-#-# 2.0 R 1.42
9 BQ00RC 3- #-55-1-2-1-1-1-1-#-#-# 1.5 R 3.00
11 (CML176/Kuleni) (F2)-3-1-1-2-1-1-1-#-#-# 2.3 MR 0.63
12 CML142 X 144-7-b (F 2)- 9-2-1-2-2-1-#-#-# 2.5 MR 1.51
13 BQ00RC3-#-354-2-1-2-2-1-1-#-#-# 1.8 R 3.07
([[[CML159/[CML159/[MSRXPOOL9]C1F2-205-1(OSU23i)-5-3-X-X-1-BB]F2-3sx]-8-1-BB/CML509]F2-
14 2.0 R 1.53
1/[GQL5/[GQL5/[MSRXPOOL9]C1F2-205-1(OSU23i)-5-3-X-X-1-BB]F2-4sx]-11-3-1-1-B*4]-12/CML373IR)-B-8-B-7-2-3-B-#-#-#
15 CML142 Short-#-#-# 1.5 R 1.37
16 CML144/KULENI(F2) 11-2-1-3-1-1-1-#-#-# 2.0 R 2.02
17 Obtanpa-14-6-1-1-1-1-#-#-# 1.5 R 1.51
18 (cml142x124-b (113) (f2) x124-b (113) (f2) x124-b (113) (f2))-B-4-1-1-1-#-#-# 1.5 R 3.47
19 CML-144 X CML-159 (F2)-20-1-1-1-2-#-# 1.5 R 2.37
20 CML-144 X CML-159 (F2)-20-1-1-1-1-#-#-# 1.5 R 1.74
21 CML142 X 144-7-b (F 2)- 9-2-2-1 -1-1-1-#-#-# 2.3 MR 2.30
22 1.8 R 3.69
23 BQ00RC3-#-356-1-1-1-1- -#-# 1.8 R 4.18
24 Obatanpa-5-4-1-1-1-#-#-# 1.5 R 2.03
26 BQ00RC#32-1-2-2-1-1-1-#-#-# 2.8 S 3.44
27 BQ00RC3-#--331-2-1-1-1-1-1-#-#-# 1.5 R 1.15
28 CML194-#-# 1.5 R 1.65
[97]
29 [[CML506/[CML205/CML176]-B-2-1-1-B] F2-1/[CML144/[CML144/CML395] F2-8sx]-1-2-3-2-B*4]-24-B-2-BBB-3-B-B-#-#-# 1.8 R 3.51
30 (CML142/144-7-B) (F 2)- 9-1-1-1-1-1-#-#-# 1.5 R 2.04
31 Obatanpa-301-4-3-1-1-#-# 2.3 MR 2.71
32 Z97SYNGLS(B)-F2-188-2-1-3-B*8-#-#-#-# 1.5 R 2.34
33 (GH-132-28)-22-1-6-1-1-#-#-# 1.3 R 1.28
34 BK02-Z -311-28(F2) B-1-#-# 1.5 R 1.92
35 CML142-#-# 2.5 MR 4.03
36 1.5 R 2.75
37 (CML176 x KULENI(F2)-4-3-1-1-1 -#-# 1.5 R 2.57
38 CML334-# 3.3 S 2.15
39 Obatanpa -5-4-1-2-1-1-#-# 1.5 R 4.05
40 (CUBA/GUADC1F27-4-3-3-B-1-BX[KIiIMAiTA94A]-30MSV-03-2-10-B-2-B-B)-306-1-B-2-B-B-B-# 1.5 R 3.68
41 (CML142/144-7-B) (F 2)- 2-2-2-2-1-1-#-#-# 1.8 R 2.69
42 CML-144 X CML-159 (F2)-19-1-1-2-2-#-#-# 2.3 MR 3.31
LSD
1.1 2.01
(0.05)
CV 29.9 32.10
1.0–2.0=Resistant (R); 2.1-2.5=Moderately Resistant (MR); 2.6 -3.0= Susceptible (S) (Roane et al., 1974)
[98]
Table 2. Mean Grey Leaf Spot severity and yield of 42 QPM inbred lines evaluated under artificial inoculation during 2018 main cropping season at Bako.
GLS sev. Grain

Entry Reaction
Pedigree (Scale 1-5) Yield t/ha
No.
1 BQ00RC3-#-28-2-1-1-1-1-1-#-#-# 3.5 HS 3.36
2 MBRC5BcF108-2-3-1-B-5-2-B-B-# 2.3 MR 2.06
3 Obatanpa-180-2-1-1-2-2-1-#-#-# 3.5 HS 4.26
4 2.3 MR 4.09
1/[CML182/[CML182/CML395]F2-3sx]-4-1-B]-3/CML390IR)-BBB-4-1-1-B-#-#-#
5 (CML142/144-7-B) (F 2)- 9-2-1-2-2-1-#-#-# 3.8 HS 2.23
6 (CML197-B/[CML144/[CML144/CML395] F2-8sx]-1-2-3-2-B*5) F2-B-46-1-3-B-#-#-# 4.0 HS 2.52
7 BQ00RC3-#-356-1-1-2-1-1-1-#-#-# 2.0 R 1.28
8 BQ00RC3-#-216-2-2-1-1-1-#-#-# 2.3 MR 1.57
9 BQ00RC 3- #-55-1-2-1-1-1-1-#-#-# 3.5 HS 4.95
11 (CML176/Kuleni) (F2)-3-1-1-2-1-1-1-#-#-# 3.1 HS 0.97
12 CML142 X 144-7-b (F 2)- 9-2-1-2-2-1-#-#-# 3.8 HS 1.65
13 BQ00RC3-#-354-2-1-2-2-1-1-#-#-# 3.3 HS 3.89
14 3.5 HS 1.52
15 CML142 Short-#-#-# 3.8 HS 1.45
16 CML144/KULENI(F2) 11-2-1-3-1-1-1-#-#-# 3.3 HS 2.09
17 Obtanpa-14-6-1-1-1-1-#-#-# 1.5 R 1.88
18 (cml142x124-b (113) (f2) x124-b (113) (f2) x124-b (113) (f2))-B-4-1-1-1-#-#-# 3.3 HS 4.52
19 CML-144 X CML-159 (F2)-20-1-1-1-2-#-# 3.3 HS 2.52
20 CML-144 X CML-159 (F2)-20-1-1-1-1-#-#-# 2.8 S 1.83
21 CML142 X 144-7-b (F 2)- 9-2-2-1 -1-1-1-#-#-# 2.5 MR 2.30
22 3.8 HS 4.68
23 BQ00RC3-#-356-1-1-1-1- -#-# 3.5 HS 4.23
[99]
24 Obatanpa-5-4-1-1-1-#-#-# 1.5 R 2.09
26 BQ00RC#32-1-2-2-1-1-1-#-#-# 3.8 HS 3.56
27 BQ00RC3-#--331-2-1-1-1-1-1-#-#-# 3.5 HS 1.20
28 CML194-#-# 2.5 MR 1.67
29 [[CML506/[CML205/CML176]-B-2-1-1-B] F2-1/[CML144/[CML144/CML395] F2-8sx]-1-2-3-2-B*4]-24-B-2-BBB-3-B-B-#-#-# 3.8 HS 4.54
30 (CML142/144-7-B) (F 2)- 9-1-1-1-1-1-#-#-# 3.8 HS 2.10
31 Obatanpa-301-4-3-1-1-#-# 4.8 HS 2.69
32 Z97SYNGLS(B)-F2-188-2-1-3-B*8-#-#-#-# 3.0 S 3.03
33 (GH-132-28)-22-1-6-1-1-#-#-# 1.5 R 1.36
34 BK02-Z -311-28(F2) B-1-#-# 3.8 HS 1.94
35 CML142-#-# 4.3 HS 3.08
36 2.0 R 2.92
37 (CML176 x KULENIi(F2)-4-3-1-1-1 -#-# 1.5 R 3.41
38 CML334-# 3.5 HS 2.29
39 Obatanpa -5-4-1-2-1-1-#-# 3.0 S 4.19
40 (CUBA/GUADC1F27-4-3-3-B-1-BX[KIiIMAiTA94A]-30MSV-03-2-10-B-2-B-B)-306-1-B-2-B-B-B-# 2.0 R 3.72
41 (CML142/144-7-B) (F 2)- 2-2-2-2-1-1-#-#-# 4.3 HS 2.84
42 CML-144 X CML-159 (F2)-19-1-1-2-2-#-#-# 2.5 MR 3.26
LSD
1.1 2.05
(0.05)
CV 17.9 30.14
GLS, Gray Leaf spot
[100]
80.00 76.19
70.00
60.00
Frequency (%)
50.00
40.00
30.00
19.05
20.00
10.00 4.76
0.00
R MR S
Reaction Categories
Figure 1. Frequencies of QPM inbred lines with resistant (R), moderately resistant (MR) and susceptible (S) reactions to
Turcicum leaf blight (TLB).
70.00
61.90
Frequency (%)
60.00
50.00
40.00
30.00
20.00 16.67
14.29
10.00 7.14
0.00
R MR S HS
Reaction Catagories
Figure 2. Frequencies of QPM inbred lines with resistant (R), moderately resistant (MR) susceptible (S), and Highly
susceptible (H S) reactions to Gray Leaf Spot (GLS).
Field screening studies indicated that there were clear cut differential response of
inbred lines to TLB and GLS (Figures 1 & 2) due to artificial inoculation and
significant variation was observed among inbred lines with respect to disease
severity Chandrashekara et al. (2014). Resistance/tolerance reactions to TLB and
GLS were recorded in 76.19% and 16.67% of the inbred lines tested, respectively.
The data revealed that the highest numbers of the inbred lines tested have shown
resistant reaction to TLB and GLS, of which five inbred lines were showed
resistant/tolerant reaction to both GLS and TLB. Inbred lines, namely
[101]
(CUBA/GUADC1F27-4-3-3-B-1-BX[KIiIMAiTA94A]-30MSV-03-2-10-B-2-B-
B)-306-1-B-2-B-B-B-#, BQ00RC3-#-356-1-1-2-1-1-1-#-#-#, Obtanpa-14-6-1-1-1-
1-#-#-#, Obatanpa-5-4-1-1-1-#-#-#, (GH-132-28)-22-1-6-1-1-#-#-#, (CML176 x
KULENI (F2)-4-3-1-1-1 -#-# were showed multiple disease resistance. It is likely
that the more susceptible inbred lines loss their active leaf tissues and resulted in
less photosynthetic leaf area, then the plant eventually produce few kernels and/or
may contribute to the overall yield loss, showing negative correlation with disease
severity. Similar research finding was reported by Singh et al. (2014), where 27
maize population and 38 inbred lines screened for resistance to Turcicum leaf
blight showed significant differences. The moderately resistant and resistant
inbred lines should further be screened for stability to TLB and GLS across
location and years at hotspot areas and/or under controlled condition. Generally,
32 inbred lines were found resistant (scored < 2.0 on 1-5 scale) to TLB, and 6 to
GLS under artificial epiphytotic condition. Thus, it can be emphasized from the
results that the identified resistant lines hold promise for resistance against E.
turcicum and C. zeae maydis causing TLB and GLS of maize, respectively. The
selected inbred lines can also be used for developing hybrids and composites in
future breeding programs for multiple-disease resistance. These findings are in
agreement with the works of Patil et al. (2000), Muiru et al. (2007) and
Pandurangegowda et al. (1994), who reported differential reaction to diseases
among the various maize germplasm. Sharma and Payak (1990) reported durable
resistance in CM-104 and CM-105 maize inbred lines against E. turcicum.
Promising maize disease-resistance sources were also reported by Dagne et al.,
(2008), who identified 143-5-I and CML-387 as resistant, Gotto LMS5, SC-22 and
CML-395 moderately resistant and A-7016 and CML-197 susceptible to GLS.

Maize inbred lines identified as resistant/tolerant in this study should also be
screened under controlled environments to properly verify the level of resistance
to TLB and GLS. It would be better to use molecular methods to locate the gene
(s) involved in the resistance and incorporating them to cultivars having desired
agronomic characteristics. Moreover, the promising lines with good yield and
other agronomic performance identified through this investigation can be
deployed in disease endemic areas for sustainable maize productivity.
Acknowledgements
The authors would like to thank EIAR for financial support provided to conduct
the experiment. We are also grateful to Bako National Maize Research Center
breeding program for delivering maize genotypes. It is also our pleasure to thank
the National Maize Research plant protection division technical assistants; Geta
[102]
Gelana, Abebech yilma & Diriba Oljira for assistance they provided in field
management and data collection.
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Identification of additional sources to Exserohilum turcicum in maize (Zea mays L.).
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[104]
Screening of Maize Inbred Lines for Gray
Leaf Spot and Turcicum Leaf Blight Resistance
Midekssa Dida, Temesgen Deressa and Debela Diro
Ethiopian Institute of Agricultural Research, Bako Agricultural Research Centre,
P.O.Box 3, Bako, Ethiopia
Abstract
Maize (Zea mays) is one of the most widely cultivated crops in the world as well in
Ethiopia. But maize production in Ethiopia is constantly threatened by the potential
outbreak of major foliar diseases such as Turcicum leaf blight (TLB) and Gray leaf
spot (GLS). These diseases are characterized by relatively rapid leaf necrosis and
premature death of foliage which eventually reduces grain yield. The diseases become
a major problem in all major maize growing areas. Development of host resistance to
these diseases can provide an important component of integrated disease
management, which is the most effective and practical method of managing maize
diseases. The study was conducted to evaluate the reaction of maize inbred lines to
TLB and GLS in the main cropping season during 2018. The inbred lines were
obtained from Bako National Maize Research Center, breeding program, and it was
arranged using alpha-lattice design with two replications. The inbred lines were
evaluated in TLB and GLS screening field under artificial inoculation at Bako
Agricultural Research Center, West Shewa, Ethiopia. Disease severity (1-5 scale) was
assessed at ten days intervals from disease onset until the maize attained
physiological maturity. All the inbred lines showed symptoms of respective diseases
during the season, but the intensity of the diseases differed significantly (P<0.05)
among the inbred lines. Accordingly, out of 66 genotypes screened forTLB; 30G19F2-
43-1-1-1-1-1-1-#, [CML312/CML442//[CML390/CML206]-BB-2-4-BBB]-1-B-3-1-1-
BBB-#-#-#, SZSYNA99F2-133-2-1-1-1-#-#, 143-5-I-#, SZSYNA 99-F2-3-6-2-1-1-1-1-
#, CIMCALI8843/S9243-BB-#-B-5-1-BB-2-3-4-B-#-#-#,(ZM-605-C2F2-428-3-B-B-B-
B-B-B-B/F7215)-2-2-2-2-1-1-#,[DTPWC8F31-4-2-1-6B2/CML395//[CML445/
ZM621B]-2-1-2-3-1-BB]-3-2-1-1-1-2-B*4-#,TZMI740 -#-#, Kulenic1-0080-4-2-4-1-
2-#-#, GIBE1-265-5-4-1-1-1-#-#and for GLS:SZSYNA99F2-133-2-1-1-1-#-#, 30H83-
7-1-2-1-1-1-#-#,ZM-605-C2F2-428-3-B-B-B-B-B-B-B/F7215)-2-2-2-2-1-1-#,
DTPWC8F31-4-2-1-6-B2/CML395//[CML445/ZM621B]-2-1-2-3-1-BB]-3-2-1-1-1-2-
B*4-#, TZMI746 -#-#, GIBE-1-158-1-1-1-1-#-#,30V53F2-20-2-1-3-3-1-1-#, 142-1-e,
CML 312BK, TZMI746, CML536, DE-38-Z-126-3-2-2-2-2-# were selected as
resistant based on disease severoity scored on 1-5 rating scale.
Introduction
Maize (Zea mays L.) is one of the most important cereal crops in Ethiopia, ranking
second in area coverage and first in total production. Currently, about 2.13 million
ha of land is covered by maize with an average production of 8.39 million tons,
and the average national yield of maize under small-scale farmers reaches 3.94
t/ha (CSA, 2018). However, its productivity is very low as compared to the
potential of maize in the high rainfall and irrigated areas (8 –11 t ha-1) and the
world average (5.78ton ha-1) (USDA-FAS, 2017). An important portion of this
[105]
low yielding in maize is attributable to abiotic and biotic stresses, poor
management practice and as well as climatic change. Among biotic factors foliar
diseases are reported to be widespread and destructive disease of maize mainly in
warm and humid growing regions of Ethiopia (Tewabech et al., 2001). Out of
various maize foliar diseases, Turcicum Leaf Blight (TLB) and Grey Leaf spot
(GLS), caused by, respectively, Excerohillum turciccum and Cercosporra zeae
maydis, are the most economically important diseases threatened maize
production (Tewabech et al., 2012). Due to the presence of conducive
environmental factors such as relatively higher humidity, moderate to high
temperature and/ or warm condition during the maize cropping season at mid-
altitude sub-humid agro-ecology of Ethiopia, Turcicum leaf blight and grey leaf
spot are considered as the most economically important, which are identified to
cause significant yield lose. If infection occurs in early developmental stage of the
plant, Turciccum leaf blight can cause leaf necrosis and premature death of foliage
which reduces grain yield of the crop (Raymundo and Hooker, (981). On the other
hand, grey leaf spot shows necrotic lesion tend to be long and individual lesion
may merge lead to leaf senescence greatly reducing the photosynthic areas leading
to poor grain filling (Darera et al., 2008). In Ethiopia, yield loss due to GLS was
estimated at about 37 % (Wegary et al., 2004) and 60% in South Africa (Ward et
al., 1997). Similarly, according to Assefa and Tilahun (1992), TLB caused the
highest mean grain yield loss of 50%, and 1000 kernel weight loss of 16.4% on
susceptible cultivars. Methods to manage TLB and GLS diseases include cultural
practices, chemical and host plant resistance (Pratt et al., 2003). Due to its
inconsistency with environment and expensiveness, use of chemical fungicides are
not effective. The most effective and cost-efficient means of managing these
diseases is the use of host plant resistance. It is, therefore, desirable to identify
resistant inbred lines from diverse sources in maize pre-breeding program in order
to improve genetic resistance to these foliar diseases. Though early research
efforts made to identify maize germplasm resistant to these diseases and utilizing
them for maize breeding program, subsequent study for additional source of maize
germplasm should be screened under artificial inoculation to obtain new and
stable resistance. Therefore, the objective of study was to select inbred lines that
are resistant/tolerant to GLS and/or TLB through evaluation from locally
developed and adapted maize inbred lines for use in maize improvement program.

The experiments were conducted at Bako national maize research center
(BNMRC) of maize disease nursery field during main season of 2018/19. Bako is
located at 370E, 090 060 N and receives the annual rain fall of about 1237 mm and
situated at an altitude of 1650 m above sea level, which represent mid altitude sub-
humid agro-ecology zone of Ethiopia. It has minimum and maximum average
[106]
temperature of 15.60C and 30.70C, respectively. Both experiments were conducted
in the field under artificial epiphytotic conditions for evaluation against TLB and
GLS.
Description of experimental materials and design

A total of sixty-six Non-QPM inbred lines were used and arranged in 6x11 alpha
lattice design with three replications. Each inbred line was planted in a plot
consisting of two rows of 3.6m long spaced at 25 and 75cm between plants with in
and between rows, respectively. The non-quality protein maize inbred lines
‘CML-197’ and ‘SC-22’ were used as susceptible check for TLB and GLS,
respectively. Plots were hand sown with fertilization of P2O5 and nitrogen at the
rate of 69 and 92 kg h-1, respectively and all agronomic management practices for
the area were applied as per the recommendations.
Inoculum preparation and inoculation

Inoculum of E. Turcicum and C. zeaemaydis was prepared a year before
experimentation by collecting from heavily infected maize fields showing distinct
TLB and GLS symptoms, respectively. The infected leaves were dried under
shade and crushed/grounded in to mill about the coarseness of wheat bran and
stored in paper bags at a temperature of 4oC until inoculation date. The pulverized
leaves then dusted in the whorls of the plants according to Dagne (2008) by
placing a pinch of leaf mill into whorl of each plant, when plant attains 6-8 leaf
stage during moist environments in order to retain long enough to permit spore
germination. A second inoculation was made ten days later after the first
inoculation to ensure adequate infection.
Assessment of disease reaction

TLB and GLS were visually assessed in the field two weeks after artificial
inoculation on a plot basis from the two rows. Data collected included date first
disease appearance, disease incidence, disease severity, and other agronomic traits
including plant height (cm) and grain yield (t/ha). The progress of severity of
foliar diseases on each inbred lines were quantified at ten days intervals starting
from onset of disease until dent stages and the highest or final severity value of
each inbred lines was used for statistical analysis. Disease incidence was measured
as percent of infected plants per total plant per plot. Disease severity was rated
based on 1-5 scoring scale (CIMMYT, 1985); where 1=no disease symptoms,
2=moderate lesion below the leaf subtending the ear,3=heavy infestation on and
below the leaf subtending the ear with few lesions above it, 4=severe lesion on all
but the uppermost leaves which may have a few lesions, 5=all leaves dead. The
categorization on each disease reactions was made on the basis of disease severity
ratings using a 1-5 scale (Roane et al., 1974) with some modifications, where;1.0–
2.0=Resistant (R); 2.1-2.5=Moderately Resistant (MR); 2.6 -3.0= Susceptible (S),
and >3.0 Highly susceptible (HS).
[107]
Statistical data analysis
Data were analyzed using PROC GLM of SAS version 9.2 (SAS Institute, 2004).
Mean separation was performed to compare treatment means using LSD-test at
5% level of significance.

A total of sixty-six maize inbred lines were screened for resistance to TLB and
GLS. The mean disease severity and yield results indicated significant (P < 0.001)
variation among the inbred lines for TLB and GLS resistance as shown in Tables 1
and 2, respectively. Disease severity ranged from 1.6 to 4.1 for TLB, and 1.3 to
4.5 for GLS. Inbred lines with mean severity values of < 2 were categorized under
resistant/tolerant to TLB and GLS. Whereas, inbred lines with mean severity
values ranging from 2.1-2.5 considered as moderately resistant, 2.6-3.0 as
susceptible and those with severity value > 3 were considered as highly
susceptible to TLB and GLS diseases (Tables 1 and 2). Accordingly, 22 inbred
lines were resistant/tolerant, 17 inbred lines moderately resistant, 17 inbred lines
susceptible and 11 inbred lines were categorized under highly susceptible to TLB
(Table 1). Similarly, 22, 11, 11 and 22 inbread lines were, respectively,
categorized under resistant/tolerant, moderately resistant, susceptible and highly
susceptible to GLS (Table 2). Those inbred lines which have showed
resistance/tolerance were compared to the susceptible checks CML 197 and SC
22, respectively, for TLB and GLS. Selected inbred lines could be used as TLB
and GLS resistance sources for use in resistance breeding programs.
Table 1. Mean Turciccm leaf blight severity and yield of 66 inbred lines evaluated under artificial inoculation during 2018
main cropping season at Bako.
No. of TLB Severity Reaction Yield
Pedigree
Entry (scale1-5) to TLB t ha-1
1 TZMI719-#-# 2.3 MR 1.18
2 30G 19F2-43-1-1-1-1-1-1-# 1.8 R 0.86
3 SZSYNA99F2-3-6-3-1-#-# 2.3 MR 0.71
4 GIBE-1-178-2-1-2-1-#-# 2.9 S 0.23
5 (CML205/CML208//CML202)-X-2-1-2-B-B-B-# 2.9 S 3.02
6 [CML312/CML442//[CML390/CML206]-BB-2-4-BBB]-1-B-3-1-1-BBB-#-#-# 1.7 R 3.13
7 DE-38-Z-126-3-2-2-2-2-# 2.6 S 1.69
8 [[CML388/CML391]-BB-5-2/CML390]-2-1-2-1-1-BBB-#-#-# 2.5 MR 0.89
9 30H83-7-1-5-1-1-1-1-# 2.0 R 1.93
10 [CML312/CML445//[TUXPSEQ]C1F2/P49-SR] F2-45-3-2-1-BBB]-1-2-1-1-2-B*4-#-# 2.6 S 3.38
11 [CML444/DRB-F2-60-1-1-1-BBB//[LZ956441/LZ966205]-B-3-4-4-B-5-B*7-#-#-# 2.0 R 1.22
12 CKL05019-# 4.1 HS 3.53
13 SZSYNA99F2-133-2-1-1-1-#-# 1.7 R 1.89
14 TZMI741-#-#-# 2.9 S 0.95
15 CML197-# 4.1 HS 3.27
16 143-5-I-# 1.9 R 1.96
17 TZMI733 -#-# 3.0 S 2.07
18 TZMI750-#-#-# 2.1 MR 0.98
19 30H83-7-3-4-1-1-1-#-# 2.8 S 2.95
20 BH-660F2-31-1-1-2-1-1-1-1-# 2.8 S 0.60
21 30H83-7-1-2-1-1-1-#-# 2.5 MR 2.54
[108]
22 POOL9A-128-5-1-1-1-2-1-# 1.8 R 0.66
23 SZSYNA 99-F2-3-6-2-1-1-1-1-# 1.7 R 1.16
24 CIMCALI8843/S9243-BB-#-B-5-1-BB-2-3-4-B-#-#-# 1.7 R 1.76
25 (ZM-605-C2F2-428-3-B-B-B-B-B-B-B/F7215)-2-2-2-2-1-1-# 1.8 R 1.37
26 CML-197 x 142-1-e(F2) 86-1-1-1-1-1-# 3.2 HS 3.52
27 DE-38-Z-126-3-2-2-2-1-1-# 2.9 S 1.39
28 139-5- j-# 2.2 MR 0.21
29 DE-147-Z-126-72-1-1-2-2-2-1-# 1.6 R 0.72
30 Gibe-1-54-3-6-1-2-1-#-# 1.9 R 0.86
31 CIMCALI8843/S9243-B-B-#-B-5-1-B-B-2-3-1-B-#-#-# 1.9 R 0.70
32 TZMI759-#-#-# 2.2 MR 1.30
33 TZMI407-short-#-#-# 1.8 R 0.93
34 POOL 9A-4-4-1-1-1-#-# 2.3 MR 1.16
[DTPWC8F31-4-2-1-6-B2/CML395//[CML445/ZM621B]-2-1-2-3-1-BB]-3-2-1-1-1-2- R
35 1.7 2.28
B*4-#
36 TZMI740 -#-# 1.8 R 1.68
37 30G 19F2-54-1-1-1-#-# 2.0 R 0.97
38 SC-715-154-1-1-# 2.1 MR 3.30
39 TZMI751-#-#-# 3.1 HS 1.05
40 Kulenic1-0080-4-2-4-1-2-#-# 1.9 R 1.24
41 MAS[MSR/312]-117-2-2-1-B*9-#-#-# 2.6 S 3.52
42 FH625-272-1-1-1-#-# 2.5 MR 0.89
43 (DRBF2-60-1-2)-B-1-B-B-B/F7215)-1-1-3-# 3.2 HS 3.03
44 POO'E 4-2-2-1-2-1-# 3.9 HS 2.16
45 (DRBF260-1-2)-B-1-B-B-B-# 2.9 S 2.87
46 GIBE-1-20-2-1-2-1-1-#-# 2.5 MR 2.34
47 SZSYNA99F2-3-6-4-1-1-1-1-#-# 2.0 R 0.52
48 35B-190-O-S-10-2-1-2-2-# 2.4 MR 1.68
49 GIBE1-265-5-4-1-1-1-#-# 1.9 R 1.31
50 KULENI 320-2-3-1-1-2-1-1-#-# 2.7 S 2.73
51 TZMI742-#-#-# 3.6 HS 1.85
52 TZMI754-#-#-# 3.1 HS 1.58
53 35B-190-O-S-10-2-1-1-1-#-# 3.1 HS 1.96
54 35B-190-O-S10-9-1-1-#-# 2.3 MR 1.57
55 ILOO'E 1-9-1-1-2-1-2-# 2.8 S 1.83
56 SC-22-430(63)-# 2.5 MR 1.58
57 TZMI746 -#-# 2.8 S 2.79
58 GIBE-1-158-1-1-1-1-#-# 2.6 S 1.34
59 30H83-56-1-1-3-1-1-#-# 2.2 MR 3.19
60 TZMI747-#-#-# 2.6 S 0.56
61 [LZ956441/LZ9662O5]-B-3-4-4-B-5-B*7-#-#-# 3.5 HS 1.28
62 SC 22 3.0 HS 0.18
63 FH625-272-1-2-1-#-# 2.6 S 1.97
64 SC-715-13-2-1-# 1.8 R 0.74
65 30V53F2-20-2-1-3-3-1-1-# 2.2 MR 0.96
66 [INTB-F2-90-2-1-1-BBB/CML395]-B-1-1-1-1-BB-# 2.4 MR 1.97
LSD
1.4 1.31
(0.05)
CV 27.9 33.9
1.0–2.0=Resistant (R); 2.1-2.5=Moderately Resistant (MR); 2.6 -3.0= Susceptible (S), and >3.0 Highly susceptible (HS).
[109]
Table 2. Mean Gray Leaf Spot severity and yield of 66 inbred lines evaluated under artificial inoculation during 2018 main cropping season at Bako.
Plant Ear Lodging Ears/ Husk Ear Reaction Ear
Entry Grain Yield (t/ha) Height (cm) Height (cm) Root (%) Stem (%) Plant (#) Cover (%) Rot (%) GLS (1-5) categries Aspect (1-5)
1 1.03 116 72 19.1 2.4 0.73 2.1 24.1 1.5 R 2.6
2 1.06 170 73 6.7 2.1 0.49 6.4 25.6 1.5 R 2.8
3 0.68 135.5 55.5 7.2 3 0.57 0 5 2 R 3.4
4 0.15 120 65 2.1 2.1 0.24 0 43.4 3.3 HS 3.9
5 0.86 143 76 47.3 0 0.81 0 17.2 1.5 R 2.4
6 1.47 134.5 60 10.1 3 1.05 0 18.7 1.5 R 2.6
7 1.19 107.5 57.5 10.6 0 0.81 0 21.4 1.5 R 2.8
8 0.65 167.5 63.5 34.7 7 0.49 0 11.1 2.8 S 1.9
9 1.44 175 85.5 22.9 2.1 0.82 8.4 26.2 3.5 HS 3.6
10 4.2 157.5 74 11.6 0 1.18 24.4 9.6 1.8 R 2
11 1.27 146.5 71 2.5 2.2 0.72 21.2 49.2 2 R 3.1
12 2.04 145 79.5 6.3 4.2 1.11 8.4 17.2 3.5 HS 2.5
13 1.91 140 79 14.6 14.6 0.86 6.3 26.6 2.3 MR 2.7
14 1.09 88.5 39 41.7 0 0.69 0 14.9 2 R 2.6
15 3.31 161.5 97 6.3 0 1.42 10.4 14.6 4.3 HS 2.5
16 2.14 136.5 109 31.3 2.1 0.8 6.3 27.5 3.3 HS 2.8
17 0.89 128 66 22.9 0 0.58 0 23.8 1.8 R 3.2
18 0.53 140.5 64.5 11.9 2.4 0.53 6.9 27.4 1.5 R 3.9
19 5.32 174.5 68 6.3 4.2 1.17 23 7.3 1.5 R 1.1
20 0.31 144 68 0 0 0.43 4.6 41.7 2.8 S 4.4
21 3.18 159 70 41.7 6.3 1.12 2.1 23 1.5 R 2.1
22 0.61 148.5 49 0 21.5 0.74 28.6 62.5 3 S 3.9
23 0.77 161.5 70 40.3 0 0.81 0 13.9 3.5 HS 3.9
24 0.67 105 39 9.4 15.7 1.07 0 0 2.5 MR 2.7
25 1.96 140.5 63 19.6 3 1.01 0 15.8 2.3 MR 2.7
26 1.51 138 66 4.2 2.1 0.65 10.5 52.8 1.5 R 2.4
27 1.06 154 68 6.3 6.3 0.71 0 25.9 2.8 S 3.2
28 0.34 143 77 0 0 0.63 8.4 77.8 1.5 R 4.1
29 0.99 130 52.5 22.9 10.4 0.71 0 19.7 1.8 R 3.9
30 0.9 135.5 62.5 0 0 0.64 0 30 4 HS 2.8
31 0.8 115 46.5 23.7 0 0.84 2.1 28 2.3 MR 3.3
32 1.18 91 43.5 42.6 0 0.79 0 7.9 1.8 R 2.6
33 1.1 96.5 45 2.8 2.8 1.25 8.4 11.8 2 R 2.3
34 1.29 146.5 62 19.8 4.8 0.81 11.9 40.6 2.8 S 3.8
35 1.94 112 48 0 0 0.85 0 17.1 1.8 R 1.9
36 1.41 83.5 38.5 8.4 0 0.86 2.1 4.9 1.5 R 2.9
[110]
37 1.47 133 71.5 16.3 3 0.79 4.2 9.6 2.5 MR 2.4
38 1.76 138.5 67.5 2.1 12.5 1.03 0 12.4 2.8 S 2.4
39 0.54 93.5 56 2.5 0 0.48 0 87.5 3.5 HS 3.3
40 1.21 148.5 74 6.3 20.9 0.67 0 17.1 3.5 HS 3.1
41 4.02 190 81 25 6.3 0.86 8.4 7.5 3.8 HS 1.7
42 0.6 122 63.5 2.1 0 0.45 0 16.7 3.8 HS 3.7
43 3.35 137 63.5 8.4 8.6 0.96 8.4 4.4 2.5 MR 1.9
44 1.61 167 71 14.6 4.2 0.86 6.3 9.9 3.8 HS 3.4
45 2.89 188.5 79 22.9 8.4 0.81 4.2 20.7 4.5 HS 2.6
46 1.7 151.5 72.5 16.7 6.3 0.96 0 17.2 3.8 HS 2.7
47 0.46 122.5 58 7.7 0 0.73 0 18.8 2.8 S 3.1
48 1.28 170.5 86.5 8.5 6.3 0.73 0 35.3 3.5 HS 3.2
49 0.96 148.5 81 2.1 0 0.59 14.2 35.9 3 S 3.1
50 2.87 162.5 89 14.6 4.2 0.94 8.4 11 3.3 HS 2.9
51 1.13 108 53.5 13.7 9.4 0.77 0 24.3 3.8 HS 3.3
52 0.43 110.5 59 2.1 4.2 0.24 6.3 52.8 3.5 HS 3.9
53 2.46 151.5 78.5 10.4 6.3 0.98 12.5 19.6 4.5 HS 2.7
54 1.63 166 100.5 54.2 8.3 1.11 0 65 2.3 MR 3.6
55 1.44 150 78.5 0 10.5 0.64 0 30.4 2.3 MR 3.5
56 1.05 164.5 82 2.7 3.9 0.81 0 63.2 3.5 HS 4.4
57 2.82 130 94.5 6.3 0 1.19 6.3 9.6 1.3 R 2.1
58 1.75 136 70 14.6 8.4 0.88 10.4 26.2 2.3 MR 2.9
59 7.76 208.5 110.5 6.3 6.3 1.42 8.4 4.8 4.3 HS 1.7
60 0.61 109.5 58.5 6.3 4.2 0.36 0 41.5 3.3 S 3.1
61 1.32 111.5 47 18.8 6.3 0.73 4.2 20.1 2.8 S 2.6
62 0.38 148.5 69 125 12.5 0.82 43.8 45 3 S 3.9
63 1.04 141 91.5 2.1 0 0.55 0 36.3 4 HS 3.1
64 0.83 105 54 4.4 12.9 0.58 0 19.9 2.5 MR 3.1
65 1.59 131 52.5 0 0 0.8 33.1 22.6 1.5 R 2.4
66 1.48 135 64.5 29.2 14.6 0.79 0 13.7 2.8 MR 2.7
Mean 1.57 139 68.2 15.2 4.7 0.79 5.8 25.5 2.7 2.9
LSD (0.05) 1.04 36.5 17.1 47.3 12.7 0.36 12.2 33.7 1.3 1.2
MSe 0.27 334.7 73.3 562 40.6 0.03 37.3 285.2 0.4 0.4
CV 33.28 13.2 12.5 156 135 22.62 106 66.3 23.8 20.2
p 0 0 0 0.16 0.13 0 0 0.001 0 0
p *** *** *** ns ns *** *** ** *** ***
Min 0.15 83.5 38.5 0 0 0.24 0 0 1.3 1.1
Max 7.76 208.5 110.5 125 21.5 1.42 43.8 87.5 4.5 4.4
1.0–2.0=Resistant (R); 2.1-2.5=Moderately Resistant (MR); 2.6 -3.0= Susceptible (S), and >3.0 Highly susceptible (HS).
[111]
35.00
R
30.00
MR
Frequency (%)
25.00
S
20.00
HS
15.00
10.00
5.00
0.00
Disease Reaction
Figure 1. Frequencies of maize inbred lines with resistant (R), moderately resistant (MR), susceptible (S) and Highly
susceptible (H S) reactions to Turcicum leaf blight (TLB).
35.00
30.00
25.00
Frequency (%)
20.00
15.00
10.00
5.00
0.00
R MR S HS
Disease Reaction
Figure 2. Frequencies of maize inbred lines with resistant (R), moderately resistant (MR), susceptible (S) and Highly
susceptible (H S) reactions to Grey Leaf Spot (GLS).
Field screening studies indicated that there were clear cut differential response of
inbred lines to TLB and GLS due to artificial inoculation (Figures 1 and 2).
Similar work done elsewhere also showed significant variation among inbred lines
[112]
for TLB and GLS severity (Chandrashekara et al., 2014). Resistance/tolerance
reactions to TLB and GLS were recorded in 31.82% and 33.33% of the inbred
lines tested, respectively. The data revealed that the highest number of the inbred
lines tested have shown resistant reactions to TLB and GLS, where some inbred
lines were showed resistant/tolerant reaction to both GLS and TLB. Inbred lines,
namely 30G 19F2-43-1-1-1-1-1-1-#, [DTPWC8F31-4-2-1-6-B2/CML395//
[CML445/ZM621B]-2-1-2-3-1-BB]-3-2-1-1-1-2-B*4-#[CML312/CML442//
[CML390/CML206]-BB-2-4-BBB]-1-B-3-1-1-BBB-#-#-#, TZMI407-short-#-#-#
were showed multiple disease resistance compared with the checks. It is likely that
the more susceptible inbred lines loss their active leaf tissues and resulted in less
photosynthetic leaf area, then the plant eventually produce few kernels and/or may
contribute to the overall yield loss, showing that yield are negativly correlated
with disease severity. A similar research finding was reported by Singh et al.
(2014). The moderately resistant and resistant inbred lines should further be
screened for stability resistance to TLB and GLS across location and years at
hotspot areas and/or under controlled condition. Generally, 22 inbred lines were
found resistant (scored < 2.0 on a 1-5 scale) to TLB and 22 to GLS under artificial
epiphytotic condition. Thus, it can be emphasized from the results that the
identified resistant lines hold promise for resistance against E. turcicum and C.
zeaemaydis causing TLB and GLS of maize, respectively. Promising inbred lines
can also be used for developing hybrids and composites in future breeding
programs for multiple-disease resistance. These findings are in agreement with the
works of Patil et al. (2000), Muiru et al. (2007) and Pandurangegowda et al.
(1994), who reported differential reaction to diseases among the various maize
germplasm. Sharma and Payak (1990) reported durable resistance in CM-104 and
CM-105 maize inbred lines against E. turcicum. Promising maize disease-
resistance sources were also reported by Dagne et al. (2008), who identified 143-
5-I and CML-387 as resistant, Gotto LMS5, SC-22 and CML-395 moderately
resistance and A-7016 and CML-197 susceptible to GLS.
Conclusion and Recomendation

Maize inbred lines identified as resistant/tolerant in this study should also be
screened under controlled environments to properly verify the level of resistance
for TLB and GLS. It would be better to use molecular methods to locate the gene
(s) involved in the resistance and incorporating them to cultivars having desired
agronomic traits. Moreover, the promising lines with good yield and other
agronomic performance identified through this investigation can be deployed in
disease endemic areas for sustainable maize production.
[113]
Acknowledgements
The authors would like to thank EIAR for financial support provided to conduct
the experiment. We are also grateful to Bako National Maize Research Center
breeding program for delivering maize genotypes. It is also our pleasure to thank
the National Maize Research plant protection division technical assistants; Geta
Gelana, Abebechyilma and Diriba Oljira for assistance they provided in field
management and data collection.
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[116]
Evaluation of Soybean Genotypes (Glycine max
(L.) Merr.) for Asian Soybean Rust (Phakopsora
pachyrhizi ) in South Western Ethiopia
Yechalew Sileshi1, Mesfin Hailemariam1, Abush Tesfaye2,
Kifile Belachew1 and Negassa Dechasa1
1
EIAR, Jimma Agricultural Research Centre, P.O. Box, 192, Jimma, Ethiopia;
2
International Institutes of Tropical Agriculture, PMB 5320, Oyo Road 200001, Ibadan, Nigeria.
Abstract
Soybean is one of the rapidly growing crops, both globally and in Ethiopia, though
its productivity in the country is very low due to several production constraints,
among which soybean rust is one of the most important one. Therefore, this study
was designed to evaluate sixty-four soybean genotypes which were introduced from
USA, including three standard check varieties to identify sources of resistance for
the disease. The entries were laid out in an 8X8 simple lattice design and grown in
the Southwestern Ethiopia at Jimma agricultural research center, during the 2017
and 2018 main cropping season. The analysis of variance revealed that significant
difference for most of the studied traits, i.e., including Area under Disease Progress
Curve (AUDPC), mean disease severity, and grain yield. Based on the field
evaluation, a total of 44 soybean genotypes showed Reddish Brown (RB) lesions
and 20 soybean genotypes showed Tan lesions; while none of the tested soybean
genotypes showed immune reaction to soybean rust. Genotype PI417089A showed
Tan lesions and higher value of AUDPC, and rust severity; however this genotype
was the highest yielding genotype, indicating the tolerance of this genotype for Asian
soybean rust. Genotypes that showed reddish brown reaction and tolerance to the
disease need to be subjected to further testing and used as parental line for
hybridization. The effort to identify very good source of resistance need to be
strengthened, with major emphasis on characterizing the different types of rust
races, evaluating large number of materials from diverse sources and understand the
mechanisms of soybean genotypes tolerance and resistance to soybean rust.
Introduction
Soybean is becoming one of economically important crop in Ethiopia, which ranks
first based on the productivity potential among all other oil crops (CSA, 2018).
Asian Soybean rust (ASR) is one of the production constraints of soybean
production and productivity, both globally and in the country. It is caused by
either of the two fungal species Phakopsora pachyrhizi and Phakopsora
meibomiae (Ono et al., 1992). Asian Soybean Rust(ASR) infects more than 95
species of plants from more than 42 genera, including soybean and related Glycine
species (Koch, 1988). The Asian soybean rust was first reported in Japan in 1902
(Bromfield et al., 1980), while it was first reported in Africa in the late 20th
century. ASR was first reported in Ethiopia in 2017 (Abush et al., 2017).
[117]
Percentage of yield losses due to ASR was reported in the range of 10% to 80%,
and in some African countries, such as Zimbabwe loss ranges from 60-80%
(Levy, pers. comm), Uganda 23-36 % ( Kawuki et al., 2003) and Nigeria 23-42%
(Akinsanmi et al., 2001) . Sometimes yield losses due to soybean rust might reach
up to 100%, depending on the weather conditions and degree of plant
susceptibility (Kawuki et al., 2003a; Mueller et al., 2009).
Asian Soybean Rust (ASR) might occur at any stage of the crop development, and
most commonly, infection starts at the reproductive stage (Faske et al., 2014).
Asian Soybean Rust might be managed using: fungicides, genetic resistance, and
cultural practices. However, host-genetic resistance remains the most
economically viable, environmentally friendly and strategically important option
for resource-constrained farmers in the developing countries, like Ethiopia. The
use of adapted soybean cultivars with resistance to P. pachyrhizi is cost-efficient
and environmentally friendly ways of controlling the disease (Goellner et al.,
2009; Hassan et al., 2014; Yamanaka et al., 2011).
To date, six major resistance genes (Rpp1, Rpp2, Rpp3, Rpp 4, Rpp5, and Rpp
Hyuuga) to soybean rust have been identified with different genes of resistance to
a specific set of rust isolates, and the effectiveness is depending on the soybean
rust-plant pathosystem (Hartman et al., 2005; Maphosa et al., 2012; Kato et al.,
2006; Hyten et al., 2007; Garcia et al., 2008). Lesion type and proportion of
sporulating lesions are important disease parameters that can be used to
extrapolate disease progress. Three different reaction types may occur on soybean
genotypes in response to ASR infection based on lesions types, i.e. Immune
reaction (IM) without visible lesions, Reddish-brown (RB) lesions a resistant
reaction and (TAN) lesions susceptible reaction (Bromfield, 1984; Goellner et al.,
2010). To plan successful breeding strategy for Asian soybean rust, identifying the
response of soybean genotypes to soybean rust is crucial stage (Miles et al., 2011).
In Ethiopia, soybean breeding program is focused on developing high-yielding
varieties. To date a total of 25 soybean varieties were released for different agro-
ecology of Ethiopia. Recently soybean is becoming the most important industrial
raw material. Since soybean rust becomes serious disease in Ethiopia, it is
important to develop soybean genotypes with high yield potential and less yield
loss from soybean rust. Therefore, this study was aimed at assessing the resistance
or tolerance of soybean genotypes to rust disease.
Materials and Mmethods

The Study area
The study was conducted at Jimma Agricultural Research Center, and its sub
center (Mettu) (Figure 1) for two years (2017 and 2018). Jimma and Mettu are
[118]
located in Oromia region, South Western Ethiopia (detail description of the
locations is presented in Table 1). These sites represent areas of high potential for
soybean production in Ethiopia with seasonal soybean rust epidemics.
Figure1. Diagrammatic sketch and descriptions of experimental sites
Table 1. Monthly weather conditions of the 2017/18 cropping seasons for the experimental site.
Jimma, 2017 Jimma, 2018

Month RF(mm) T 0c T 0c RH Dew RF(mm) T 0c T 0c RH
total Max Min mean mean total max Min mean
June 148.2 26.0 10.2 74.5 14.9 201.80 26.27 9.97 78.97
July 181.8 25.5 10.7 78.6 14.9 190.20 26.11 10.45 77.21
August 169.1 26.8 11.0 80.3 13.8 268.30 26.29 10.21 76.74
September 359.0 26.4 11.2 73.1 15.9 147.70 26.37 9.95 76.41
October 319.1 26.4 10.9 76.18 14.7 133.40 25.89 10.19 79.27
November 27.6 28.4 10.2 75.0 15.8 83.40 26.09 10.43 78.92
RF= rainfall, RH =relative humidity; T0c = temperature. Source: Jimma agricultural research center
Experimental layout and design

Sixty-one introduced soybean genotypes, including two sources of resistance,
G7955-C3RPP and MKSOY-2Nwhit and three released varieties (Table 2) were
laid out in a simple lattice design. Each plot was four rows of 4 m long. Out of the
total four rows, one border row was planted with a released Asian rust susceptible
variety (Clark-63k), as a spreader row in each plots. The spacing between rows
and plants were 60 cm and 5 cm, respectively. Di Ammonium Phosphate fertilizer
[119]
at the rate of 100 kg ha-1 was applied at planting and all other cultural practices
were applied as per the recommendation for the crop.
Data collection and statistical analyses

Asian soybean rust disease severity score was graded by employing 1 to 9 scale
(Mayee and Datar, 1986). Data was collected from five randomly marked plants at
10 days interval starting from the 4th week from planting to R7 (maturity). Where;
1 = nil (no visible disease symptom = Immune);
3 = slight (some small discrete and a few large lesions = Resistant);
5 = medium (some coalesced lesions, many spotting and some defoliation =
Moderately resistant);
7 = severe (large coalesced lesions with about 50% defoliation, few dead stems
per plant = Susceptible); and
9 = very sever (extensive lesions, sever defoliation, stem girdling, many dead
plants = Highly Susceptible.
The percentage severity index (PSI) for disease was calculated using the formula:
PSI result was transformed using arc sin transformation method for statistical
analysis.
Based on the percent severity index level for the disease, varieties categorized as;
0% =absolutely resistant; 0.01-11.11% = Highly resistant; 12.22-33.33% =
Moderately resistant (MR); 34.44-55.55% = Moderately susceptible (MS), 56.66-
77.77% = Susceptible; 78.88-100.0%= Highly susceptible.
Severity data were converted into area under the disease progress curve (AUDPC),
as proposed by Campbell and Madden, using standard interactive procedures
(Shaner and Finney, 1977). The AUDPC is calculated as:
AUDPC = ∑
Where: X= is the time in days; Y = is the proportion of diseased leaves

(cumulative severity, = xct), i= is the ith observation, i+1 = is the
next observation, n = is the total number of observations, Xi = is the
time on first observation (xi = 0 time).
The soybean genotypes were also assessed for types of rust reaction based on
lesion color observed on leaves, and sporulation were evaluated at the R6 stage
(Fehr et al., 1971), the stage at which the disease symptoms were conspicuous on the
[120]
three trifoliate leaves of the mid-canopy. Soybean genotypes without any visible
rust symptoms were considered to be immune; and those with reddish brown (RB)
lesions were resistant or partially resistant; while genotypes with tan-colored
lesions (TAN) were susceptible (Bromfield, 1984; Miles et al., 2005). Level of
sporulation was also assessed based on 0 - 3 scale score on the most diseased
leaves; where 0 = no pustule, 1 = no sporulating pustules, 2 = sporulating pustules and 3
highly sporulating pustules (Shanmugasundaram, 1977). Other important quantitative
traits were also recorded, including; days to flowering, days to maturity, plant height
(cm), pod per plant, seed per plant, hundred seed weight (g) and yield (t/ha). Weather data
(temperature, rainfall and relative humidity) was collected for the test sites from planting
to the final harvesting.
Table 2. Details of the soybean genotypes used in this study
Seed Seed Seed Seed

Designation Source Designation Source Designation Source Designation Source
PI567099A USA PI605891A USA PI567025A USA PI567189A USA
G7955-C3RPP(C1) USA PI567104B USA PI605838 USA PI594538A USA
MKSOY-2Nwhite(c2) USA PI567054C USA PI567090 USA AFGAT(C5) ET r
PI605823 USA PI605891B USA PI605773 USA PI230970 USA
pi567020A USA PI567069A USA PI416810 USA PI615437 USA
PI567102B USA PI417089A USA PI605854B USA PI416886 USA
PI471904 USA PI606379B USA PI594767A USA PI417085 USA
PI567039 USA PI567056A USA PI566989A USA PI203398 USA
PI567058D USA PI628932 USA PI200466 USA PI423972 USA
PI605824A USA PI587905 USA PI635999 USA PI423960B USA
PI578457A USA PI567061 USA PI423960A USA PI507004 USA
PI567046C USA PI567024 USA PI417208 USA PI340898A USA
PI615445 USA PI605865B USA PI567059 USA PI416873B USA
PI567180 USA PI416826A USA PI567053 USA PI506677 USA
PI606405 USA PI506939 USA PI567068A USA Clark-63k ET r
PI594760B USA PI587880A USA PI567034 USA Nyala ET r
Statistical analysis
Data was subjected to analysis of variance (ANOVA) using General Linear Model
(GLM) of SAS. Least significant difference (LSD) at p<0.05 was employed to
separate genotypes that are significantly different from each other.

The mean square of the pooled analysis of variance over years revealed that the
genotypes showed highly significant difference (P≤ 0.01) for all the studied traits,
except plant height which was significant only at P<0.05 level of significance
(Table 3). Similarly, the mean squares due to genotypes revealed significant
(P≤0.01) differences for all of the yield and yield related traits, except plant
height, number of pods per plant, number of seeds per plant, which was significant
[121]
only at P<0.05 level of significance. This indicates the existence of varying
different genotypic response for yield and rust resistance. The combined ANOVA
also revealed highly significant genotype x year interaction for days to flowering,
hundred seed weight and grain yield. The genotype X year interaction for rust
severity and rust AUDPC was non-significant indicating the response of the
genotypes for rust was consistent, irrespective of seasonal differences.
Table 3. Combined analysis mean squares of rust severity score and other traits at Jimma over year
Rust AUDPC Yield

Source DF DM PH PP SP HW severity Rust
MSY 7634.50** 6952.15** 964.11* 8856.19** 280316.80 914.83** 466.11** 259197.5** 87.54**
MSG 347.92** 441.32** 2001.64* 1184.98* 7929.15* 49.62** 139.32** 64369.5** 0.65**
MSGXY 51.32** 98.12ns 221.63ns 570.81ns 6408.25ns 4.67** 84.48ns 1765.3NS 0.58**
MSE 30.82 80.99 165.44 741.90 4746.39 1.88 67.08 8428.29 0.17
Where DF = days to 50% flowering, DM = days to maturity, PH = plant height, PP = number of pods per plant, SP= seed
per plant, HW =hundred seed weight, AUDPC=area under disease progress curve,* = significant at (P≤0.05), and **=
significant at (P≤ 0.01), MSY = mean Squares of years, MSG = mean squares of genotypes, MSG x Y = mean square of
genotype x year interaction, MSE = mean squares of error,
The genotypes PI340898A (92 days), PI567056A (90 days) and PI567090 (86
days) took the longest days to flower (Table 3); while the earliest flowered entries
were PI200466 (50 days), PI416873B (50 days), PI423960B (50 days),
PI416826A (51 days), PI507004 (52 days), PI416886 (53 days), PI506677 (56
days), PI628932 (56 days), PI567034 (57 days), and PI594538A (57 days).
Similarly, the entries that took long to mature were PI340898A, PI567104B,
PI578457A, PI567056A, PI567069A, MKSOY-2N, PI567020A, PI230970, and
PI605823 with respective days to flowering of 150, 149, 147, 146, 143, 140, 140,
139 and 138. The genotypes that showed early maturity were PI615437 and
PI507004 which, respectively, took 96 and 109 days for maturity. Such early
maturing genotypes might be desired for production in areas where the growing
period is short, and to help escape some of the biotic stresses, such as diseases and
insect pest damages. Plant height ranged from 42.8 cm (PI416886) to 148.4 cm
(PI567104B) with mean value of 90.9 cm. The highest number of pods per plant
was produced by the genotype PI567102B (148.3); while the lowest number of
pod was obtained from the genotype PI416826A (21.8). Similarly, the highest
number of seed per plant was observed from the genotype PI567068A (284.8);
while the smallest seed per plant was found in the genotype PI507004 (34.2).
Maximum value of hundred seed weight was recorded on the genotype PI506677
(24g); while the smallest from genotype PI567068A (7.8g). The maximum amount
of yield was obtained from the genotype PI417089A (3.43 t/ha) and the smallest
from the genotype PI566989A (1.35 t/ha). Thirty one Soybean genotypes yielded
more than the mean value which is 2.25 tons per hectare and seven genotypes
(PI417089A, G7955-C3RPP(C1), PI471904, PI567104B, PI567054C, PI594538A
and PI230970) gave more than the released variety Clark-63k (2.66), other
released varieties AFGAT and Nyala yielded 2.44 and 2.27 t/ha, respectively.
[122]
Table 4. Mean performance of 64 soybean genotypes for the studied traits over year at Jimma
No Treatment name DF DM PH PP PS HW AUDPC PDI Reaction Level of YD/t/h

type sporulation
1 PI567099A 79 129 118.8 65.6 141.6 8.6 226.4 30.4 RB 2.0 1.89
2 G7955-C3RPP(C1) 68 134 103.4 43.5 84.0 18.7 266.5 33.6 RB 1.0 3.34
3 MKSOY-2Nwhite(c2) 72 140 94.6 65.8 110.5 13.3 505.9 35.3 T 3.0 1.53
4 PI605823 75 138 80.4 41.8 100.7 14.1 557.4 21.0 RB 0.0 2.52
5 PI567020A 75 140 126.9 44.4 114.9 11.3 686.3 41.3 RB 1.0 2.49
6 PI567102B 82 149 142.7 148.3 241.9 11.4 493.9 32.0 RB 0.0 2.10
7 PI471904 72 133 107.1 51.6 108.0 12.7 697.6 34.7 RB 1.0 2.84
8 PI567039 74 132 111.2 28.7 71.2 13.5 408.7 33.4 RB 0.0 2.55
9 PI567058D 72 136 107.5 40.2 111.3 12.9 509.1 31.2 T 3.0 1.89
10 PI605824A 63 125 100.8 31.3 67.3 12.8 506.4 36.2 RB 1.0 1.82
11 PI578457A 71 147 113.0 33.8 67.1 11.7 533.5 40.0 T 3.0 1.61
12 PI567046C 71 130 99.5 43.2 163.9 12.1 427.2 30.1 RB 1.0 2.58
13 PI615445 70 126 87.3 34.6 70.9 12.8 804.5 30.6 RB 0.0 2.01
14 PI567180 60 121 79.5 28.4 52.8 15.2 226.4 32.1 T 3.0 1.57
15 PI606405 64 128 73.7 29.2 63.8 15.3 402.8 31.1 T 2.0 1.80
16 PI594760B 65 126 97.0 43.7 98.6 13.9 340.1 29.1 RB 1.6 2.37
17 PI605891A 65 124 85.9 28.0 65.5 13.9 595.7 37.0 RB 1.0 2.33
18 PI567104B 83 149 148.4 76.4 147.1 11.3 583.0 34.1 T 2.0 2.94
19 PI567054C 69 132 100.1 45.2 98.2 12.4 354.4 31.9 RB 1.0 2.99
20 PI605891B 69 132 96.2 47.7 103.1 13.8 245.1 37.7 RB 0.0 2.44
21 PI567069A 77 143 109.3 66.8 134.2 8.6 510.0 40.4 T 3.0 2.09
22 PI417089A 62 128 87.9 35.9 69.7 18.7 611.4 44.8 T 3.0 3.43
23 PI606379B 68 128 78.9 38.5 66.5 14.7 478.2 46.0 T 3.0 2.63
24 PI567056A 90 146 121.2 58.5 122.4 8.7 714.3 45.7 RB 0.0 1.86
25 PI628932 56 128 64.1 26.2 46.5 19.1 799.6 35.4 RB 1.0 2.27
26 PI587905 60 119 59.2 59.3 245.4 14.2 365.2 24.2 RB 1.0 2.08
27 PI567061 73 136 114.6 51.5 123.9 13.9 465.6 29.9 RB 1.0 2.35
28 PI567024 68 127 98.3 43.7 187.4 14.8 494.9 40.2 RB 1.6 1.96
29 PI605865B 65 122 89.7 36.7 70.8 15.1 540.3 38.9 RB 2.0 2.59
30 PI416826A 51 113 55.8 21.8 55.8 13.8 384.1 34.3 RB 1.0 1.53
31 PI506939 69 131 104.1 40.6 100.1 12.4 378.0 25.9 T 3.0 2.63
32 PI587880A 57 132 70.8 43.8 90.7 20.5 582.2 22.3 RB 0.0 1.60
33 PI567025A 70 131 105.2 42.3 94.7 12.4 318.5 35.3 RB 1.0 2.25
34 PI605838 75 137 120.5 44.5 104.0 11.8 386.0 31.4 T 2.0 2.19
35 PI567090 87 140 133.1 52.0 109.1 9.1 338.0 44.7 RB 1.0 2.02
[123]
36 PI605773 59 133 84.5 46.2 105.0 15.4 589.6 33.9 RB 1.0 1.89
37 PI416810 59 120 117.2 37.6 63.3 18.4 524.0 37.7 RB 2.3 2.63
38 PI605854B 65 131 83.2 36.8 62.8 14.4 638.8 36.0 RB 1.0 2.35
39 PI594767A 66 115 56.2 36.9 74.4 11.5 741.8 28.3 RB 0.0 2.29
40 PI566989A 79 137 92.5 50.7 104.7 11.2 603.1 44.1 RB 1.0 1.35
41 PI200466 50 119 69.3 42.6 98.8 19.1 555.3 38.3 RB 2.0 2.46
42 PI635999 64 132 90.2 30.9 82.7 19.0 456.0 26.0 RB 0.0 2.02
43 PI423960A 59 121 84.8 36.8 80.4 13.6 589.0 32.0 RB 1.0 1.96
44 PI417208 62 113 63.7 30.5 124.0 21.1 319.7 39.0 RB 2.0 2.55
45 PI567059 61 117 88.5 38.4 86.8 15.2 471.1 29.9 RB 1.0 2.06
46 PI567053 81 134 106.5 47.9 113.2 10.0 713.8 36.9 RB 0.0 1.92
47 PI567068A 78 128 104.3 61.7 284.8 7.8 766.1 31.1 T 3.0 1.59
48 PI567034 57 114 79.1 27.7 65.7 9.2 566.6 33.3 RB 0.0 2.18
49 PI567189A 66 130 85.6 40.1 88.8 13.8 513.1 33.0 T 3.0 2.18
50 PI594538A 57 115 49.8 25.4 70.5 19.1 597.5 17.5 RB 1.0 2.73
51 AFGAT(C5) 64 132 97.4 46.5 113.2 13.9 373.3 37.7 T 3.0 2.44
52 PI230970 61 139 94.7 58.0 141.1 11.4 484.9 24.4 T 3.0 2.66
53 PI615437 65.2 96 70.7 36.8 92.6 15.2 551.6 31.5 T 3.0 2.24
54 PI416886 52.6 111 42.8 25.1 66.1 19.9 544.0 32.4 RB 0.0 2.31
55 PI417085 65.3 126 88.1 37.3 77.5 21.2 219.7 36.6 T 2.0 2.55
56 PI203398 61.6 128 86.4 41.2 81.4 14.2 509.3 30.8 RB 2.0 2.48
57 PI423972 71.8 132 93.3 44.0 92.3 16.5 215.7 41.2 RB 2.0 2.33
58 PI423960B 50.4 119 76.0 38.9 85.9 18.4 458.4 35.0 RB 1.0 2.27
59 PI507004 52.2 109 46.5 21.8 34.2 20.6 475.7 28.4 RB 2.0 2.25
60 PI340898A 92.2 150 122.7 48.3 96.5 10.7 524.5 37.5 T 1.0 1.53
61 PI416873B 50.2 110 52.8 29.1 119.4 20.3 530.3 33.4 RB 0.0 1.94
62 PI506677 56.1 113 58.6 23.3 56.3 24.0 341.7 35.0 RB 1.0 2.36
63 Clark-63k 63.9 124 75.2 41.7 96.5 14.9 484.3 50.5 T 3.0 2.66
64 Nyala 59.9 119 63.3 32.1 65.6 12.7 625.2 47.1 T 3.0 2.27
Min 50.1 96 42.8 21.8 34.2 7.8 215.7 17.5 0.0 1.35
Max 92.2 150 148.4 148.3 284.8 24.0 804.5 50.5 3.0 3.43
Mean 66.9 128 90.9 44.2 102.2 14.4 496.1 34.4 1.5 2.25
LSD 7.7 13 18.02 38.16 96.51 1.92 11.47 12.6 0.57
CV % 8.31 7 14.16 63 68.63 9.53 18.2 23.77 18.11
[124]
Reaction of soybean genotypes to soybean rust disease
The response of soybean genotypes to ASR varied in relations to the number of
pustules, lesion types and the disease progress curve (AUDPC). The analysis of
variance indicated that there was significant (P<0.05) difference for AUDPC value
of Asian soybean rust among the 64 soybean genotypes (Table 3). This result
might indicate the presence of variation among the soybean genotypes with
respect to resistance or tolerance to Asian soybean rust. The AUDPC value ranged
from 215.67 (PI423972) to 804.45 (PI615445) with a mean value of 496.07
percent days. The AUDPC value of 30 of the soybean genotypes was less than the
mean value. The AUDPC values of released soybean varieties in Ethiopia, namely
Nyala, Clark and AFGAT were, respectively, 625.23, 484.2 and 373 percent-days,
the highest and lowest values recorded being, in order, on Nyala and AFGAT. The
AUDPC value of the other two checks G7955-C3RPP (C1) and MKSOY-2Nwhite
(c2) were 505.9 and 226.3, respectively.
The mean percent rust severity index of 64 soybean genotypes to Asian soybean
rust presented in table 4. The differences in mean rust severity among the
genotypes were highly significant (p<0.001). The lowest percent rust severity
index value was observed from the genotypes PI594538A(17.490) and the highest
rust severity value was observed from the genotypes Clark - 63k (50.50%) and
Nyala (47.12%), while the mean severity index value of the two resistant checks
‘G7955-C3RPP (C1)’ and ‘MKSOY-2Nwhite(c2)’ were 33.6 and 35.2
respectively. Based on percentage of rust severity index, out of the total 64
soybean genotypes 36 genotypes showed moderately susceptible reaction, while,
the remaining 28 genotypes showed moderately resistant.
Based on the reaction types, a total of 44 soybean genotypes produced Reddish
Brown (RB) lesions, and had less sporulation and sparse uredinia, implying that
they were moderately resistant to soybean rust. Reddish Brown (RB) lesions are
formed as a result of hypersensitive response of the crops to P. pachyrhizi, and its
formation inhibits the pathogen development or sporulation (Singh and Thapliyal,
1977). Lesion color is known to be controlled by resistance genes of Rpp1- Rpp6,
and thus should be considered when selecting resistant genotype (Yamanaka et al.,
2010; 2013). While, only 17 PI materials and three released soybean genotypes
(PI567058D, PI578457A, PI567180, PI606405, PI567069A, PI417089A,
PI606379B, PI506939, PI567104B, PI605838, PI567189A, AFGAT, PI230970,
PI340898A, Nyala, PI567068A, PI615437, PI417085 and Clarck-63k) showed
TAN reaction. Similarly, it was reported that the genotypes PI567069A, MKSOY-
2Nwhite, PI 417089A, PI 340898A and PI 606405 showed TAN lesions, while PI
230970 (Rpp2) and PI 567104B showd RB lesion in Vietnam (Pham et al., 2010)
and G7955-C3RPP (C1 Rpp3) in Uganda (Oloka et al., 2008). This indicates the
existent of diverse race of rust and instable resistance nature of the genotype
which is controlled by a single gene.
[125]
Several screening trials have been conducted by different workers for
identification of resistant sources. In Brazil, Santa Rosa, FT-1 and Uniao were
identified as resistant cultivars and all the varieties and germplasm from US were
found to be susceptible during screening trials (Ribeiro et al. 1985). Asian
Vegetable Research and Development Centre (AVRDC) had also screened over
9000 soybean accessions against rust but no immune cultivars have been
identified (Tschanz et al., 1985; Hartman et al., 2005).
Genotypes which exhibited TAN reaction also had high level of sporulation,
which indicates the susceptibility of these genotypes to soybean rust. It is
worthwhile to note that all the tested genotypes were not immune, indicating that
all the tested genotypes lack complete resistance. The genotype with characteristic
resistance to ASR had a lower number of rust severity, AUDPC value, and reddish
brown (RB) lesion type, but in the present study the increased value of AUDPC
was not linearly followed by the maximum reduction of the yield in some soybean
genotypes. Soybean genotypes having high value of AUDPC, rust severity and tan
reaction but still gave higher yield were categorized as tolerant.

For the occurrence of soybean rust, a continuous dew period, temperatures of
between 17-25 0C and relative humidity of above 70% are required for optimum
germination of the urediniospores, with rainfall being the most important factor
causing rapid increase in severity and disease spread (Melching et al., 1989).
Infection of Asian soybean rust might occur at any stage of crop development,
however, symptoms and severity increases after flowering under field conditions.
Hence, evaluating genotypes during the reproductive growth stage is ideal (Miles
et al., 2006; Walker et al., 2011). From this study there was a significant
difference for mean severity and AUDPC value for Asian soybean rust among the
tested soybean genotypes. A total of 49 soybean genotypes showed Reddish
Brown (RB) lesion which indicates presence of partial resistance gene while only
19 showed TAN reaction, and no soybean genotype classified as immune or
complete resistant to the disease.
The genotypes PI417089A, G7955-C3RPP (C1), PI471904, PI567104B,
PI567054C, PI594538A and PI230970 can be advanced for further testing and the
genotypes which have been identified as having partial resistance may used as
parental material for hybridization. Genotypes which showed RB reaction to soya
bean rust may or may not show a similar reaction in other locations due to the
variation of soybean rust race. Therefore, in the future, identifying the types of
rust race is one of the key stages to validate such research results. Similarly,
emphasis needs to be given to screening of large number of soybean genotypes
including locally released varieties. Moreover, conducting study based on cultivar
[126]
maturity group for identification of resistant genotypes which can be used as
source in breeding programs or for direct release if found suitable under diverse
climatic situation. A study on the tolerance and resistance mechanism of soybean
genotypes to soybean rust is also essential.
Acknowledgements
The authors thank the Ethiopian Institutes of Agricultural Research and soybean
innovation laboratory (SIL) for the financial support provided.
References
Akinsanmi OA, Ladipo JL and Oyekan PO. 2001. First report of soybean rust (Phakopsora
pachyrhizi) in Nigeria. Plant disease, 85(1), pp.97-97.
Bromfield KR and Hartwig EE. 1980. Resistance to soybean rust and mode of inheritance. Crop
Science, 20(2), pp.254-255.
Bromfield KR, Melching JS and Kingsolver CH. 1980. Virulence and aggressiveness of
Phakopsora pachyrhizi isolates causing soybean rust. Phytopathology, 70(1), pp.17-21.
CSA (Central Statistical Authority). 2018. Agricultural sample survey, Area and production of
temporary crops, private holdings for the 20017/18 Meher season.
Faske T, Kirkpatrick Zhou J, Tzanetakis I. 2014. Asian soybean rust. In: Arkansas Soybean
Production Handbook. Cooperative Extension Service, University of Arkansas Division of
Agriculture, Little Rock,
Garcia A, Calvo ÉS, Kiihl RAdS, Harada A, Hiromoto DM, Vieira LGE. 2008. Molecular
mapping of soybean rust (Phakopsora pachyrhizi) resistance genes: discovery of a novel
locus and alleles. Theoretical Applied Genetics 117, 545-553.
Goellner K, Loehrer M, Langenbach C, Conrath U, Koch E, Schaffrath U. 2010. Phakopsora
pachyrhizi, the casual agent of Asian soybean rust. Molecular Plant Pathology 11, 169-177.
Hartman GL, Miles MR, Frederick RD. 2005. Breeding for resistance to soybean rust. Plant
Disease 89, 664-666.
Hartman GL, West ED, Herman TK. 2011. Crops that feed the World 2. Soybean- worldwide
production, use and constraints caused by pathogens and pests. Food Security, 1-13.
Hyten DL, Hartman GL, Nelson RL, Frederick RD, Narvel JM, Cregan PB. 2007. Map location of
Rpp1 locus that confers resistance to soybean rust in soybean. Crop Science 47, 837-841.
Hyten DL, Smith JR, Frederick RD, Tucker ML, Song Q, Cregan PB. 2009. Bulk segregant
analysis using the GoldenGate assay to locate the Rpp3 locus that confers resistance to
soybean rust.Crop Science 49, 265-271.
Kawuki RS, Adipala E and Tukamuhabwa P. 2003. Yield loss associated with soybean rust
(Phakopsora pachyrhizi Syd.) in Uganda. Journal of Phytopathology. 151:7-12.
Koch E, and Hoppe HH. 1988. Development of infection structures by the direct-penetrating
soybean rust fungus (Phakopsora pachyrhizi Syd.) on artificial membranes. J. Phytopathol.
122:232-244.
Levy C, Techagwa JS and Tattersfield JR. 2002.The status of soybean rust in Zimbabwe and South
Africa. Paper read at Brazilian Soybean Congress, at Fozdo Iguacu, Prarana, Brazil.
Melching JS, Dowler WM, Koogle DL, and Royer MH. 1989. Effects of duration, frequency, and
temperature of leaf wetness periods on soybean rust. Plant Dis. 73:117-122.
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Mueller TA, Miles MR, Morel W, Marois JJ, Wright DL, Kemerait RC, Levy C, Hartman GL.
2009. Effect of fungicide and timing of application on soybean rust severity and yield. Plant
Diseases 93:243–248.
Ono Y, Buritica P, and Hennen JF. 1992. Delimitation of Phakopsora, Physopella and Cerotelium
and their species on Leguminosae. Mycol. Res. 96:825-850.
Oloka HK, Tukamuhabwa P, Sengooba T, Shanmugasundram S. 2008. Reaction of exotic
germplasm to Phakopsora pachyrhizi in Uganda. Plant Disease 92, 1493-1496.
Pante E, Esker P. 2008. Meteorological factors and Asian soybean rust epidemics-A systems
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[128]
Screening of Linseed (Linum usitatissimum)
Accessions for Resistance to Fusarium
oxysporum f. sp. Lini., at Holetta, Ethiopia
Habtewold Kifelew, Misteru Tesfaye and Birhanu Mengistu
Ethiopian Institute of Agricultural Research, Holetta Agricultural Research Center, P.O.box. 31, Holetta
Abstract
Reaction of linseed accessions against Fusarium oxysporum wilt was evaluated in
field sick plot and greenhouse condition at Holetta Agricultural Research Center. A
total of 144 genotypes from Holetta High and midland oil seeds research program
was used for filed screening and 60 for greenhouse study in the controlled
environment. Out of the 144 genotypes tested 88 genotypes were found highly
susceptible as the disease incidence in these cultivars was above 75%, 33 genotypes
fell in susceptible category, 16 genotypes including the released variety CI-1525 were
found moderately susceptible which were showed 25.1-50% disease intensity, 4
genotypes (Acc. 207789, Acc. 233993, Acc. 241824, and Acc. 10053) were under
moderately resistance (MR) and shown 10.1-25% diseases incidence and 3 genotypes
(Acc. 241820, Acc. 241822, and Acc. 10061) were found resistant (R) with incidence
between 0.1-10%. From greenhouse study the test genotypes were grouped as;
moderately resistance (MR) (1 genotype); Moderately Susceptible (MS)( 6 genotype);
Susceptible (S) (9 genotype) and Highly Susceptible (HS) (44 genotypes). The methods
used were found effective in screening linseed for Fusarium wilt, however, variable
results were found when comparing sick plot and controlled greenhouse tests.
Introduction
Linseed is one of the oldest plant species cultivated for food and fiber. In Ethiopia,
it is the third most important oilseed crop next to Sesame and Noug (CSA, 2017)
and cultivated solely by subsistent farmers at high-elevations (Adefris et al.,
1992). Ethiopia is considered center of diversity and ranks 7th among linseed
producing countries in the world. In 2017, a total of 96, 863 tons of linseed
produced from 92, 305 hectares of land (FAOSTAT, 2017), where it is mainly
used for domestic consumption.
Research on crop improvement, although started three decades ago, was

intensified only in the past 10 years, with the primary objective of identifying lines
or cultivars with stable, high seed and oil yields, better nutritional value and better
resistance/tolerance to major pests.
Teklemariam et al. (1985), Yitbarek (1992) and Geremew et al. (2009) reported
more than nine type of diseases attacking linseed in Ethiopia, among which , wilt
caused by Fusarium oxysporum causes up to 58.6% yield loss, and still remained
[129]
as the major challenge in linseed production. According to Kommedahl et al.
(1970), the disease can infect linseed at any growth stage and may result in 100%
disease incidence in certain cultivars.
Due to the ability of the pathogen to survive as a saprophyte for many years either
in plant debris or on the soil, it is difficult to eradicate the disease once the field
gets infected. The most commonly practiced disease control method is the use of
resistant varieties, while crop rotation for at least three years helps to lower the
inoculum level, thus reducing disease incidence and severity. However, the
resistance among varieties can differ due to the variability of pathogen races in
different geographical regions with varying temperatures and environmental
conditions (Rashid and Kenaschuk, 1993).
In case of soil-borne disease such as wilt, use of resistant varieties is the most
economic and eco-friendly method of diseases management. Screening for
resistance in sick plot in the field and greenhouse condition is a mandatory to
investigate the true resistance nature of the germplasm. The present study is
focused on screening of linseed accessions resistant/tolerant to wilt disease.

The study was conducted under field and greenhouse condition in the wilt sick
plot of of Holetta Agricultural Research Center. A total of 144 genotypes derived
from Holetta high and midland oil seeds research program was used for filed
screening and 60 for greenhouse study in the controlled environment. The growth
medium used was sterilized soil, which is sterilized using Harter Erddämpfer
Sterilo soil sterilizer, thermostatic control 30-95° C, and recommended 80 ° C for
24 hours. Eight seeds were sown per pot and thinned to six plants 7 days after
seeding. The accessions were planted in completely randomized design in a
growth chamber. Plants were watered daily and the trays covered with lids to
preserve the moisture. The experiment consisted of three replications and repeated
once. Within each replication, inoculated and un-inoculated (with sterile distilled
water only) was included for comparison.
In sick plot, each accession under test was sown in 2 meters long, single row, 20
cm apart, in rod row design. Fertilizer application, irrigation, weeding and other
intercultural operations were done as per the normal practice recommended. The
total numbers of plants in each row was counted for initial plant stand, wilting of
plants was carefully monitored right after emergence of seedlings to crop maturity.
The final wilt incidence (Table 1) was calculated by deducting the number of
plants survived from the initial crop stand. The percentage of wilting was
calculated using the following formula:
[130]
Percentage wilt incidence= (number of wilted plants/total plant population )*100
To produce mass of inoculums for augmenting the sick-plot, pure culture of the
pathogen was multiplied on autoclaved wheat seed.
Table.1. Rating scale used in grouping of resistance and susceptibility of the genotypes
S.N. Wilting percentage Scale Abbreviation
1 No wilting Immune I
2 0.1-10 Resistant R
3 10.1-25 Moderately resistant MR
4 25.1-50 Moderately susceptible MS
5 50.1-75 Susceptible S
6 75% and above Highly susceptible HS
Source: Kumar et al. (2014)

A total of 144 accessions were screened for resistance/ tolerance to Fusarium wilt.
Accordingly, 88 accessions were grouped under highly susceptible (HS), 33
susceptible (S), 16 moderately susceptible (MS), 4 (Acc. 207789, Acc. 233993,
Acc. 241824, and Acc. 10053) moderately resistant (MR) and 3 (Acc. 241820,
Acc. 241822, and Acc. 10061) accession were found resistant (R).
Table 2. Reaction of different linseed genotypes against wilt in sick plot under field condition, 2018
Disease
Name of Rating Name of Disease Rating
incidence S.N
genotypes Scale genotypes incidence (%) Scale
S.N (%)
1 Acc. 10101 72.72 S 73 Acc. 234012 84.29 HS
2 Acc. 13503 88.39 HS 74 Acc. 233996 98 HS
3 Acc. 13504 95.89 HS 75 Acc. 234002 99.13 HS
4 Acc. 13506 91.82 HS 76 Acc. 234004 97.75 HS
5 Acc. 13507 87.69 HS 77 Acc. 234005 90.00 HS
6 Acc. 13508 90.68 HS 78 Acc. 234008 48.24 MS
7 Acc. 13509 95.83 HS 79 Acc. 234009 43.93 MS
8 Acc. 13511 98.99 HS 80 Acc. 234010 85.25 HS
9 Kassa-2 74.07 S 81 Acc. 234011 50.00 MS
10 Acc. 13515 94.02 HS 82 Acc. 234012 63.85 S
11 Acc. 13517 93.94 HS 83 Acc. 235019 79.29 HS
12 Acc. 13519 98.24 HS 84 Acc. 235161 56.41 S
13 Acc. 13537 83.08 HS 85 Acc. 235163 61.18 S
14 Acc. 13548 83.87 HS 86 Acc. 235165 87.39 HS
15 Acc. 13568 34.02 MS 87 Acc. 235166 100.00 HS
16 Acc. 13578 48.53 MS 88 Acc. 235170 33.02 MS
17 Acc. 13598 54.79 S 89 Acc. 235171 96.88 HS
18 Acc. 13620 65.93 S 90 Acc.235175 97.16 HS
19 Acc. 13649 66.45 S 91 Acc. 235177 93.28 HS
20 Acc. 13653 79.46 HS 92 Acc. 236997 60.00 S
21 Acc. 13654 52.94 S 93 Acc. 236998 65.91 S
22 Acc.13682 75.83 HS 94 Acc. 237000 54.17 S
23 Acc. 207786 78.71 HS 95 Acc. 237001 92.42 HS
24 Acc. 207789 11.43 MR 96[131]
Acc. 237491 86.30 HS
25 Acc. 207791 85.61 HS 97 Acc. 237493 39.25 MS
26 Acc. 208360 54.17 S 98 Acc. 237501 39.02 MS
27 Acc. 208425 46.43 MS 99 Acc. 238278 95.06 HS
28 Acc. 208664 96.77 HS 100 Acc. 238279 76.71 HS
29 Acc. 208794 93.85 HS 101 CI- 1525 47.29 MS
30 Acc. 212512 93.08 HS 102 Acc. 241816 79.31 HS
31 Acc. 212517 99.05 HS 103 Acc. 241817 91.73 HS
32 Acc. 212625 99 HS 104 Acc. 241818 88.81 HS
33 Acc. 212747 98.26 HS 105 Acc. 241820 0 R
34 Acc. 212854 83.87 HS 106 Acc. 241821 50.74 S
35 Acc. 212857 87.5 HS 107 Acc. 241822 0 R
36 Acc. 215716 93.18 HS 108 Acc. 241823 54.76 S
37 Acc. 216012 88.52 HS 109 Acc. 241824 10.85 MR
38 Acc. 219963 98.13 HS 110 Acc. 241825 55.03 S
39 Acc. 219964 89.23 HS 111 Acc. 241826 38.57 MS
40 Acc. 219969 90.91 HS 112 Acc. 241827 89.80 HS
41 Acc. 223229 98.36 HS 113 Acc. 241828 52.63 S
42 Acc. 223231 91.80 HS 114 Acc. 241829 41.67 MS
43 Acc. 226025 100.00 HS 115 Acc. 241831 61.21 S
44 Acc. 226026 100.00 HS 116 Acc. 242588 72.31 S
45 Acc. 226028 97.04 HS 117 Acc. 242589 94.49 HS
46 Acc. 226032 81.31 HS 118 Acc. 242590 86.17 HS
47 Acc. 230029 99.10 HS 119 Acc. 242591 91.67 HS
48 Acc. 230033 87.22 HS 120 Acc. 242592 94.55 HS
49 Acc. 230034 69.05 S 121 Acc. 242593 98.26 HS
50 Acc. 230658 45.56 MS 122 Acc. 242594 92.47 HS
51 Acc. 230660 90.55 HS 123 Acc. 242595 100.00 HS
52 Acc. 230819 42.50 MS 124 Chilalo 63.08 S
53 Acc. 230822 96.74 HS 125 Acc. 243797 99.09 HS
54 Acc. 230825 87.37 HS 126 Acc. 243798 71.74 S
55 Acc. 230826 57.78 S 127 Acc. 243799 82.61 HS
56 Acc. 230827 96.47 HS 128 Acc. 243800 69.86 S
57 Acc. 231252 55.36 S 129 Acc. 243801 78.21 HS
58 Acc. 232215 6.73 MS 130 Acc. 243802 64.84 S
59 Acc. 233987 59.46 S 131 Acc. 243803 85.43 HS
60 Acc. 233989 87.43 HS 132 Acc. 10052 63.64 S
61 Acc. 233992 87.85 HS 133 Acc. 10053 14.41 MR
62 Acc. 233993 10.84 MR 134 Acc. 10054 88.30 HS
63 Acc. 233994 68.53 S 135 Acc. 10055 93.10 HS
64 Acc. 233995 97.30 HS 136 Acc. 10056 67.83 S
65 Acc. 233996 90.00 HS 137 Acc. 10058 58.62 S
66 Acc. 234002 96.49 HS 138 Acc. 10059 72.62 S
67 Acc. 234004 98.10 HS 139 Acc. 10061 9.02 R
68 Acc. 234005 97.14 HS 140 Acc. 10062 47.37 MS
69 Acc. 234008 38.95 HS 141 Acc. 10064 71.60 S
70 Acc. 234009 96.00 HS 142 Acc. 10066 83.80 HS
71 Acc. 234010 97.35 HS 143 Acc. 10067 89.69 HS
72 Acc. 234011 96.30 HS 144 Acc. 10068 89.66 HS
*S, susceptible; HS, highly Susceptible; MS, moderately susceptible; MR, moderately resistant and R, Resistant.
The reaction of linseed accessions against F. oxysporum f. sp. lini was presented
in table 2. The result revealed that out of the 144 genotypes tested, 88 were found
highly susceptible (as the disease incidence in these cultivars was above 75%), 33
[132]
fell in susceptible category, 16 genotypes including the released variety CI-1525
were found moderately susceptible (with disease intensity between 25.1-50%), 4
(Acc. 207789, Acc. 233993, Acc. 241824, and Acc. 10053) genotypes were under
moderately resistant (MR) (with incidence of 10.1-25%) and 3 (Acc. 241820, Acc.
241822, and Acc. 10061) accession were found resistant (R) with incidence ranges
of 0.1-10%.
Tested genotypes were categorized into resistant, moderately resistant, moderately

susceptible, susceptible and highly susceptible as per wilt incidence recorded.
However, some of the high yielding improved varieties like Chilalo, and Kassa-02
were found susceptible whereas, CI-1525 the released and highly performed
variety in wilt prone areas of western and northern Shewa, Arsi, and Bale known
as resistant/tolerant to the disease were found moderately susceptible in the
present study. The disease was characterized by rolling and withering of
cotyledons when disease appeared at seedling stage, and young seedlings
collapsed on ground. In grown up plants, dark green to brown patches appeared on
leaves. The leaves shriveled later on dropping off symptoms from tip of the plant
and drying of leaves and stem, discoloration of the vascular tissues were similar
symptoms to wilt caused by Fusarium oxysporum f. sp. Lini as reported by Kumar
et al. (2014).
Table 3. Reaction of linseed genotypes against wilt under greenhouse condition, 2018
S.N. Genotype Disease incidence Rating S.N. Genotype Disease incidence Reaction
%age scale %age
1 Acc. 10101 83.33 HS 31 Acc. 212625 66.67 S
2 Acc. 13503 100.00 HS 32 Acc. 212747 100.00 HS
3 Acc. 13504 83.33 HS 33 Acc. 212854 100.00 HS
4 Acc. 13506 100.00 HS 34 Acc. 212857 66.67 S
5 Acc. 13507 50.00 HS 35 Acc. 215716 100.00 HS
6 Acc. 13508 83.33 HS 36 Acc. 216012 66.67 S
7 Acc. 13509 100.00 HS 37 Acc. 219963 100.00 HS
8 Acc. 13511 100.00 HS 38 Acc. 219964 66.67 S
9 Kassa-2 50.00 MS 39 Acc. 219969 66.67 S
10 Acc. 13515 100.00 HS 40 Acc. 223229 83.33 HS
11 Acc. 13517 100.00 HS 41 Acc. 223231 100.00 HS
12 Acc. 13519 100.00 HS 42 Acc. 226025 100.00 HS
13 Acc. 13537 100.00 HS 43 Acc. 226026 83.33 HS
14 Acc. 13548 100.00 HS 44 Acc. 226028 100.00 HS
15 Acc. 13568 16.67 MR 45 Acc. 226032 100.00 HS
16 Acc. 13578 100.00 HS 46 Acc. 230029 66.67 S
17 Acc. 13598 66.67 S 47 Acc. 230033 66.67 S
18 Acc. 13620 83.33 HS 48 Acc. 230034 83.33 HS
19 Acc. 13649 83.33 HS 49 Acc. 230658 50.00 MS
20 Acc. 13653 83.33 HS 50 Acc. 230660 100.00 HS
21 Acc. 13654 83.33 HS 51 Acc. 230819 100.00 HS
22 Acc.13682 83.33 HS 52 Acc. 230822 100.00 HS
23 Acc. 207789 50.00 MS 53 Acc. 230825 100.00 HS
24 Acc. 207791 66.67 S 54 Acc. 230826 100.00 HS
25 Acc. 208360 33.33 MS 55 Acc. 230827 100.00 HS
26 Acc. 208425 100.00 HS 56 Acc. 231252 100.00 HS
27 Acc. 208664 100.00 HS 57 Acc. 232215 50.00 MS
28 Acc. 208794 100.00 HS 58 Acc. 233987 50.00 MS
29 Acc. 212512 100.00 HS 59 Acc. 233989 100.00 HS
30 Acc. 212517 100.00 HS 60 Acc. 233992 100.00 HS
[133]
Linseed screening for resistance/tolerance in greenhouse have been done on 60
genotypes. Result indicated that the tested genotypes were grouped as; moderately
resistant (MR) (1 genotype); Moderately Susceptible (MS) (6); Susceptible (S) (9)
and Highly Susceptible (HS) (44) accessions (Table 3).
The two methods used were found effective in screening linseed for Fusarium
wilt; however, variable results were found when sick plot and controlled
greenhouse tests were compared. Some genotype which is susceptible at field sick
plot condition found highly susceptible in greenhouse test.
Recommendations
Promising genotypes with resistance gene need to be included in crossing program
to develop resistant variety with good oil and seed yield. It is also recommended
that crossing need to start to improve the already released variety Chilalo and
Kassa-02. As future line of work, the remaining 70 genotypes and segregating
materials should also pass through the screening test for resistance/tolerance to
wilt.
References
Adefris Tekle Wold, Getinet Alemaw and Tesfaye Getachew. 1992. Linseed Breeding in Ethiopia.
http://agris.fao.org/agris-search/search.do?recordID=ET9300005
Central Statistical Authority (CSA). 2017. The Federal Democratic Republic of Ethiopia.
Agricultural Sample Surveys Report on area and production of Major crops for Meher season
crops for the year 2017/18.
FAOSTAT. 2017. http://www.factfish.com/statistic country/ethiopia/linseed,+production+ quantity).
Geremew Terefe, Dereje Gorfu, Dawit Tesfaye and Fekede Abebe. 2009. Review of research on
diseases of oil crops in Ethiopia In: Abraham T (ed) Increasing crop production through
improved plant protection Vol. II. PPSE and EIAR, Addis Ababa, Pp.253-274
Kommedahl T, Christensen JJ and Frederiksen RA. 1970. A half century of research in Minnesota
on flax wilt caused by Fusarium oxysporum. Tech. Bull. agric. Exp. Stn., (273).
Kumar M, Tripathi UK, Tomer A, Kumar P and Singh A. 2014. Screening of Linseed Germplasm
for Resistance/Tolerance against Fusarium oxysporum F Sp. Lini (Bolley) Disease. J Plant
Pathol Microb 5: 235. doi:10.4172/2157- 7471.1000235
Newkirk R. 2008. Flax Feed Industry Guide. Flax Canada, 2015.
Rashid KY and Kenaschuk EO. 1993. Effect of trifluralin on fusarium wilt in flax. Can. J. Plant
Sci. 73: 893-901
Teklemariam W, Asfaw Tulu and Mesfin Tessera. 1985. A review of research on oil crop disease
in Ethiopia. In:Tsedeke Adate (ed) proceeding of first crop protection symposium, 4-7
February, 1985. IAR, Addis Ababa. Pp. 291-312
Yitbarek S. 1992. Pathological Research in niger, linseed, gomenzer and rapeseed in Ethiopia. In:
oilseeds research and development in Ethiopia. Proceedings of the first national oilseed
workshop, 3-5 December, 1991, Addis Ababa, Pp.151-162
[134]
Development of Integrated Management of
Faba Bean Gall (Olpidium viciae Kusano)
in West Shewa Highland
Tajudin Aliyi, Alemayehu Hailu, Bayoush Birke and Gudisa Hailu
Ethiopian Institute of Agricultural Research, Ambo Agricultural Research Centre, Ambo, Ethiopia
Abstract
Faba bean gall (FBG) caused by Olpidium viciae Kusano, locally known as “Qormid
or Kormid” has become a serious threat to faba bean production in some parts of
Ethiopia causing a yield loss up to 100%. The spread of the disease has been very fast
and expanding from year to year in all faba bean-growing areas of the country. This
study was designed to develop integrated disease management option to control FBG.
The investigation carried out for three consecutive years to test Trichoderma species
for the management of FBG and integrate the effective species with other alternative
option. Randomized complete block design (RCBD) with three replications through
all experiments in all growing seasons was used in spite of the treatment
arrangements and numbers were differ from season to season depending on the
treatment combinations. Six Trichoderma species and two controls (positive and
negative) were used in pot experiment comprising a total of eight treatments. FBG
infected soil was used as positive control while sterile soil without any treatment used
as negative control in the first year experiment. Susceptible faba bean cultivar to FBG
was planted at hot spot area in west Shewa, Midakegn district to enhance the
inoculum source of the pathogen in the soil for subsequent years’ field trials along
with the first year’s experiment. In the second year two effective T. viride and T.
harzianum selected from first year result and then coupled with five foliar fungicides.
Totally eleven treatment combinations arranged in factorial setting. From the second
year results, the effective treatment combinations of fungicides and bio control agents
were selected. Thus, four treatment combinations involving two effective fungicides
(Sabozeb 80%WP and MORE 720 WP), two biocontrol agents (T. viride and T.
harzianum) and one untreated control were used in RCBD with three replications in
third year experiment. Data were recorded on disease incidence and severity. Severity
scores were converted to percent severity index (PSI) and used to calculate area
under disease progress curve (AUDPC). Grain yield and yield components were
measured after harvest. Disease parameters varied markedly across treatments,
especially biological agents-foliar sprays of fungicide combinations had a highly
significant variation (p<0.01) in disease parameters. Among six Trichoderma species
tested in the first year, T. viride and T.harzianum showed promising results against
the FBG. The results of the 2016/17 main cropping season revealed that the
integrated effect of seed treatment with T. harzianum and twice foliar sprays of
Sabozeb 80% WP reduced the terminal PSI of FBG by 18.67% as a result increased
the grain yield per hectare by 93.9% compared to untreated control. The highest
terminal PSI (75%) and AUDPC value (1125.0 %-days) were recorded on untreated
control plots. On the other hand the highest grain yield (3.144 t ha-1) obtained from
treatment of T.harzianum and twice foliar sprays of Sabozeb 80%WP fungicide while
the lowest grain yield (1.272 t ha-1) recorded from untreated plots at the final
experimental year of the study. Based on current results, seed treatment with T.
viridae or T. harzianum at the rate of 10 g powder per 1kg of seed before sowing
[135]
combined with twice foliar sprays of Sabozeb 80%WP or MORE 720 WP fungicides
at the rate of 2 kg ha-1 starting from seedling (five leaves) stages were effective to
manage FBG. Host resistance integrated with other cultural practices applicable in
all faba bean growing areas, and verification of the integrated disease management
(IDM) in the current study across locations and seasons should be given due
attention. In addition soil application of effective bio control agents that comprised
other available alternative FBG management option should be studied.
Introduction
Faba bean (Vicia faba L.) is a cultivated species belonging to the wild pea genus
(Vicia) of the Leguminosae family. Vicia faba, also known in the culinary sense as
the broad bean, fava bean, or faba bean is a species of flowering plant in the pea
and bean family Fabaceae. China is the leading faba bean producer of the world
followed by Ethiopia, Australia, Morocco, Egypt and France. Ethiopia is
considered as the secondary center of diversity and one of the nine major agro-
geographical production regions of faba bean (Asfaw et al., 1994). Faba bean
takes the largest share of the area and production of the pulses grown in Ethiopia.
It occupies 538,458.21 ha of land with annual production about 991,700.283 tons
and productivity of 1.842 t ha-1. It also covers about 41,747.00 ha of land with
annual production of 87,785.275 tons in North Shoa Zone (CSA, 2014). Even
though Ethiopia is the world’s second largest producer of faba bean next to China,
its share is only 6.96 % of world production and 40.5 % within Africa (Chopra et
al., 1989). Moreover, despite the availability of high yielding varieties (> 2 t ha -1)
(MoA, 2011), the average national yield of faba bean under small-holder farmers
is not more than 1.89 t ha-1 (CSA, 2014). The low productivity of the crop is
attributed to susceptibility to biotic and abiotic stresses (Sahile et al., 2008; Mussa
et al., 2008). Among economically important diseases, faba bean gall (which is
also locally known as ‘Qormid or Kormid’) incited by the pathogen Olpidium
viciae Kusano, causing up to complete crop failure over wide areas within short
period of time and aggravates the diminution of yield to maximum nationwide
(Dereje et al., 2012). This disease is recently reported from different faba bean
growing areas of Ethiopia, increasingly devastating the crop.
Different experiments were conducted to control faba bean diseases using
Trichoderma species and reported that the biological agents known to enhance
growth, nodulation, physiological activities and yield of faba bean (Saber et al.,
2009). In an attempt to manage faba bean gall using different fungicides, Bitew et
al. (2016) demonstrated that Bayleton, Ridomil and Mancozeb at manufacturers’
recommendation with thrice application resulted in minimized disease severity
and maximized grain yield.
Among seventeen varieties evaluated for their reaction to faba bean gall, four
varieties namely Dosha, Tumsa, Hachalu and Walki were found high yielder and
[136]
resistant to the disease, while Gabelcho and local check showed moderately
resistant reaction (Mastewal et al., 2018).
For better management of faba bean gall, study on integrated disease management
involving integration of various individual control options was carried out over the
last three years on which results of this paper is based. Therefore, the objective
this study was to develop integrated disease management option against faba
bean gall.

Experimental site, design and treatments
Field experiments were conducted on-farm under rain-fed conditions at
Barobidaru specific location of Midakegn district supposed “hot-spot” for faba
bean gall “Qormid”. Barobidaru located between 8°08' 61'' N and 37°28' 94'' E
and at an altitude of 2885 m.a.s.l. The study was conducted for three consecutive
main cropping seasons (2017-18/19). The experimental design was randomized
complete block (RCBD) with three replications through all experiments in all
seasons in spite of the treatment arrangements were different from season to
season depending on factors and treatment combinations used. Mass
multiplications and formulation of bio-agents (i.e. Trichoderma spp.) have been
done at Ambo Agricultural Research Center (AmARC), Mycology laboratory. The
seeds to be planted were coated/dressed on the bowl until fully covered with the
formulated Trichoderma using coffee husk (Habtegebriel and Boydom, 2016;
Abd-El-Khair et al., 2018). The Trichoderma spp. (T. harzianum) applied at the
rate of 10 g powder per 1kg of seed before sowing.
The study in the first year aimed at screening of Trichoderma spp. used as seed
treatment against faba bean gall (FBG) in pot experiment using faba bean seeds of
susceptible variety. Eight treatments comprised of six Trichoderma species and two
controls (positive and negative) were used in RCBD with three replications. FBG
infected soil was used as positive control while sterile soil without any treatment
used as negative control.
Two effective Trichoderma spp. viz. T. viride and T. harzianum were selected and
coupled with five fungicides, namely Triadmefon 250 g/l (Bayleton 25% WP) at 1
kg ha-1, Sabozeb 80%WP (Mancozeb 80%WP) and More 720WP at the rate of 2
kg ha-1, Propiconazole (Tilt 250EC) at the rate of 500 ml ha-1 and Dadozeb 80% at
the rate of 2.3 kg ha-1 in the second season. Fungicide application was started at
the onset of FBG (for systemic) and before onset of the disease (for contact),
repeated once after 7 and 10 days interval of the first application for contact and
systemic, respectively. A total of eleven treatment combinations were used in
factorial arrangement in RCBD and replicated three times. A plot size of 4.8m2
[137]
(2.4m x 2m) was used. Inter and intra row spacing was 40 cm and 10 cm,
respectively.
From the second year results, effective fungicides and bio control agents were
selected, integrated and evaluated in the third year. The treatments comprised of
four combinations involving two effective fungicides (Sabozeb 80%WP and
MORE 720 WP), two bio control agents (T. viride and T.harzianum) and one
untreated control was used in RCBD with three replications. Each experimental
unit area was 60m2 (10m x 6m). The distance between blocks and plots were,
respectively, 1.5 m and 1 m. Twenty-five rows per plot were used, and each row
accommodated 60 plants with an inter- and intra-row spacing of 40 cm and 10 cm,
respectively.
Recommended rate (100 kg ha-1) of phosphorous was applied once at planting in

the form of di-ammonium phosphate (DAP) and all other agronomic practices
were applied uniformly throughout all consecutive main cropping seasons.
Data collection
Disease assessment
Days to disease onset: Measured as the date on which the disease was first
noticed.
Disease incidence (%): Measured as proportion (%) of plants showing disease
symptom out of the total plants assessed at ten days interval starting from the
onset of the disease.
Disease severity (%): Disease severity was recorded by estimating the percentage
of leaf and stem area diseased using the 0-9 disease scoring scale (Ding et al.,
1993). The severity scales were converted in to percentage severity index (PSI) for
analysis using the following formula (Wheeler, 1969; Kumar et al., 2011):
Area under disease progress curve (AUDPC): The disease percent severity
index scores were used to calculate AUDPC for each treatment. The area under
disease progress curve was used to quantify the beginning of epidemic and the
time until disease reached maximum stage. AUDPC was calculated with the
formula suggested by Campbell and Madden (1990):
[138]
∑
Where, xi = is the cumulative percent severity index expressed as a proportion at

the ith observation, ti = is the time (days after sowing) at the ith observation, and n
= is total number of observations. Since the percent severity index was expressed
in percent and time (t) in days, AUDPC values were expressed in %-days
(Wilcoxson et al., 1975).
Grain yield and yield component parameters

The average number of pods per plant was determined from randomly selected
plants and the mean number of seeds per pod was also obtained from randomly
taken plants per plot at harvest. Similarly, the weight (in gram) of randomly taken
thousand seeds was recorded from each plot while the grain yield per plot was
recorded from the central rows. Then, grain yield per plot was calculated and
converted into yield per hectare basis.
Data analysis
Data on disease incidence, terminal percent severity index, AUDPC, yield and
yield component parameters were subjected to the analysis of variance (ANOVA)
to determine the treatment effect using SAS software version 9.4 ® (SAS, 2015).
All the disease reaction for each treatment was evaluated by averaging the data
from the individual plants. Data on disease incidence and severity were subjected
to square root transformation before analysis. The least significant difference
(LSD) values were used to separate differences among treatments means at 5%
probability level.

Screening of Trichoderma species against Faba Bean Gall (FBG) at
Midakegn
The pot experiment of the first year revealed that T. viride and T. harzianum
showed promising result against faba bean gall (FBG) throughout plant growth
stages (Table 1). Therefore, these two biological control agents can be
recommended for use in combination with fungicides.
[139]
Table 1. Effect of Trichoderma species against faba bean gall on pots experiment at Midakegn, 2016/2017 main cropping
season
Disease incidence (%)
Treatments⃰ 1st 2nd 3rd
T. hamatum against FBG 0 75 75
T. harzianum against FBG 0 8.3 25
T.viride against FBG 0 8.3 25
T. atroviride against FBG 0 0 100
T. asperellum against FBG 0 100 100
T. longibrachiatum against FBG 0 0 100
Positive control (FBG infected only) 100 100 100
Negative control (sterilized soil) 0 0 0
*FBG=faba bean gall
Integrated management of faba bean gall using fungicides and

Trichoderma species at Midakegn
Percent severity index
Percent severity index (PSI) calculated at the last date of disease assessment
showed significant difference (p≤0.05) among treatment combinations. The least
mean terminal percent severity index was recorded on plots received treatment
combinations; T. viride coupled with MORE 720 WP (9.67%) and T. harzianum
with Sabozeb 80% WP (8%), whereas the highest PSI (26.67%) was recorded on
the control plots (Table 2). The current result indicated that seed treatment of faba
bean with two aforementioned bio control agents and two times foliar spray of
Sabozeb 80%WP and MORE 720 WP fungicides had marked differences over
other treatment combinations. For instance, seed coated with T. harzianum before
planting and twice spray of Sabozeb 80%WP as well as with T. viride and two
times application of MORE 720 WP reduced PSI by 18.7% and 17%, respectively
(Table 2).
Yield and yield component
Thousand seed weight (TSW) grain yield in kilogram per plot and per hectare
showed significant (P≤0.05) variation among treatment combinations (Table 2).
Consequently, the highest mean TSW (0.87 kg plot-1) was obtained from plots
treated with T. viride with twice foliar sprays of MORE 720 WP fungicide. The
lowest mean TSW (0.47 kg plot-1) recorded on untreated control plots.
Significantly the highest grain yield (4.417 t ha-1) was obtained from the treatment
combination of T. harzianum with foliar spray of Sabozeb 80%WP at the rate of 2
kg ha-1. However, this yield was not significantly different from plots of T.
harzianum coupled with foliar spray of Triademifon, Propiconazole and MORE
720 WP fungicides. This indicated that we can use any of these fungicides with T.
harzianum to minimize the severity of faba bean gall then increase the yield of the
crop. The lowest (2.278 t ha-1) recorded on untreated control followed by
treatment combination of T. viride with foliar spray of Dadozeb 80% fungicide
(2.556 t ha-1), (Table 2).
[140]
The increment of the yield might be due to the role of biological agents and
fungicides. In nature application of biological agents is not only for the purpose
of control of disease, but they can also promote the growth of the crop hence
increases the vigorousness of the plants (Mastouri et al., 2010). This enable the
plant to defend against disease then produce more yield than the weak and
diseased/stressed plant. As observed from this study, seed dressed with T.
harzianum at planting and twice foliar spray of Sabozeb 80%WP fungicide before
disease onset significantly (p≤0.05) increased faba bean yield (by 93.9%) as
compared to untreated plot followed by T. viride and twice spray of MORE 720
WP which increased the grain yield by 87.8%. In contrary, the lowest yield
increased (by 12.2%) on seed treated with T. viride and twice sprays of Dadozeb
80% fungicides compared to untreated.
Table 2. Integrated effect of Trichoderma species and foliar sprays of fungicides on mean disease incidence, per cent severity index,
grain yield and yield component parameters at Midakegn, 2017/2018 main growing season
DI TSW Yield Yield
Treatments (%) PSI (%) PoPP SPPo (kg plot-1) (kg plot-1) (t ha-1)
100 18.67 bc 12.9 abc 2.9 ab 0.57 bc
1.7cde 2.833cde
T1(T.viride + Triadimefon)
T2(T.viride + Sabozeb 80%WP) 100 16.00bcd 16.4ab 2.9ab 0.6bc 1.87bcde 3.111bcde
T3 (T.viride + MORE 720 WP) 100 9.67fg 15.0abc 3.1a 0.87a 2.57ab 4.278ab
T4 (T.viride + Propiconazole) 100 15.33bcde 13.7abc 2.5b 0.53bc 2.17abcd 3.611abcd
T5 (T.viride + Dadozeb 80%) 100 19.33b 13.8abc 3.1a 0.73ab 1.53de 2.556de
T6(T.harzianum + Triadimefon) 100 13.33def 12.1bc 2.9ab 0.63bc 2.33abc 3.889abc
T7(T.harzianum + Sabozeb 80%WP) 100 8.00g 17.0a 2.9ab 0.73ab 2.65a 4.417a
T8(T.harzianum + MORE 720 WP) 100 11.00efg 11.3c 2.7ab 0.6bc 2.47abc 4.111abc
T9 (T.harzianum + Propiconazole) 100 14.00cdef 14.3abc 3a 0.63bc 2.20abcd 3.667abcd
T10 (T.harzianum + Dadozeb 80% WP) 100 15.67bcde 13.7abc 3a 0.63bc 2.03abcde 3.389abcde
T11 (Control, Untreated) 100 26.67a 12.3bc 3.1a 0.47c 1.37e 2.278e
Mean 100a 15.24 13.86 2.9 0.65 2.08 3.467
CV (%) 8.69 18.03 19.8 10.7 19.45 21.74 21.74
LSD (5%) 16.23 4.68 4.7 0.53 0.21 0.77 1.2838
* Means with the same letter are not significantly different, PoPP = Pods per plant, SPPo= Seeds per pod, TSW = Thousand seed weight, DI =
Disease incidence, PSI=per cent severity index, CV=Coefficient variation, LSD=least significant difference
Evaluation of effective Trichoderma species and fungicides
Percent Severity Index (PSI) and Area under disease progress curve (AUDPC)
The integrated effect of the two biological agents and fungicides tested in this
study showed significantly (p≤0.05) different response compared to the untreated
plots with regard to the final per cent severity index and area under disease
progress curve values. The significantly highest mean PSI (75%) recorded on
untreated control plots (Table 3). In contrast, the lowest (18.33%) recorded on
plots treated by Trichoderma harzianum and twice foliar sprays of Sabozeb
80%WP fungicide. Similarly, combined treatment of T. harzianum and twice
foliar sprays of Sabozeb 80%WP fungicide reduced the terminal percent severity
index by 56.67%.
According to El-Mougy and Abdel-Kader (2018) who conducted an experiment
under field condition using T. harzianum, antioxidants and fungicides stated that
the combined effect of T. harzianum with fungicides and antioxidants reduced the
[141]
disease incidence and severity on faba bean plants. The disease severity reduced
by up to 38.7 - 75.1% when antioxidants used alone, and 20.9% when combined
with T. harzianum and fungicide.Treatment combination of T. harzianum with
either one or all of the used antioxidants had superior effect in reducing disease
incidence and severity on faba bean plants compared to fungicide and untreated
control.
The result of analysis of variance (ANOVA) showed that area under disease
progress curve (AUDPC) showed highly significant difference (p≤0.01) among
treatment combinations. The significantly (p≤0.05) mean maximum of AUDPC
value (1125.0 %-days) recorded on untreated control plots whereas the mean
minimum AUDPC value (356.7%-days) was on plots treated with T. harzianum
and Sabozeb 80%WP fungicide (Table 3).
The area under disease progress curve (AUDPC) is a very convenient summary of
plant disease epidemics that incorporates initial intensity, the rate parameter, and
the duration of the epidemic, which determines the final disease intensity (Madden
et al., 2008). Therefore, it was used to quantify the beginning of epidemic and the
time until disease reached maximum stage.
The results of this study demonstrated the integrated effect of the two bio control
agents used as seed treatment before planting combined with fungicides as foliar
spray were effective to reduce faba bean gall intensity. This might be due to
Trichoderma species out compete fungal and fungal-like plant pathogens for
nutrient and space resources on plant roots, resulting in reduced pathogen success
and plant disease; recognize and attack harmful fungal and fungal-like plant
pathogen by dissolving their cell walls and absorbing the released nutrients, i.e.
mycoparasitism and promote general plant disease defences, increasing the
“immunity” (acquired resistance) of the plant (Hermosa et al., 2012).
Yield and yield components
Pods per plant and Seeds per pod: The number of pods per plant was
significantly(p≤0.05) different among the treatment combinations and untreated
plots, while the number of seeds per pod showed significant variation not within
treatments but compared to untreated control plots.
The highest (20.58) mean number of pods per plant recorded on the treatment
combination of T. harzianum with Sabozeb 80%WP fungicide followed by
(17.595) T. viride seed treatment integrated with two foliar sprays of MORE 720
WP fungicide. The mean lowest number of pods per plant (10.08) obtained from
untreated control.
The highest (3.50) mean number of seeds per pod recorded on plots treated with T.
harzianum and fungicide Sabozeb 80%WP followed by (3.01) plots treated with
[142]
T. viride combined with foliar sprays of fungicide MORE 720 WP (Table 3).
Grain yield and thousand seed weight (TSW): The analysis of variance for grain
yield (t ha-1) and thousand seeds weight (g pot-1) showed a significant (p ≤ 0.05)
difference between treatment combinations and untreated control. But, there were
no significant difference observed among each combination of bio control agents
with foliar fungicide application. Compared with untreated plots, the mean
maximum (3.144 t ha-1) grain yield was obtained from seed coated by T.
harzianum and twice foliar sprays of fungicide Sabozeb 80%WP, while the mean
minimum (1.272 t ha-1) grain yield recorded from control plots (Table 3). More or
less the same is true for the result of thousand seed weights in the current study.
From this study it can be observed that significantly better yield of faba bean
obtained from treatment combinations compared to the untreated control plots.
Yield increases were computed relative to the average grain yield of the unsprayed
control plots (i.e. the lowest yield and the highest disease severity). For instance,
treatment combinations of T4, T1, T3 and T2 increased yield significantly by
47.2%, 37.9%, 32.4% and 19.2%, in that order, compared to untreated control
plots (Table 3).
It has been reported that, under stressful conditions, plants often mobilize nutrients
and redirect their metabolism to support active defence mechanisms to the
detriment of growth and final yield (Smith and Moser, 1985). However, the
application of fungicides might have prevented the triggering of active defence
mechanisms by killing the inoculum prior to infection and/or attempted tissue
colonization. This would allow the allocation of more metabolic resources to the
sink organ and lead to a better yield. On the other hand, research has also shown
that Trichoderma species promote general plant disease defences, increasing the
“immunity” (acquired resistance) of the plant and its colonization of crops induced
higher photosynthetic rates and systemic increases in CO2 uptake in leaves thereby
can lead to increased yield under stress conditions (Vargas et al., 2009; Guler et
al., 2016). Therefore, Trichoderma colonize the rhizosphere of many plants,
resulting in beneficial effects such as increased resistance to pathogens and greater
yield and productivity.
[143]
Table 3. Integrated effect of T. species and fungicides on mean incidence, terminal per cent severity index, Area under disease progress curve values, grain yield and yield component
parameters at Midakegn, in 2018/2019 main growing season⃰
AUDPC TSW Yield

Treatments DI (%) PSI (%) (%-days) PoPP SPPo (g plot-1) (t ha-1)
T1 (Trichoderma viride + MORE 720 WP fungicide) 90.00 30.667b 413.3b 17.595ab 3.012a 557.33a 3.028a
T2 (Trichoderma viride + Sabozeb 80%WP fungicide) 100.00 25.667 b 503.3b 15.282 b 2.800 a 554.00a 2.789a
T3 (Trichoderma harzianum + MORE 720 WP fungicide) 100.00 23.333 b 425.0b 16.128 ab 2.804 a 547.33a 2.956a
T4 (Trichoderma harzianum + Sabozeb 80%WP fungicide) 96.67 18.333b 356.7b 20.582a 3.501a 566.67a 3.144a
T5 (Untreated Control) 100.00 75.00a 1125.0a 10.082c 2.00b 423.33b 1.272b
Mean 97.33 34.60 564.67 15.43 3.819 529.73 2.438
CV (%) 8.697 30.05 25.36 17.65 14.48 3.60 24.56
LSD (5%) 15.939 19.579 269.59 5.1281 0.702 35.916 1.1275
* Means with the same letter are not significantly different
N.B.:- PoPP = Pods per plant, SPPo= Seeds per pod, TSW = Thousand seed weight, DI = Disease incidence, PSI=percent severity index, CV=Coefficient of variation, LSD=least
significant difference
[144]
Among six Trichoderma species tested for the management of FBG, T. viride and
T.harizanum showed promising results compared to others. The results of the
2017/18 main cropping season revealed that the integrated effect of seed treatment
with T. harzianum and foliar sprays of Sabozeb 80% WP reduced the terminal
percent severity index of faba bean gall by 18.67%, as a result the grain yield (ton
per hectare) increased by 93.9% compared to the untreated control. This
investigation demonstrated the two Trichoderma species coupled with either
MORE 720 WP or Sabozeb 80% WP with two times foliar spray managed faba
bean gall under natural infection. Consequently, the highest (3.144 t ha-1) mean
grain yield was obtained from plots treated with T.harzianum and twice foliar
sprays of fungicide Sabozeb 80%WP while the lowest (1.272 t ha-1) mean grain
yield recorded from untreated control plots.
The current results revealed that faba bean seed treatment of T. viridae or T.
harzianum at the rate of 10 g (coffee husk based formulation containing spores)
powder per 1kg of seed before sowing combined with twice foliar sprays of
Sabozeb 80%WP or MORE 720 WP fungicides at the rate of 2 kg ha-1 at seedling
stage (i.e. starting from five leaves and before flowering) were effective to manage
FBG. However, the following recommendation was made to come up with
effective, affordable and sustainable FBG management strategies.
 Verification of the developed integrated disease management (IDM) options reported in this
study across locations and seasons should be given due attention.
 Host resistance integrated with other cultural practices applicable in all faba bean growing
areas should also be sought and verified
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American Phytopathological Society. St. Paul, Minnesota, USA.
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[146]
Screening of Fungicides for the Control of
Aloe vera Rust under Field Condition
Mihiret Mekonnen1 and Begashaw Manahlie2
1
Ethiopian institute of Agriculture Research, Wondo Genet Agricultural Research Center, P.O.Box.198,
Shashemene, Ethiopia; 2Debre Berhan University, College of Natural and Computational Science,
Department of Biotechnology, Debre Berhan, Ethiopia
Abstract
Aloe vera belonging to the Asphodelaceae family is a perennial succulent medicinal
plant which grows in tropical and subtropical areas. It is used to preserve foods and as
medicine all over the world. Its leaf possesses many pharmaceutical compounds with
antimicrobial, anticancer, antioxidant, antidiabetic, antiulcer, hepatoprotective and
immunomodulatory activities. Commercially, Aloe vera can be found in pills, sprays,
ointments, lotions, liquids, drinks, jellies, and creams. However, it is severely affected by
rust disease, a very serious limiting factor to Aloe vera production. Therefore, the
objective of this study was to screen fungicides for the control of Aloe vera rust in field
condition. Five fungicides were tested in randomized complete block design (RCBD) with
three replications during 2018. All the five tested chemicals significantly (p = 0.05%)
reduced disease severity at least by 15% compared to untreated plants. Progress 250
EC, Panazole 25% EC and Nativo SC 300 at recommended rates showed significantly
superior performance with 78.33, 75 and 71.67% disease control efficacy, respectively,
compared to Bless (66.67%) and Noble 25 WP* (55%). Therefore, Progress 250 EC,
Nativo SC 300 and Panazole 25% EC can be recommended for the control of Aloe vera
rust.
Introduction
Aloe vera, which belongs to the Asphodelaceae family, is a perennial succulent
medicinal plant with very short stem or stemless long leaves. Aloe vera,
cultivated in tropical and subtropical areas of the world, is one of the 250
species of Aloe. The plant has lance-shaped, sharp pointed, and jagged and
edged leaves (Knowledge Base Script, 2009). Aloes are often thought to only
grow in hot and dry climates but they actually grow in a variety of climates
including desert, grassland, and coastal or even alpine locations (Davis, 2009).
Aloe vera is mainly used as a food preservative and medicine all over the
world. According to findings of various studies Aloe vera leaf possesses many
pharmaceutical compounds with antimicrobial (Bashir et al., 2011), anticancer
(Naveena et al., 2011), antioxidant (Miladi and Damak, 2008), antidiabetic
(Jones, 2007), antiulcer (Borra et al., 2011), hepatoprotective (Chandan et al.,
2007) and immunomodulatory (Atul et al., 2011) activities. Commercially,
aloe can be found in pills, sprays, ointments, lotions, liquids, drinks, jellies,
and creams (Hosseini and Fakhraee, 2014). Many of the health benefits
associated with Aloe vera have been attributed to the polysaccharides
[147]
contained in the gel of the leaves (Josias et al., 2008). Aloe vera is the most
commercialized aloe species, and processing of the leaf pulp has become a
large worldwide industry. In the food industry, it has been used as a source of
functional foods and as an ingredient in other food products, for the production
of gel-containing health drinks and beverages. In the cosmetic and toiletry
industry, it has been used as base material for the production of creams,
lotions, soaps, shampoos, facial cleansers and other products.
However, according to various literatures, Aloe vera cultivation has been
adversely affected by rust (Uromyces aloes) in different parts of the world viz,
Africa (Ethiopia, Kenya, Lesotho, Madagascar, Malawi, Rhodesia, Tanzania
and Uganda), Asia (India), and United Kingdom. Similarly, its production is
severely affected by rust at Wondo Genet Agricultural Research Center pilot
sites (Wondo Genet and Hawassa) with 70% disease incidence and 100%
disease severity. This causes both qualitative and quantitative losses of the gel.
The disease is characterized by the appearance of golden yellow big pustules,
arranged concentrically on both sides of the leaves which upon maturity turn
yellow to brown. The leaves turned yellow and finally die depending on the
severity of infection. Sometimes, severe infection may lead to premature death
of the plant. However, no appropriate management option was developed for
the disease so far. Therefore, the objective of this study was to screen synthetic
fungicides for effective control of Aloe vera rust in field condition.

Five fungicides were tested against Aloe vera rust at Wondo Genet
Agricultural Research Center experimental field during 2018, Ethiopia. The
site is located at 7° 192′ N latitude and 38° 382′ E longitude with an altitude of
1780m above sea level. The site receives a mean annual rainfall of 1000 mm
with minimum and maximum temperatures of 10 and 30°C, respectively. The
soil textural class is clay loam with an average pH of 7.2. The tested fungicides
were Progress 250 EC at 0.5L/ha in 250L water, Noble 25 WP* at 0.5 kg/ha in
150-400 L water, Nativo SC 300 at 0.5 L/ha, Bless at 250 ml/ha and Panazole
25% at 500 ml/ha in 400 L water. Untreated plants were also included as a
control.
The experiment was laid out in randomized complete block design (RCBD)
with 3 replications. A plot size of 3m × 3m with 60cm spacing between plants
was used. Spacing between plots and blocks were 1.5m and 2m, respectively to
minimize the spread of the disease by wind. Healthy planting materials were
planted. The chemicals were sprayed four times at 15 days interval since the
time of disease occurrence.
[148]
Data collection and statistical analysis
Data were recorded on disease severity, disease control, plant height, leaf
number per plant and gel weight. Final recording of disease severity was taken
after 4 weeks of spraying using 0 – 4 rating scale, where 0 = no disease
(healthy); 1 = 1 – 25%, 2 = 26 – 50%; 3 = 51- 75%; 4 = 76 –100% leaf areas
infected using the following formula:
Disease severity = Area of plant tissue affected x 100
Total area
From each plot, five plant samples were measured for each parameter
Percent disease control (PDC) = Disease in control plot - Disease in treated plot x 100
Disease in control plot
Data were statistically analyzed using analysis of variance (ANOVA) and

difference between means was assessed using LSD at P= 0.05 probability level
using SAS software version 9.2.

The results of the present study revealed that all the five tested chemicals
significantly (p = 0.05%) reduced disease severity at least by 15% compared to
untreated plants. Yet, Progress 250 EC, Panazole 25% EC and Nativo SC 300
at recommended rates showed significantly superior performance with 78.33,
75 and 71.67% disease control efficacy, respectively, compared to Bless
(66.67%) and Noble 25 WP* (55%). Therefore, Nativo SC 300, Progress 250
EC and Panazole 25% EC were most effective fungicides for the control of
Aloe vera rust. Similar finding was reported by Masson et al., 2011, where
Progress 250 EC and Panazole 25% EC at recommended rates showed 92.01
and 74.23% disease control efficiency.
Significantly higher plant height (50.47cm), leaf number (54.53) and fresh leaf
weight (3976.10 g/p) were recorded from Progress 250 EC treated plants
compared to all the rest treatments (Figure 1). However, no significant
difference was observed for gel weight among all chemical treatments (Table
1). The result in this study agrees with the finding of Singh and Dwivedi
(1990) who reported that application of Progress 250 EC at 0.1% on wheat rust
resulted in significant yield enhancement. As explained by Balardin et al.
(2010) fungicide application prior to any contact between pathogen and host is
considered to be preventative.
[149]
Table 1. Efficacy of different fungicides for the control of Aloe vera rust in field condition at Wondo Genet ARC
Trade name Constituent PH LNPP FLW GELW DS (%) DC (%)
Progress 250 EC 25% Propiconazole + 75% inert 50.47a 54.53a 3976.1a 946.9a 6.67b 78.33a
ingredients
Noble 25 WP* 25%wp Triadimefon 47.47ba 44.40ab 2570.2b 1041.0a 10.00b 55.00ab
Nativo SC 300 100g/L Trifloxystrob+ 200g/L 49.47ba 54.80a 2953.4b 963.4a 8.33b 71.67a
Tebuconazole
Bless 430g/L Tebuconazole + 570g/Linert 46.00b 45.93ab 3297.5ba 1001.0a 8.33b 66.67ab
materials
Panazole 25% EC 250g/L EC Propiconazole 49.87ba 56.13a 3399.7ba 940.5a 6.67b 75.00a
Control - 48.53ba 44.40ab 3000.2ba 781.0b 25.00a 0.00c
CV - 2.23 2.23 2.23 2.23 2.234 2.232
LSD - 4.4315 17.029 19.23 18.79 4.598 24.446
Means with the same letter within the same column are not statistically different (P < 0.05). Where, PH = plant height; LNPP = leaf number per plant;
FLW = fresh leave weight; GELW = gel weight; DS = disease severity and DC = disease control.
Figure 1. Effect of different fungicides on Aloe vera rust under field condition; where: A = Progress 250 EC treated plants; B = Bless treated plants;
C = Nativo SC 300 treated plants and D = control
[150]
Among investigated fungicides Progress 250 EC, Panazole 25% EC and Nativo
SC 300 at recommended rates showed significant Aloe vera rust reductions and
had great effect on plant height, leaf number per plant, fresh leaf weight and gel
weight. Therefore, these chemicals can be good candidates for the control of Aloe
vera rust in field conditions. Further study should be conducted under typical
conditions by varying rate of application.
Acknowledgements
Authors would like to acknowledge Wondo Genet Agricultural Research Center
and Aromatic and Medicinal plants Research project for providing all the
necessary facilities and support.
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[151]
[152]
Evaluation of Fungicides Efficacy against Rice sheath
Rot (Sarocladium oryzae) in Rain-fed Lowland Rice
(Oryzae sativa L.) in Fogera Hub
Muluadam Berhan1, Desalegn Yalew1, Tekalgn Zeleke2
1
Ethiopian Institute of Agricultural Research (EIAR), Fogera National Rice Research and Training Center
(FNRRTC), P. O. Box 1937, Bahir Dar, Ethiopia; 2EIAR, Ambo Agricultural Research Center, P.O. Box 37,
Ambo, Ethiopia.
Abstract
Rice (Oryzae sativa L.) is by far the most stable food crop for over half of the world
society, they are dependent for half of their calorie, and its production is enhancing
gradually even with in developing nations. Rice sheath rot is a disease complex which
can be caused by fungi (Sarocladium oryzae) pathogen and it infects the upper most
important plant part (panicle). The study was aimed to evaluate the efficacy of different
fungicides and recommend the most effective ones. The experiment was conducted at
Fogera National Rice Research and Training Center experimental station for two
consecutive years (2017-2019) using X-Jigina cultivar. Nine treatments, four seed
dressing (carboxin+ thiram + imidacloprid, imidacloprid + tebuconazol,
thiamethoxam 20% + metalaxyl- 20 % + ifenoconazol 2% and imidaclopride 250
gm/kg) and four foliar applied fungicides (epoxiconazole + thiophanate-methyl,
mancozeb + metalaxyl WP, tebuconazole, and mancozeb 80% WP were used including
untreated check. The treatments were arranged in randomized complete block design
with three replications. Disease incidence and severity data were recorded before
maturity of the crop on 0-9 scale. The result revealed that seed dressing fungicides
were by far better over foliar fungicides. Proseed plus 63 WS (carboxin+ thiram +
imidacloprid), Joint 246 FS (imidacloprid + tebuconazol) and ImidalmT 450 WS
(imidaclopride 250 gm/kg) fungicides were effective against rice sheath rot disease
control, respectively. Furthermore, the study had proved that the highest marginal rate
of return was obtained from application of Imidaclopride 250 gm/kg fungicide which
was 32.17 extra birr beyond covering cost of investment. In a net shell, effective seed
dressing fungicides were much better to manage rice sheath rot disease since its source
of transmission is seed.
Introduction
Rice (Oryzae sativa L.) is an important crop worldwide, serving as the staple food
for half of humanity and additionally being used in industry and for animal feed.
Rice is grown in various agro-ecological zones in tropical and subtropical areas,
especially in Asia, the continent accounting for 90% of the world production
(IRRI, 2015). Sixty percent of the world`s population are depending on rice for
their half of the calorie intake (Deepmala, 2012). In Ethiopia, it is considered as
"millennium crop", believing it ensures food security. Even though total rice
production and area coverage have been increasing year to year since its
recognition by government in 2005, rice productivity has stayed almost stagnant
below three tones over the years, which is very low as compared to rice yield
potential.
[153]
There are several factors that are limiting its productivity in the country. Rice
disease and insect pests have been identified as leading constraints (Nigussie and
Alemu, 2011). Rice sheath rot is a disease complex that can be caused by various
fungal pathogens. Major pathogens associated with rice sheath rot are fungi such
as Sarocladium oryzae and Fusarium species belonging to the Fusarium
fujikuroi complex (Dileep, 2016). A variety of other pathogens have been
associated with rice sheath rot. The various described sheath rot agents all cause
very similar disease symptoms (Cottyn et al., 1996). This explains why there are
practically no comprehensive studies mentioning the link between the presence
and quantity of disease inoculum and yield loss (Mew and Gonzales, 2002). The
unpredictable nature of the factors acting in the pathosystem explains why losses
attributed to Sarocladium oryzae can be as variable as in the range of 20–85%
(Sakthivel et al., 2002).
The disease is prevalent in all rice-growing areas in Ethiopia causing significant

damage to rice crop. Sheath rot is now considered to be a major destructive
disease which is common in rain-fed lowland rice ecosystems. The pathogen
attacks flag leaf sheaths and grains and yield losses result mainly from poor
panicle formation and exertion, spikelet sterility (80-100%), reduced grain filling,
and losses in milling (Simon, 2016). Panicle exertion depends on the time of
infection and severity of the disease. Under severe conditions, the panicle remains
inside the leaf sheath (Naeimi et al., 2003). The booting stage of the rice plant is
the most vulnerable stage for infection, causing maximum damage (Nasu, 2004).
While emerging from the infected sheath, the young panicles are affected,
increasing the number of chaffy, discolored, and shriveled grains and reducing the
weight and number of healthy grains. Yield loss incurred by sheath rot infection
was found to be as high as 85% (Bigirimana et al., 2015). Caused yield losses are
variable from 10 to 85%, depending on the pathosystem conditions (Sakthivel,
2001).
According to 2007/2008 E.C production year rice disease survey, rice sheath rot is
found to be one of the most serious diseases which bring significant yield loss on
the popular cultivar X-Jigina at different sub-districts of the Fogera, Dera and Libo
kemkem districts. The assessment shows that the disease prevalence, incidence
and severity in the above areas were 100, 47 and 44 %, respectively. However,
there is no report regarding the management options in Ethiopia.
Use of fungicides is an option to manage rice sheath rot, in which an integration of

cultural practices with chemical control is the most effective. Therefore, this study
was conducted to evaluate fungicides efficacy against rice sheath rot at rain fed
lowland rice growing areas of the country and recommend it to be included in the
rice package.
[154]
The study was conducted for two consecutive years (2017-2019) in rain fed lowland
rice ecosystem at Fogera National Rice Research and Training Center (FNRRTC),
South Gondar zone of Ethiopia, located at 610 Km to north-west of Addis Ababa
and about 55 Km north of Bahir Dar. Geographically the experimental site is
located at latitude of 11° 58′ N and longitude of 37° 41′ E with an altitude of 1819
meter above sea level and it receives average annual rainfall of 1230 mm. Mean
minimum and maximum temperature of the area is 12 and 280C, respectively. The
soil is brown clay (vertisol) which is rich in underground water (Getahun, 2015).
Experimental design and treatments

The treatments were arranged using randomized complete block design with three
replications. The plot size was 3 meters with 2.4 meter (7.2 m2) and the block size
was 25.6 meter with 11 meters (281.6 m2). A spacing of 0.5 meter and 1 meter
was used between plots and blocks, respectively. In this experiment nine
treatments were evaluated, of which four were seed dressing fungicides while the
other four were foliar fungicides and one control check. Randomization was held
independently for each replication by which treatments were assigned completely
at random as described by Gomez and Gomez (1984) and Poduska (2008).
The seed dressing fungicides were carboxin + thiram + imidacloprid,
imidacloprid + tebuconazol, thiamethoxam 20% + metalaxyl- 20 % +
difenoconazole 2% and imidaclopride 250gm/kg + thiram 200 gm/kg. On the
other hand, epoxiconazole + thiophanate-methyl, mancozeb + metalaxyl WP,
tebuconazole, and mancozeb 80% WP were applied through foliar spray. The list
of applied fungicides, dose and their method of application are indicated as in
table 1.
Table 1. List of applied fungicides, dose and their method of application
Code⃰ Trade name Active ingredient name Method of Recommended

application rates per hectare
T1 Proseed Plus 63 WS Carboxin+ Thiram + Imidacloprid Seed treatment 200 gm/100kg seed
T2 Rex® Duo Epoxiconazole +thiophanate- methyl Foliar 0.5 L+ 200 Lt water
T3 Joint 246 FS Imidacloprid + Tebuconazol Seed treatment 200 ml/100kg seed
T4 Mancolaxyl 72 % WP Mancozeb + metalaxy WP Foliar 3.5 L + 550 L water
T5 Natura 250 EW Tebuconazole Foliar 0.5 L+ 250 Lt water
T6 Apron Star 42 WS Thiamethoxam 20% + metalaxyl Seed treatment 200 gm/100kg seed
- 20 % + ifenoconazol 2%
T7 Indofil M-45 Mancozeb 80% WP Foliar 2 kg + 750 Lt water
T8 Imidalm T 450 WS Imidaclopride 250 gm/kg + thiram Seed treatment 100 gm/100kg seed
T9 Untreated check 200 gm/kg
⃰T1= Treatment one, T2= Treatment two, T3= Treatment three, T4= Treatment four, T5= Treatment five, T6: Treatment six,
T7= Treatment seven, T8= Treatment eight, T9= Treatment nine
[155]
Experimental procedures
The experimental site was ploughed four times and precise leveling was done. Sheath
rot infected rice seeds were planted as a spreader row which could be a source of
inoculums for the disease to occur uniformly on test genotypes. The X-Jigina rice
cultivar was used at recommended seed rate (80 kilogram per hectare).
Accordingly, 58 gram seed was used for each plot. Furthermore, 60.5 kilogram per
hectare nitrogen, phosphorus and sulfur in the form of NPS with grade of 19N-
38P-7 S and urea fertilizer at the recommended rate of 130 kilogram per hectare
were applied. All plots were received 43.56 gram of Nitrogen, phosphorus and
sulfur (NPS with grade of 19N-38P-7S), and urea fertilizer at the rate of 93.6 gram
per plot.
The seeds were dressed with fungicides one week before sowing. Seeds were
thoroughly dressed with the fungicides through adding an ample amount of water
to coat it well. Foliar fungicides were applied at booting stage of the crop, once in
a season as per the recommended dose. All agronomic management practices were
kept the same for each treatment except fungicides application which were the
target to determine their efficacy.

Disease incidence and severity data of rice sheath rot were recorded regularly at
weekly intervals from ten randomly selected plants in each plot and the degree of
infection occurred in each treatment were visually scored and averaged to obtain
the mean of the ten selected plants, and done similarly for each plot. Data on days
to emergence, days to heading, plant height, panicle length, number of spikes per
plant, number of fertile tillers per plant, number of filled grains per panicle,
number of unfilled grains per panicle, thousand grain weight, grain yield and grain
moisture content were taken.
Moreover, disease data viz., incidence and severity were scored for both pre and
post application of treatments. Scoring was made four times so as to have
confidence in the efficacy of fungicides evaluation. Disease intensity was
recorded at crop maturity using 0-9 scales following standard evaluation
procedure developed by International Rice Research Institute (IRRI, 2013).
The collected data were analyzed using version 9.0 Statistical Analysis Software
(SAS) and mean separation was done at 5% least significant difference. The
analysis of variance was employed for the analysis of treatments variation in terms
of sheath rot control among variables.
[156]
The result of analysis of variance showed that there was highly significant
difference among seed dressing and foliar fungicides in terms of disease incidence
and severity (Table 3). In addition, treatments had shown significant variation on
grain yield and panicle length of the crop. It clearly revealed that application of
fungicides in various forms had provided different level of sheath rot control. Plots
that were treated with Proseed plus 63 WS (carboxin+ thiram + imidacloprid),
Joint 246 FS (imidacloprid + tebuconazol) and ImidalmT 450 WS (imidaclopride
250 gm/kg + thiram 200 gm/kg) fungicides gave the best grain yield, since they
effectively inhibited the disease progress. Viswanathan and Narayanasamy (1992)
also reported that the use of fungicides at different concentration effectively
inhibited spore germination, germ tube elongation, mycelial growth and
sporulation of Sarocladium oryzae.
The lowest disease incidence was recorded on plots sown to seeds treated with
fungicides ‘imidalm T 450 WS (Imidaclopride 250 gm/kg + thiram 200 gm/kg),
proseed plus 63 WS (Carboxin+ Thiram + Imidacloprid) and joint 246 FS
(Imidacloprid + Tebuconazol)’, which had 11.02, 16.44 and 18.5 % incidences,
respectively. Similarly, the lowest disease severities of 8.69, 12.01 and 16.84%
were recorded, respectively, on plots sown to seeds treated with fungicides
‘imidalm T 450 WS (imidaclopride 250 gm/kg + thiram 200 gm/kg), proseed plus
63 WS (carboxin+ thiram + imidacloprid) and Apron Star 42 WS (thiamethoxam
20% + metalaxyl- 20 % + difenoconazole 2%) (Table 2).
On the other hand, joint 246 FS (imidacloprid + tebuconazol), imidalm T 450 WS

(imidaclopride 250 gm/kg) and Apron Star 42 WS (thiamethoxam 20% +
metalaxyl- 20 % + difenoconazole 2%) gave the highest thousand grain weight
(29.92, 29.82 and 29.60 gram), respectively (Table 2). This implies that the grain
quality obtained from plots treated with these fungicides were excellent. The
current findings further substantiated results of Painkra (2016) who reported,
under the in-vivo conditions, fungicides significantly reduced the disease intensity
over control and increased the grain yield of rice.
In the same fashion, seed dressing fungicides had direct effect on the panicle
length since the pathogen mainly infects the upper most flag leaf sheaths that
enclose the emerging young panicle during the boot stage and causes empty head.
Moreover, seed dressing fungicides were by far better than foliar ones since the
pathogen is seed born and can be transmitted by infected crop residue. This
realizes that these fungicides had controlled the pathogen which can be available
in the soil and on the seed. Furthermore, pre-sowing treatments were effective in
the enhancement of plant height due to the fact that it inhibits clogging the panicle
[157]
exertion. Similarly, Ramabadran and Velazhahan (1990) confirmed that seed
treatment with seed dressing fungicides improved seed germination.
Table 2. The effect of seed dressing and foliar fungicides on the rice sheath rot⃰
Treatments PH PL TPP SPP TGW GY DI DS

Proseed plus 63 WS 79.77 17.83 4.96 9.8 28.77 4.02481a 16.44de 12.01f
Rex® Duo 74.36 17.56 4.70 9.4 28.24 3.13085bc 33.33ab 32.73b
Joint 246 FS 79.86 18.23 5.73 9.5 29.92 3.97978 ab 18.50 cde 17.59e
Mancolaxyle 72% WP 79.40 17.93 4.90 10.1 27.97 3.29156abc 24.50bcd 22.68d
Natura 250 EW 74.03 16.90 5.53 9.3 27.94 2.70119c 41.50a 39.13a
Apron star 42 WS 76.16 17.40 5.43 9.26 29.60 3.4039 abc 20.44 cde 16.84e
Indofil M-45 75.13 18.16 5.30 9.8 28.78 3.00916c 24.83bcd 21.67d
ImidalmT 450 WS 77.60 18.53 4.93 9.43 29.82 3.94371ab 11.02e 8.69g
Untreated check 76.63 18.40 5.17 10.3 27.70 2.87170 c 27.06 bc 25.82c
Grand mean 76.99 17.88 5.18 9.65 28.74 3.3733 24.18 21.90
LSD (5%) 5.75 1.09 0.98 0.98 1.77 882.55 10.58 2.49
CV(%) 6.42 5.23 16.33 8.78 5.29 22.45 37.57 9.78
p-value 0.24 0.06 0.50 0.27 0.08 0.01 0.0003 0.0001
⃰PH= plant height (cm), PL= Panicle length (cm), TPP= Number of tillers per plant, SPP= Number of spikelet per panicle,
TGW= Thousand grain weight (gram), GY= Grain yield in ton/ha, DI= Disease incidence percentage, DS= Disease
severity percentage, LSD= Least significant difference at 5% significance level, CV= Coefficient of variation in percent.
[158]
GY
4.5
3.5
Grain yield in ton

2.5
1.5
0.5
DI DS
DI and DS in %
Figure 1. Effect of fungicides (seed dressing and foliar application) on grain yield (kg/ha) and sheath rot severity and
incidence in percent. DI= Disease incidence in percent, DS= Disease severity in percent, GY= Grain yield in
ton/ha
[159]
As shown in Figure 1, the lowest grain yield was recorded from the untreated plot,
Natura 250 EW and Indofil M-45 sprayed plots. Thus, grain yield is directly
proportional with sheath rot severity. When the severity increased the yield
becomes decreased and vice versa. However, this may not always true as the
disease affects the flag leaf and leaf sheath. This is due to the fact that, the disease
record was mostly undertaken from panicle initiation to flowering crop stages and
the fungicides were applied early in the season following the appearance of
disease incidence. The trend in figure 1 revealed that fungicides efficacy was
varied with respect to sheath rot incidence and severity. All seed dressing
fungicides by far better controlled the sheath rot than foliar applied fungicides.
This clearly revealed that the disease is seed borne and most likely transmitted via
the seed.
Table 3. The analyses of variance in terms of different variables mean squares with the interaction of inputs across year⃰
SV DF PH PL TPP SPP TGW GY DI DS

Block 2 20.67 1.77 0.09 9.70 4.71 745678.84 528.58 55.72
Year 1 897.92** 92.3** 142.11** 45.92** 544.54** 475387.31ns 54.12ns 21.18**
Treatment 8 31.53 ns 1.64 * 0.69 ns 0.8 ns 4.40 ns 1514989.57 * 503.76 ** 560.27**
Year * Trt 8 31.11 ns 0.52ns 0.61 ns 1.15 ns 2.51 ns 46778.46ns 10.47ns 1.42ns
Error 34 22.82 0.95 0.74 0.61 2.27 598689.31 99.53 5.34
* = significant at p<0.05, ** = significant at p<0.01, SV = source of variation, DF = degree of freedom, PH = plant
height(cm), PL= panicle length(cm), TPP = number of tillers per plant, SPP = number of spikelet per panicle, TGW =
thousand grain weight, GY = grain yield, DI = disease incidence, DS = disease severity, Trt = treatment
Economic analysis
As the decision which disease management method to be applied depends on the
effectiveness and economic feasibility of the method, partial budget analysis was
done to identify the rewarding treatments. Yield from experimental plots was
adjusted downward by 10% for management and plot size difference, to reflect the
difference between the experimental yield and the yield that farmers could expect
from the same treatment (CIMMYT, 1988).
The partial budget analysis includes only production costs that vary across
treatments while common costs for all treatments were excluded. Average market
grain price of rice (14 ETB per kg) and labor valued at ETB 50 per person day
were used. The result of the partial budget analysis is described in Table 4. The
economic analysis revealed that the highest net benefit of (birr 39462.12 ha-1) was
obtained from the application of proseed plus 63 WS, followed by joint 246 FS
and imidalm T 450 WS as seed dressing fungicides. The marginal rate of return
showed that when a farmer invest one birr on imidalm T 450 WS, Proseed Plus 63
WS and Joint 246 FS to control sheath rot, he/she will get one birr + extra 3217,
2877 and 2760 %, respectively, whereas natura 250 EW and indofil M-45 have
provided negative rate of return which were -265 and -32 %, respectively (Table
4).
[160]
The change in total variable inputs cost (total variable input cost minus the control
check) was highest in Mancolaxyle 72% WP, Indofil M-45 and Rex® Duo inputs
which was 2710, 1990 and 1550 ETB per hectare, respectively. On the contrary,
proseed plus 63 WS (11,032.29 ETB ha-1), joint 246 FS (10,586.49 ETB ha-1) and
imidalm T 450 WS (10,292.90 ETB ha-1) showed highest change in net benefit
(net benefit minus the control check). Change in net benefit implies that the
benefit or profit that one obtains after he/she recover the application cost of
fungicides to control rice sheath rot. Whereas, natura 250 EW and indofil M-45
were resulted in the lowest change in net benefit (-2708.05 and -629.15 ETB ha-1),
respectively. It showed that with the application of these fungicides, one lost an
extra 2708.05 and 629.15 ETB ha-1. On the other hand, the untreated plot has
provided better yield than the above foliar fungicides as shown in the table below.
[161]
Table 4. Partial budget analysis and marginal rate of return of fungicides (CIMMYT, 1988)
Inputs GY AGY kg/ha CPR GFB Birr/ha TVIC NB Birr/Ha CNB Birr/Ha CTVIC Birr/Ha MRR=I
(ton/ha) = A (A*0.9)=B (ETB)=C (B*C)=D /Ha=E ((B*C)-E)=F (F-control)=G (E-con trol)=H
Proseed plus 63 WS 4.02481 3.62233 11.00 39845.62 383.5 39462.12 11032.29 383.5 2877
Rex® Duo 3.13085 2.81777 11.00 30995.42 1550 29445.42 1015.59 1550 66
Joint 246 FS 3.97978 3.58180 11.00 39399.82 383.5 39016.32 10586.49 383.5 2760
Mancolaxyle 72% WP 3.29156 2.96240 11.00 32586.44 2710 29876.44 1446.61 2710 53
Natura 250 EW 2.70119 2.43107 11.00 26741.78 1020 25721.78 -2708.05 1020 -265
Apron star 42 WS 3.4039 3.06351 11.00 33698.61 390 33308.61 4878.78 390 1251
Indofil M-45 3.00916 2.70824 11.00 29790.68 1990 27800.68 -629.15 1990 -32
ImidalmT 450 WS 3.94371 3.54934 11.00 39042.73 320 38722.73 10292.90 320 3217
Untreated check 2.8717 2.58453 11.00 28429.83 0 28429.83 0.00 0 0.00
Note: GY= grain yield, AGY= Adjusted grain yield, CPR (ETB): current price of rice in Ethiopian birr, GFB= gross farm get benefit, TVIC/Ha= total variable input cost per hectare, NB
Birr/Ha= net benefit in birr per hectare, CNB Birr/Ha= change in net benefit in birr per hectare, CTVIC Birr/Ha= change in total variable input cost in birr hectare, MRR= marginal rate of
return (%)
[162]
In conclusion, the results of this experiment showed that three seed dressing
fungicides were the most effective in the control of rice sheath rot. It is clearly
shown that the highest yields were recorded in plots sown to seeds treated with
fungicides ‘Proseed plus 63 WS (carboxin+ thiram + imidacloprid), Joint 246 FS
(imidacloprid + tebuconazol) and ImidalmT 450 WS (imidaclopride 250 gm/kg +
thiram 200 gm/kg)’ with grain yield of, respectively, 4.02481, 3.97978 and
3.94371-ton ha-1. However, the partial budget analysis proved that Imidalm T 450
WS (imidaclopride 250 gm/kg + thiram 200 gm/kg) was the most promising seed
treatment fungicide in terms of disease severity, incidence, and change in net
benefit as well as marginal rate of return.
Seed treatment fungicides are by far better than foliar ones since the rice sheath rot
is most likely transmitted by infected seed and crop residue. In other words, the
above most effective fungicides are ideal for the aquatic fungi as compared with
the foliar applied fungicides, possibly because they are applied early in the season
when there is no stand water.
Therefore, based on the economic analysis, fungicides efficacy and the grain yield
obtained, ImidalmT 450 WS (imidacloprid 250 gm/kg + thiram 200 gm/kg),
Proseed plus 63 WS (carboxin+ thiram + imidacloprid) and Joint 246 FS
(imidacloprid + tebuconazol) fungicides can be recommended for the management
of rice sheath rot at the doses specified in table 1. However, it is worthwhile to
note that environment friendly and sustainable disease management can only be
achieved through integrated disease management. Thus, the effective fungicides
just identified should be used integrated with cultural and resistant/tolerant rice
varieties.
Acknowledgements
Our sincere gratitude goes to Fogera National Rice Research and Training Center
and Ethiopian Institute of Agricultural Research for financial support provided and
facilitation to do the work. The authors would like to acknowledge field assistants
for their vital role played without which this success is unattainable.
References
Bigirimana VP, Hua GKH, Nyamangyo OI and Höfte M. 2015. Rice sheath rot: An emerging
ubiquitous destructive disease complex. Front Plant Science. 6: 1066.
CIMMYT (International Wheat and Maize Improvement Program). 1988. From Agronomic Data
to Farmer Recommendations: An Economic Training Manual. Completely Revised Edition,
Mexico D.F.
[163]
Cottyn B, Cerez MT, Van Outryve MF, Barroga J, Swings J, Mew TW. 1996. Bacterial
diseases of rice. Pathogenic bacteria associated with sheath rot complex and grain
discoloration of rice in the Philippines. Plant Disease. 80: 429 - 437.
Deepmala K. 2012. Studies on management of sheath rot disease of rice. Master of science in plant
pathology.1: 15-17.
Dileep KP. 2016. A study on management of sheath rot disease of rice caused by Sarocladium
oryzae (Sawada) Games and Hawksworth. Master of science in plant pathology,1-122.
Getachew Agegnehu and Taye Bekele. 2005. On-farm integrated soil fertility management in
wheat on nitisols of central Ethiopian highlands. Ethiopian Journal of Natural Resources. 7: 141-
155.
Getahun Dessie. 2015. Open Field Performance of Tomato (Lycopersicon esculentum Mill.)
Cultivars in the Rainy Season at Woreta, South Gondar, Ethiopia. International Journal of
Scientific Research in Agricultural Sciences, 2(5): 117-125.
Gomez K. and Gomez A.1984. Statistical procedure for Agricultural Research. 2nd Edition.
John Wiley and Sons, Inc. Canada. Pp:138-170.
IRRI (International Rice Research Institute). 2013. Standard Evaluation System for rice. Injuries
caused by diseases; 5th edition. Pp:18-33.
IRRI (International Rice Research Institute). 2015. World Rice Statistics in 2013. Available
at: http://ricestat.irri.org
Mew TW and Gonzales P. 2002. A Handbook of Rice Seedborne Fungi, IRRI. 35-36.
Naeimi S, Okhovvat SM, Hedjaroude GA and Khosravi V. 2003. Sheath rot of rice in Iran.
Commun. Agriculture Applied Biological Science. 68: 681-684.
Nasu H. 2004. Seed transmission of sheath rot fungus of rice and its infection site. Japan Journal
of Phytopathology. 70: 18-21.
Poduska, J. 2008. Building Split-Split-plot Designs in Statistical Software. Internet document.
Rajendra Mane Sachin. 2014. Integrated Weed Management in Upland Rice (Oryza sativa L.).
Master of science thesis in India.
Ramabadran, R. Velazhahan, R. 1990. Management of sheath rot of rice. Basic research for crop
disease management. 367-375.
Sakthivel N, Amudha R, Muthukrishnan S. 2002. Production of phytotoxic metabolites
by Sarocladium oryzae. Mycology Research. 106: 609 - 614.
Simon MM. 2016. Increasing the resilience of elite rice cultivars to sheath rot (Sarocladium oryzae
[(Sawada) W. Gams & D. Hawksw]) in Rwanda through breeding for resistance . PhD
dissertation. Pp 157.
Teshome Nigussie and Dawit Alemu. 2011. An overview of national rice research and
development strategy and its implementation. Oromia Bureau of Finance and Economic
development, Ethiopian Institute of Agricultural Research; Addis Ababa Ethiopia. FRG II
Project; Empowering Farmers Innovation. 2: 1-2.
Vidhyasekaran P, Ranganathan K, Raja Manikam B and Radhakrishnan J. 1984. Quality of rice
grain from sheath rot affected plants. International Rice Research Newsletter. 9: 19.
[164]
PART II
Weed Science
[165]
[166]
Survey and Identification of Major
Weeds of Seed Spices in Ethiopia
Merga Jibat1, Wakjira Getachew2, Abukiya Getu1, Habetewold Kifelew3
1
Ethiopian Institute of Agricultural Research, Tepi Agricultural Research Centre, P. O. Box 34, Tepi
2
EIAR, Jimma Agricultural Research Centre, P. O. Box: 192, Jimma
3
EIAR, Holetta Agricultural Research Centre, P. O. Box 2003, Addis Ababa
Abstract
In Ethiopia, coriander (Coriandrum sativum L.), fenugreek (Trigonella foenum-
graecum) and black cumin (Nigella sativa L.) are economically important seed
spices. They are highly affected due to weed infestation especially from competition
for water and nutrient. However, before estimating yield losses due to weeds and
develop weed control strategies, identification and quantification of weeds
associated to these spice plants were fond very important. Weed survey was
conducted in East Shewa, Arsi and Bale zones during 2016 and 2018 main cropping
seasons to identify most common and prevalent weeds associated with the seed
spices. Weed species characteristics, density, frequency, relative density, relative
frequency, summed dominant ratio over locations and seasons were calculated. The
result showed that a total of 22, 37 and 21 weed species were identified in coriander,
fenugreek and black cumin fields, respectively. The frequency of individual weed
species in coriander, fenugreek and black cumin field ranged from 0.14% to 1%,
0.13% to 1% and 0.25% to 5% while the dominance value ranged from 0.14 to
49.1%, 0.25 to 26.5% and 0.25 to 4.5%, respectively. The most frequent and
dominant weed was Chenopodium album in coriander field whereas, the most
frequent species was Chenopodium album and weed was Drymaria cordata in
fenugreek field. In black cumin field Cynadon dactylon and Solanum nigrum were
the most dominant and frequent weed species. Therefore, integrated weed
management should be developed targeting mainly those weed species associated to
specific seed spice plants more frequently and dominantly.
Introduction
Spices are well known for their flavoring, culinary uses and essential oil
derivatives. They are also valued for their coloring, as preservatives and
fumigants, in pharmaceutical, textile and other industry (Purseglove et al., 1991).
They do fetch premium price for they are highly concentrated by nature and are
considered as an alternative cash generating commodity to stabilize the fluctuating
price of coffee in the international trade (Girma et al., 2008). Production of
essential oil in an agro-industry venture can effectively fits the national
development endeavors of most developing countries, including Ethiopia.
Development of these spices does not only benefit producing farmers but also
other citizens who are in one way or another involved in spices marketing and
utilization, and at large benefit the country to earn foreign currency.
[167]
The annual global economic loss caused by weeds has been estimated at more than
$100 billion U.S. dollars (Appleby et al., 2000).Weeds are undesirable plants,
which infest different crops and inflict negative effect on crop yield through
competition for water, nutrients, space and/or light (Reddy and Reddi, 2011).
There are innumerable reports on the inhibitory effects of weeds on crop plants
(Javaid et al., 2007). Weeds are notorious yield reducers that are, in many
situations, economically more important than insects, fungi or other pest
organisms. At low density, weeds do not affect yield and certain weeds can even
stimulate the crop growth (Thijssen, 1991).
Weeds, considered as obnoxious plants, are one of the pests associated with any
agriculture endeavor and compete with coriander, fenugreek and black cumin
plants for sunlight, space, water and nutrients. Weeds may also act as alternate
hosts to insect pests and pathogens attacking seed spices. Research indicated that,
worldwide, over 10% of agricultural production is lost as a result of crop weed
competition for the resources light, water and nutrients (Parker and Fryer, 1975).
According to Akobundu (1987) when weeds are left uncontrolled, yield losses
range from 20-100%, depending on the crop and its environment. The author
reported loss estimates of 5% in developed countries, 10% in the less developed
countries and 25% in the least developed countries.
Weeds surveys are commonly used to characterize weed populations in cropping

systems (Uddin et al, 2010). Therefore, to develop an effective weed management
strategy, weed survey is necessary to address the current weed problems on seed
spices. In addition, survey information is important in developing target oriented
research and control measure. So far, no related study has been conducted
regarding the occurrence and distribution of common weeds associated with
coriander, fenugreek and black cumin in Ethiopia.
Therefore this survey was conducted with two main objectives: (1) to identify and
document the kinds of weed species and (2) to characterize and classify common
weeds relative distribution associated with coriander, fenugreek and black cumin
in the major production areas of Ethiopia.

Weed survey was conducted in East Shewa, Arsi and Bale zones of Oromiya
Regional Administrative State. The survey was conducted during 2016 and 2018
main cropping seasons using random sampling technique. Samples were taken
using 0.5 m × 0.5 m size quadrat at 2–3 m interval in cross diagonal line, by
randomly placing in weed infested areas of the crops production fields. All weed
species obtained within a quadrat were uprooted, cleaned, and separately placed in
plastic bags. There were five sampling spots considered per a given field. After
[168]
completing the weed collection from the fields, the specimens were identified to
species level and counted using the ‘Flora of British India’ by Hooker (1872-
1897);‘Flora of Andhra Pradesh’ by Pullaiah and Chennaiah (1997); and Weed
Identification and Control Guide (Donaldson and Bowers, 1998).
Data analysis
After collecting the quantitative data of the weed species MS Excel was used to
compile the data initially. Then, their density, relative density, frequency, and
relative frequency, summed dominant ratio (SDR) were calculated using the
following formula:
a. Density (D) = Total number of individuals of a species in all quadrats / Total number of
quadrats used
b. Frequency (F)= Number of quadrates in which a given species occurs/Total number of
quadrates used
c. Relative density (RD) = (Density of a given species / Total density for all species) × 100
d. Relative frequency (RF)= (Frequency of a given species / Total frequency for all species)
× 100
e. Summed dominant ratio (SDR) = (Relative density/Relative frequency) × 100

Coriander
A total of 22 weed species were identified from coriander field (Table 1). The
most important families according to the number of represented species were
Amaranthaceae, Caryophyllaceae, Primulaceae, Fabaceae and Polygonaceae. The
frequency of occurrence of individual weed species ranged from 0.14% up to 1%
while, the infestation level (dominance) ranged from 0.14% up to 49.1%. The
most frequent and dominant weed was Chenopodium album.
[169]
Table 1. Weed composition, dominance, frequency, relative density, relative frequency, summed dominant ratio in
coriander field, 2016 and 2018
S.N Name of Weeds Family Weed occurrence status in coriander*

D F RD RF SDR
1 Avena fatua Poaceae 0.71 0.43 0.44 3.8 11.6
2 Phalaris minor Poaceae 2.86 0.29 1.76 2.53 69.6
3 Chenopodium album Amaranthaceae 49.1 1 30.3 8.86 342.1
4 Vicia hirsutum Fabaceae 8.71 0.86 5.37 7.59 70.76
5 Vicia sativa Fabaceae 10.4 1 6.43 8.86 72.59
6 Anagalis arvensis Primulaceae 15.1 1 9.34 8.86 105.4
7 Solanum nigrum Solanaceae 2.86 0.57 1.76 5.06 34.8
8 Oxalis corniculata Oxalidaceae 8.14 0.86 5.02 7.59 66.12
9 Medicago denticulate Fabaceae 0.71 0.43 0.44 3.8 11.6
10 Fumaria parviflora Papaveraceae 5.57 0.71 3.44 6.33 54.29
11 Cynadon dactylon Chlorideae 1.71 0.43 1.06 3.8 27.84
12 Gnaphalium affine Asteraceae 0.57 0.29 0.35 2.53 13.92
13 Polypogon fudax Poaceae 2.43 0.43 1.5 3.8 39.44
14 Polygonum plebijum Polygonaceae 15.1 0.86 9.34 7.59 123
15 Drymaria cordata Caryophyllaceae 26.1 0.57 16.1 5.06 318.4
16 Euphorbia spp Euphorbiaceae 0.14 0.29 0.09 2.53 3.48
17 Rumex crispus Polygonaceae 7 0.57 4.32 5.06 85.26
18 Spergula arvensis Caryophyllaceae 2.29 0.14 1.41 1.27 111.4
19 Melilotus indica Fabaceae 0.43 0.14 0.26 1.27 20.88
20 Ammania baccifera Lythraceae 0.71 0.14 0.44 1.27 34.8
21 Other number - 0.71 0.14 0.44 1.27 34.8
22 Galinsoga spp Asteraceae 0.57 0.14 0.35 1.27 27.84
*D-Density, F-Frequency, RD-Relative density, RF-Relative frequency, SDR-Summed dominant ratio
Fenugreek
From the surveyed fenugreek fields a total of 37 weed species were identified
(Table 2). The most important families according to the number of represented
species were Amaranthaceae, Fabaceae and Polygonaceae. The frequency of
occurrence of individual weed species ranged from 0.13% up to 1% while, the
infestation level (dominance) ranged from 0.25% up to 24.9%. Similar to in
coriander, Chenopodium album was the most frequent and dominant weed in
fenugreek too.
[170]
Table 2. Weed composition, dominance, frequency, relative density, relative frequency and summed dominant ratio in
fenugreek fields, 2016 and 2018.
S/N Name of weeds Family Weed occurrence status in fenugreek

D F RD RF SDR
2 Chenopodium album Amaranthaceae 24.9 1 13.7 7.02 195.7
3 Vicia hirsutum Fabaceae 3.88 0.75 2.14 5.26 40.65
4 Vicia sativa Fabaceae 10.8 1 5.94 7.02 84.58
5 Anagalis arvensis Primulaceae 8.38 0.63 4.62 4.39 105.4
6 Solanum nigrum Solanaceae 15.1 0.38 8.35 2.63 317.3
8 Medicago denticulate Fabaceae 1 0.75 0.55 5.26 10.49
10 Gnaphalium affine Asteraceae 6 0.63 3.31 4.39 75.53
12 Polygonum plebijum Polygonaceae 19.1 0.63 10.6 4.39 240.7
13 Galinsoga ciliate Asteraceae 2.38 0.38 1.31 2.63 49.83
14 Drymaria cordata Caryophyllaceae 26.5 0.63 14.6 4.39 333.6
15 Stelaria media Caryophyllaceae 5.88 0.5 3.24 3.51 92.44
16 Euphorbia spp Euphorbiaceae 0.63 0.25 0.35 1.75 19.67
17 Cyperus esculentus Cyperaceae 2 0.38 1.1 2.63 41.96
18 Rumex crispus Polygonaceae 3.75 0.38 2.07 2.63 78.67
19 Convolvulus arvensis L Convolvulaceae 0.63 0.13 0.35 0.88 39.34
22 Echinochloa colona Poaceae 0.5 0.13 0.28 0.88 31.47
23 Caesulia axillaris Asteraceae 0.13 0.13 0.07 0.88 7.867
24 Lathyrus aphaca Fabaceae 0.25 0.25 0.14 1.75 7.867
25 Amaranthus viridis Amaranthaceae 0.63 0.25 0.35 1.75 19.67
26 Ageratum Spp Asteraceae 0.5 0.13 0.28 0.88 31.47
27 Malva parviflora Malvaceae 1.25 0.13 0.69 0.88 78.67
28 Ludwigia hissopifolia Onagraceae 1.38 0.38 0.76 2.63 28.85
29 Spergula arvensis Caryophyllaceae 5.75 0.5 3.17 3.51 90.48
30 Ammania baccifera Lythraceae 0.5 0.13 0.28 0.88 31.47
31 Polygonum hydropiper Polygonaceae 1.38 0.13 0.76 0.88 86.54
32 Dopatrium junceum Scrophulariaceae 1.88 0.13 1.04 0.88 118
33 Coronopus didymus Brassicaceae 0.25 0.13 0.14 0.88 15.73
34 Other number - 9.88 0.63 5.45 4.39 124.3
35 Stellaria aquatiica Caryophyllaceae 0.5 0.13 0.28 0.88 31.47
36 Galinsoga spp Asteraceae 4.38 0.38 2.42 2.63 91.79
37 Dactyloctenium Poaceae 0.38 0.13 0.21 0.88 23.6
aegypticum
Black cumin
From surveyed black cumin fields a total of 21 weed species were identified
(Table 3). The most important families according to the number of represented
species were Chlorideae and Scrophulariaceae. The frequency of occurrence of
individual weed species ranged from 0.25% up to 5% while, the infestation level
(dominance) ranged from 0.25% up to 4.5%. The most frequent weed was
Solanum nigrum whereas the most dominant weed was Cynadon dactylon.
[171]
Table 3. Weed composition, dominance, frequency, relative density, relative frequency, summed dominant ratio in black
cumin fields, 2016 and 2018.
S/N Name of weeds Family Weeds occurrence status in black cumin*

D F RD RF SDR
2 Chenopodium album Amaranthaceae 2 0.75 5.63 5.77 97.65
3 Vicia sativa Fabaceae 1.25 0.25 3.52 1.92 183.1
4 Anagalis arvensis Primulaceae 0.25 0.25 0.7 1.92 36.62
5 Solanum nigrum Solanaceae 0.75 5 2.11 38.5 5.493
7 Medicago denticulate Fabaceae 2 0.75 5.63 5.77 97.65
9 Gnaphalium affine Asteraceae 1 0.5 2.82 3.85 73.24
11 Polygonum plebijum Polygonaceae 2.5 0.5 7.04 3.85 183.1
12 Galinsoga ciliate Asteraceae 1.5 0.25 4.23 1.92 219.7
13 Stelaria media Caryophyllaceae 3.5 0.25 9.86 1.92 512.7
14 Commelina benghalensis Commelinaceae 1.25 0.25 3.52 1.92 183.1
15 Ageratum Spp Asteraceae 1.25 0.75 3.52 5.77 61.03
16 Ludwigia hissopifolia Onagraceae 1.5 0.25 4.23 1.92 219.7
18 Ammania baccifera Lythraceae 1 0.25 2.82 1.92 146.5
19 Dopatrium junceum Scrophulariaceae 3.75 0.25 10.6 1.92 549.3
20 Other number - 2 0.25 5.63 1.92 293
21 Stellaria aquatiica Caryophyllaceae 1 0.25 2.82 1.92 146.5
Dominant weeds were those species which occurred in relatively greater number
than the other species. Most of the weeds found associated with coriander,
fenugreek and black cumin production areas are weeds that emerge with or before
the crops. Weeds that emerge later than the crops are much less competitive in
terms of crop yield loss but still may be considered problematic if they influence
crop harvest ability or reduce crop quality.
Weed density is an important factor in the control of weed species as explained by

Wicks et al. (2003). Most of common weeds identified in this survey were found
to be grouped under annual weeds. This can be explained by the fact that seeds of
annual weeds survive in unfavorable conditions and they are able to complete their
life cycle from seed to seed in one season (Singh et al., 2008). Moreover, the
weeds which appeared common in seeds spice production areas with the highest
mean field densities indicate that these weeds were more difficult to control. So,
these species should be carefully monitored.
Since weed succession and distribution patterns in seeds spice fields are dynamic
in nature, the composition of the weed flora may differ depending on location
(Uddin et al., 2010; Moody et al., 2014). The weed vegetation of a particular area
is determined not only by the environment but, also edaphic and biological factors
that include soil structure, pH, nutrients and moisture status, associated crops,
[172]
weed control measures and field history especially in local geographical variation
(Hakim et al., 2010).

Survey and identification of weeds were done in the coriander, fenugreek and
black cumin production areas of Ethiopia during 2016 and 2018 main cropping
seasons. The results of the present survey provided a quantitative comparison of
the common weed species in seeds spice production of major growing areas of
Ethiopia. The frequency of individual weed species in coriander, fenugreek and
black cumin fields ranged from 0.14% to 1%, 0.13% to 1% and 0.25% to 5%
while, the dominance value ranged from 0.14 to 49.1%, 0.25 to 26.5% and 0.25 to
4.5%, respectively.
The most frequent and dominant weed species was Chenopodium album in
coriander fields, whereas in fenugreek fields the most frequent weed was
Chenopodium album and the most dominant weed was Drymaria cordata. In
black cumin fields Cynadon dactylon and Solanum nigrum were found to be the
most dominant and the most frequent weed species, respectively. This survey has
ranked the most abundant and troublesome weed species in coriander, fenugreek
and black cumin growing areas of Ethiopia. Therefore, in the future integrated
weed management options should be developed mainly targeting those identified
major weed species.
Acknowledgment
We thank the Ethiopian Institute of Agricultural Research for financial support.
Our sincere thanks go to Debre Zeit, Kulumsa and Sinana Agricultural Research
Centers spice research team for their unreserved support during survey and data
collection.
Reference
Akobundu IO. 1987. Sugarcane. Weed Science in the Tropics: Principles and Practices (IO
Akobundu, ed.). John Wiley and Sons, New York, pp.414-416.
Appleby AP, Muller F, Carpy S. 2000. Weed Control. Ullmann's Encyclopedia of Industrial
Chemistry, New York: Wiley. pp 687–707.
Donaldson, S and Bowers G. 1998. Weed Identification and Control Guide, Educational Bulletin- 98-
01, University of Nevada Cooperative Extension.
Girma H, Digafie T and Edossa E. 2008. Spices Research achievements, revised edition, Ethiopian
Institute of Agricultural Research, pp.12: 23.
Hakim MA, Juraimi AS, Ismail MR, Hanafi, MM and Selamat A. 2010. Distribution of weed
population in the costal rice growing area of kedah in peninsular Malaysia. J. Agron, 9, pp.9-16.
Hooker JD. 1897. The Flora of British India (Vol. 7). L. Reeve.
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Javaid AR, Bajwa R, Rabbani N and Anjum T. 2007. Comparative tolerance of rice (Oryza sativa
L.) genotypes to purple nutsedge (Cyperus rotundus L.) allelopathy. Allelopathy Journal,
20(1), pp.157-166.
Moody K, Munroe CE, Lubigan RT. 2014. Major Weeds of the Philippines. Second Edition.
UPLB, Laguna. Philippines
Parker C and Fryer JD. 1975. Weed control problems causing major reductions in world food
supplies (No. 76-120049. CIMMYT.).
Pullaiah T and Chennaiah E. 1997. Flora of Andhra Pradesh (India) (Vol. 2, pp. 910-911).
Scientific Publication.
Purseglove JW, Green EG and CL Robbins SRJ. 1991. Spices.
Reddy TY and Reddi GHS. 2011. Principles of Agronomy. Kalyani Publishers, Noida, India.
P.527.
Singh A, Sharma GP and Raghubanshi AS. 2008. Dynamics of the functional groups in the weed
flora of dry land and irrigated agro ecosystems in the Gangetic plains of India. Weed biology
and management, 8(4), pp.250-259.
Thijssen R. 1991. Agro forestry and soil organic matter, about green manure, mulch,
decomposition and nutrient release. ICRAF, Nairobi. In: S. Manandhar, B.B. Shrestha and
H. D. Lekhak. 2007. Weeds of paddy field at Kirtipur, Kathmandu. Scientific World 5(5).
Uddin MK, Juraimi AS, Ismail MR and Brosnan JT. 2010. Characterizing weed populations in
different turfgrass sites throughout the Klang Valley of Western Peninsular Malaysia. Weed
Technology, 24(2), pp.173-181.
Wicks GA, Popken DH, Mahnken GW, Hanson GE and Lyon DJ. 2003. Survey of winter wheat
(Triticum aestivum) stubble fields sprayed with herbicides in 1998: weed control. Weed
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[174]
Critical Weed Competition Period on Roselle (Hibiscus
sabdariffa L.) at Wondo Genet, Southern Ethiopia
Ano Wariyo and Negasu Guteta
Wondo Genet Agricultural Research Center, EIAR, P. O. Box. 198, Shashemene, Ethiopia
Abstract
A study was carried out to determine critical weed competition period for growth,
yield and yield components of roselle under rain fed condition at Wondo Genet during
2017 to 2018 main cropping seasons. The weed competition duration had significant
influence on numbers of the crop primary branches, capsules and yields of the crop;
fresh and dry calyx, thousand seed weight and seed yield. The highest fresh calyx
yield, dry calyx yield and seed yield, 15348.60, 1389.42 and 39.03 kg/ha were
recorded from weed-free check. The highest fresh and dry calyx yield losses were
recorded in weedy-check (93.42%) and weedy for 90 DACE (82.54%). Therefore,
roselle should be kept weed free between 15 to 45 DACE to obtain higher economic
return at 5 to 10% acceptable yield loss levels.
Introduction
Roselle (Hibiscus sabdariffa L.) belongs to Malvaceae family and is one of the
common flower plants grown in tropical and subtropical regions of different parts
of the world (Morton 1987; Abu-Tarboush et al., 1997; Babatunde and Mofoke
2006). Even though the origin of the crop is unknown, it is generally believed to
be originated from India to Malaysia (Morton 1987) and Saudi Arabia (Abu-
Tarboush et al. 1997). Roselle is a versatile plant found in almost all warm
countries (Ismail et al. 2008). Particularly, African countries like Benin, Sudan,
Cote D’Ivoire, Ghana, Niger, Burkina Faso and Nigeria were reported as major
growers of the flowering plant (Oyewole, and Mera, 2010).
Calyx is one of the most economical part of the plant that is obtained by removing
the petals of the flower from its capsule containing the seeds. The calyx is high in
calcium, niacin, riboflavin, vitamin C and iron. It contains three and nine times
more vitamin C than blackcurrant (Ribes nigrum L) and citrus (Citrus sinesis L)
fruit, respectively (Ismail et al. 2008). The plant is used as valuable component of
food products; wine, jelly, beverages, jam, color and flavor ingredients (Tsai et al.
2002; Ismail et al. 2008). The use of the plant in non-food products includes the
pharmaceutical use of flower and fruits to treat cases of bronchitis and cough,
hypertension, diarrhea and many other diseases (Chewonarin et al., 1999; Haji
Faraji and Haji Tarkhani 1999).
Roselle plant is mainly used to produce healthy juice and herbal tea with full of
flavor and tart, due to its high contents of vitamin C and anthocyanins that are
[175]
found in the calyces in Ethiopia. Ethiopia has a suitable environment for the
production of roselle plant. Besides, the country has a diverse agro-ecology that
allows growing various medicinal crops both in rain fed and irrigation conditions.
It is predominantly produced by small scale farmers in their homestead garden.
Despite these potentials and economic importance, the overall yield and quality of
the crop are lower both in small scale and private farms mainly due to biotic and
abiotic factors. These limiting factors include insect pests, diseases and weeds, and
poor cultural practices and harvest conditions during the growing periods. Among
the biotic factors, weeds cause significant effects on yield of roselle (Ahmed and
Salaheldeen 2010). About 10 and 25% of the crop yield losses from weed
competition have been estimated in less developed and least developed countries,
respectively (Akobundu 1991).
To determine appropriate time and methods of weed management including the

efficient uses of herbicide for a given crop, identification of critical period is
essential in the growing period of the crops; it is helpful in determining the crop
growth stages most sensitive to weed competition (Evans et al., 2003; Otto et al.,
2009). The critical period is defined by Hall et al. (1992) and Knezevic et al.
(1994) as a number of weeks after crop emergence during which a crop must be
weed-free with the intention of preventing yield losses greater than 5%. Thus,
timing of weed control measures is important to maintain optimum crop yield.
Critical period for weed control (CPWC) is a fundamental constituent of any

management strategies in weed control options. This determining the critical
period of weed completion could help minimize yield losses due to weed
interference (Knezevic et al. 2002). However, Rajcan and Swanton (2001)
reported that competition between crop and weeds, and thus CPWC, are dependent
on-site specific factors: climatic conditions, management strategies, composition
of weed flora and weed emergence time. At the same time, CPWC tends to be
varying widely with other agricultural crops. Zimdahl (1987) reported that delayed
weeding until late stages could result in irreversible damage to the crop due to
weed competition. Similarly, Ahmed and Salaheldeen (2010) and Upadhyay et al.
(2011) reported that losses in economic yield of roselle due to weed infestation in
Sudan were ranged 45 to 90.17%.
There has not been experiment targeted to generating information on critical

period of weed control in roselle in Ethiopia. Determining the critical weed
competition period is the initial step in tailoring responses for management of the
weeds. Therefore, the objectives of this study were to determine the effect of weed
competition periods on growth and yield performance, and the critical period of
weed competition on roselle at Wondo Genet, southern Ethiopia.
[176]
Study area
The experiment was carried out at Wondo Genet Agricultural Research Center
experimental site in Southern Ethiopia during 2017 to 2018 main cropping
seasons. Wondo Genet is located at 7°19' N latitude, 38°38' E longitude and an
altitude of 1780 m.asl. (meters above sea level). It receives mean annual rainfall of
1128 mm with minimum and maximum temperatures of 11 °C and 26 °C,
respectively. The soil textural class is sandy loam with pH of 6.4 (Abayneh et al,
2006).
Experimental treatments and design

The experiment was laid out in RCBD with three replications having plot size
measuring 3.6 × 3.6 m. The crop genotype, WG-Hibiscus-Sudan was sown in June
of 2017 and 2018. Spacing both between rows and plants was 60 cm. Plants to
plant distances were maintained by thinning extra plants at early growth stage.
The experimental treatments consisted of a quantitative series of both the
increasing duration of weedy periods and the length of the weed free periods.
Timing of weed removal was based on the number of days after crop emergence.
Weeds were removed with a hand hoe from respective plots at prescribed duration
and kept weed free till the harvesting time.
To determine the beginning of CPWC, an increase in length of weed-free periods

was established by maintaining weed-free condition for 15, 30, 45, 60, 75 and 90
days after crop emergence, referred as weed- free plots before allowing
subsequent emergence of weeds to compete with the crop for the remaining
growing season. While to evaluate the end of the CPWC, an increase in duration
of weed interference was established by allowing the weeds to compete with the
roselle for 15, 30, 45, 60, 75 and 90 days after crop emergence, referred as weedy
plots after which the plots were maintained weed-free until the harvesting time. In
addition, weedy check and weed-free check were included as negative and
positive control, respectively.

Weed flora and density: Data on weed types and density were collected by
randomly and repeatedly throwing sampling quadrat (0.25 m×0.25 m size). The
data were taken at four spots in each plot at the time of weed removal for early
competition and at 10 days interval before the expected harvest time in the case of
late competition to avoid possible foliage and seed shading.
Weed aboveground dry biomass (g): The weeds found within the quadrat were cut
near the soil surface and kept separately in paper bags. The samples were sun-
[177]
dried for 3-4 days and thereafter placed into an oven at 65 °C temperature and the
dry weight was taken using sensitive balance.
Crop data: Data on height, number of branches and capsules, weight of fresh and
dry calyx, weight of seed yield and number of seeds were recorded by randomly
selecting four plants from each plot. Thousand seed weight data were taken using
an electric balance.
Data collected on growth and yield parameters of the crop were analyzed using
Fisher’s analysis of variance technique. Least significant difference (LSD) test at
0.05 probability level was mployed to compare the treatment means (Steel et al.,
1997).
On the other hand, to analyze the critical period of weed control, the relative roselle
yields (Y) of each treatment was computed as follow:
Y = 𝐻. 𝑠𝑎𝑏𝑑𝑎𝑟𝑖𝑓𝑓𝑎 yield in treatment /𝐻. 𝑠𝑎𝑏𝑑𝑎𝑟𝑖𝑓𝑓𝑎 yield in weed free check
x 100
Determination of yield loss of roselle to the maximum level due to weed competition
wwas premeditated using the following formula:
Yield losses (%) = [1- ] x 100

Weed species
During the two years of experimentation, 24 weed species belonging to 10
families were identified occurring in roselle growing plots of which broadleaf
weed species recorded greatest number than grass and sedge weeds (Table 1).
Moreover, annual weeds were the most common group than perennial weeds in
the crop field. The findings were in line with Adjun (2003), Mahadevan et al.
(2009) and Ahmed and Salaheldeen (2010) who reported that the weed species
associated with roselle were composed of a wide range of annual, biennial and
perennial broad leaved, grass and sedge weeds.
Weed density (plants m-2)

The present result revealed that total weed density increased significantly with an
increase in competition duration and the increase was significant at each
increasing step in the different competition durations. The average maximum
(211) and minimum (19) total weed density were obtained in plots with season
long weed-infestation (weedy) check and weed free check, respectively. The
weedy check showed the maximum weed density because there was longer period
available for weeds to continue germinating throughout the growing period. This
[178]
result was agreed with earlier report of Tunio et al. (2004). The weed free plots
showed minimum density because weeds were considerably reduced by repeated
hand hoeing (Table 2). In these plots the higher number of weed density was
recorded in year the two (110) as compare to the year one (88) of the experimental
period.
Table 1. List and biological characteristics of weed species recorded in roselle field, 2017 and 2018.
S/N Scientific name Family name Category Life cycle

1 Bidens pilosa L. Asteraceae Broad leaf Annual
2 Commelina benghalensis L. Commelinaceae Grass Perennial
3 Commelina latifolia L. Commelinaceae Grass Perennial
4 Guizotia scabra L. Asteraceae Broad leaf Annual
5 Ageratum conyzoides L. Asteraceae Broad leaf Annual
6 Amaranthus hybridus L. Amaranthaceae Broad leaf Annual
7 Amaranthus spinosus L. Amaranthaceae Broad leaf Annual
8 Galinsoga parviflora L. Asteraceae Broad leaf Annual
9 Plantago lanceolate L. Plantaginaceae Broad leaf Perennial
10 Cyperus esculentus L. Cyperaceae Sedge Perennial
11 Datura stramonium L. Solanaceae Broad leaf Annual
12 Xanthium strumarium L. Asteraceae Broad leaf Annual
13 Xanthium spinosum L. Asteraceae Broad leaf Annual
14 Solanum anguivi L. Solanaceae Broad leaf Perennial
15 Parthenium hysterophorus L. Asteraceae Broad leaf Annual
16 Biden pachyloma L. Asteraceae Broad leaf Annual
17 Phalaris paradox L. Poaceae Grass Annual
18 Digitaria ternate L. Poaceae Grass Annual
19 Oxalis corniculate L. Oxalidaceae Broad leaf Annual
20 Tribulus terrestris L. Zygophyllaceae Broad leaf Annual
21 Portulaca olerace L. Portulacaceae Broad leaved Annual
22 Nicandra physalodes L. Solanaceae Broad leaf Annual
23 Rottboallia cochinchinesis L. Poaceae Grass Annual
24 Anagallis arvensis L. Primulaceae Broadleaf Annual
Weed dry weight (g m-2)

The dry weight of the weed species was increased with an increase in competition
period and vice versa, being maximum dry weight (116.87) recorded by weedy
check and the minimum (7.49) by weed free check (Table 2). The recorded weeds
dry weight and weed density amounts were found directly and positively
proportional to the increasing weedy period (Table 2) that the longer growth
period resulted more photosynthates accumulation and greater biomass. In
agreement with this result, Stagnari and Pisante (2011) and Smitchger et al. (2012)
reported that the weight of dry weeds and weed density were inversely
proportional to the increase in weed removal periods. Similarly, but in the other
way Akhtar et al. (2000) found that by increasing weed crop competition duration
one increased weed biomass.
[179]
50% flowering date
The results of the experiment indicated that days to 50% flowering were increased
significantly with an increase in competition duration and the increase was
significant at each step of the increasing competition durations (Table 2).
Table 2. Effect of weeding times on weed biomass and density, and on 50% flowering date of roselle, 2017 and 2018
Treatment WDB (gm) WD 50% FD
IDWFP at DACE
15 65.23d 130.59c 144.00e
30 42.22e 86.54e 133.83f
45 30.15f 81.58e 128.67fg
60 13.79ghi 46.75g 125.00gh
75 17.99gh 45.26g 124.33gh
90 10.08hi 33.98g 119.17h
WC 116.87 a 211.19 a 157.50a
IDWP at DACE
15 14.08ghi 63.77f 149.67bcde
30 20.09g 112.67d 146.17de
45 38.04 ef 129.95 c 147.67cde
60 69.87d 134.76c 154.83ab
75 88.49c 176.54b 154.00abc
90 102.62b 179.99b 152.33abcd
WFC 7.49i 18.73 h 130.33fg
lsd0.05 8.18 13.61 6.48
CV (%) 15.54 11.33 3.98
IDWFP= Increasing duration of weed free period, IDWP= Increasing duration of weedy period, DACE= Days after
crop emergence, WC= Weedy-check, WFC= Weed free check, WDB= Weight of dry biomass, WD= Weed
density, FD = Flowering date
The longest days of 50% flowering (157.50) was recorded from weedy-check that
statistically at par with weedy competition duration for 60 DACE but significantly
varied from other treatments while the shortest days (119.17) was from weed-free
for 90 DACE. This might be due to the severe competition of the weeds with the
crop for the limited environmental resources when weeds were allowed to
germinate and grow for prolonged periods. The competition in turn may aggravate
the stress for the plant which compels the crop towards physiological response for
stressful environment that enables the crop to mature earlier to escape the stressful
conditions before drying. This is in line with the observation of Desclaux and
Roumet (1996) who reported that limited water supply triggers a signal to cause
an early switching of plant development from the vegetative to reproductive phase.
Roselle growth and yield parameters

Plant height
The present study revealed that there was no significant difference observed
among tested treatments on roselle height (Table 3).
Number of primary branches per plant

There was a significant effect of weed-crop competition duration on number of the
[180]
roselle primary branches per plant. A gradual and progressive decrease in number
of primary branches per plant was recorded with the increasing competition
duration. The maximum number (29.50) of primary branches per plant was
recorded by weed-free check followed by weed free competition durations for 90,
75 and 60 DACE, and significantly different from the rest of the treatments. This
might happen due to the available less time during the competition for resources
between the crop and weed plants in such the short competition duration. The
result are in accordance with the findings of Almarie (2017) who obtained that the
increase of soybean branches continued significantly between the weed removal
time and then after no significant increase was obtained. Similarly, Singh et al.
(2015) also described that number of primary branches per plant in field pea
increased when the weed-free days were prolonged.
Number of capsules per plant

The result revealed that there is a significant effect of weed-roselle competition
durations on the number of the crop plant capsules per plant. Weed-free check,
weed free for 90 and 75 DACE were produced maximum number of capsules per
plant respectively, 186.50, 184.50 and 177.33, (Table 3).
Absence of weeds in weed-free check might have been enabled the crop to make
best use of growth resources that resulted in a greater number of capsules per
plant. In conformity with Mengesha et al. (2013) report that in common bean the
number of pods per plant significantly increased with an increase in duration of
weed-free period and decreased with increase in durations of weedy period.
Table 3. Effect of weed competition and control time on yield components of roselle plant, 2017 and 2018
Treatment PH (cm) NPB NCPP WFCYPP WFCY WFC RYL

(gm) (kg/ha) (%)
IDWFP at DACE
15 131.83abc 9.67fg 39.50de 188.58d 5238.20d 65.87
30 128.83abc 12.33e 51.00d 237.29c 6591.30c 57.06
45 133.5abc 17.83c 81.17c 327.84b 9106.70b 40.67
60 122.00bc 19.83b 123.00b 521.08a 14474.40a 5.70
75 122.83bc 26.67b 177.33a 530.47a 14735.30a 4.00
90 120.50c 26.50b 184.50a 546.09a 15169.20a 1.17
WC 145a 6.00h 10.00g 36.75f 1009.70f 93.42
IDWP at DACE
15 133.83abc 10.00ef 42.67d 246.94c 6859.40c 55.31
30 141.5ab 15.00d 52.17d 250.63c 6961.90c 54.64
45 131.67abc 9.33fg 27.67ef 134.53e 3737.00e 75.65
60 137.17abc 9.00fg 25.50f 142.64e 3962.30e 74.18
75 127.33abc 7.50fgh 19.33fg 118.04e 3278.90e 78.64
90 141.33ab 7.17gh 16.00fg 115.96e 3220.90e 79.02
WFC 124.83bc 29.50a 186.50a 552.55a 15348.60a 0.00
lsd0.05 19.74 2.66 13.71 35.34 981.62
CV% 12.96 15.63 16.00 10.82 10.82
PH= Plant height, NPB= Number of primary branches, NCPP= Number of capsules per plant, WFCYPP= Weight of fresh calyx yield per
plant, WFCY (kg/ha) = Weight of fresh calyx yield per hectare, WFC YRL (%) = Weight of fresh calyx relative yield loss
[181]
Yield of fresh and dry calyx
The weight of fresh and dry calyx yield per plant was significantly influenced by
the duration of weed competition. The maximum fresh calyx yield per plant
(552.55 g m-1) was found in weed-free check. This was statistically at par with
competition duration of 90, 75 and 60 DACE and significantly different from the
rest of the treatments (Table 4). Similarly, the maximum dry calyx yield per plant
(50.02 g m-1) was recorded in weed-free check followed by weed-free for 90
(47.76 g m-1) and 75 (46.51 g m-1) DACE (Table 4). Likewise, the highest total
dry calyx yield/ha was also recorded from weed-free check (1389.42 kg/ha)
followed by weed free for 90 (1326.76 Kg/ha) and 75 (1291.96 Kg/ha) DACE
(Table 4). In general, an increased duration of the weed-free periods attributed to
an increase in fresh and dry calyx yield per area. This result is in line with Singh et
al. (2015) report that the yield attributes were highest in season-long weed-free
period and at par with weed-free for initial 40 days or plots kept weedy only for
initial 20 days.
Thousand seed and yield weight

The study revealed that thousand seed weight of roselle was significantly
influenced by the duration of weed competition. The maximum thousand seed
weight (28.14g) was recorded in weed free check followed by treatments kept
weed-free beyond 75 DACE while the minimum (6.12g) was in weedy-check. It
was found that 1000 seed weight of roselle was inversely related to the increase in
the duration of weedy period and directly proportional to the increase in duration
of weed free periods. The accumulation of adequate dry matter by the crop
through the proper utilization of available above and belowground growth
resources at absence or reduced weed competition help the highest harvestings.
Similarly, Singh et al. (2015) stated that yield attributes, including 1000 seed
weight of field pea increased with an increase in weed-free duration and decrease
in weedy periods.
[182]
Table 4. Effect of weed competition and control time on yields of roselle, 2017 and 2018.
Treatments WDCYPP WDCYPH WDC RYL TSW WSYPH

(gm) (Kg) (%) (Kg)
IDWFP at DACE
15 12.87ef 357.36ef 74.28 18.74cd 11.44ef
30 18.41d 511.36d 63.20 20.85c 12.88e
45 31.82c 884.03c 36.37 23.72b 19.79c
60 45.36b 1259.81b 9.33 24.57b 35.61b
75 46.51ab 1291.96ab 7.01 25.04b 35.87b
90 47.76ab 1326.76ab 4.51 25.52b 37.63ab
WC 9.84efg 273.37efg 80.32 6.12g 4.76h
IDWP at DACE
15 12.93e 359.03e 74.16 16.93d 10.45efg
30 13.61e 378.10e 72.79 18.07d 16.27d
45 12.05efg 334.65efg 75.91 14.37e 11.07ef
60 10.93efg 303.57efg 78.15 12.86e 7.81g
75 8.73g 242.57g 82.54 10.08f 8.05g
90 8.91f 247.38fg 82.20 9.73f 9.50fg
WFC 50.02a 1389.42a 0.00 28.14a 39.03a
lsd0.05 4.00 111.19 2.45 2.74
CV% 14.68 14.68 11.62 12.72
WDCYPP= Weight of dry calyx yield per plant, WDCYPH= Weight of dry calyx yield per hectare,
TSW= Thousand seed weight, WSYPH= Weight of seed yield per hectare, RYL (%) = Relative yield loss
Yield losses
The losses that showed due to each of the different weed competition period were
considered relative to the yield of weed-free check for each treatment. The data in
losses indicated that the fresh calyx yield per hectare was higher in beyond 45
weedy days after emergence including weedy-check as compared to the WFC. The
fresh calyx yield per hectare losses ranged from 54.64 to 93.42% along increasing
duration of weedy periods while from 0 to 65.87% with the increasing duration of
weed-free periods (Table 3). Likewise, dry calyx yield losses were higher in
weedy for 75 and 90 DACE followed by WC as compared to the WFC. In this
case the dry calyx yield per hectare losses ranged from 72.79–82.54% along
increasing duration of weedy periods while with the increasing duration of weed
free periods ranged from 0 to 74.28 (Table 4). Thus, the losses come through
weed-crop competitions regarding the nearby resource utilization in the growing
period. Prolonged crop-weed competition resulted in reduced dry biomass
accumulation which ultimately rendered the yields of parameters considered and
the correspondingly higher yield losses.
Critical period of weed control

The critical period of weed control for roselle was estimated based on the relative
yields at 5% to 10% acceptable yield loss. The beginning of the critical periods of
weed competition was obtained from the late weed-crop competition (from the
increasing duration of weedy periods) while the end of the critical periods of weed
[183]
control was obtained from the early crop weed competition (from the increasing
duration of weed-free periods). Based on the current result, the roselle should kept
weed-free from 15 to 45 DACE. Thus, the weeds have to be managed during these
periods through appropriate methods to prevent more than 10% yield loss of the
crop. This finding had followed previous studies. Le Bourgeois and Marnotte
(2002) located the critical period between 30 and 90 DAS for long-cycle crops
(yams, cassava, sugarcane, etc.). Another finding by Zuhal et al. (2010) reported
that the critical period of weed control in faba bean started at 30 days and ends 45
days after crop emergence at 10% acceptable yield loss. However, according to
Knezevic et al. (2002), critical periods of weed control varied with weed species
composition, weed density and intensity, climatic conditions, frequency of tillage
operation and soil type of the area that need to considered at final
recommendation.

There is an overall sensitivity of roselle to infestation by weeds, which
demonstrates the needs for weed control in the crop field. The highest weed
density and biomass at harvest actually associated with the lowest values of seed
yield and yield components. From this study, it can be concluded that to obtain
better yield which is more than 90% of the potential yield, roselle has to be weed-
free for periods between 15 to 45 days after emergence of the crop as it is found to
be the critical period of weed-roselle competition at Wondo Genet Agricultural
Research Center experimental site and other similar agro-ecology.
Acknowledgement
The authors are grateful to Wondo Genet Agricultural Research Center and
Ethiopian Institute of Agricultural Research for providing the research fund and
vehicles. Our gratitude also extends to field assistants at the research center for
their support during the whole study period.
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Critical Weed Competition Period on Basil
(Ocimum basilicum L.) at Wondo Genet and
Alage, Southern Ethiopia
Wondo Genet Agricultural Research Center, EIAR, P.O.BOX. 198, Shashemene, Ethiopia
Abstract
A study was carried out at Wondo Genet Agricultural Research Center experimental
site under rain fed condition and at Alage site under supplementary irrigation to
determine critical weed competition period for growth, yield and yield components of
basil (Ocimum basilicum L.) during 2017 and 2018. At both locations, the weed
competition and weeding time had significant influence on per plant height, number
of primary branches, fresh stem weight yield, fresh leaves weight yield, fresh flower,
and also per hectare products of fresh leaves, fresh flower, yield and essential oil
content of the crop. The highest fresh and dry herbage (leaves and inflorescences)
yield (kg/ha) were recorded from weed-free for 60 DACE followed by weed-free
check at both locations. Similarly, the maximum essential oil content and yield were
obtained from weed-free check followed by weed-free for 60 DACE. A yield loss of
basil was estimated based on the fresh and dried herbage, and the essential oil yield
per hectare. Correspondingly, the highest fresh and dried herbage yield (kg/ha)
losses were recorded from weedy-check while the lowest from weed-free check.
However, the highest essential oil yield loss was recorded from weedy-check
(95.02%) at Wondo Genet but from weedy for 60 DACE (98.35%) at Alage. Thus, to
gain higher economic return from basil plant and reduce the yield losses due to weed
competition to less than 10%, weeds must be controlled in between 10 to 30 DACE at
both Wondo Genet and Alage, and similar agroecologies.
Introduction
Basil (Ocimum basilicum L.) is an important aromatic and medicinal plant
belonging to Lamiaceae family (Tucker et al., 2000; May et al., 2008; Sullivan,
2009). The crop plant is usually referred as the “King of the herbs”, being widely
utilized due to its economic, culinary, industrial and medicinal importance.
Fresh or dried leaves can be used in soups, stews, sauces, meat, fish, and egg
dishes, and in salads and vegetables. The leaves are used in flavored vinegar, teas
and steeped in oil to produce flavored oils. Cinnamon and lemon basil add a final
touch to special dishes with their delicate overtone flavors. They also used for
making jellies, jams, vinegars, and sorbets. Its blends with tomatoes used to
season pizza, spaghetti sauce, sausage, soup, tomato juice, dressings, salads and
pesto. While basil is included in many peppery main dishes, it also is used in fresh
fruit desserts (Plasters, 2006).
[187]
Moreover, it has been used as a folk remedy for an enormous number of ailments
including boredom, cancer, convulsion, deafness, diarrhea, epilepsy, gout, hiccup,
impotency, insanity, nausea, sore throat, toothaches and whooping cough, insect
repellent, anti-viral, anti-microbial, antioxidant, and anti-cancer properties of the
oils (Sullivan, 2009). In addition, the extract of the herb used in preventing
cardiovascular diseases through improved diet and several antioxidant compounds
display a high antioxidant power (Erum et al., 2011).
There are many factors that influence agronomic characters, biomass and essential
oil yield of medicinal and aromatic plants. Among these, weed competition is a
major constraint. As with all commonly grown crops, also in medicinal and
aromatic plants, weeds function as crop competitors, create problems for
mechanized harvest, and may alter the end quality when mixed with the harvested
product.
The well-known interference of weeds takes additional relevance for medicinal

and aromatic plants for several reasons. Firstly, the synthesis of secondary
metabolites in plants is linked to many genetic and environmental factors
(Sangwan et al. 2001). Buyers often grade such plants according to their specific
quality features, which are primarily determined by their content in essential oils
or other secondary metabolites, which on their turn can be reduced in presence of
weeds (Carrubba and Catalano, 2009). Although sparse specific experiments have
been conducted, the presence of weeds is therefore believed to exert a significant
effect on plant metabolic pathways, in this way acting negatively on the market
end value of such crops (Gil et al. 1998). Some experiments carried out on
suitability of medicinal and aromatic plants to field conditions have confirmed the
importance of weed competition; de la Fuente et al. (2003) demonstrated that on
coriander, especially under poor soil conditions, weeding had a greater effect than
did N fertilization.
In Ethiopia, basil is commonly grown plant in most regions as natural plant and
cultivated crop at homestead areas. Yimer (2010) reported that Ethiopia has
exported 68,786 kg of basil essential oil to Sudan and USA from which a total
foreign currency of $ 54,991.20 and 746.00, respectively was obtained in 2009.
The export volume accounted 19.77% in 2006-2009 and exhibit 0.15 and 0.14%
both volume and value share of the total spice export. However, with such export
demand the productivity of basil is much below its potential. The limiting factors
could be lack of adaptable, high yielder and pest resistant variety; and presence of
inadequate agronomic practices for crop production and systematic oil analyzing
laboratory. Among these, weed competition and lack of appropriate weeding time
were found as critical problem limiting basil production and productivity in the
country.
[188]
The concept of critical periods of weed competition, during which weeds have the
greatest impact on crop growth, was verified by Nieto et al. (1968). It is a specific
minimum period of time during which the crop must be free of weeds in order to
prevent loss in yield and represents the overlap of two separate components
(Weaver and Tan, 1983). The first component is the length of time weeds can
remain in a crop before interference begins. The second component is the length
of time that weed emergence must be prevented so that subsequent weed growth
does not reduce crop yield.
The critical period is the prime period most suitable for conducting weeding
operations considering the following factors: the environment (climate and soil),
the period of weed infestation in the field, the weed species, the cultural practices
including crop rotation, fertilization, density and methods of seeding (broadcast,
hill seeding or transplanting), and the relative growth rates of the crop and its
associated weeds. For example, according to Le Bourgeois and Marnotte (2002),
the critical period is generally located between 15 and 60 days after seeding
(DAS) for short-cycle annual crops (cotton, corn, sorghum, rice, etc.) and between
30 and 90 DAS for long-cycle crops (yams, cassava, sugarcane, etc.). Critical
period of competition varies from crop to crop depending on weed emergence
time, weed type, weed density and management practices.
There is no a crucial period so far established during which weed infestation is

particularly harmful to basil plant and at what growth stage weeding practice can
become more effective in Ethiopia. Thus, to reduce basil yield loss resulting from
weed competition and improve the efficacy of the producers’ weed management
practices an investigation found crucial. The objectives of these studies were
therefore, to determine the critical period for weed competition and control time in
basil both under rain fed and supplementary irrigation conditions.

Study areas
experimental site and Alage during the year 2017 to 2018 cropping seasons.
Wondo Genet is located at 7° 192′ N latitude and 38° 382′ E longitudes with an
altitude of 1780 m above sea level. The site receives a mean annual rainfall of
1000 mm with minimum and maximum temperatures of 10 and 30 °C,
respectively. The soil textural class is clay loam with an average pH of 6.4
(Beemnet et al., 2010). Alage is located at longitude of 38o 30’East and a latitude
of 7o30’ North, with an altitude of 1600 masl. The site is characterized by mild
tropical weather with a minimum and maximum temperatures ranging from 11 0c
to 29 0c and experiences a bimodal rainfall distribution with an annual average of
[189]
800 mm (NMSA, 2010).

The experiment was laid out in RCBD with three replications having net plot size
measuring 2.4 m × 3.6 m. The genotype basil-05 was sown in June month of the
years 2017 and 2018 with 60 cm and 40 cm space between rows and plants,
respectively. The plant to plant distance was maintained by thinning extra plants at
early growth stage. The experimental treatments consisted of a quantitative series
of both the increasing duration of weed interference and the length of the weed
free period. Timing of weed removal was based on the number of days after crop
emergence. Weeds were removed with a hand hoe from respective plots at
prescribed duration and kept weed free till harvest.
To determine the beginning of the CPWC, increasing length of weed-free period

was established by maintaining weed-free condition for 10, 20, 30, 40, 50 and 60
days after crop emergence, referred to as weed- free plots before allowing
subsequent emerging weeds to compete for the remainder of the growing season.
To evaluate the end of the critical period of the CPWC, the second component,
increasing duration of weed interference, was established by allowing the weeds to
compete with the basil for 10, 20, 30, 40, 50 and 60 days after crop emergence,
referred to as weedy plots after which, plots were maintained weed-free until
harvest. In addition, season long weedy and weed-free check were included as
control.

Weed flora and density: In the experimental fields, data on weed flora type and
density were recorded randomly thrown sample quadrat (0.25 m × 0.25 m) at four
places in each plot. The weed density data were taken at the time of weed removal
for early competition and about 10 days before the expected harvest time in the
case of late competition to avoid possible foliage and seed shading. The weed
population found within the sample quadrat were converted and expressed as
number m-2.
Weed aboveground dry biomass (g): The weeds encountered within the quadrat
were cut near the soil surface immediately after recording data on weed count and
placed separately into paper bags treatment wise. The samples were sun-dried for
3-4 days and thereafter were placed into an oven at 65 °C temperature till a
constant weight and subsequently, their dry weight (g m-2) was taken. Selecting
four plants at random data were record on per plant height, number of primary
branches, fresh stem yield, fresh and dry herbage yield, and also per hectare
products of fresh and dry her bage yield, yield and essential oil content. Data
collected on growth and yield parameters of the crop were analyzed using Fisher’s
analysis of variance technique. Least significant difference (LSD) test at 0.05
probability levels was employed to compare the treatment means (Steel et al.,
[190]
1997). On the other hand, to analyze the critical period of weed control, the
relative basil yields (Y) of each treatment were computed as followed፡
x 100
Wondo Genet experimental site:

Weed species
There were 23 annual and perennial weed species belonging to 7 families of which
more number grouped under broadleaf type than grass and sedge weeds (Table 1).
The annual weeds were found the most common and frequently occurring weed
group during the two years of study. The result was in line with Adjun (2003),
Mahadevan et al. (2009) and Ahmed and Salaheldeen (2010) who reported that
the weed species were composed of a wide range of annual, biennial and
perennial, and broad leaved, grass and sedge weed types.
Table 1. Category of weed species occurring in basil field, 2017 and 2018.
S/N Scientific name Family name Leaf type Life cycle
1 Bidens Pilosa L. Asteraceae Broad leaf Annual
11 Cyperus rotundus L. Cyperaceae Sedge Perennial
14 Phalaris paradox L. Poaceae Grass Annual
17 Portulaca Olerace L. Portulacaceae Broad leaf Annual
20 Annagallis arvansis L. Primulaceae Grass Perennial
21 Cynodon dactylon L. Poaceae Grass Annual
22 Solanum nigrum L. Solanaceae Broad leaved Annual
23 Viala riviniana L. Violaceae Broad leaved Perennial
[191]
Weed Density (m-2)
Total weed density increased significantly with an increase in competition
duration and the increment was significant at each the competition duration. The
maximum (147.00) and minimum (0) total weed densities were obtained from
plots with season long weed-infestation (weedy check) and season long weed free
(weed free check) respectively (Table 2). Weedy check was showed the maximum
weed density that longer period available for weeds to germinate and continue to
grow throughout the study period. This result well agreed with Tunio et al. (2004)
who reported that weed free plots showed minimum density because weeds
eradicated by repeated hand hoeing.
Weed Dry Weight(gm-2)

The aboveground weed biomass dry weight was increased with an increase in
competition period (Table 2). The maximum biomass dry weight (125.67) in
weedy check could be due to higher weed density and longer growth period
resulting in more accumulation of photosynthates and greater biomass. Similarly,
Akhtar et al. (2000) found that by increasing weed crop competition duration
increased weed biomass. The result further can be explained that the weeds
emerging at the later growth stages offer less competition to crops as it
accumulates lower weed biomass (Uremis et al., 2009).
Table 2. The effect of competition duration and weeding time on weed biomass and total density, 2017 and 2018.
Treatment WDB (gm-2) WD

IDWFP at DACE
10 89.14b 60.67de
20 18.94e 47.67ef
30 36.84d 45.00ef
40 17.17e 30.50f
50 11.96ef 29.33f
60 0.00f 0.00g
WC 125.67a 105.50c
IDWP at DACE
10 11.23ef 80.67d
20 14.56e 106.67bc
30 32.80d 102.17c
40 67.03d 109.17bc
50 37.27d 126.50ab
60 58.03bc 147.00a
WFC 0.00f 0.00g
lsd0.05 12.28 20.84
CV% 20.80 17.56
DACE=Days after crop emergence
[192]
Growth and yield parameters
Plant height
Basil plant height was significantly influenced by weed competition period. An
increased length of weed interference up to 30 DACE was reduced significantly
the plant height. The tallest plants were found in weed-free check (47.13cm)
treatment. The results agreed to the previous research results, where the taller rice
plants were found in all weed-free treatments (Begum et al., 2008). The height of
plants is often associated with their interspecific competitive ability (Freckleton
and Watkinson 2001).

The result revealed that significant difference was observed on number of primary
branches per basil plant at different weed competition periods (Table 3). Gradual
and progressive decreases in number of primary branches per plant were recorded
with increasing competition duration. The highest number of primary branches per
plant (21.21) was found in weed-free check. This was statistically at par with
competition duration of 50 and 60 DACE but significantly different from the rest
treatments. The higher number of primary branches per plant from short
competition durations treatments could be due to the less time availability for
weeds competition with the crop plant for growth resources. The less time
availability for weeds competition enable crop plants to achieve good vegetative
growth rate and maximum assimilates. Accordingly, Almarie (2017), obtained that
an increase of soybean branches continued significantly between weed removal
and then after no significant increase was obtained. Similarly, Singh et al. (2015)
also described that number of primary branches per plant in field pea increased
when the weed-free days were prolonged.
Fresh stem, fresh and dry herbage yield

The present study was indicated the significant effect of weed-crop competition
durations on fresh stem, and fresh and dry herbage yield. Weed-free check and
competition free for early 60 DACE were produced maximum fresh stem weight
per plant (163.02 g m-1) and (159.38 g m-1), respectively (Table 3).
[193]
Table 3. The mean effect of weed competition and control time on basil yield components, 2017 and 2018.
TFHY TDHY
Treatment PH NPB FSWPP FHPP DHPP (kg/ha) (kg/ha)
IDWFP at DACE
10 39.21bcd 9.75efg 57.32ef 27.78de 14.19e 1157.60de 591.43e
20 39.00bcd 13.13d 103.10d 33.10d 18.64d 1379.30d 776.56d
30 40.92bcd 16.64c 129.17c 42.57c 21.93c 1773.60c 913.59c
40 41.88a-d 18.60bc 135.51bc 51.74b 25.09b 2155.70b 1045.28b
50 42.38abc 19.67ab 143.12b 67.41a 28.15a 2808.70a 1172.79a
60 43.13ab 20.03ab 159.38a 67.51a 28.36a 2813.00a 1181.49a
WC 32.54e 6.20h 20.80h 11.73g 7.87h 488.60g 328.00h
IDWP at DACE
10 36.63de 11.25de 65.09e 27.82de 17.05d 1158.90de 710.50d
20 37.17cde 10.88ef 47.19f 20.86ef 14.25e 869.10ef 593.74e
30 38.42bcd 9.02fg 32.98g 19.11f 13.27ef 796.10f 552.86ef
40 39.08bcd 8.91fg 28.14gh 17.93fg 12.44efg 747.00fg 518.32efg
50 40.79bcd 8.18gh 28.05gh 17.84fg 11.87fg 743.30fg 494.75fg
60 38.75bcd 6.37h 25.95gh 13.77fg 10.66g 573.70fg 444.14g
WFC 47.13a 21.21a 163.02a 67.90a 28.63a 2829.10a 1193.06a
Lsd0.05 5.65 2.02 11.15 7.25 2.07 302.15 86.02
CV (%) 12.27 13.58 11.84 18.01 9.90 18.01 9.90
PH = Plant height, NPBPP = Number of primary branches per plant, FSWPP= Fresh stem weight per plant, FHYPP =
Fresh herbage yield per plant, DHYPP= Dried herbage yield per plant
Correspondingly, Highest fresh (67.90 g m-1) and dry herbage (28.63 g m-1) yield
per plant and also yield (kg/ha) were recorded from weed-free check. These were
statistically at par with competition free duration for initial 60 and 50 DACE. At
all, increasing the weed free periods was enabled to harvest higher weight for each
crop yield components due to the relatively better control of weeds and favorable
growing conditions to accumulate more amount of the crop dry biomass.
Similarly, Singh et al., (2015) reported that the yield attributes were highest in
season-long weed-free period and at par with weed-free for initial 40 days or plots
kept weedy only for initial 20 days.
Essential oil content and yield

Interaction effects of accessions and location exerted a significant influence on
days to second flowering. Essential oil content and yield of basil were
significantly influenced by the duration of weed competitions (Table 4).
Maximum essential oil content (1.05 v/w) and yield (12.66 kg/ha) were obtained
from weed-free check. It was at par with the treatments kept weed-free for early
60 and 50 DACE. An increasing of duration of the weed-free period might
allowed accumulation of adequate dry matter by basil plants through utilization of
available aboveground and belowground growth resources promoting of the crop
essential oil content and yield. In general, essential oil content and yield of basil
was inversely related to the increasing duration of weedy period and directly
proportional to the increase in weed free periods. In conformity with this result,
[194]
Welsh et al. (1999) and Ahmad and Shaikh (2003) found that wheat yield
decreased as the weed infested duration increased. Zuhal et al. (2010) also
reported that the yield of faba bean significantly varied when weeds were allowed
to grow for different durations and about 46% yield loss was recorded from weedy
check plot.
Yield losses
Yield losses due to the different weed competition periods were calculated as
compared to the yield from weed-free check treatment. The results indicated that
the fresh and dry herbage and essential oil yield were become higher with the
increased duration of weedy periods than the increased duration of weed-free
periods (Table 4).
Table 4. The effect of weed-basil competition and controlling time on the crop yield components
Treatment EOC EOY YL% YL% YL%

(v/w) (kg/ha) TFHY TDHY EOY
IDWFP at DACE
10 0.41e 2.38ef 59.08 50.43 81.20
20 0.60d 4.65d 51.25 34.91 63.27
30 0.76c 6.95c 37.31 23.42 45.10
40 0.90b 9.43b 23.80 12.39 25.51
50 0.96ab 11.30a 0.72 1.70 10.74
60 1.01ab 12.06a 0.57 0.97 4.74
WC 0.19g 0.63g 82.73 72.51 95.02
IDWP at DACE
10 0.53d 3.81de 59.04 40.45 69.91
20 0.40e 2.36ef 69.28 50.23 81.36
30 0.33ef 1.79fg 71.86 53.66 85.86
40 0.24fg 1.27fg 73.60 56.56 89.97
50 0.22fg 1.19fg 73.73 58.53 90.60
60 0.24fg 1.11fg 79.72 62.77 91.23
WFC 1.05a 12.66a 0.00 0.00 0.00
Lsd0.05 0.12 1.48
CV (%) 18.87 25.07
The fresh herbage yield losses ranged from (59.04– 82.73%) for the increased
duration of weedy periods while (0 – 59.08%) for increased duration of weed-free
periods. Likewise, the dry herbage yield losses ranged from (40.45– 72.51%) for
the increased duration of weedy periods while for the increased duration of weed
free periods it ranged from (0- 50.43%). For the essential oil yield, the maximum
loss was found in weedy-check (95.02%) while the minimum was in weed-free
check (0%). The crop product loss due to the weed-crop competitions for growth
resources takes the highest share in the early growing period. Prolonged crop-
weed competition was resulted the reduced dry biomass accumulation which
ultimately rendered the yields parameters considered and associated yield losses.
[195]
The critical period of weed control for basil was estimated based on the relative
yields at 5% to 10% acceptable yield loss. The beginning of the critical periods of
weed competition was obtained from the late weed-crop competition, from the
increasing duration of weedy periods. While the end of the critical periods of weed
control was obtained from the early crop weed competition, from the increasing
duration of weed-free periods. Based on the current result, the critical period of
weed control in basil found to be from 10 to 30 DACE. Thus, the weeds must be
managed in these time range using appropriate methods to prevent more than 10%
yield loss by the crop. According to Knezevic et al. (2002), critical periods of
weed control varied with weed species composition, weed emergence pattern,
weed density and intensity, ecological variations, climatic conditions, frequency of
tillage operation and soil type of the area. However, Le Bourgeois and Marnotte
(yams, cassava, sugarcane, etc.). This was in line with the finding of Zuhal et al.
(2010) who reported that the critical period of weed control in faba bean started at
30 days and ends 45 days after crop emergence at 10% acceptable yield loss.
Alage site:
Weed species
A total of 12 annual and perennial weed species belonging to 7 families were
recorded in basil field (Table 1). Out of which the broadleaf species number were
found more than grass and sedge weeds. In addition, the annual weeds were found
to be the most common and frequent group. The finding was in line with Adjun
(2003), Mahadevan et al. (2009) and Ahmed and Salaheldeen (2010) who reported
that the weed species found were composed of a wide range of annual, biennial
and perennial, and also broad leaved, grass and sedge weed types.
Table 1. Category of weed species found in basil field, 2017 and 2018.
Scientific name Family name Category Life cycle

Commelina benghalensis L. Commelinaceae Grass Perennial
Cyperus rotundus L. Cyperaceae Sedge Perennial
Guizotia scabra L. Asteraceae Broad leaf Annual
Ageratum conyzoides L. Asteraceae Broad leaf Annual
Amaranthus hybridus L. Amaranthaceae Broad leaf Annual
Galinsoga parviflora L. Asteracea Broad leaf Annual
Plantago lanceolate L. Plantaginaceae Broad leaf Perennial
Bidens Pilosa L Asteraceae Broad leaf Annual
Cynodon dactylon L. Poaceae Grass Annual
Datura stramonium L. Solanaceae Broad leaf Annual
Digitaria ternate L. Poaceae Grass Annual
Nicandra physalodes L. Solanaceae Broad leaf Annual
[196]
Weed Density (number of plants m-2)
Weed density was increased significantly with increasing competition duration
and the increase was significant with the increasing competition durations. The
maximum (148.67) and minimum (2.00) weed density was obtained in treatments
with weedy-for 60 DACE and season long weed free (weed free check),
respectively (Table 2). Increasing weedy competition duration was showed
proportional increment in weed density that the weeds gain longer period for
germination and continued growth throughout the study period. These results
agree with Tunio et al. (2004) who reported that weed free plots showed minimum
density because weeds eradicated by repeated hand hoeing.
Table 2. The effect of weed competition and controlling time on biomass and density of weed, 2017 and 2018
Treatment Weed density

IDWFP at DACE
10 56.33e
20 48.67ef
30 45.67ef
40 34.17ef
50 32.67f
60 2.67g
WC 99.17cd
IDWP at DACE
10 85.67d
20 106.67bcd
30 99.17cd
40 110.83bc
50 126.83ab
60 148.67a
WFC 2.00g
lsd0.05 22.29
CV% 18.61
Growth and yield parameters of basil plant

Plant height
Basil plant height was significantly influenced by weed competition periods, an
increment of the length of weed interference drastically reduced height of the crop
plant (Table 3). The tallest plants were obtained from weed-free for 50 DACE
(55.12 cm) while the shortest plants from weedy for 60 DACE (21.10cm). The
height of basil plants was increased with prolonged weed-free conditions and
decreased with extended weed infested periods. The present study revealed a
greater reduction in plant height with weed competition which might indicate the
presence of more competition for growth factors than for light. The height of
plants is often associated with their interspecific competitive ability (Freckleton
and Watkinson, 2001). Accordingly, the taller rice plants were found in all weed-
free treatments as reported by Begum et al. (2008).
[197]
Significant difference was observed on number of primary branches per basil plant
due to different weed competition periods (Table 3). A gradual and progressive
decrease in the number of primary branches per plant was recorded with an
increasing competition duration. The highest number of primary branches per
plant (21.50) was found in weed-free check. It was statistically at par with weed
free duration for initial 50 and 60 DACE. The short durations for weed
competition with the crop for growth resources meant effective utilization of
available resources and proper vegetative growth including number of primary
branches per plant. The result is in accordance with the findings of Almarie
(2017), that an increase of soybean branches continued significantly between
weed removal and then after no significant increase was obtained. Similarly,
Singh et al. (2015) also described that number of primary branches per plant in
field pea increased when the weed-free days were prolonged.
Fresh stem, and fresh and dry herbage yield

There was significant effect due to weed competition durations on fresh stem, and
fresh and dry herbage yield of basil (Table 3). Weed-free check, and weed free for
initial 60 and 50 DACE were produced maximum fresh stem weight per plant
(265.20, 258.69 and 256.03 g m-1), respectively. The highest fresh (74.45 g m-1)
and dry (27.57 g m-1) herbage yield per plant were recorded in weed-free check. It
was statistically at par with weed free for initial 60, 50 and 40 DACE but
significantly different from the rest treatments. Similarly, the highest fresh
(3101.87 kg/ha) and dry (1148.78 kg/ha) herbage yields per hectare were recorded
in weed-free check and at par with weed free for initial 60, 50 and 40 DACE. An
increasing weed competition periods was exerted an extended severe competition
for available resources which led to less dry biomass accumulation as the yield
components. Similarly, Singh et al. (2015) reported that the yield attributes were
highest in season-long weed-free period and at par with weed-free for initial 40
days or plots kept weedy only for initial 20 days.
[198]
Table 3. The mean effect of weed competition periods on basil yield components
Treatment PH NPB FSWPP TFHPP TDHPP TFHYHA TDHYHA

IDWFP at DACE
10 42.85d 9.17d 176.83d 46.35c 17.29d 1931.15c 720.52d
20 47.89bcd 13.67c 230.10c 60.60b 19.65c 2525.17b 818.66c
30 45.38cd 13.67c 236.30bc 63.92b 22.82b 2663.13b 951.00b
40 50.02abc 17.00b 247.88abc 73.16a 26.43a 3048.28a 1101.24a
50 55.12a 20.33a 256.03ab 73.03a 27.44a 3042.94a 1143.32a
60 54.00ab 21.17a 258.69ab 74.34a 27.35a 3097.33a 1139.43a
WC 23.95fg 1.67g 2.72h 22.81g 11.34fg 950.29g 472.60fg
IDWP at DACE
10 50.62abc 9.33d 157.56de 44.29cd 16.74de 1845.18cd 697.41de
20 46.24cd 8.67d 152.13e 41.51d 15.37e 1729.68d 640.46e
30 34.29e 5.83e 81.76f 33.98e 13.10f 1415.83e 545.84f
40 28.28ef 3.00f 30.59g 31.06e 11.87fg 1294.10e 494.49fg
50 22.53fg 2.00fg 4.60h 27.06f 10.25gh 1127.64f 427.14gh
60 21.10g 1.33g 5.06h 24.67fg 9.04h 1027.70fg 376.60h
WFC 49.18abc 21.50a 265.20a 74.45a 27.57a 3101.87 a 1148.78a
Lsd0.05 6.23 1.30 22.65 3.80 1.80 158.94 74.82
CV (%) 13.19 10.52 13.01 6.67 8.48 6.67 8.48
Essential oil content and yield

Interaction effects of accessions and location excreted a significant influence on
days to second flowering. Essential oil content and yield were significantly
influenced by the durations of weed competition (Table 4). The maximum
essential oil content (1.27 v/w) and yield (14.58 kg/ha) were obtained from weed-
free check. It was at par with the treatments kept weed-free for early 60, 50 and 40
DACE. The high essential oil content and yield with an increasing duration of
weed-free periods might be due to the accumulation of adequate dry matter
through the utilization of available aboveground and belowground growth
resources by the crop. The obtained essential oil content and yield of basil were
inversely related to the increase in the duration of weedy period and directly
proportional to the increase in weed free periods. In conformity with this result,
Zuhal et al. (2010) reported that the yield of faba bean significantly varied when
weeds were allowed to grow for different durations and about 46% yield loss was
recorded from weedy check plot. Similarly, Welsh et al. (1999) and Ahmad and
Shaikh (2003) found that wheat yield was decreased as the weed infested duration
increased.
Yield losses
Yield losses due to different weed competition periods were considered relative to
the yield of weed-free check (Table 4). The fresh and dry herbage, and essential
oil yield losses were higher with the increasing duration of weedy periods. The
fresh herbage yield losses ranged from 40.51 to 69.36% in increased duration of
weedy periods. Likewise, the dry herbage yield losses ranged from 39.29 to
67.22% in increased duration of weedy periods. The maximum essential oil yield
[199]
loss was found in weedy for 60 DACE (98.35%). The prolonged crop-weed
competition was resulted the reduced dry biomass accumulation which ultimately
rendered the yield parameters considered and then also caused higher yield losses.
Table 4. The effect of weed competition periods on basil yield components
Treatment EOC EOYPHA YL % TFH YL % TDH YL % EOY

IDWFP at DACE
10 0.73c 5.29d 37.74 37.28 63.72
20 0.80c 6.60c 18.59 28.74 54.73
30 0.90b 8.63b 14.14 17.22 40.81
40 1.25a 13.82a 1.73 4.14 5.21
50 1.25a 14.38a 1.90 0.48 1.37
60 1.26a 14.46a 0.15 0.81 0.82
WC 0.13fg 0.63f 69.36 58.86 95.68
IDWP at DACE
10 0.64d 4.46d 40.51 39.29 78.74
20 0.48e 3.10e 44.24 44.25 78.74
30 0.52e 2.87e 54.36 52.49 80.32
40 0.18f 0.90f 58.28 56.96 93.83
50 0.14fg 0.61f 63.65 62.82 95.82
60 0.07g 0.24f 66.87 67.22 98.35
WFC 1.27a 14.58a 0.00 0.00 0.00
Lsd0.05 0.08 0.96
CV (%) 10.52 12.82

The critical period of weed control for basil was estimated based on the relative
yield at 5% to 10% acceptable yield loss. The beginning of the critical periods of
weed competition was obtained from the late weed-crop competition or from the
increasing duration of weedy periods while the end of the critical periods of weed
control was obtained from the early crop weed competition or from the increasing
duration of weed-free periods.
Based on the current result, the critical period of weed control for basil could be
from 10 to 30 DACE at Alage site. Thus, the basil weeds need to be managed
during these periods through appropriate methods to prevent more than 10% yield
loss of the crop. In this regard, Knezevic et al. (2002) reported that critical periods
of weed control varied with weed species composition, weed emergence pattern,
weed density and intensity, ecological variations, climatic conditions, frequency of
tillage operation and soil type of the area. However, Le Bourgeois and Marnotte
(yams, cassava, sugarcane, etc.). Accordingly, Zuhal et al. (2010) who reported
that the critical period of weed control in faba bean started at 30 days and ends 45
days after crop emergence at 10% acceptable yield loss.
[200]
There was an overall sensitivity of basil plants to infestation by weeds, which
demonstrates the need for implementing weed management. The highest weed
biomass and density at harvest associated with the lowest values of essential oil
yield and yield related parameters. It can be concluded from the results that to
obtain better yield, more than 90% of the potential crop yield, the basil weeds
need to be controlled in between 10 to 30 DACE both at Wondo Genet and Alage.
Acknowledgement
Authors are grateful to Wondo Genet Agricultural Research Center and Ethiopian
Institute of Agricultural Research management staff for providing the research
fund and vehicles service. Our gratitude also extends to field assistants at the
Wondo Genet for their assistance during the study.
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[202]
Critical Weed Competition Period on
Chamomile (Matricaria chamomilla L.)
Wondo Genet Agricultural Research Center, EIAR, P. O. Box. 198, Shashemene, Ethiopia
Abstract
A study was carried out to determine critical weed competition period for growth,
yield and yield components of chamomile plant at Wondo Genet during 2017 and
2018 main cropping seasons. The weed competition duration had significant
influence on the per plant height, primary branches number, fresh stem yield, fresh
leaves yield, fresh and dry flower, and also on fresh and dry flower yield/ha, yield/ha
and essential oil content(v/w). The highest fresh and dried flower yields/ha,
respectively 3780.10 and 798.32 were recorded from weed-free check followed by
weed-free for 60 DACE; 3652.10 and 789.43. The maximum essential oil content and
yield respectively, 0.79 v/w and 6.25 kg/ha were recorded from weed-free check. The
yield losses of chamomile were estimated based on the fresh and dry flower, and
essential oil yield per hectare. Thus, the highest fresh and dry flower yield losses,
respectively 83.58% and 81.05% were recorded from weedy-check. The highest
essential oil yield loss was also recorded from weedy for 60 DACE (97.92%)
followed by weedy-check (97.60%). Therefore, to earn higher economic return of
chamomile from its maximal yielding potential, weeds must be kept free within 10 to
30 DACE at 5 to 10% acceptable yield loss levels.
Introduction
Chamomile (Matricaria chamomile L.) is an annual medicinal and aromatic herb
that belongs to the family Asteraceae (Baghalian, 2000; Shams et al., 2012). It is
native to southern and eastern Europe and often referred as “the star among
medicinal species” (Salamon, 1992). The crop grows widely in various ecological
regions of the world (Pourohit and Vyas, 2004). It is cultivated commercially in
Europe, former USSR, North Africa, Asia, North and South America and New
Zealand (Salamon, 2004).
The chamomile drug is included in the pharmacopoeia of 26 countries (Pamukov

and Achtardziev, 1998). This herb has been believed by Anglo-Saxons as 1 of 9
sacred herbs given to humans by the lord (Crevin and Philpott, 1990). Chamomile
is used mainly as an anti-inflammatory and antiseptic, also antispasmodic and
mildly sudorific (Mericli, 1990). It has been used in herbal remedies for thousands
of years, known in ancient Egypt, Greece, and Rome (Issac, 1989). Hungary is the
main producer of the plant biomass and also grows abundantly in poor soils and it
is a source of income to poor inhabitants of these areas. Flowers are exported to
Germany in bulk for distillation of the oil (Svab J., 1979). There is no demand for
[203]
blue oil as such in India. However, flowers of chamomile are in great demand.
Presently, two firms, namely, M/s Ranbaxy Labs Limited, New Delhi and M/s
German Remedies are the main growers of chamomile for its flowers.
There are many factors that influence agronomic characters, biomass and essential
oil yield of medicinal and aromatic plants. Among these, weed management is a
major constraint. As with all commonly grown crops, also in medicinal and
aromatic plants, weeds function as crop competitors, create problems for
mechanized harvest, and may alter the end quality when mixed with the harvested
product.
The well-known interference of weeds takes additional relevance for medicinal

and aromatic plants for several reasons. Firstly, the synthesis of secondary
metabolites in plants is linked to many genetic and environmental factors
(Sangwan et al., 2001). Buyers often grade such plants according to their specific
quality features, which are primarily determined by their content in essential oils
or other secondary metabolites, which on their turn can be reduced in presence of
weeds (Carrubba and Catalano, 2009). Although sparse specific experiments have
been conducted, the presence of weeds is therefore believed to exert a significant
effect on plant metabolic pathways, in this way acting negatively on the market
end value of such crops (Gil et al., 1998).
Furthermore, especially in those crops where seeds are collected, weed

competition has to be avoided to guarantee certain yield. Some experiments
carried out on suitability of medicinal and aromatic plants to field conditions have
confirmed the importance of weed competition; de la Fuente et al. (2003)
demonstrated that on coriander, especially under poor soil conditions, weeding
had a greater effect than did N fertilization. Weeding efforts become more
effective in a crop when done at a proper growth stage i.e., critical weeding time.
The concept of critical periods of weed competition, during which weeds have the
greatest impact on crop growth, was verified by Nieto et al. (1968). It is a specific
minimum period of time during which the crop must be free of weeds in order to
prevent loss in yield and represents the overlap of two separate components
(Weaver and Tan, 1983). The first component is the length of time weeds can
remain in a crop before interference begins. The second component is the length
of time that weed emergence must be prevented so that subsequent weed growth
does not reduce crop yield.
It was reported that weed removal in chamomile plant during 5–11 weeks after
planting was necessary to obtain a higher yield of the flower and oil (Singh, 1997).
The uncontrolled weed growth caused 34.4% reduction in the dry flower yield as
compared with the weed-free condition. In Ethiopia, where diverse ecological
conditions are predominant and the crop plant become important no attempt made
[204]
so far in determining proper weed removal duration in chamomile field.
Determination of the critical period of weed control for chamomiles is a
prerequisite to get optimum yields and also to maintain its proper agronomic and
chemical characters. The objective this study was, therefore, to determine the
critical period for weed control in chamomile under rain fed condition, at Wondo
Genet.

Study area
experimental site during 2017 to 2018 main cropping seasons. Wondo Genet is
located at 7°19' N latitude, 38°38' E longitude and an altitude of 1780 m asl.
(meters above sea level). It receives mean annual rainfall of 1128 mm with
minimum and maximum temperatures of 11 °C and 26 °C, respectively. The soil
textural class is sandy loam with pH of 6.4 (Abayneh et al, 2006).

The experiment was laid out in RCBD with three replications having plot size
measuring of 2.4 × 1.8 m. The German chamomile, genotype was sown in July
month of the two experimental years with 30 and 40cm space between rows and
plants, respectively. Distance between plants was maintained by thinning extra
plants at early growth stage. The experimental treatments consisted of a
quantitative series of both the increasing duration of weed interference and the
length of the weed free period. Timing of weed removal was based on the number
of days after crop emergence. Weeds were removed with a hand hoe from
respective plots after prescribed duration and kept weed free till harvest.
To determine the beginning of the CPWC, an increase in length of weed-free

periods was established by maintaining weed-free condition for 10, 20, 30, 40 ,50
and 60 days after crop emergence, referred as weed- free plots before allowing
subsequent emergence of weeds to compete with the crop plant for the remain
growing season. While to evaluate the end of the CPWC, an increase in duration
of weed interference was established by allowing the weeds to compete with the
roselle for 10, 20, 30, 40, 50 and 60 days after crop emergence, referred as weedy
plots after which, plots were maintained weed-free until the harvesting time. In
addition, season long weedy and weed-free check was included respectively as
negative and positive control.

Weed flora and density: In the experimental plots, data on weed flora type and
population found within randomly and repeatedly thrown sampling quadrat (0.25
m×0.25 m) were recorded. The data were taken at four spots in each plot at the
[205]
time of weed removal for early competition and about 10 days interval before the
expected harvest time in the case of late competition to avoid possible foliage and
seed shading. The collected weed plants population count values from sampling
quadrat were converted and expressed as number of plants per m2 bases.
Weed aboveground dry biomass (g): The weeds obtained within the quadrat were
cut near the soil surface and placed separately into paper bags the treatment wise.
The samples were sun-dried for 3-4 days and thereafter placed into an oven at 65
°C temperature till a constant weight, and subsequently their dry weight was taken
using sensitive balance. The weed plants dry weight values per the quadrat were
converted and expressed in gram per m2 bases.
Crop data: Four plants per plot selected at random and data on per plant height,
number of primary branches, weight of fresh stem and leaf, weight of fresh and
dry flower, and also fresh and dry flower per hectare, essential oil yield per
hectare and essential oil were collected.
Data collected on growth and yield parameters of the crop were analyzed using
Fisher’s analysis of variance technique. Least significant difference (LSD) test at
0.05 probability level was employed to compare the treatment means (Steel et al.,
1997).
x 100

Weed species
There were 20 annual and perennial weed species identified belonging to 9
families in which more numbers of broadleaf species found than grass and sedge
weeds (Table 1). In the experimental field, the annual weeds were found as most
common group during the two years study. The findings are in line with Adjun
(2003), Mahadevan et al (2009) and Ahmed and Salaheldeen (2010) who reported
that the crop associated weed species were composed of a wide range of annual,
biennial and perennial and also broad leaved, grasses and sedges weed groups.
[206]
Table 1. List and characteristics of weed species associated with chamomile field, 2017 and 2018.
No Scientific name Family name Category Life cycle

2 Bidens pilosa L. Asteraceae Broad leaf Annual
11 Cyperus rotundus L. Cyperaceae Sedge Perennial
14 Phalaris paradox L Poaceae Grass Annual
16 Portulaca olerace L. Portulacaceae Broad leaf Annual
18 Annagallis arvansis L. Primulaceae Broad leaf Annual
Weed Density (m-2)

Total weed density increased significantly with the increase in competition
duration and the increase was significant for each the increasing duration. The
maximum (89.26) and minimum (0) total weed densities were obtained in plots
with season long weed-infestation (weedy check) and weed free (weed free check)
respectively (Table 2). Weedy check showed the maximum weed density because
there was longer period available for weeds to germinate and weeds continued to
germinate throughout the growth period. These results agree with Tunio et al.
(2004) report that weed free plots showed minimum density because weeds were
reduced by repeated hand hoeing.
Weed Dry Weight (gm-2)

The dry weight of aboveground weed biomass was increased with an increase in
competition period, being the maximum biomass (113.53) recorded by weedy
check treatment (Table 2). This indicates that higher weed density and longer
growth period were resulted more accumulation of photosynthates and the greater
biomass. Akhtar et al. (2000) found that by increasing weed crop competition
duration increased the weed biomass. Similarly, but in the other way the weeds
emerging at the later growth stages offer less competition to crops as it
accumulates lower weed biomass (Uremis et al., 2009).
[207]
Table 2. The effect of weeding times on weed biomass and weed density, 2017 and 2018.
Treatments WDB (gm-1) WD
IDWFP at DACE
10 90.15b 72.23b
20 31.03e 29.39de
30 22.32ef 22.55ef
40 10.14fg 10.83gh
50 10.72fg 11.28fgh
60 8.52fg 9.513gh
WC 113.53a 89.26a
IDWP at DACE
10 11.62fg 19.49efg
20 22.54ef 24.55de
30 35.85e 34.20d
40 67.03d 56.78c
50 74.79cd 61.83bc
60 87.90 bc 72.57b
WFC 0.00g 0.00h
lsd0.05 15.17 11.4
CV% 21.6 18.48
IDWFP= Increasing duration of weed free period, IDWP= Increasing duration of weedy period, DACE= Days after
crop emergence, WC= Weedy-check, WFC= Weed free check, WDB= Weight of dry biomass, WD= Weed density
Chamomile growth and yield parameters

The present result revealed that the effect of different weed-chamomile
competition durations on the plant height was significantly varied. The tallest
plant height (73.33cm) was recorded from weedy check plot (Table 3). This was
due to the fact that weeds were left to grow for longer periods and the weed plant
population per unit area increased which resulted in severe competition between
the crop and weed plants for light and space. In agreement with the present study,
Freckleton and Watkinson (2001) reported that the height of plants is often
associated with their interspecific competitive ability. Similarly, Singh et al.
(2015) also reported that the field pea plant height increased with increase in
duration of weed interference and decreased with increase in weed free periods.
There was also significant effect of weed-crop competition durations on number of

primary branches per plant. A gradual and progressive decrease in number of
primary branches per plant was recorded with the increasing competition
durations. The highest number of primary branches per plant (31.00) was obtained
from weed-free check. This was statistically at par with weed free competition
duration for 60 DACE but significantly different from the rest of the treatments
(Table 3). This mean than when there is less time for competition between the
crop and weed plants for available resources, it gives a chance for obtaining the
higher number of primary branches per plant. When weeds are removed, the crop
plants gain good growth rate and maximum assimilates that allow good vegetative
growth and higher number of branches per plant. The results are in accordance
with the findings of Almarie (2017) who obtained that the increase of soybean
[208]
branches continued significantly between weed removals and then after no
significant increase was achieved. Similarly, Singh et al. (2015) also described
that number of primary branches per plant in field pea increased when the weed-
free days were prolonged.
The present study indicated that the weight of fresh and dry stem, and yield of
leaves and flower were significantly influenced by the duration of weed
competition. The result revealed that weed-free check (211.25 gm-1) followed by
weed-free for 60 DACE (210.35 gm-1) were produced maximum fresh stem per
plant (Table 3). Likewise, the maximum fresh leaf per plant was recorded in weed-
free for 60 DACE (77.62 gm-1). This was statistically at par with the weed- free
check and weed free competition duration for 50 and 40 DACE but significantly
different from the rest of the treatments.
The maximum fresh flower yield (45.36 gm/plant and 3780.10 kg/ha) was
obtained from weed-free check. It is statistically at par with the weed free
competition durations for 60, 50 and 40 DACE and significantly different from the
rest treatments (Table 3). Similarly, the maximum dry flower yield (9.58 gm/plant
and 798.32 kg/ha) was harvested from weed-free check which statistically at par
with the weed free competition durations for 60, 50, 40 and 30 DACE, and
significantly different from the rest treatments (Table 4). The absence of weeds in
weed-free plots beyond 40 DACE enable the crop to make best use of growth
resources that result in a greater fresh and dry stem, leaves and flower per given
areas. The results were in line with Singh et al. (2015) report that the yield
attributes of field pea were highest in season-long weed-free period and at par with
weed-free for initial 40 days or plots kept weedy only for initial 20 days. The
authors also stated that yield attributes of the crop increased with an increase in
weed-free duration and decrease with the decreased weedy periods.
[209]
Table 3. Effect of weed competition and weeding periods on yield components of chamomile, 2017 and 2018.
Treatment PH (cm) NPB FSWPP FLWPP FFWPP FFW

(gm-1) (gm-1) (gm-1) (Kg-1 ha)
IDWFP at DACE
10 62.50cde 17.50e 68.53f 45.01d 32.98c 2748.30c
20 63.67bcd 20.67d 145.74d 58.64b 39.49b 3290.40b
30 60.25 de 21.83 cd 161.58 c 59.25 b 39.49 b
3290.30b
40 56.75 defg 23.50 c 203.34 ab 74.77 a 42.44 ab 3536.50ab
50 52.46fgh 26.17b 197.96b 74.97a 43.39ab 3615.60ab
60 50.21gh 29.50a 210.35a 77.62a 43.83ab 3652.10ab
WC 73.33 a 5.33 g 18.28 h 11.17 h 7.45 g 620.80g
IDWP at DACE
10 56.58efg 21.17d 155.85cd 51.04c 27.39cd 2282.30cd
20 58.92def 16.83e 104.35e 28.73e 23.39de 1949.40de
30 62.46cde 8.33f 57.66f 26.05ef 17.87ef 1488.80ef
40 67.75abc 6.67fg 38.63g 22.07f 15.37f 1280.90f
50 67.79abc 5.17g 31.67g 22.01f 9.03g 752.30g
60 70.50 ab 4.67 g 29.03 gh 16.23 g 8.17 g 680.40g
WFC 47.50h 31.00a 211.25a 77.25a 45.36a 3780.10a
Lsd0.05 6.94 2.05 11.68 5.20 5.67 472.87
CV (%) 9.86 10.42 8.65 9.58 17.35 17.35
PH=Plant height, NPB=Number of primary branches, FSWPP=Fresh stem weight per plant, FLWPP=Fresh leaf weight
per plant, FFWPP=Fresh flower weight per plant, FFW(Kg/ha) =Fresh flower weight per hectare
Essential oil content and yield were significantly influenced by the duration of
weed competition. The maximum essential oil content and yield (0.79 v/w and
6.25 kg/ha) were recorded from weed-free check which is at par with the
treatments kept weed-free beyond 40 DACE and the lowest (0.10 v/w and 0.1
5kg/ha) found in weedy for 60 DACE (Table 4). With the increasing duration of
weed-free period, the accumulation of adequate dry matter like number of
branches per plant, and fresh and dry flower per plant through the utilization of
available aboveground and belowground growth resources help on obtaining the
highest essential oil content and yield by the crop. In general, essential oil content
and yield of chamomile was inversely related to the increase in the duration of
weedy period, and directly and positively proportional to the increase in weed free
periods. In conformity with this result, Zuhal et al. (2010) reported that the yield
of faba bean significantly varied when weeds were allowed to grow for different
durations and about 46% yield loss was recorded from weedy check plot. Welsh et
al. (1999) and Ahmad and Shaikh (2003) were also found that wheat yield
decreased as the weed infested duration increased.
[210]
Table 4. Effect of weed competition and weeding periods on yield components of chamomile, 2017 and 2018
Treatment DFWPP DFW EOC EOY

(gm-1) (Kg-1 ha) (V/W) (Kg -1 ha)
IDWFP at DACE
10 7.24bc 603.20b 0.43cd 2.56cd
20 7.40b 617.00b 0.47c 2.93c
30 8.87a 739.23a 0.58b 4.27b
40 8.99a 749.48a 0.78a 5.89a
50 9.17a 763.89a 0.76a 5.84a
60 9.47a 789.43a 0.76a 5.91a
WC 1.82f 151.30f 0.10g 0.15h
IDWP at DACE
10 6.35cd 528.72cd 0.39cde 2.06de
20 5.54d 461.69d 0.34def 1.55ef
30 4.28e 356.11e 0.30ef 1.04fg
40 3.79e 315.62e 0.26f 0.80gh
50 2.16f 179.67f 0.12g 0.21h
60 1.94f 161.64f 0.08g 0.13h
WFC 9.58a 798.32a 0.79a 6.25a
Lsd0.05 0.99 82.66 0.10 0.74
CV (%) 13.86 13.86 19.67 22.46
DFWPP=Dry flower weight per plant, DFW(Kg/ha) =Dry flower weight per hectare, EOC=Essential oil content,
EOY=Essential oil yield
Yield losses
The losses in crop yield due to the different weed competition periods were
considered relative to the yield of weed-free check. The fresh flower yield losses
were ranged from 39.32 – 53.58% with the increasing duration of weedy periods
(Table 5). The loss in the fresh flower yield per hectare was higher from weedy
beyond 40 DACE. Likewise, dry flower yield losses were higher from WC plot
followed by weedy for 50 and 60 DACE. The dry flower yield losses ranged from
(33.77 – 81.05%) with the increasing duration of weedy periods (Table 5). The
maximum essential oil yield loss was found in weedy for 60 DACE (97.92%)
followed by weedy-check (97.60%) and weedy for 50 DACE (96.64%) while the
minimum was found in weed-free check that ranged from (0-59%) with an
increasing duration of weedy periods as shown in Table 5. Prolonged crop-weed
competition period results reduced dry biomass accumulation which ultimately
rendered the yield parameters and actual yield considered. In conformity with this
result, Singh (1997) reported that weed removal in chamomile plant during 5–11
weeks after planting was necessary to obtain a higher yield of the flower and oil.
The uncontrolled weed growth was caused 34.4% reduction in the dry flower yield
as compared with the weed-free condition.
[211]
Table 5. Yield losses of chamomile due to the weed competition and weed control time, 2017 and 2018.
Treatment RYL (%) FFW RYL (%) DFW RYL (%) EOY
IDWFP at DACE
10 27.30 24.44 59.04
20 12.95 22.71 53.12
30 12.96 7.40 31.68
40 6.44 6.12 5.76
50 4.35 4.31 6.56
60 3.39 1.11 5.44
WC 83.58 81.05 97.60
IDWP at DACE
10 39.62 33.77 67.04
20 48.43 42.17 75.20
30 60.61 55.39 83.36
40 66.11 60.46 87.20
50 80.10 77.49 96.64
60 82.00 79.75 97.92
WFC 0.00 0.00 0.00
RYL (%) FFW= Relative yield loss of fresh flower, RYL (%) DFW= Relative yield loss of dry flower,
RYL (%) EOY= Relative yield loss of essential oil yield

The critical period of chamomile weed control was estimated based on the relative
yields at 5 to 10% acceptable yield loss. The beginning of the critical period of
weed competition was obtained from the late weed-crop competition (from the
increasing duration of weedy periods) while the end of the critical period of weed
control obtained from the early crop weed competition (from the increasing
duration of weed-free periods).
Based on the harvested yield data, the critical period of weed control for
chamomile ranges from 10 to 30 DACE. This result follows previous studies. Le
Bourgeois and Marnotte (2002) located that the critical period between 30 and 90
DAS for long-cycle crops (yams, cassava, sugarcane, etc.). This was also in line
with the finding of Zuhal et al. (2010) that the critical period of weed control in
faba bean started at 30 days and ends at 45 days after crop emergence at 5 to 10%
acceptable yield loss.

There was an overall sensitivity of chamomile to the infestation of weeds, which
demonstrates the need for weed control measures. The highest weed biomass and
density at harvest actually were associated with the lowest values of essential oil
yield and yield related components. From this result it can be concluded that to
obtain better yield of chamomile, greater than 90% of the crop genetic potential
[212]
yield, it needs to keep weed-free between 10 to 30 days after crop emergence at
Wondo Genet Agricultural Research Center and similar agro-ecologies.
Acknowledgement
The authors are grateful to Wondo Genet Agricultural Research Center and
Ethiopian Institute of Agricultural Research management bodies for the research
fund support and field vehicles provision. Our gratitude also extends to field
assistants at the research center for their unreserved assistance during the study
period.
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[214]
Critical Weed Competition Period
on Roselle (Hibiscus sabdariffa L.) at
Arba Minch, Southern Ethiopia
Getachew Gudero1 and Ano Wariyo2
1
Arba Minch Agricultural Research Center, SARI, P.O.BOX. 2228, Arba Minch, Ethiopia
2
Wondo Genet Agricultural Research Center, EIAR, P.O.BOX. 198, Shashemene, Ethiopia
Abstract
In major roselle (Hibiscus sabdariffa) growing areas of Ethiopia, high weed
competition is among the greatest constraints to its production. Controlling weeds at
the critical period can be a means for effective weed control, and can help in reducing
the strikes of weeds on yield of roselle and increasing the benefit from its production.
Thus, a field experiment was conducted in 2017 cropping season to determine the
effect of weed competition periods on growth and yield performance, and the critical
period of crop-weed competition and yield losses in roselle. The experiment was
consisted of 14 treatments in two series: increasing duration of weedy period (IDWP)
and increasing duration of weed-free period (IDWFP) compared with weedy check
(WC) and weed-free check (WFC). The treatments were arranged with RCBD with
three replications. Weeds were allowed to compete for 15, 30, 45, 60, 75 and 90 days
after the crop transplanting (DAT) and then after avoided throughout the growing
season along with a WFC. The results exhibited that a total of 69 weed species were
competing vigorously with roselle, of which large numbers belong to the dominant
families 17 species under Poaceae followed by 14 species under Asteraceae. In IDWP
series, weed density and weed dry biomass were increased correspondingly, whereas
crop phenology and yield attributes were decreased. The reverse was true in IDWFP
series for both weed and crops parameters. The highest fresh and dried calyx yields
were obtained from WFC followed by weed free for the initial 90 DAT in IDWFP
series, whereas the lowest was gathered from WC followed by weedy for 75 and 90
DAT. The high weed interference significantly reduced fresh and dried calyx yield by
68.69 and 65.93%. Overall, as it has been found to be the critical period of weed-crop
competition in roselle, to reduce the yield losses to less than 10% and obtain higher
economic return, weeds must be kept free within 15 to 45 DAT.
Introduction
Roselle (Hibiscus sabdariffa L.) is a shrub crop plant belonging to family
Malvaceae. It is one of important medicinal crops growing in different countries of
the world. The crop is specifically well known in the tropical countries where it
was native to Southeast Asian or tropical Africa and distributed to many countries
in tropical and sub-tropical regions including eastern Africa (Crane, 1949;
Mordock, 1959; Cobley, 1975; Leung and Foster, 1996; Mohamed et al., 2012).
It is grown mainly as medicinal crop in traditional farming system and also as

important cash crop and source of income for small scale farmers in the growing
[215]
areas. The economic part of the crop is the fleshy calyx (sepals) and the
surrounding fruit (capsules). In addition, in many parts of the world leaves is
consumed as green vegetable, the stem is served as a source of pulp for paper
industry, seeds used as a poultry feed and as an aphrodisiac coffee substitute and
raw material in industrial and domestic cottages (Khidir 1997; Aziz et al., 2007;
Mahadevan et al., 2009; Ahmed and Salaheldeen 2010). The worldwide
consideration of roselle is acknowledged since their enormous unexploited
economic advantages have been seen by the producers and end users (Aziz et al.,
2007; Mahadevan et al., 2009; Ahmed and Salaheldeen 2010).
Despite the fact that now a day worldwide production of roselle becomes high,
under Ethiopia condition it is at infant stage and grown primarily for commercial
purpose in limited private farms (Tamirat et al., 2014). This is because of little or
no recognition of its importance, and it has got diminutive reflection to embark on
the production for commercial purpose only. Consequently, the contribution of
this locally called “karkade” crop is not important as other cultivated crops to the
country growth and development although in recent time few cooperatives are up-
and-coming in the production and supplying it as essential oils and medicinal
herbs for national and in few amounts for foreign market (Endeshaw 2007;
Tamirat et al. 2014).
Besides, the country has a diverse agro-ecology that can grow various medicinal
crops with a vast impending for roselle production as well both in rain fed and
irrigation conditions. Also have a huge potential in internal and external markets if
possibly produce by the farming communities of the country. Despite these
potentials and economic importance, the overall yield and quality are lower both
in farmer and private farms mainly due to biotic and abiotic factors. These factors
included increased pressure of insects, diseases and weeds, and also poor cultural
practices and harvest constraints during the growing periods. Among the biotic
factors, weeds cause significant effects on yield of roselle and play a greatest role
in the production system (Ahmed and Salaheldeen 2010).
Though not so far studied in Ethiopia, Ahmed and Salaheldeen (2010) and
Upadhyay et al. (2011) reported that losses in economic yield of roselle due to
weed infestation in the neighboring country Sudan were ranged from 45.00 to
90.17%. Moreover, in the study areas and in the country as well the growers spend
about 70 to 80% of their total available farm labor on weed control and this is
usually carried out by hoe-weeding.
To determine appropriate time of weed management, the identification of critical

competition period is essential in the growing period of the crops (Evans et al.
2003; Otto et al. 2009). Rajcan and Swanton (2001) reported that competition
between crop and weeds, and thus critical period for weed control (CPWC), are
[216]
dependent on-site specific factors: climatic conditions, management strategies,
crop types, composition of weed flora and weed emergence time.
There is no study committed to find out the critical period for weed control and
victims resulting from the antagonism concerning this crop in Ethiopia. Therefore,
the objectives of this study were to determine the effect of weed competition
periods on growth and yield performance, and the critical period of crop-weed
competition and yield losses in roselle at Arba Minch, southern Ethiopia.
Material and Methods

Description of experimental site
The study was carried out at Chano mile sub-station of Arba Minch Agricultural
Research Center during the 2017 main cropping season with supplementary
irrigation. It is geographically located at 06O 06’ 841’’ latitude, 037O 35’ 122’’
longitude and an altitude of 1216 m asl in the plateau of southern part of Ethiopia
and in the environs of Abaya and Chamo lakes about 476 km south of Addis
Ababa and 280 km from Hawasa. The area is characterized by a bimodal rainfall
pattern where short rainy season occurs during the months of March and April,
and the main long season rain starts in mid-August and extends to late October.
The late August and September months are known with very high amount of
rainfall. Thus, in the last decade the area had received an average annual rainfall
and temperature of 750 mm and 27.5 oC, respectively. The soil is characterized by
alluvial, black sandy-loam and clay-loam. The area is among the major suitable
areas for the production of roselle in the southern region of the country.
Seedling raising and transplanting

Following the standard method, roselle seedlings growing was made on well
prepared seedbeds. Roselle ‘Sudan type’ seed was sourced from Wondo Genet
Agricultural Research Center. Seedlings were raised in the nursery on raised beds
with 1 m width and 10 m length using polyethylene pots filled with recommended
proportion of forest, sand and top soil i.e., 1:2:1 ratio, respectively. The seeds were
sown on polyethylene pots at a depth of 0.5 cm on 04 August 2017 cropping
season. Grass mulch was applied on well-arranged polyethylene pots on the bed
and removed after seven days when the seedlings emerged. The pots were weeded
and irrigated as deemed necessary.
Field plots were meticulously tilled, leveled and armed ridges were then prepared.
At appropriate growth stage, usually at 30 days after sowing healthy looking,
vigorous and uniformly sized roselle seedlings were selected for transplanting to
the field. The seedlings were transplanted to sides of armed ridges of experimental
field on 05 September 2017.
[217]
Treatments and design of experiment
The experiment was consisted of 14 treatments that laid out in a randomized
complete block design with three replications as described by Gomez and Gomez
(1984). The treatments were arranged following the method described by Nieto et
al. (1968) in two sequences such as weed interference up to 15, 30, 45, 60, 75 and
90 days after transplanting (DAT) i. e, increasing duration of weedy period
(IDWP) then after reserved weed free awaiting harvesting date and weed-free up
to 15, 30, 45, 60, 75 and 90 DAT i. e, increasing duration of weed-free period
(IDWFP) afterward permitted to grow until harvesting date, and compared with
two checks namely complete weed free and weedy check in the growing period.
The experimental field was lied out on71.40 m length x 16.80 m width plot size
with a unit plot of 3.60 x 3.60 m. It was laid on five years fallow period land after
clearing the existing plant population. Plots were spaced apart by 1.5 m and blocks
separated by 2.5 m. The plot was consisted of a total of six rows, but avoiding the
border effect four harvestable middle rows remained for data collection. Spacing
between plants and rows were maintained at 60 cm apart. In each row there were
six plants with a total of 24 crop plants per plot excluding the border rows. The
recommended agronomic practices for growing of roselle were applied uniformly
during the field management, except for treatment application. Due to the
available soil moisture and favorable environment, the weeds were started
emerging five DAT and get satisfactorily developed after one week. The weeds
appeared in plots were removed by hand pulling and hoeing following the days set
for each treatment.
Weed parameters
Mixed weed species were considered to determine critical period of weed
completion. The weed species found at each plot were sampled from three spots
diagonally using 0.5 m x 0.5 m quadrat, identified, counted and recorded prior to
weeding practice. The collected weed data were summarized following the
procedures designed by Akobundu and Agyakwa (1987), Akobundu (1991) and
Shabbir and Bajwa (2006).
After recording data on the weed count, the weed plants within the quadrat were
harvested at ground level and placed separately into kenaf made bags treatment
wise for dry biomass weighing. For each treatment on plots of IDWFP, weeds
were harvested at the time of crop maturity, while on plots of IDWP weed
harvested at the initial weed control time after which kept clean of weeds until the
harvesting time. The samples were sun dried for five to six days and subsequently
put into an oven at 70 °C until a constant reading maintained to measure the dry
biomass. The dry weight per plot was converted and expressed in kg ha-1 base.
[218]
Crop parameters
Data on the crop growth, phenology and yield parameters were determined from
each plot. Number of days to 50% flowering was recorded as number of days from
DAT of roselle to the first flower appeared on 50% of the plants in each plot. Plant
height (m) at physiological maturity was taken with a measuring rope from the
base to the apex of the main stem of five randomly sampled and pre-tagged plants
in each net plot. Number of branches was recorded as the average number of
reproductive branches per plant of the five sampled plants in the middle rows
during harvesting. Fresh weight of calyx (kg) was recorded from the four middle
rows for each treatment and converted to kilogram per hectare base. Dry weight of
calyx was obtained from randomly selected 10 fresh calyxes for each treatment
and put into oven at 106 OC for 48 hours and then converted to kilogram per
hectare base. Fresh leaf weight (g) was taken from five pre-tagged plants from
each plot prior to physiological maturity and measured using electrical balance.
Hundred seed weight (g) was recorded as weight of dried 100 seeds from
randomly selected harvested pod. Seed weight (kg) was recorded from the four
middle rows for each treatment and converted to kilogram per hectare base. It was
measured after lamming the harvested and sun-dried pods from each net plot and
it was adjusted at 10% seed moisture content.
Data analysis
Data on weed and crop parameters were subjected to analysis of variance
(ANOVA) to determine the treatment effects using general linear model procedure
of the SAS software version 9.1 (SAS 2009). To separate differences among
treatment means Fisher’s protected least significant difference (LSD) test at p ≤
0.05 was used during the analysis following the procedure developed by Gomez
and Gomez (1984). On the other hand, the critical period of weed control, the
relative roselle yields (Y) of each treatment were computed as follow.
x 100
[219]
Weed Parameters
Weed communities
The weed community found in the investigational plots was identified to species
levels (Table 1). The weed species in family Asteraceae (7 species) followed by
Poaceae (3 species) were found richer than the rest of top most families in number
and got greater area coverage. In this regard, Commelinaceae, Amaranthaceous,
Euphorbiaceae and Cyperaceae correspondingly had two species per each.
Table 1. Weed community found in the investigational plots
Rank Weed species Family Common name

1 Xanthium strumarium Asteraceae Cocklebur
2 Galinsoga parviflora Asteraceae Gallant soldier
3 Bidens pilosa Asteraceae Black jack
4 Brachiaria eruciformis Poaceae Signal grass
5 Digitaria ternate Poaceae Crabgrass
6 Amaranthus hybridus Amaranthaceae Green amaranth
7 Cyperus esculentus Cyperaceae Yellow nut sedge
8 Hyptis suaveolens Lamiaceae Pignut
9 Amaranthus graecizans Amaranthaceae Mediterranean amaranth
10 Ageratum conyzoides Asteraceae Goat weed
11 Euphorbia heterophylla Euphorbiaceae Milkweed
12 Digitaria abyssinica Poaceae Blue couch grass
13 Crassocephalum rubeno Asteraceae Rag leaf
14 Commelina benghalensis Commelinaceae Wandering Jew
15 Acalypha crenata Euphorbiaceae Copperleaf
16 Commelina latifolia Commelinaceae Water maker
17 Cyperus rotundus Cyperaceae Nut siege
18 Portulaca oleracea Portulacaceae Purslane
19 Launaea cornuta Asteraceae Wild lettuce
20 Parthenium hysterophorus Asteraceae Congress weed
Weed density and dry biomass

Durations of weed competition periods were significantly (p < 0.01) affected weed
density and dry matter production in the growing period of roselle (Table 2). Due
to violation of ANOVA assumption the weed density (population m-2) data were
transformed using square root (𝑋 = √ +0.5) transformation as described by Gomez
and Gomez (1984) before the statistical analysis.
Significance reductions of weed density were observed in IDWFP series. The

density of weeds was increased significantly with each increased competition
periods. However, there was no significant distinction among the IDWP at 45, 60
and 75 DAT for the reduction of weed density. Likewise, there was no statistically
significance difference on weed density among IDWFP at 30, 45, 60 and 75 DAT
although they had lower density than IDWP at the same dates. The highest
[220]
reduction of weed density was observed from WFC and IDWFP at 90 DAT (Table
2).
Weed dry biomass (kg ha-1) accretion speculates the growth performance of weeds
and gives better signal of weed-crop competition. It was increased with the IDWP
and decreased with the IDWFP. The highest weed dry biomass (kg ha-1) was
obtained from WC followed by weeding at 90, 75 and 60 DAT under the series of
IDWP. The likely reasons for the higher weed dry weight at the investigational
areas could be the relatively better rainfall and temperature, and supplementation
of irrigation during the shortage of precipitation during the cropping period, which
might have brought on more buildup of weed dry biomass. On the contrary, the
lowest weed dry weight was found on WFC followed by the weed free treatment
for the initial 90 DAT under IDWFP series (Table 2).
Table 2. Effect of increasing duration of weedy and weed free periods on weed density and dry weight at Arba Minch,
2017
Treatment Weed density (m2) Weed dry biomass (kg/ha)

IDWP series
15 7.14 (51.11)cd 850.50fg
30 7.27 (52.91)bcd 916.30f
45 7.41 (55.17)abcd 1529.50d
60 7.44 (55.48)abcd 1757.60c
75 7.63 (58.90)abc 1810.00c
90 7.92 (62.94)ab 2209.40b
WC 8.05 (64.97)a 2779.00a
IDWFP series
15 7.47 (55.92)abcd 1653.10cd
30 7.06 (49.90)cd 1309.70e
45 7.19 (51.78)cd 944.50f
60 7.11 (50.72)cd 880.50f
75 7.21 (52.06)cd 836.80fg
90 6.98 (48.79)d 672.30g
WFC 6.98 (48.79)d 189.50h
LSD (0.05) 0.65 0.65
CV (%) 5.34 9.48
Means followed by the same letters within each column are not significantly different. DAT = Days after transplanting;
IDWP = Increasing duration of weedy period; WC = Weedy check; IDWFP = Increasing duration of weed-free period;
WFC = Weed-free check; CV = Coefficient of variation and LSD = Least significant difference at 0.05 probability level.
Crop parameters
Phenological and growth parameters
Days to 50% flowering: Significant (p < 0.0001) difference was observed among
the duration of weed competition effect on days to 50% flowering (Table 3). It
was increased with the IDWFP and decreased with the increasing IDWP. In the
IDWP the weeds might have exercised a severe competition and utilized the
nearby resources for a longer period of time and as result the probable reason
[221]
could be the crop might need to perpetuate themselves and getting to flowering
forcedly. On the other hand, in IDWFP the weeds might have less chance to
compete with the crop due to the interference of weeding practices and as result
the crop got a favorable condition to grow appropriately and undertake normal
physiological function with no or little competition of resources with weeds during
the growing period. The highest days to 50% flowering was observed in IDWFP
although statistical correspondence with the values obtained little difference
among the treatments. Weed free check and weed free for initial 90 DAT in
IDWFP category had longer days to 50% flowering than the other periods (Table
3).
Plant height (M): It was significantly (p < 0.001) affected by the different
durations of crop-weed completion (Table 3). Increasing duration of weedy
periods was resulted in decreasing of plant heights but the opposite was true for
IDWFP. Higher plant height was associated with the lower the weed-crop
competition period. The highest plant height was recorded on WFC and in the
category of IDWFP for 60, 75 and 90 DAT. The probable reason in minimum
plant height in IDWP, the weeds might have caused a strict competition with the
crop and utilized the nearby resources for a longer period and thus ensued in
lowering of the heights.
Number of branches: They were important characters of roselle which greatly

influenced by nutrient supply, soil moisture and environmental stress. Significant
difference (p < 0.01) was observed on number of branches per plant at different
crop-weed competition periods (Table 3). Holding the investigational plots weed
free from 15 to 60 DAT resulted in a higher number of branches per plant that was
statistical at par with the number of branches per plant from WFC. Overall, the
number of branches per the crop plant was increased as weed interference
decreased and the vice versa. Thus, the higher number of branches per plant was
because of less or no competition of the weeds with the growing crop during the
growing season.
[222]
Table 3. Effect of increasing duration of weedy and weed-free periods on phenological parameters of roselle at Arba
Minch, 2017
Treatment Phenological and growth parameters
50% date of flowering Plant height (m) Number of branch per plant
IDWP series (DAT)
15 34.46d 80.42bc 3.62 (3.00)abcd
30 30.13 ef 63.67 cde 3.27 (2.42)bcdef
45 30.06 ef 60.83 cde 2.73 (3.00)cdef
60 28.06 f 60.83 cde 2.39 (2.17)def
75 29.60ef 51.83de 2.27 (1.58)def
90 27.40f 50.67de 1.89 (0.67)ef
WC 27.86 f 43.75 e 1.70 (0.25)f
IDWFP series (DAT)
15 29.53f 53.83de 2.71 (2.42)cdef
30 28.73 f 67.50 cd 3.39 (2.83)bcde
45 32.60de 62.67cde 3.58 (2.92)bcd
60 35.06cd 91.17ab 4.03 (11.42)abc
75 37.60 bc 91.58 ab 4.19 (12.17)abc
90 40.46 ab 94.67 ab 4.49 (9.25)ab
WFC 42.60a 111.67a 5.21 (13.42)a
LSD (0.05) 3.05 23.32 1.58
CV (%) 5.64 19.84 29.14
Means followed by the same letters within each column are not significantly different. IDWFP = Increasing duration of
weed-free period; IDWP = Increasing duration of weedy period; WFC = Weed-free check; WC = Weedy check; CV =
Coefficient of variation and LSD = Least significant difference at 0.05 probability level
Yield parameters and yield losses estimation

Harvesting of the calyx was undertaken on 134 DAT. Fresh and dry weight of
calyx, seed weight, hundred seeds weight and fresh leaves weight were
significantly (p < 0.05) affected by crop-weed competition durations (Table 4). In
IDWP series, fresh and dry weight of calyx, seed weight and hundred seed weight
highest values were obtained from weeding at 15 to 45 DAT and statistical at par
with the values obtained from the WC and weeding at 75 and 90 DAT. In the
same line, in IDWP the highest fresh leaf weight was obtained from weeding at 15
DAT followed by weeding at 30 DAT. Overall, in IDWP series the weeds were
exerted a severe competition and utilized environmental resources for a longer
period of time that led to low amount of dry biomass accumulation for the crop
yield parameters.
On the other hand, in IDWFP fresh weight of calyx was significantly differed only
by WFC, whereas dry weight of calyx was influenced by the weed free plots for
long period of time (WFC) and weed free for early 90 DAT. Regarding the seed
and fresh leaves weight of the crop plant, the effect of competition durations were
increased from 45 up to 90 DAT including WFC which statistically at par with
each other. The highest hundred seeds weight was obtained from WFC compared
to the rest of treatments in both categories. Overall, in IDWFP series the likely
reasons for the higher weight for each parameter could be the relatively better
[223]
control of weeds and favorable growing conditions to the crop, which were
persuaded more accruement of available dry biomass (Table 4).
The losses that were inflicted due to the different weed competition periods
including WC were considered and compared relative to the yield of WFC for the
two categories of an experiment. The results in lose analysis were exhibited that
higher fresh weight of calyx obtained from weeding at 75 and 90 DAT including
WC. Likewise, dry weight of calyx loss was higher in the WC followed by
weeding at 75 and 90 DAT. Thus, the more calyx losses were come through the
weed-crop completions for prolonged durations regarding the nearby resource
utilization in the growing period (Table 4). Prolonged crop-weed competition was
resulted in reduced dry biomass accumulation which ultimately rendered the
yields of parameters considered and higher yield losses for them. However, yield
losses found out could not be solely attributed to weed competition as some
damages were recorded due to other pests like insect pests and disease.
Table 4. Effect of increasing duration of weedy and weed-free periods on yield parameters and yield loss of roselle at
Arba Minch, 2017
Yield parameters Yield loss (%)

Treatment FWC (Kg/ha) DWC (Kg/ha) SW (Kg/ha) HSW (g) FLW (g) FWC DWC
IDWP series (DAT)
15 222.62bcd 40.39bc 49.88abcd 3.46b 15.91abc 40.15 38.37
30 189.30 bcde 34.85bcd 43.36bcd 3.27bc 14.34bcd 49.11 46.83
45 171.79bcde 33.89bcd 42.86bcd 3.35bc 14.08cd 53.82 48.29
60 162.16cde 29.33bcd 41.74bcd 2.72cd 13.86cd 56.41 55.25
75 121.07e 26.68cd 36.17cd 1.98e 12.58def 67.45 59.30
90 116.57e 26.57cd 32.83cd 2.34de 10.76ef 68.66 59.47
WC 116.46f 22.33d 24.90d 1.97e 10.39f 68.69 65.93
IDWFP series (DAT)
15 157.98de 30.98bcd 39.90bcd 3.23bc 13.76cde 57.53 52.74
30 217.97 bcd 34.89bcd 47.38bcd 3.42bc 14.41bcd 41.45 46.76
45 222.12bcd 34.78bcd 53.38abc 3.46b 15.58abcd 40.29 46.93
60 237.02bcd 38.72bcd 63.01ab 3.49b 16.38abc 36.28 40.92
75 242.10bc 43.09bc 63.20ab 3.64b 16.39abc 32.32 34.26
90 251.76b 45.06b 74.00a 3.79ab 17.31ab 34.92 31.25
WFC 371.99a 65.54a 74.94a 4.47a 18.19a 0.00 0.00
LSD (0.05) 74.99 16.19 26.17 0.72 3.08
CV (%) 24.66 29.41 31.86 13.66 12.68
Means followed by the same letters within each column are not significantly different. DWC= Dry weight of calyx; FWC=
Fresh weight of calyx; FLW= Fresh leaf weight; HSW= Hundred seeds weight; SW= Seed yield weight; IDWFP =
Increasing duration of weed-free period; IDWP = Increasing duration of weedy period; WFC = Weed-free check; WC =
Weedy check; CV = Coefficient of variation and LSD = Least significant difference at 0.05 probability level.
Critical periods of weed control

Estimating the begging and end of the critical period was made based on fresh and
dried calyx yield of roselle at 10% tolerable yield loss level. The beginning of
[224]
CPWC was found to be 15 DAT whereas the end supposed to be 45 DAT.
Accordingly, weeds should be controlled starting on 15 DAT until 45 DAT in the
study areas so as to reduce the yield losses in roselle production.
Discussion
The weed species identified and characterized in the present study were in line
with Adjun (2003), Mahadevan et al (2009) and Ahmed and Salaheldeen (2010)
who reported that the weed species were composed of a wide range of annual,
biennial and perennial broad leaved, grass and sedge weed types. There were
significance variations in weed species composition under different weed
competition periods. The high frequency, abundance, dominance and evenness
values were indicated the high economic importance of these particular weed
species. The diversity and dominancy of weed species were found somewhat
variable from plot to plot indicating the variability of weed management intensity
following the trends of critical period of weed competition. According to
Izquierdo et al (2009) investigation the diversity and dominancy of weed species
were found variable from crop field to field even in the same field with different
places demonstrating the different weed management intensity among crops. The
high abundance, dominancy and diversity values of weed species in roselle can be
accredited to better rainfall distribution and constant irrigation supplement during
shortage of rainfall that make favorable condition for the growth and high
infestation of the weeds that thrive long period under the IDWP series. There were
positive and significance correlations among the weed parameters measured in
that the higher the weed frequency, the higher would be its abundance,
dominancy, evenness and diversity index, and vice versa.
In the study area, weeds were found causing serious damage on the target crop,
roselle. Previous researchers were also reported that weeds pose the most
important threat to roselle production (Mahadevan et al 2009; Ahmed and
Salaheldeen, 2010; Bahaeldeen et al. 2012). To combat the influence of weeds on
agricultural production and reduce yield losses, proper management of weeds
through cultural, herbicidal or combination of different tactics, at the right time is
needed. The principles of integrated weed management (IWM) provided the basis
for mounting most advantageous weed control methods and efficient use of
herbicides (Gaffer 1984; Mahadevan et al., 2009). Several researchers reported
that the critical period of weed control is a fundamental component of an IWM
program (Hall et al., 1992; Halford et al., 2001; Kavaliauskaite and Bobinas,
2006).
An increasing of days of weedy period (IDWP) associated with an increased

number of weed species. Whereas increasing frequency of weeding had lowered
the number of weed species infesting the experimental plots. Under prolonged
[225]
weed free conditions, starting at the late growth stages of the crop the importance
of individual weed species numbers was decreased, whereas under an extended
date of weedy periods the relative important of the number of weed species were
increased. Akobundu and Agyakwa (1987), Adjun (2003), Ahmed and
Salaheldeen (2010) and Peer et al. (2013) reported that an extended weeding time
interval could have resulted from the germination and regeneration capability of
weed species within such a growth time.
In IDWP category, the weeds might have a number of species that can exert a
severe competition and utilize the surrounding resources for a longer period of
time and thus resulting accumulation of more weed dry biomass. Conversely, in
IDWFP series, the weeds come forth and grew after the respective weed free
periods under stress and thus resulted in less buildup of the dry biomass. The
lowest weed density and dry weight were found from weed free for initial 90 DAT
and WFC, while the highest were recorded from weedy for early 90 DAT and
WC. An extended weeding practice did not encourage much weed growth.
Several researchers reported that the weed parameters had influenced different
weed control timings, (Nieto et al. 1968; Knezevic et al. 2002; Knezevic et al.
2003; Norsworthy and Oliveira 2004; Ahmadi et al. 2007; Mahadevan et al 2009;
Ahmed and Salaheldeen 2010; Mukhtar 2012; Getachewu et al. 2017; Imoloame
and Omolaiye 2017).
In the current study, the results exhibited that weed competition and control time
had influenced the roselle growth and yield parameters. The crop fields left
interfered with weeds for more than 30 DAT including WC were considerably
reduced the size of crop parameters, whereas those avoided of weeds during the
initial growth stage after 30 DAT including WFC measured relatively higher crop
parameters. Prolonged weedy periods were lowered the measurable crop
parameters due to higher weed infestation, lower weed control efficiency and
severe competition with roselle for growth resources. Previous researchers
reported that in many crops including roselle keeping the crop with weed infested
condition for long periods resulted in significant growth depression and as a
consequence yield attributes were reduced (Nieto et al. 1968; Knezevic et al.
2002; Knezevic et al. 2003; Norsworthy and Oliveira 2004; Ahmadi et al. 2007;
Mahadevan et al 2009; Ahmed and Salaheldeen 2010; Mukhtar 2012; Getachew et
al. 2017; Imoloame and Omolaiye 2017).
There is a period during the life cycle of a crop when it is most sensitive to the
presence of weeds. Management of weeds supported with critical period of weed
competition is the most suitable approach to optimize weed control applications
(Mahmoodi and Rahimi, 2009). The period in which weeds possibly compete with
[226]
the crop and devoid of more than 10% yield was ranged from 15 to 45 DAT.
Previous researchers reported in different crops, the length of the critical period of
weed control might vary depending on site specific factors and the acceptable
yield losses (Nieto et al. 1968; Swanton and Weise, 1991; Hall et al., 1992;
Rajcan and Swanton, 2001; Knezevic et al. 2002; Evans et al. 2003; Mahmoodi
and Rahimi, 2009; Getachew et al. 2017). The current result agreed with finding
of many researchers like Amador-Ram´ırez (2002), Evans et al. (2003), Uremis et
al. (2009), Norsworthy and Oliveira (2004), who investigated on irrigated and
non-irrigated crops and indicated that the end of CPWC was variable and highly
dependent on density, competitiveness and emergence periodicity of the weed
population.

Even though worldwide consideration of roselle is high, in Ethiopia it is found at
infant stage. In the country it is grown primarily for commercial purpose on
limited areas, and its production is constrained by both biotic and abiotic factors.
Among the biotic factors, weeds are the number one problem in roselle
production. The results of this study indicated that a total of 69 different weed
species identified and the importance of each species determined by scheming
with the frequency, abundance, dominance, evenness and diversity, and
importance values. The dominant families according to the number of weed
species were Poaceae with 17 species followed by Asteraceae 14 species. Among
the key components in the development of IWM, critical period of weed
completion plays a major role for optimum weed control methods and efficient use
of herbicides. The variation of weed density and dry matter weight had the inverse
association with variation in the crop parameters. Under IDWFP the minimum
weed interference at critical weed completion period of roselle had potential
impact on weed growth and development.
The results of this study showed that the highest fresh and dried calyx yields were
obtained from WFC followed by weed free for initial 90 DAT, whereas the lowest
gathered from WC followed by weedy treatments for early 75 and 90 DAT. About
65.93% yield losses due to the highest weed interference were registered from the
WC. The accurate measurement of crop-weed interference period is an
indispensable part for setting up of an effective weed control time in cropping
systems and, hence, can be conceived the primary step to devise weed control
strategy for roselle production. For both fresh and dried calyx maximal yield
preventing more than 10% yield loss, the efficient weed control methods could be
carried out by keeping the crop weed-free between 15 to 45 DAT. In general, the
study suggested that to reduce the yield losses to less than 10% and obtain higher
economic return, weeds must be controlled within 15 to 45 DAT as it has been
[227]
found to be the critical period of weed-crop competition in roselle in the study
areas and elsewhere with similar agro ecology in Ethiopia.
Acknowledgement
The study was financed by Ethiopian Institute of Agricultural Research (EIAR)
through Wondo genet Agricultural Research Center, Ethiopia. The institute and
center are duly acknowledged for funding the study. The authors are also
acknowledged the staff of Arba Minch Agricultural Research Center and Arba
Minch Crop Protection Clinic who contributed in one way or another for the
success of this study.
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[229]
[230]
Herbicidal Control of Chrysanthemum segetum
in Wheat Field at Cheliya District
Shashitu Bedada, Nigusie Hundesa, Gectachewu Bekele,
Tesfaye Abdissa and Tadele Gudeta
EIAR, Ambo Agricultural Research Center. P. O. Box 37. Ambo, Ethiopia
Abstract
A field experiment was conducted between 2017 and 2018 to evaluate effectiveness of
different herbicides against C. segetum in wheat in Cheliya District of West Shewa,
Ethiopia. Experimental treatments were included 2,4-D 720 g/l A.E, Pyroxsulam,
Aminopyralid + Florasulam, Florasulam 75 g/l + Flumetsulam 100g/l,
Pendimethalin, S-Metolachlor, S-metolachlor 290 g/l + atrazine 370 g/l and twice
hand weeding. The experimental design was randomized complete design with three
replications. Data were collected on crop and weed related parameters. Results
obtained indicted that treatments significantly affected wheat plant height, spick
length, biomass yield, grain yield and thousand seed weight. The highest mean grain
yield; 3861.4 kg/ha was obtained from Aminopyralid + Florasulam treated plot followed
by Pyroxsulam treated one; 3729.4 kg/ha. Similarly, the highest crop biomass yield;
9554.2 kg/ha was obtained from Aminopyralid + Florasulam treated plot followed by
Pyroxsulam treated one; 9395.8 kg/ha. Weed count, height and biomass were
significantly influenced by the treatments. The lowest number of the weed plants count
(11.33 per m2) was recorded from twice hand weeding; followed by pendamentalin,
Aminopyralid + Florasulam, s-metolachlor and S-metolachlor 290 g/l + atrazine 370 g/l
2
with 14, 15.33, 22.67 and 26 plants/m , respectively. The lowest C. segetum height
was recorded in twice hand weeding; followed by Pyroxsulam and Florasulam 75 g/l
+ Flumetsulam 100 g/l treated plots. The lowest biomass was recorded in twice hand
weeding followed by Aminopyralid + Florasulam. Then, twice hand weeding,
Aminopyralid + Florasulam and Pyroxsulam were found effective in reducing C.
segetum height and fresh biomass. Therefore, Aminopyralid + Florasulam and
Pyroxsulam found effective in increasing yield and yield components of wheat and
reducing C. segetum count, height and biomass. This indicted that both herbicides
found equally effective for C. segetum management. Hence, both herbicides
recommended as a component of integrated C. segetum management.
Introduction
Chrysanthemum segetum L also called corn marigold is native to Eastern
Mediterranean and North Africa. It is leafy, very vigorous and aggressive annual
weed, and can easily be distinguished by its grayish green toothed leaves; stems
are simple to somewhat branched and golden yellow florets (Howart and
Williams, 1958). It can grow up to 1-meter height depending on the fertility status
of the soil.
Its introduction to Ethiopia and distribution status is not documented, but its wide
infestation has been reported in the highlands of northern Ethiopia (Asres, 2011).
It was observed very recently but invading crop lands at alarming rate in Cheliya
[231]
district of West Shewa Zone (Shashitu, 2019). It has become a serious problem in
major crops in Cheliya district.
In spite of the recognized importance of C. segetum in the Cheliya district almost

no control alternative measures are available. Manual weeding is the only control
practice used by farmers to manage the weed in crop lands. But hand weeding is
costly, as it requires frequent operations owing to the emergency pattern of weed.
Therefore, other alternative management options which can reduce the impact of
this weed is essential for the farmers of the district. Among the alternatives,
chemical could offer a valuable option for the economical control of this
troublesome weed. Thus, the purpose of this study was to screen registered
herbicides with the attributes of economically cheap, persistent but having lesser
residual effects in the environment. Therefore, the objective of the study was to
evaluate chemical herbicides which can best perform against C. segetum in wheat
field at Cheliya district.

Description of study area
The study was conducted at Cheliya district in West Shewa Zone of Oromiya
Regional Administrative State on farmer’s field during the main cropping season
of 2017/18 and 2018/19. In major wheat growing area of the district, testing site
was selected after field observation on the availability of naturally infested heavy
and wide population C. segetum.
Treatments and experimental design

The experiment was laid out in RCBD with three replications. The treatments
were post emergence herbicides namely Agro-2,4-D amine 720 g/l A.E (2,4-D 720
g/l A.E) at 1 litter ha-1, Pallas 45 OD (Pyroxsulam ) at 0.5 liter ha-1, Lanceolat
(Aminopyralid + Florasulam) at 30 gram ha-1, Derby 175 SC (Florasulam 75 g/l +
Flumetsulam 100g/l ) at 80 milliliter ha-1 and pre- emergence herbicides Stomp
455 CS (Pendimethalin) at 3 liter ha-1, Dual Gold 960 EC (S-Metolachlor) at 1-
liter ha-1, Primagram Gold 660 Sc (S-metolachlor 290 g/l + atrazine 370 g/l ) at 3
liter ha-1, two times hand weeding and weedy check as standard and weedy
control, respectively.
Thus, the total number of experimental treatments was eight. Pre-emergence

herbicides were applied within two days after seeding, post emergence herbicide
were applied at 30-35 days after sowing, hand weeding was done at 30-35 and 55-
60 days after sowing, and weedy check plot was kept un weeded. The size of each
plot was 5 m x 4 m (20 m2) with a distance of 1-m between plots and 1.5 m
between blocks left to avoid spray drift from adjacent plots. The bread wheat
[232]
cultivar “Kakaba” was used as experimental crop plant. The land was prepared to
fine tilt with oxen plough. The crop was established in 25 cm distant rows using
150 kg/ha seeding rate and fertilizers DAP and Urea were applied at 100 and 50
kg ha-1 rates, respectively. Urea was split into two that half amount applied at
sowing time while the remaining half applied at tillering stage of the crop.
Land was prepared with oxen plough to fine tilt soil bed which supports easy
contact of the seed with soil. All other agronomic practices required for the crop
were applied according to recommendation given for the crop.
Method of application: The required quantity of the herbicide was calculated and
measured out into manual knapsack sprayer with a pre-determined water volume
for each plot and each herbicide was applied separately and under non windy
conditions.

Data on yield components and yield of wheat like plant height, spick length, crop
biomass, grain yield and thousand seed were recorded. In addition, data on C.
segetum plant like height, density/count after herbicide application and biomass at
harvest was recorded. The data collected were subjected to analysis of variance by
SAS software. Means that showed a significant difference were separated by least
significant difference at 0.05 probability level (SAS, 2004).

Effect of different herbicides on yield and yield components of
wheat:
Plant height and spick length
Wheat plant height showed significant (P <0.05) difference due to herbicides. The
highest plant height (96.57 cm) was recorded from the treatment applied with
Aminopyralid + Florasulam at 30 grams ha-1 at 30-35 days after sowing. The
lowest plant height (89.93 cm) was recorded from a treatment applied with S-
metolachlor 290 g/l + atrazine 370 g/l at 3 liters ha-1 at two days after sowing
(Table 1). Similarly, the effect of treatments had significant (P <0.05) effect on
wheat spick length. The highest spick length (14.1cm) was recorded from the
treatment applied with 2-4, D at 1 litter ha-1 at 30-35 days after sowing. However,
the lowest spick length (12.73 cm) was recorded from the weedy check (Table 1).
Biomass, grain yield and thousand seed weight

Biomass and grain yield of wheat as affected by treatments is presented in Table
1, As shown in the table highly significant effect was observed on wheat biomass.
[233]
Maximum biomass yield (9554.2 kg/ha) was recorded from the plots applied with
Aminopyralid + Florasulam at 30 grams ha-1 at 30-35 days after sowing.
Moreover, higher biomass yield (9395.8 kg/ha), 8816.7 kg/ha) and (8558.3 kg/ha)
were obtained from plots treated with Pyroxsulam at 0.4 liter ha-1 at 30-35 days
after sowing, twice hand weeding at 30-35 and 55-60 days after sowing and
Florasulam 75 g/l+ Flumetsulam 100g/l at 80 milliliters ha-1 at 30-35 days after
sowing, respectively which was statistically similar with Aminopyralid +
Florasulam (Table 1). The increased above ground biomass yield in these
treatments might be due to the crop plants utilized the resources more efficiently
that resulted in higher final crop stand. On the other hand, significantly low crop
biomass yield was recorded from weedy check (5045.8 kg/ha), S-metolachlor 290
g/l + atrazine 370 g/l at 3 liters ha-1 at 2 days after sowing and S-metolachlor at 1
liter ha-1at 2 days after sowing (Table 1). This might be due to severe competition
for growth resources resulting in lower availability of nutrients for the crop thus
causing reduction in number of tillers thereby leading to low crop biomass yield.
Wheat grain yield was significantly (P <0.05) affected by different treatments

(Table 1). Maximum grain yield (3861.4) was recorded from the plots applied
with Aminopyralid + Florasulam at 30 grams ha-1 followed by Pyroxsulam at 0.5
liters ha-1 at 30-35 days after sowing (3729.4 kg/ha). However, lowest wheat
grain yield (1703.3 kg/ha) was recorded from the weedy check. Lower grain yield
(2104.2 kg/ha and 2436.8 kg/ha) were obtained from plots treated with S-
metolachlor 290 g/l + atrazine 370 g/l at 3 liters ha-1 at 2 days after sowing and S-
metolachlor at 1-liter ha-1 respectively which was statistically similar with weedy
check (Table 1). Generally, application of Aminopyralid + Florasulam at 30 grams
ha-1 and Pyroxsulam at 0.5 liters ha-1 at 30-35 days after sowing provided
significantly better grain yield compared with other treatments. The increased
grain yield in these treatments might be due to the proper utilization of moisture,
nutrients, light and apace by the crop under reduced weed competition.
Thousand seeds weight

Thousand seed weight of wheat as affected by different weed control treatments is
presented in Table 1. Highly significant (P <0.05) difference was observed on
wheat thousand grains weight due to the applied treatments. Heavier thousand
seeds weight was recorded from the plots treated with twice hand weeding at 30-
35 and 55-60 days after sowing, Aminopyralid + Florasulam at 30 gram ha-1,
Pyroxsulam at 0.5 liter ha-1 at 30-35 days after sowing, Florasulam 75 g/l +
Flumetsulam 100 g/l at 80 mill liter ha-1 and 2-4, D at 1 liter ha-1 at 30-35 days
after sowing (Table 1). This might be due to the fact that the crop grown under
weed free environment utilized available resources to the maximum benefit
leading to increased seed weight. Moreover, the more and vigorous leaves under
weed free environment might have improved the supply of assimilate to be stored
in the seed, hence, the weight of thousand grain increased. Meanwhile,
[234]
significantly lower thousand seeds weight (36.82, 37.23, 37.45 and 37.67 gram)
was recorded from Pendimethalin at 3 liter ha-1 at 2 DAS, S-metolachlor 290 g/l +
atrazine 370 g/l at 3 liter ha-1 at 2 DAS, S-metolachlor at 1 liter ha-1 at 2 DAS and
weedy check, respectively.
Table 1. Effect of treatments on yield and yield components of wheat at Cheliya district, 2017-2018
Treatments Wheat Wheat spike Wheat Wheat grain 1000

plant height length biomass yield (kg/ha) Seed
(cm) (cm) (kg/ha) Weight (g)
Year 2017 83.5 b 14.28 a 4680.6b 1721.94 b 38.7741
Year 2018 104.28 a 12.60b 10247.2 a 3966.67 a 39.3926
LSD 1.6111 0.3505 508.05 162.21 ns
2-4, D at 1litter ha-1 at 30-35 DAS 95.033ab 14.1a 8466.7b 3216.1c 39.8333a
Pyroxsulam 0.5 litter ha-1 at 30-35 DAS 91.683bc 13.4750abcd 9395.8 ab 3729.4ab 40.5833a
Aminopyralid+ Florasulam at 30 grams 96.575a 13.4833abc 9554.2a 3861.4 a 40.7667a
ha -1
Florasulam 75 g/l + Flumetsulam 100 g/l 94.208ab 13.55abc 8558.3ab 3345.9c 40.2250a
at 80 mill liter ha-1 at30-35 DAS
Pendimethalin at 3 liters ha-1 at 2 DAS 95.058ab 12.9917cd 7116.7c 2436.8d 36.825b
S-Metolachlor at 1-liter ha-1 at 2 DAS 94.908ab 13.5abc 4783.3d 1721e 37.45b
S-metolachlor 290 g/l+ atrazine 370 g/l 89.933c 13.8750ab 5437.5d 2104.2d 37.2333b
at 3 liters ha-1 at 2 DAS
Twice HW at 30-35 and 55-60 DAS 93.225abc 13.2667 bcd 8816.7ab 3480.6bc 41.1583 a
Weedy check 94.367ab 12.7333d 5045.8d 1703.3e 37.6750b
CV 4.49 6.82 17.79 14.91 5.76
LSD 3.4178 0.7435 1077.7 344.09 1.8256
Note: CV= Coefficient of Variation, LSD=Least significance difference, DAS=Days after sowing, Means followed
by the same letter within a column are not significantly different at 0.05p
Effect of different herbicides on Chrysanthemum segetum:

Count
Maximum C. segethum number was found in the weedy check. The highest
number of weed count was obtained from Pyroxsulam 0.5 litter ha-1 at 30-35 DAS
followed by Florasulam 75 g/l + Flumetsulam 100 g/l at 80 mill liters ha-1 at 30-35
DAS and 2-4,D at 1-liter ha-1 at 30-35 DAS, respectively which was statistically
similar with the weedy check. The lowest number of weed (11.33 and 14 plant
per m2) were counted in twice hand weeding at 30-35 and 55-60 days after sowing
and Pendimethalin at 3 liters ha-1 at two DAS treated plot, respectively. But non
significantly Aminopyralid + Florasulam at 30 gram ha-1 at 30-35 DAS, S-
metolachlor at 1 liter ha-1 at 2 DAS, S-metolachlor 290 g/l + atrazine 370 g/l at 3
liter ha-1 at two DAS and 2-4,D at 1-liter ha-1 at 30-35 DAS showed lower number
of C. segetum count (Table 2). This indicted that twice hand weeding at 30-35 and
55-60 days after sowing followed by Aminopyralid + Florasulam at 30 gram ha-1
at 30-35 DAS was effective in lowering weed count compared with other
treatments. As weed population decreases in a crop-weed environment, the
competitive power of crop increases due to the fact that the crop gets advantage of
absorbing the available limited nutrient and moisture. Moreover, the crop can get
[235]
ample space and light which are required for the growth and development of the
crop plants.
Height
In the present study, treatments showed significant (p <0.05) difference on C.
segetum height (Table 2). Taller heights were recorded from the weedy check, S-
metolachlor at 1 liter ha-1 at 2 DAS, Pendimethalin at 3 liter ha-1 at 2 DAS, and 2-
4, D at 1 litter ha-1 at 30-35 DAS treated plot (Table 2). The shortest height was
recorded from the plots hand weeded twice at 30-35 and 55-60 days after sowing.
Shorter height was recorded from the plots treated with Pyroxsulam at 0.5 liter ha-
1
and Florasulam 75 g/l + Flumetsulam 100 g/l at 80 mill liter ha-1 at 30-35 DAS,
which was significantly at par with Aminopyralid + Florasulam at 30 gram ha-1 at
30-35 DAS and S-metolachlor 290 g/l + atrazine 370 g/l at 3 liter ha-1 at two DAS
(Table 2). This indicted that, twice hand weeding at 30-35 and 55-60 days after
sowing followed by Pyroxsulam at 0.5 liter ha-1 and Florasulam 75 g/l +
Flumetsulam 100 g/l at 80 mill liters ha-1 at 30-35 DAS has affected C. height as
compared to other treatments.
Biomass
Biomass of C. segetum was showed significant (P <0.05) differences due to
treatments. Highest biomass recorded by twice hand weeding at 30-35 and 55-60
days after sowing and Aminopyralid + Florasulam at 30 grams ha-1 at 30-35 DAS.
The lower biomass (4162.5 kg/ha, 4062.5 kg/ha, 3925 kg/ha and 3479.2 kg/ha)
were recorded from S-metolachlor 290 g/l + atrazine 370 g/l at 3 liter ha-1 at 2
DAS, 2-4,D at 1-liter ha-1 at 30-35 DAS, S-metolachlor at 1 liter ha-1 at two DAS
and weedy check, respectively. But non significantly, Pendimethalin at 3 liter ha-1
at two DAS, Pyroxsulam 0.5 litter ha-1 at 30-35 DAS and Florasulam 75 g/l +
Flumetsulam 100 g/l at 80 mill liters ha-1 at 30-35 DAS showed higher C. segetum
biomass (Table 2). The higher weed biomass might be due to higher weed density
that provided an opportunity to the weeds to compete vigorously for nutrients,
space, light, water and carbondioxaide resulting in higher biomass production.
However, the lowest C. segetum biomass (1316.7 kg/ha) was recorded from twice
hand weeding at 30-35 and 55-60 days after sowing; followed by Aminopyralid +
Florasulam at 30 grams ha-1 at 30-35 DAS gave 1775 kg/ha. This indicated that C.
segetum biomass was highly reduced in twice hand weeding at 30-35 days after
sowing and Aminopyralid + Florasulam at 30 grams ha-1 at 30-35 DAS treated
plot as compared to other treatments (Table 2).
[236]
Table 2. Effects of herbicides on C. segetum weed count /m2, height and biomass, 2017-2018
Fresh
Treatments Count /m2 Height (cm) biomass (kg/ha)
Year 2017 38.926 a 46.243 b 4475.0 a
Year 2018 16.148 b 81.906 a 1650 b
LSD 10.822 4.2688 795.56
2-4,D at 1litter ha-1 at 30-35 DAS 29.33 abcd 70.842ab 4062.5a
Pyroxsulam 0.5 litter ha-1 at 30-35 DAS 42.50 ab 58.983c 3108.3ab
Aminopyralid+ Florasulam at 30 grams 15.33 cd 62.742bc 1775bc
ha-1 at 30-35 DAS
Florasulam 75 g/l+ Flumetsulam 100g/l at 37.33 abc 61.358c 2491.7abc
80 mil-liter ha-1 at30-35 DAS
Pendimethalin at 3 liters ha-1 at 2 DAS 14.00d 73.650a 3241.7ab
S-metolachlor at 1-liter ha-1 at 2 DAS 22.67 bcd 73.783a 3925.0a
S-metolachlor 290g/l+ atrazine 370 g/l at 26.00 bcd 63.133bc 4162.5a
3 liters ha-1 at 2 DAS
Twice hand weed at 30-35 and 55-60 11.33 d 35.908d 1316.7c
DAS
Weedy check 49.33 a 76.267a 3479.2a
CV 71.55 17.41 67.9
LSD 22.957 9.0554 1687.6
Note: CV= Coefficient of Variation, LSD=Least significance difference, DAS=Days after sowing, Means followed
by the same letter within a column are not significantly different at 0.05p

From the present field experiment it can be concluded that application of the
herbicides Aminopyralid + Florasulam at 30 grams ha-1 at 30-35 days after sowing
and Pyroxsulam at 0.5 liters ha-1 significantly reduced the C. segetum height and
fresh biomass. These treatments also significantly increased yield and yield
components of wheat. Hence, these two herbicides can be recommended to be
used as a component of integrated C. segetum management.
Reference
Asmare Yallew, Alemu Hailye, Alemayehu Assefa, Melkamu Ayallew, Tessema Zewdu, Birhane
Assayehegn and Halima Hassen. 1995. Barley production practice in Gojam and Gonder. In:
Chilot Yirga, Fekadu Alemayehu and Woldeyesus Sinebo (eds.). Barley-based fanning
systems in the highlands of Ethiopia. Ethiopian Agricultural Research Organization. Addis
Ababa, Ethiopia
C1MMYT. 1988. From Agronomic Data to Farmers Recommendations: An Economic Training
Manual. Completely revised edition. Mexico, D.F.
Howart SE and JT Williams.1955. Chrysanthemum segetum L. In: Biological flora of the British
Isles. Department of Biology, Oxford High School, Ilford, England.
Yeshanew Ashagrie, Alemayehu Asefa and Minale Liben. 1993. Screening of herbicides for the
control of broad leaf weeds in wheat. Adet Research Center Progress Report, April 1992-
March 1993, IAR, Addis Abeba.
[237]
[238]
Herbicides Efficacy on Weeds of Upland Rice
(Oryza sativa L.) in Fogera, Ethiopia
Muluadam Berhan1, Desalegn Yalew1, Tekalgn Zeleke2 and Esuyawukal Demis1
1Ethiopian Institute of Agricultural Research (EIAR), Fogera National Rice Research and Training Center (FNRRTC),
2EIAR, Ambo Agricultural Research Center
Abstract
Rice (Oryza sativa L.) is by far the most important stable food crop for over half of
the world population for both developing and developed nations, where its
consumption has increased considerably due to food diversification and its calorie
demand. However, its productivity and quality are highly limited by the occurrence of
diseases, insect pests and high weed infestations. Weeds can reduce rice yield by over
30% and this makes farmers to incur 70% of their cost to manage it because of the
availability of ample moisture within the rice field and long period of infestation. The
objective of this study was to evaluate the efficacy of broadleaf and grass weed
targeted herbicides of upland rice and recommend the most effective ones. An
experiment was conducted at Fogera National Rice Research and Training Center
experimental station for two consecutive years (2017-2019) using Nerica 4 variety. It
was carried out in aerobic soil condition. Eight treatments, two pre-emergences
herbicides (S-metolachlor 290 g/l + atrazine 370 g/l and S-metolachlor 960 g/l), four
post-emergence herbicides (Bispyribac-sodium 10% SC, Pyroxsulam 45 gm/l,
Lodosulfuron-methyl-sodium 7.5 g/l + Mesosulfuron methyl 7.5 g/l and 2,4-D
dichlorophenoxy acetic acid 720 g/l acid), two times manual weeding and control
check (weedy check) were used for evaluation. The treatments were arranged in
randomized complete block design with three replications. Weed population and the
crop agronomic parameters data were recorded. The result revealed that post-
emergence herbicides were highly effective over pre-emergence herbicides in
managing upland rice weeds. Twice manual hand weeding followed by Bispyribac-
sodium 10% EC and Pyroxsulam 45 gm/l treatments gave highest grain yield; 3243.4
and 3063.6 kg ha-1 respectively. Therefore, it can be concluded that Bispyribac-
sodium 10% EC followed by Pyroxsulam 45 gm/l could be recommended for upland
weeds control as component of integrated management system.
Introduction
Rice (Oryza sativa L.) is an important staple food for the people of Asian
countries. It provides 35-80% of total calorie requirement and it occupies an
enviable prime place among the food crops cultivated around the world, it is
grown in an area of 167 million hectares with a production of 678 million tons
(Anon, 2011). Rice is the second most common staple food in East Africa, after
corn. By 2014, annual consumption had reached 1.8 million metric tons.
Production, however, stood at 1.25 million metric tons (FAO, 2014).
Among the target commodities that have received due emphasis in promotion of
agricultural production, rice is expected to contribute to ensure food security in
[239]
Ethiopia. Even though, it is a recent introduction to the country, rice is considered
as the “Millennium crop” and has shown promising adoption and advantage to be
among the major crops that can immensely contribute towards ensuring food
security in Ethiopia. The country has vast suitable ecologies for rice production
along with the possibility of growing it where other food crops do not do well.
Based on geographic information system techniques and agro-ecological
requirements of rice, the potential rain-fed rice production area in Ethiopia is
estimated to be about thirty million hectares (Teshome and Dawit, 2011). Rice is
compatible with various traditional food recipes like bread, soup, “enjera”, and
local beverages like “tela” and “areki”. The country has also a comparative
advantage of producing rice due to the availability of huge and cheap rural labor as
the crop is labor intensive. The importance of rice as a food security crop, source
of income and employment opportunity due to its relative high productivity as
compared to other cereals is recognized by farmers as well as private investors
who frequently request for improved varieties for different agroecologies
(Teshome and Dawit, 2011) with better diseases, insect pests and weed
management.
Rice production is constrained by several factors: technological, bio-physical,

socioeconomic, institutional and financial limitations. Of these constraints, biotic
and abiotic factors are considered to be the most important rice production
limiting ones. Abiotic stresses include: variable rainfall, with drought and flooding
occurring in the same season; poorly-drained soils of the coastal lowlands, and
alkalinity in dry areas. Biotic stresses include: weeds, insect pests (stem borers
such as stalk-eyed fly, African rice gall midge and rice bugs), diseases (blast,
brown spot, and viral diseases), rats and birds (Hadush, 2015).
Weeds have been identified as leading constraint of rice production in Ethiopia.

The infestation of weeds is mostly high in upland rice which has paramount
importance as compare to lowland rice potential areas in Ethiopia. Weed
infestation and competition is more severe in direct seeded aerobic rice as
compared to transplanted rice, because of the reasons viz., land is exposed during
initial crop growth stages, aerobic soil conditions, dry tillage practices and
alternate wetting and drying make the conditions more conducive for germination
and growth of weeds. Direct seeded aerobic rice is highly infested with grasses,
broad leaved weeds and sedges (Madhukumar, 2011). Farmers have incurred 70%
of their cost for rice weed management in Fogera, Dera and Libokemkem districts
and the weeding is done by hand weeding which ranges from 3 to 5 times in a
season (unpublished). The upland rice production is highly reduced by weed
infestation since the weed’s species are not suppressed by water and compete
throughout the crop growth period.
[240]
Unlike transplanted rice early and timely weed control is essential in aerobic rice;
otherwise yield loss can reach as high as 82% due to crop weed competition
(Thimme, 2006). Among the agronomic practices to improve the yield levels of
aerobic rice, timely weed control plays an important role. Traditional method of
weed management practices are widely adopted for control of weeds in aerobic
rice. These practices are tedious, time consuming, labor intensive, costly and not
possible to practice over an extensive area. Further, due to high labor cost as a
result of limited income, rapid industrialization and urbanization, traditional weed
management practices are becoming impractical.
Addition of safe and effective herbicide in integrated weed management is

believed to increase efficiency in weed management (Madhukumar, 2011). Access
to herbicides for rice weed management is very less in the country due to
unavailability of registered herbicides specifically for rice weed management.
However, many herbicides have been registered for wheat, barley and tef weed
management in Ethiopia. Because of similar morphological character of rice with
the above-mentioned cereal crops, the registered herbicides may work for weed
management in upland rice. Therefore, the major objective of this study was to
evaluate some pre and post emergence herbicides and identify the most effective
herbicide that can be integrated with other management practices in managing
upland rice weeds.

Description of the study area:
Fogera district is one of the 151 districts of the Amhara Regional Administrative
State found in South Gondar Zone. It is situated at 11º46 to 11º59 latitude North
and 37º33 to 37º52 longitude East. Altitude ranges from 1774 to 2410 meters
above sea level (m asl) and is predominantly classified as mid altitude or 'Woina-
Dega'. Based on the existing digital data, mean annual rainfall was 1216 mm
ranging from 1103 to 1336 mm from both the short (March and April) and long
rains (June to September). Farmers depend on long rainy season 'Kiremt' for crop
production (Kibebew et al., 2011).
The study was conducted at Fogera National Rice Research and Training Center
(FNRRTC) South Gondar zone of Amhara region, Ethiopia, located 610 km away
from the capital city of Addis Ababa and about 55 km north of Bahir Dar; the
capital city of Amhara Regional state. Geographically the experimental site is
located at latitude of 11° 58′ N and longitude of 37° 41′ E with an altitude of
1817m above sea level and it receives average annual rainfall of 1230 mm. Mean
minimum and maximum temperature of the area is 12 and 280C, respectively. The
soil is brown clay (vertisol) rich in underground water (Dessie, 2015).
[241]
An experiment was conducted in aerobic rice ecosystem under rain fed condition
using Nerica 4 rice variety. The treatments (Table 1) were arranged in randomized
complete block design with three replications. The plot size was 3 meters with 2.4
meter (7.2 m2) and the block size was 28.5 meter with 11 meters (313.5 m2).
Spacings of 0.5 m and 1 m were used between plots and blocks, respectively and
the space between rows was 20 cm. In this experiment, eight treatments were
evaluated, of which four were post emergence herbicides, the other two were pre
mergence herbicides and two times hand weeding as well as weedy check were
included as standard and control checks. Randomization was held independently
for each replication by which treatments were assigned completely at random as
described Gomez and Gome (1984) and Poduska (2008). The pre emergence
herbicides were Primagram gold 660 SC (S-metolachlor 290 g/l + atrazine 370 g/l)
and Dual gold 960 EC (S-metolachlor 960 g/l) whereas the post emergence
herbicides were Pride 100 SC (Bispyribac-sodium 10% SC), pallas 45 OD
(Pyroxsulam 45 gm/l), Atlantis OD 37.5 (Lodosulfuron-methyl-sodium 7.5 g/l +
Mesosulfuron methyl 7.5 g/l) and Agro 2,4-D amin 720 SL (2,4-D
dichlorophenoxy acetic acid 720 g/l acid). In addition, two times hand weeding,
farmers practice; and weedy control were used for comparison with the above
herbicides.
Table 1. Type of herbicides used, their time of application, dose and target weeds
Herbicides
methods Dose of herbicide Time of
Trade name Common name application Target weeds & water/ ha Application
Pride 100 SC Bispyribac-sodium 10 Post emergence Broad leaf in wheat 250 ml + 200 lit H2O At 5 – 6 leaf stage
% SC of the crop
Primagram S- Metolachlor 290g/ l + Pre-emergence Broadleaf and grass 3.5lit+300 lit H2O After one day of
Gold 660 SC atrazine 370 g/l weeds in maize and sowing
sugarcane
Pallas 45 OD Pyroxsulam 45 gm/l Post emergence Grass weeds, annual 500ml+220 lit H2O 30 to 35 days after
broad leaf weeds on seedling
wheat and tef emergence
Dual Gold 960 S-metolachlor 960 g/l Pre-emergence Broad leaf weeds in 1.3lit+200 lit After one day of
EC haricot bean sowing
Atlantis OD Lodosulfuron-methyl- Post emergence Grass & broad leaf 1lit+120lit H2O At 2-4 leaves
37.5 sodium 7.5 g/l+ weeds in wheat stage of the
Mesosulfuron methyl weeds
7.5 g/l
Two times Farmer practice All (grass and At tillering and panicle
hand weeding broad leaves) nitiation stage
Agro 2,4-D 2,4-D dichlorophenoxy Post emergence Broad leaves 1.5 L + 200 L H2O At 15-25 cm
amin 720 SL acetic acid 720 g/l acid height of the crop
Untreated Unsprayed
check
The study was conducted during the rainy season. The land was prepared a head of
the onset of rain and it was ploughed more than three times. The Nerica 4 variety
[242]
with the recommended seed rate of 80 kilograms per hectare was used.
Furthermore, 60.5 kilogram per hectare Nitrogen, phosphorus and sulfur (NPS
with grade of 19N-38P-7S) fertilizer and 130 kilograms per hectare recommended
granular urea fertilizer rates were applied. All plots were received 43.56 gram of
Nitrogen, phosphorus and sulfur (NPS with grade of 19N-38P-7S) and 93.6 gram
of granular urea fertilizer rates per plot. The seed was sown by drilling in a row
and the fertilizers were also applied as soon as the seed was drilled.
The pre emergence herbicides were applied one day after seed sowing in between
each row. A line was made by hoe in the space between rows and the prepared
herbicides solution was sprayed on it. The herbicides were applied by hand
sprayers by measuring the distance by tape meter. The post-emergence herbicides
were applied based on the recommendation. All herbicides were sprayed once in
the season and two times hand weeding was carried out at the initial of tillering
stage (30-35 days after sowing) and the end of panicle initiation or start of heading
stage (65-70 days after sowing).

Data on weed species, agronomic and phenological parameter were collected after
both the pre and post emergence herbicides were applied. The number of a weed
species in a quadrat was counted and recorded at three spots per each plot before
and after chemical spray. The weed species frequency, dominance and abundance
were calculated based on the collected count data in addition, days to emergency,
days to heading, plant height, panicle length, number of spikes per plant, number
of fertile tillers per plant, number of filled grains per panicle, number of unfilled
grains per panicle, thousand grain weight, grain yield and grain moisture content
data were recorded.
Moreover, the weed species sample plants were documented and preserved. It was
held at the crop tillering and panicle initiation stages at which the intensity, type of
weed species and infestation was clearly variable. Other parameters were taken at
critical time of each data supposed to be collected.
Weed population
The weed population count of grasses, broad leaved weeds and sedges, and their
total weed population was recorded at 35 days after sowing (DAS) and at harvest
time from 0.5 m X 0.5 m size quadrat permanently marked in gross plot area and it
was converted to square meter.
Fresh and dry matter of weeds

The fresh and dry matter biomass of all weeds were recorded in each plot with a
sample area of 0.25 m2 (0.5m* 0.5m) at 35 days after sowing and at maturity stage
[243]
of the crop which was 130 days after sowing. Fresh weeds were collected from the
specified plot sample area and weighed as soon as collected whereas the dry
matter was weighed after the weeds had been dried through solar radiation for
three weeks.
Weed index
Weed index denotes the reduction of crop yield due to crop weed competition as
compared to weed free plots. It was calculated using the following formula as Gill
and Vijaya kumar (1969).
Weed index (%) = x 100

Where, X = grain yield of weed free plot; Y = grain yield from treatment for
which weed index is to be worked out
Crop toxicity ratings

The visual observations for toxicity ratings were taken at 14 and 35 days after the
herbicides application. The scores were taken using 0 to 10 scales by comparing
crop tolerance to a particular herbicide treatment and weedy check plot. It was
carried out as per European Weed Research Society (EWRS) classification scale
as described below in Table 2 (Anon, 1981).
Table 2. Scale of crop toxicity ratings (EWRS classification scale)

No. Level of injury Scale Grade
1 None 0.00 None
2 Slight discoloration, few plants lost, stunted growth, injury 1.0-3.0 Slight
more pronoun but effect short period
3 Moderate injury, recovery possible, non-recoverable injury 4.0-6.0 Moderate
4 Severe injury with some plants lost 7.0-9.0 Severe
5 Complete destruction 10.0 Complete
The collected data were analyzed using version 9.0 Statistical Analysis Software
(SAS) and means least significant difference separated at 5% probability level.

The result of the study revealed that the post emergence herbicides were by far
better in controlling upland rice weeds (Table 3). There was significance
difference among the two types of herbicides in inhibiting rice weeds growth and
have direct effect on yield and yield related characteristics of the crop plant. The
highest grain yield was obtained from the plot that was weeded twice manually
followed by Bispyribac-sodium 10% EC and Pyroxsulam 45 gm/l, 3243.4, 3063.6
and 2933.2 kg ha-1, respectively. This was the result of good weed control which
emerged early in the seedling and tillering as well as panicle initiation to booting
stages. On the other hand, plots treated with Bispyribac-sodium 10% EC (post
[244]
emergence) herbicide provided the best biological yield even if it has provided
ideal comparative grain yield. Two times hand weeding and Pyroxsulam 45 gm/l
also gave preferable crop biomass yield as compared to the other treatments.
On the contrary, pre emergence herbicides were not effective since they had
phytotoxic effect both on the crop and weed seeds. Almost all herbicides were not
effective in controlling weeds emerged after the standing water has been dried.
The number of tillers per square meter was high for those plots sprayed with pre-
emergence herbicides (S-metolachlor 290 g/l + atrazine 370 g/l and S-metolachlor
960 g/l). This was due to the fact that undamaged rice plants had more space and
there was no competition for nutrients to produce high number of tillers because of
the rice and weeds seeds were seriously affected by the toxicity of the herbicides.
Plant height and panicle length of the two pre emergence herbicides were also
greater than plots treated with post emergence herbicides due to the reason
mentioned above (see the numerical data below).
The weedy control showed better percent of harvest index over herbicides treated
and manually weeded plots. It was as a result of high weed competition for
nutrients, water, and no aeration in the plot and this led to stunted plant growth.
The time of application for pre germination herbicides was not appropriate as it
observed and approved in three parameters (time of application, row interval and
dose of the chemical) separately. The observation showed that application of
herbicides in four rows interval was relatively effective to control weeds
regardless of the crop damage. In general, the study showed that two times hand
weeding was the most effective rice weed management option and
environmentally ideal to the aquatic life in the swampy area.
[245]
Table 3. Effect of pre and post emergence herbicides on upland rice agronomic parameters
Treatments TPP PH (cm) PL (cm) NSPP DM TGW (g) BY (kg/ha) GY (Kg/ha) HI (%)
Bispyribac-sodium 10 % EC 4.13b 55.03c 15.03b 10.53abc 131.67b 25.84 6481.5a 3063.6a 47.40abc
S-metolachlor 290g/l + atrazine 370 g/l 10.97a 62.27ab 18.03a 11.56a 144.33a 25.06 4175.9c 1530.5c 37.94c
Pyroxsulam 45gm/l 4.77b 56.08 14.66b 8.50d 133.33b 24.94 6101.9a 2933.2a 48.68abc
S-metolachlor 960g/l 9.43a 66.27 17.66a 10.83ab 142.00a 25.03 3486.1c 1539.2c 43.26bc
Lodosulfuron-methyl-sodium 7.5g/l + Mesosulfuron 3.87b 53.67 14.43b 9.00cd 134.83b 26.92 5375.0b 2289.3b 43.40bc
methyl 7.5g/l
Two times hand weeding 4.7b 56.10c 9.20 bcd 14.60b 133.33b 24.92 6210.1a 3243.4a 52.60ab
2,4-D dichlorophenoxy acetic acid 720 g/l acid 3.63b 57.23bc 15.56b 9.40 bcd 135.17b 26.63 4953.7b 2112.2b 42.90bc
Untreated check 3.33b 61.67ab 15.80b 8.48d 131.67b 25.80 4067.9c 2194.6b 54.93a
LSD (p<0.05) 2.51 5.05 1.49 1.83 4.34 2.69 697.7 344.6 11.04
CV (%) 38.32 7.36 8.11 16.17 2.74 8.96 11.68 12.46 20.35
Total mean 5.60 58.54 20.60 9.68 135.80 25.64 5106.51 2363.25 46.39
P-value 0.0001 0.0001 0.0001 0.0092 0.0001 0.66 0.0001 0.0001 0.066
Note: PH: plant height (cm), PL: Plant length (cm), TPP: Number of tillers per plant, NSPP: Number of spikelet per panicle, TGW: Thousand grain weight (gram), BY: biological yield, GY:
Grain yield in ton/ha, HI: Harvest index, LSD: Least significant difference at 5% significance level, CV: Coefficient of variation in percent
Weed population
As it is described below the weed population was minimum in plots treated with
S-metolachlor 290 g/l + atrazine 370 g/l active ingredient (a.i.), S-metolachlor 960
g/l a.i and Lodosulfuron-methyl-sodium 7.5 g/l + Mesosulfuron methyl 7.5 g/l a.i,
respectively. On the other hand, the weed population was high in plots sprayed
with Pyroxsulam 45 gm/l a.i (326.67), control check (315.33) and Bispyribac-
sodium 10% EC a.i (312) consecutively. This indicated that pre emergence
herbicides were by far better than post emergence herbicides in reducing weed
population (Table 4).
However, the weed population before and after post emergence herbicides varied
due to difference in the moisture status and natural characteristics of the weed
species (Tables 4 and 5). Sedge type of weeds, for instance infested the plots early
in the season where as Echinochloa colonum, Dactyloctenium aegyptium and
Digitaria marginata infested later in the season. Furthermore, the weed types and
number varied from location to location depending on the weed seed bank of the
soil and also on the farm management practices during the previous cropping
seasons.
Table 4. The effect of herbicides on weed population
Trade name Common name Weed density (m-2)

Pride 100 SC Bispyribac-sodium 10% EC 312.00
Primagram gold 660 SC S-metolachlor 290 g/l + atrazine 370 g/l 92.67
Pallas 45 OD Pyroxsulam 45 gm/l 326.67
Dual Gold 960 EC S-metolachlor 960 g/l 159.33
Atlantis OD 37.5 Lodosulfuron-methyl-sodium 7.5 g/l + Mesosulfuron
methyl 7.5 g/l 178.67
Manual weeding Two times hand weeding 245.33
Agro 2,4-D amin 720 SL 2,4-D dichlorophenoxy acetic acid 720 g/l acid 271.33
Untreated check 315.33
Table 5. Common weed species observed and collected during the experiment
Local name Scientific name

Wuha anqur Commelina diffusa
Guaro atifa Ageratum conyzoides
Wonberet Launaea cortuna
Getetie Ipomoea aquatica
Korach Bolboschoenus maritimus (L.)
Chanfa Cyperus postulatus
Gicha Cyperus esculentus
Zuraha Oryza barthii
Molalie Sackiolepis africana
Dry matter production and weed index

The dry crop biomass production was high in plots that had been sprayed with
bispyribac-sodium 10% EC followed by two times hand weeding and pyroxsulam
[247]
45 gm/l; 6481.5, 6210.1 and 6101.9 kg ha-1, respectively (Table 6, Figure 1).
Whereas the pre-emergence herbicides had negative impact for the dry matter
productivity as shown below.
Table 6. The effect of herbicides on rice biological yield and yield reduction due to weed competition
Treatments Dry matter yield Weed index (%)

Bispyribac-sodium 10 % EC 6481.5 5.54
S-metolachlor 290 g/l + atrazine 370 g/l 4175.9 52.81
Pyroxsulam 45 gm/l 6101.9 9.56
S-metolachlor 960 g/l 3486.1 52.54
Lodosulfuron-methyl-sodium 7.5g/l + 5375.0 29.42
Mesosulfuron methyl 7.5 g/l
Twice hand weeding 6210.1 0
2,4-D dichlorophenoxy acetic acid 720 g/l acid 4953.7 34.87
Untreated check 4067.9 32.34
Figure 1. The impact of herbicides on Nerica 4 rice variety grain yield and plant dry matter
However, the maximum grain yield reduction 52.81 and 52.54 percents due to
weed competition were recorded from plots treated with pre-emergence
herbicides; S-metolachlor 290 g/l and S-metolachlor 960 g/l, respectively. This
study has revealed that the two pre-emergence herbicides had great yield reduction
impact since they seriously damaged the crop and weeds seeds in the soil before
germination. This led to the occurrence of few rice plants and high weed
infestation in the experimental plots. On the other hand, two times hand weeding
which had the lowest grain yield reduction was considered as weed free treatment.
[248]
Moreover, bispyribac-sodium 10% EC and pyroxsulam 45 gm/l post-emergence
herbicides had low effect on weed index in percent.
Crop toxicity ratings

The pre-emergence herbicides were not toxic to rice plants since they were applied
in the soil and their residual effect was not observed on the crop. But the post
emergence herbicides had various ratings of toxicity (Table 7).
Table 7. The toxicity levels of pre and post-emergence herbicides on rice plants
Treatments Toxicity rating Grade

Bispyribac-sodium 10 % EC 1.00-3.00 Slight
S-metolachlor 290g/l + atrazine 370 g/l 0.00 None
Pyroxsulam 45 gm/l 4.00-6.00 Moderate
S-metolachlor 960 g/l 0.00 None
Lodosulfuron-methyl-sodium 7.5 g/l + Mesosulfuron methyl 7.5 g/l 7.00-9.00 Severe
Twice hand weeding 0.00 None
2,4-Dichlorophenoxy acetic acid 720 g/l acid 4.00-6.00 Moderate
Untreated check 0.00 None
Out of which, Lodosulfuron-methyl-sodium 7.5 g/l + Mesosulfuron methyl 7.5 g/l

had severe toxicity on the rice plants growth especially when there was moisture
stress in the soil. In addition, Pyroxsulam 45 gm/l and 2, 4-Dichlorophenoxy acetic
acid 720 g/l acid had been moderately limited the crop plants growth (a scale of 4
to 6) and the most critical period of vegetative growth was inhibited and stunted
due to its effect. As a result, the yield and yield components were reduced as per
its dose and time of application.
Partial budget analysis

As farmers attempt to evaluate the economic benefits of weed management
practices, partial budget analysis was done to identify the most economical
treatments. Yield from experimental plots was adjusted downward by 10% for
management and plot size difference, to reflect the difference between the
experimental yield and the yield that farmers could expect from the same
treatment (CIMMYT, 1988).
Two years’ average market grain price of rice (14 ETB per kg) and labor valued at
ETB 50 per person day were used. The rice field management and the fertilizers
application were the same for all treatments (28 person-days per hectare and the N
and P fertilizers price were 11 ETB per kg and 12.6 ETB kg-1, respectively. The
result of the partial budget analysis is described in (Table 8). The economic
analysis revealed that the highest net benefit was obtained from the application of
post emergence herbicides Bispyribac-sodium 10% EC (birr 29292.34 ha-1)
followed by Pyroxsulam 45 gm/l (27886.16-birr ha-1) and two times hand weeding
(26109.66-birr ha-1).
[249]
Table 8. Partial budget analysis and marginal rate of return of herbicides efficacy on weeds of Upland rice (Oryza sativa L.) at Fogera (CIMMYT, 1988).
Inputs GY AGY CPR GFB TVIC /ha NB (Birr/ha) CNB CTVIC MRR
(kg/ha) kg/ha (ETB) (ETB/ha) (Birr/ha) (Birr/ha)
Bispyribac-sodium 10 % EC 3066.6 2759.94 11.00 30359.34 1067 29292.34 7565.80 1067 709
S-metolachlor 290g/l + atrazine 370 g/l 1530.5 1377.45 11.00 15151.95 2127.5 13024.45 -8702.09 2127.5 -409
Pyroxsulam 45gm/l 2933.2 2639.88 11.00 29038.68 1152.5 27886.18 6159.64 1152.5 534
S-metolachlor 960g/l 1539.2 1385.28 11.00 15238.08 1394.5 13843.58 -7882.96 1394.5 -565
Lodosulfuron-methyl-sodium 7.5g/l+ 2289.3 2060.37 11.00 22664.07 1065.00 21599.07 -127.47 1065.00 -12
Mesosulfuron methyl 7.5 g/l
Two times hand weeding 3243.4 2919.06 11.00 32109.66 6000.00 26109.66 4383.12 6000.00 73
2,4-D dichlorophenoxy acetic acid 720 g/l acid 2112.2 1900.98 11.00 20910.78 1397.50 19513.28 -2213.26 1397.50 -158
Untreated check 2194.6 1975.14 11.00 21726.54 0.00 21726.54 0.00 0.00 0.00
Note: GY: grain yield, AGY: Adjusted grain yield, CPR (ETB): current price of rice in Ethiopian birr, GFB: gross farm gets benefit, TVIC/Ha: total variable input cost per hectare, NB
Birr/Ha: net benefit in birr per hectare, CNB Birr/Ha: change in net benefit in birr per hectare, CTVIC Birr/Ha: change in total variable input cost in birr per hectare, MRR: marginal rate of
return (%)
As the marginal rate of return showed that investing one birr for the spray of
bispyribac-sodium 10% EC and pyroxsulam 45 gm/l to control weeds, yield
reduction has been enabled to gain one birr plus 709 and 534% extra birr,
respectively. This was more than seven and five times of birr that invested. Thus,
it was highly profitable as compared to manual weeding, whereas, the post-
emergent herbicides (S-metolachlor 960 g/l and S-metolachlor 290 g/l + atrazine
370 g/l) have been proven to provide negative rate of return (-5.65 and -4.09).
Moreover, the application of 2, 4-Dichlorophenoxy acetic acid 720 g/l acid and
lodosulfuron-methyl-sodium 7.5 g/l + mesosulfuron methyl 7.5 g/l herbicides
were coasted 158 and 12 % extra ETB apart from the invested money,
respectively.

In general, the study showed that the highest grain yield was obtained from two
times manual weeding (3243.4 kg ha-1) followed by Bispyribac-sodium 10% EC
(3063.6 kg ha-1) and Pyroxsulam 45 gm/l (2933.2 kg per hectare), respectively.
Even though pre emergence herbicides damage seeds in the soil, it does not mean
that they were not poor to provide good yield and crop dry matter; to which their
application method should be changed. Bispyribac-sodium 10% EC was best in
terms of crop biomass yield and weed index (yield reduction due to weed
competition). On the other hand, the highest yield reduction due to weed
competition were recorded in plots sprayed with 2, 4-Dichlorophenoxy acetic acid
720 g/l acid and Untreated check (34.87% and 32.34 %), respectively.
However, the partial budget analysis proved that Bispyribac-sodium 10% EC (it
enables to gain 709% extra birr apart from production cost cover) and Pyroxsulam
45 gm/l (534%) were promising post emergence herbicides in terms of marginal
rate of return to cultivate upland rice. Thus, pre emergence herbicides were not
effective to manage upland rice weeds though the application row interval and
time had its own effect for their inhibition to the crop. Therefore, based on all
parameters assessed post emergence herbicides, Bispyribac-sodium 10% EC is
recommended for upland weeds management followed by Pyroxsulam 45 gm/l
herbicide at the dose specified in Table 1. These herbicides have limitations in
controlling the weed plants since they have been applied once a season and this
enables some weeds flora to come up lately. Thus, very effective weed control,
sustainable productivity and eco-friendly weed- crop management can be achieved
by the use of integrated management system.
[251]
Acknowledgement
The great gratitude goes to Fogera National Rice Research and Training Center
and Ethiopian Institute of Agricultural Research for supporting the study work in
terms of resources. Technical support of field assistants at the research and
training center who had played a vital role to implementing the study also kindly
acknowledged.
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[252]
Herbicidal Control of Weeds in Turmeric at
Tepi, South western Ethiopia
1Wakjira Getachew, 2Merga Jibat and 3Habetewold Kifelew
1
Ethiopian Institute of Agricultural Research (EIAR), Jimma Agricultural Research Centre, P. O. Box: 192,
Jimma, Ethiopia; 2EIAR, Tepi ARC, P. O. Box: 34, Tepi, Ethiopia; 3EIAR, Holeta ARC,
P. O. Box 2003, Addis Ababa, Ethiopia
Abstract
Turmeric (Curcuma longa L.) which belongs to the family Zingiberaceae, is one of
the major exported spice in Ethiopia. A field experiment was conducted during the
year 2016 at Tepi Agricultural Research Center with the objective of evaluating
registered herbicides for their efficacy in controlling weeds in turmeric. Treatments
were Fenopax + hand weeding, Fenopax + 2,4-D, 2,4-D + hand weeding, Fenopax,
2,4-D and Glyphosate with weed free and unweeded control checks. The treatments
were arrangd in RCBD with three replications. The lowest weed density was observed
on weed free whereas the highest was recorded on unweeded treatment. Leaf width
and number of tillers per plant were the least but plant height and leaf length were the
highest for the weedy treatment. The highest yield was recorded from weed free on the
contrary the least was recorded from weedy treatments. There was 70% yield
reduction due to unweeding. Greater than 30% phytotoxic effect on turmeric was
recorded due to Glyphosate and Fenopax herbicides. Even if the standard check gives
highest yield than the other treatment, 2,4-D + hand weeding can be used as an
option to control turmeric weeds. In the future the cost benefit analysis and the
application of pre-emergence herbicide should be considered during future
investigation for alternative chemical herbicide.
Introduction
Weeds are the most underestimated pest in tropical agriculture, but they have
influenced human activities more than other crop pests. Although crop yield
losses from weeds vary from crop to crop and from region to region, because of
various biotic and abiotic factors, it has been estimated that weeds can cause a
yield loss of about 10% in the developed countries and 25% in the developing
countries (Akobundu, 1987). There is no detailed information available about
the amount of yield losses on turmeric due to weeds in Ethiopia.
Turmeric (Curcuma longa L.) which belongs to the family Zingiberaceae, is one
of exported spice in Ethiopia. In Southwest part of the country, turmeric is
produced as a cash crop and the lively hood of the farmers in the area is depend
on it. According to Habetewold and Wakjira (2017), from the weed survey
fields of turmeric a total of 27 weed species were identified. The most important
families according to the number of represented species were Poaceae,
Asteraceae and Amaranthaceae. The frequency of occurrence of individual
weed species was ranged from 1.25% to 58.75%, while the infestation level
ranged from 0.04% up to 21.99%. The most frequent weed was Sida alba,
[253]
whereas the most dominant weed was Galinsoga parviflora. These two species
were contributed up to 37.6% of the infestation of weed in Turmeric fields at the
study area. Due to fast growing nature of the weeds, labor shortage and cost
implication manual control of turmeric weed found difficult in Tepi area. Weed
control using herbicides seems important and can serve as component of
integrated weed management. Therefore, this trial was conducted to screen
effective, environmentally safe and less costly herbicide for the control of
turmeric weeds in the study area.

Tepi Agricultural Research Center is located in southwestern part of Ethiopia at
35008’ longitude, 7008’ latitude and an altitude of 1200 m asl. It is situated at
about 600 km from Addis Ababa of Ethiopia. The annual average rainfall of the
area is 1630 mm and the mean minimum and maximum temperatures of the area
are 15 0C and 30 0C, respectively.
Experimental material and design

Turmeric variety ‘Dame’ was used as experimental crop for the study during
2017. Treatments included were herbicides like broad leaf selective 2,4-D at
0.72 kg/ha, narrow leaf selective Fenopax at 1 kg/ha and non-selective
Glyphosate at 0.75 kg/ha, and also weed free and un weeded control checks. The
experiment was laid out using randomized complete block design (RCBD) with
three replications. The plot size was 1.8*1.65 (2.97 m2). The herbicides were
applied using knapsack sprayer during the critical stage of weed competition,
when the turmeric was at the 4 leaves growth stage, and also the most frequent
weed Sida alba and the most dominant weed Galinsoga parviflora reached 4 to
5 leaves stage. Herbicide efficacy and toxicity were recorded visually using 1-5
scoring scale at 7, 14 and 21 days after application for the herbicide treatments.
Acceptable weed control was measured as better than 80% control and
acceptable crop safety was measured as less than 30% injury.
Data collection
Crop vegetative parameters (plant height, number of tillers per plant, leaf length,
leaf width and number of leaves per tiller), and rhizome yield parameters
(length, width, number of secondary fingers, number of primary fingers and
yield weight in kg/plot) were collected. Moreover, weed parameters like density,
biomass and control efficiency were also recorded. Weed control efficiency was
calculated according to Chinnusamy et al., (2013) procedure:
WCE= WBMC-WBMT*100
WBMC
[254]
Where WCE= Weed control efficiency (%), WBMC= Weed biomass in
weedy check plot, WBMT=Weed biomass in treated plot
Data analysis
The data collected were subjected to SAS version 9.0 software for analysis and
LSD at 0.05 was used for mean separation.

Effect of different weed management practices on weed density, biomass
and control efficiency
There was significant difference among treatments effect on weeds density and
biomass (Table 1). This finding is in line with the result of Asmare et al. 2016.
2,4-D + Fenopax was found considerably reducing weed density and dry
biomass next to weed free check. These results were in line with the findings of
Erulan and Prabhakaran (2015).
Table 1. Weed control efficiency (WCE) of different weed management practices at Tepi, 2017
Treatments Weed density in Weed biomass Weed control efficiency

number /m2 in g/m2 in %
Fenopax 461b 1262b 44
2,4-D + hand weeding 111e 467e 79
Fenopax + hand weeding 190d 689d 69.5
2,4-D 218c 952c 57.8
Glyphosate 360 96f 488f 78.4
Check 78h 274h 87.9
2,4-D + Fenopax 92g 472g 79.1
Control 762a 2258a -
CV (%) 18 23 -
LSD (5%) 11 34
Effect of herbicide application on vegetative parameters of turmeric

Plant height and leaf length data were slightly affected with herbicide application,
and increased in line with weed infestation. Conversely, number of tillers per plant
and number of leaves per plant were significantly increased with the application of
2,4-D and 2,4-D + HW (Table 2).
[255]
Table 2. Effect of herbicide application on vegetative parameters of turmeric at Tepi, 2017
Treatments PH NTPP LL LW LPT
Fenopax 61.27b 4.3b 29.8b 10.27a 6.27ba
2,4-D + hand weeding 63ba 5.07ba 29.17b 10.27a 6.4ba
Fenopax + hand weeding 59.4b 4.4b 32ba 9.97a 6.57ba
2,4-D 56.5b 5.6a 31.6ba 10.2a 7a
Glyphosate 360 62ba 3.4c 31.47ba 10a 6.8ba
Check 58.7b 4.47b 30.27b 10.1a 6.4ba
2,4-D + Fenopax 57.87b 4.6b 28.13b 9.97a 6.4ba
Control 76.8a 2.87c 38.8a 7.67a 5.4b
CV (%) 13.8 10.3 13.9 21 13
LSD (5%) 15 0.78 7.68 Ns 1.49
PH= Plant height, NTPP= Number of tillers per plant, LL= Leaf length, LW= Leaf width, LPT= Leaf per tiller
Effect of herbicide treatments on yield and yield component of turmeric

There was a significant difference among treatment effects on yield and yield
components. The highest yield was recorded using 2,4-D + Fenopax followed by
Glyphosate 360. There was 70% yield reduction in control plot as compared to
the standard check plot (Table 3). Moreover, Channappagoudar et al. (2013) also
reported that maximum turmeric yield was recorded in weed free check. In line
with this finding, Krishnamurthy and Ayyaswamy (2000) were reported that the
magnitude of yield loss varies from 30 to 75 per cent, depending upon the growth
and persistence of weed density in the standing turmeric crop. 2,4-D application
was found effective exceptionally in promoting number of secondary fingers per
plant and number of primary fingers per plant.
Table 3. Effect of herbicide application on yield and yield components of turmeric at Tepi, 2017
Treatments Yield/plot (kg) RW RL NSFPP NPFPP

Fenopax 3.9c 11.4a 9.67a 10.4ba 3.2bc
2,4-D + hand weeding 3.9c 11.4a 10.37a 11.27ba 3.6ba
Fenopax + hand weeding 3.87c 11.67a 9.3a 8.27b 3.4bc
2,4-D 4.0c 10.6ba 9.47a 12.8a 4.6a
Glyphosate 360 5.29a 12.4a 9.2ba 11.67ba 4.2ba
Check (weed free) 4.47b 12.57a 10.37a 10.5ba 3.27bc
2,4-D + Fenopax 5.7a 12.5a 10.8a 11ba 4.07ba
Control (Un-weeded) 1.7d 7.47b 7.5b 3.8c 2.4c
CV (%) 6.17 16.8 10.45 20.3 18.9
LSD (5%) 0.44 3.3 1.75 3.5 1.19
RW= Rhizome width, RL= Rhizome length, NSFPP= Number of secondary fingers per plant, NPFPP= Number of
primary fingers per plant

Six herbicides: Fenopax + hand weeding, Fenopax + 2,4-D, 2,4-D + hand
weeding, Fenopax, 2,4-D, Glyphosate were used as the treatment with weed free
and un weeded check plots. The lowest weed density was observed on check plot
(always free of weeds) and the highest weed density was recorded on control plot
[256]
(un weeded). Leaf width and number of tillers per turmeric plant were
considerably reduced with weed infestation under control plot. There was 70%
crop yield reduction in control plot as compared to check plot. There was greater
than 30% phytotoxic effect due to application of Glyphosate and Fenopax on the
test crop. The application of 2,4-D has no phytotoxicity on turmeric. Even if the
check gives high yield than the other treatment, 2,4-D + two times hand weeding
can be used as an option to control turmeric weeds. Therefore, in the future cost
benefit analysis and application of pre-emergence herbicide should be done for
alternative chemical herbicide use.
Acknowledgements
We would like to thank Ethiopian Institute of Agricultural Research for financial
support. Also, we thank all technical and field assistants of crop research process
of Tepi Agricultural Research Centre for their unreserved support on field works
from land preparation up to data collection.
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[257]
[258]
Management of Striga (Striga hermonthica) in
Sorghum (Sorghum bicolor) at Jeldu District
Shashitu Bedada, Dessalegne Gella, Niguse Hundessa, Getachew Bekele,
Tesfaye Abdisa, Dereje Geremew and Tadele Gudeta
Ethiopian Institute of Agricultural Research (EIAR), Ambo Agricultural Research Center,
P. O. Box 37. Ambo, Ethiopia
Abstract
A field experiment was conducted from 2017 to 2018 to evaluate herbicides which can
best perform against Striga in sorghum fields of Jeldu district West Shewa Zone.
Treatments were 2, 4-D, Primagram, Dual Gold, Primagram + 2, 4-D; Dual Gold +
2, 4-D; Kerosene; farmers practice and weedy check. The experiment was carried
out in randomized complete block design with three replications. Data was collected
on crop and weed related parameters. Result obtained indicted that sorghum plant
height, stock biomass, grain yield and thousand seed weight showed significant (P<0.05)
difference due to treatments. The highest mean plant height (240.92 cm), stock
biomass (13967 kg/ha), grain yield (886.2 kg/ha) and thousand seed weight (24.3 gram)
were recorded from the sequential application of Dual gold + 2, 4-D as compared to
weedy check. The lowest Striga height (31.917 cm) was recorded from sequential
application of Dual gold + 2, 4-D. Similarly, the lowest Striga biomass was recorded from
farmers practice followed by kerosene (for seed dressing) and sequential application of
Dual gold + 2, 4-D. Therefore, sequential application of Dual gold + 2, 4-D was found
effective against Striga weed in sorghum. Hence, sequential application of Dual gold + 2, 4-
D can be recommended as one component of integrated Striga management in sorghum
field.
Introduction
Ethiopia is the center of origin for sorghum varieties cultivated worldwide.
Sorghum is one the most important cereal crops in the world, only exceeded by
wheat and rice as staple food in the tropics and is a source of raw materials for
many industrial products in Ethiopia.
Striga hermonthica which belongs to Scrophulariaceae family is one of the cereal

crops root parasite often causing 30-100% crop losses on farmers’ fields
throughout the world (Ouedraogo, 1992). It is an important biotic constraint to
sorghum and maize crop production, and the weed that the farmers fear most
(Ramaiah, 1985). Striga species threaten the lives of over 100 million people in
Africa and infest 40% of arable land in the Savannah region arable land may loss
of US$ 7 to 13 million. According to Baguma and Bigirwa (1996) the weed can
cause an economic loss of US$ 8 million a year. Worldwide Striga causes yield
losses that range between 20% and 100% (Kim, 1991).
The herbicidal control of Striga prior to flowering and seed setting can contribute
to reduction of the soil seed bank and subsequent Striga infestations. However,
[259]
chemical control of Striga is not widely practiced due to its high cost and the need
for specialized equipment, unavailability on market; and environmentally
unfriendly. In northern Cameron and Nigeria 2.4-D and triclopyr, Dual Gold
(Metolachlor 960 EC), and Primagram have been applied at rate of 1 l/ha. a.i. and
3 l/ha, each as pre-emergence herbicides respectively. These herbicides reduced
the number of striga species emerged leading increased sorghum yield (Carsky et
al., 1994; Lagoke et al., 1994). On the other hand Carson (1993) reported that 2.4-
D was not effective for controlling of Striga in maize in Gambia. Therefore, this
experiment was designed with the objective of determining effective herbicides
which can best perform against Striga in sorghum fields of Jeldu district, West
Shewa administrative Zone.
Material and Methods

The study was conducted at Jeldu District, West Shewa administrative Zone;
Oromia Regional State on naturally Striga infested sorghum fields. The
experiment was laid out in Randomized Complete Block Design (RCBD) with
three replications. Treatments were application 2, 4-D at 85th days after sowing,
Primagram at 3 l/ha; Gold (Metolachlor 960 EC) at 3 l/ha; 2, 4-D at 1 l/ha +
Primagram at 3 l/ha + 2, 4-D at 1 l/ha; Dual Gold (Metolachlor 960 EC) at 3 l/ha +
2.4-D at 1 l/ha. Post emergence herbicide 2, 4-D; was applied at 85th day after
sowing and pre emergence herbicide Dual Gold (Metolachlor 960 EC) and
Primagram were applied one day after sowing. Improved sorghum variety
“Alemaya ETS-2752” was used as test crop. A plot size of 6m x 5m with a distance
of 1 m between plots was used whereas 1.5m between block were left to avoid
spray drift from adjacent plot.
Data collection
Data on yield and yield component of Sorghum such as stand count, plant height,
stock biomass, grain yield and 1000 seed weight were recorded. In addition to
crop data, Striga plant count per plot, height, and biomass were collected.
Analysis of variance was done using PROC GLM of SAS version 9.2 (SAS
Institute, 2004). Mean separation was performed to compare treatment means
using LSD-test at 5% level of significance.

Effect of herbicides on yield and yield components of sorghum
Sorghum plant height, stock biomass, grain yield and thousand seed weight showed
significant (P<0.05) difference due to herbicides (Table 1). The highest mean
plant height, Stock biomass and grain yield were recorded from the sequential
application of Dual gold+ 2, 4-D as compared to weedy check. The highest
[260]
thousand seed weight was also recorded from sequential application of Dual gold+
2, 4-D and 2, 4-D. Therefore, this indicated that sequential application of Dual
gold+ 2, 4-D was effective in increasing plant height, crop biomass and grain yield
of sorghum as compared to other treatments. However, Sorghum plant population
showed non-significant due to herbicide treatment.
Effect of herbicides on striga population, height and biomass

Striga population and biomass showed no significant difference among treatments.
However, the highest Striga population (16,333 plant counts per hectare) was
recorded from the application of 2, 4-D followed by sequential application of Dual
gold + 2, 4-D. Striga height showed significant (P<0.05) difference due to
herbicides (Table 2). The lowest Striga height (31.917 cm) was recorded from
sequential application of Dual gold + 2, 4-D. Similarly, the lowest Striga biomass were
recorded from farmers practice followed by Kerosene (for seed dressing) and sequential
application of Dual gold + 2, 4-D. This finding indicated that Striga population, height
and biomass were influenced by application of herbicides. Therefore, sequential
application of Dual gold + 2, 4-D was effective in controlling Striga in sorghum field
and can be recommended as integrated Striga management in sorghum fields.
Table 1. Effect of herbicides on sorghum height, population density, stock biomass, thousand seeds weight and grain
yield at Jeldu district, 2017-2018
Plant height population biomass yield 1000 seeds

Treatments (cm) /ha (kg/ha) (kg/ha) weight (g)
Year 2017 236.083 a 29817 b 11067 716.8 a 24.2167 a
Year 2018 209.042 b 66967 a 9518 473.5 b 22.5125 b
LSD 13.787 6138.8 Ns 204.55 1.2547
2,4-D 225.08 ab 48000 12800 ab 666.6 ab 24.383 a
Primagram 228.58 ab 50200 9933 abc 564.3 ab 23.950 ab
Dual gold 217.50 ab 47533 7967 bc 497.2 ab 21.617 b
Dual gold+ 2,4-D 240.92 a 51000 13967 a 886.2 a 24.300 a
Primagram+2,4-D 222.67 ab 44067 12200 ab 609.2 ab 23.583 ab
Kerosene (seed dressing) 227.67 ab 47667 11367 ab 674.8 ab 23.933 ab
Farmers practice 211.75 b 50667 8140 bc 510.4 ab 22.833 ab
Un treated 206.33b 48000 5967 c 352.4 b 22.317 ab
LSD (0.05) 27.573 Ns 4886.5 409.11 2.5094
CV% 10.50 21.517 40.26 58.3 9.108
Note: Means followed by the same letter within a column are not significantly different at P= 0.05 Ns= non significantly
different
[261]
Table 2. Effect of herbicides on striga population and height and biomass at
West Shewa Zone of Jaldu district, 2017-2018
Treatments Population /ha Height (cm) Biomass (kg/ha)

Year 2017 23000 43.717 a 278 b
Year 2018 29742 33.119 b 60348 a
LSD Ns 3.7304 35717
2,4-D 16333 41.700 a 213.3
Primgram 38333 40.694 a 466.7
Dual gold 32000 39.856 a 386.7
Dual gold+ 2,4-D 10067 31.917 b 200.0
Primagram+2,4-D 28767 38.265 ab 253.3
Kerosene (seed dressing) 18000 38.183 ab 200.0
Farmers practice 17000 39.246 ab 146.7
Un treated 50467 37.480 ab 360.0
LSD (0.05) Ns 7.4608 Ns
CV% 130.69 16.47 199.856
Note: Means followed by the same letter within a column are not significantly different at P=0.05p;
ns= non significantly different

Sorghum height, biomass, grain yield, thousand seeds weight and striga height
showed significant (P<0.05) difference due to the application of herbicides. The
highest sorghum height, biomass, grain yield and thousand seeds weight were
recorded from sequential application of Dual gold + 2, 4-D. Similarly, significantly
lowest striga height was recorded from sequential application of Dual gold + 2, 4-D as
compared to the other treatments. Therefore, sequential application of Dual gold + 2,4-
D was found effective against striga weed and in improving sorghum productivity. Hence,
sequential application of Dual gold + 2,4-D can be recommended as one component for
integrated striga management in sorghum fields.
References
Carsky RJ, Singh, I and Nidkawa R.1994.Effect of herbicide and hand weeding on current and
subsequent season striga hermonthica, density on sorghum. International Journal of pest
management.
Carson AG.1993.Studies on Bio-and Chemical control of striga hermonthica (Del.) Benth.In the
Gamibia.Nuisibles-pests-pragas 1:71-81.
Kim SK. 1991. Breeding maize for Striga tolerance and the development of a field infestation
technique. pp. 96-108. In: S.K. Kim (ed.) Combating Striga in Africa. Proceedings of
International Work on Striga, Organized by IITA, ICRISAT and IDRC, Ibadan, Nigeria, 22 -
24 August, 1998.
Lagoke STO, Shebayan JY, Weber GK, Emechebe AM, Singh BB, Zaria A, Awad A, Nagawa L,
Olaniyan GO, Olafare SO, Adeoti AA and Odion C. 1994. Survey of striga problem and
evaluation of striga control methods and packages in crops in the Nigerian savannah. In:
improving striga management in Africa, Proceedings of the 2 nd General workshop
(PASCON).23-29 June 1991, Nairobi Kenya. Lagoke, S.T.O., Hoevers, R.M Book. S.S. and
Traboulsi, R. (Eds.). pp. 91-120. FAO. Accra.
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Ouedraogo O. 1992.Quelques methods detudes du problem du striga au Burikinafaso
DESU.Biologie Vegetale tropicale.universite pirre et Marie curie Paris VI.41 pp.
Ramaiah KV and Parker C. 1982. Striga and other weeds in sorghum. In: Sorghum in the Eighties.
Vol. I proc. Int. Symp, on Sorghum.2-7 November 1981. ICRISAT, Patancheru, A.P.502
324, India.pp.291-302.
Ramaiah KV. 1985. Hand pulling of Striga hermonthica in pearl millet. Tropical pest management
31:326-327.
[263]
[264]
Integrated Management of Wild Radish
(Raphanus raphanistrum) in Faba Bean in South
West Shewa Zone, Ethiopia
Shashitu Bedada, Niguse Hundessa, Shugute Addisu, Dessalegne Gella,
Getachew Bekele, Tesfaye Abdissa and Tadele Gudeta
Ethiopian Institute of Agricultural Research (EIAR), Ambo Agricultural Research Center,
P. O. Box 37. Ambo, Ethiopia
Abstract
A field experiment was conducted between 2017 and 2018 to evaluate effective and
economical integrated management package for the control of Raphanus
raphanistrum in faba bean in Weliso District of South West Shewa, Ethiopia. The
experimental design used was spilt plot with three replications. Two levels of
plowing frequency assigned as main plot and eight weed management practices as
sub plot. Data were recorded on crop and weed related parameters. Results
indicated that the interaction effect of plowing frequency and combination of
herbicides with hand weeding frequency were significantly affected faba bean plant
height, number of pods per plant, seeds per pod, biomass and grain yield. The
highest mean grain yield (1541 kg/ha) was obtained as a result of thrice plowing +
application of pendimethalin at 3 litter ha -1 supplemented with once hand weeding at
30-35 DAS followed by thrice plowing + application of S-Metolachlor at 1 liter ha-1
supplemented with twice hand weeding at 30-35 and 50-55 DAS (1463kg/ha). The
interaction effect of plowing frequency and combination of herbicides with hand
weeding frequency also significantly affected R. raphanistrum population, height
and biomass. The lowest population, height and biomass were obtained as a result of
thrice plowing + application of S-metolachlor at 1 litter ha-1 supplemented with twice
hand weeding at 30-35 and 50-55 DAS. Economic analysis showed that trice
plowing combined with application of Pendimethalin at 3 liter ha -1supplemented
with once hand weeding at 30-35 DAS resulted the highest net benefit 23174 ETB
followed by trice plowing combined with application S-Metolachlor at 1 liter ha-1
supplemented by twice hand weeding; 22352 ETB. Therefore, thrice plowing and
application of S-Metolachlor supplemented with twice hand weeding at 30-35 and
50-55 DAS or thrice plowing and application of Pendimethalin at 3 litter ha-
1
supplemented by once hand weeding at 30-35 DAS was recommended for
R. raphanistrum management and gain economical profit from faba bean
production in Weliso district of South West Shewa Zone, Ethiopia.
Introduction
Wild radish (Raphanus raphanistrum) is a major weed of winter crops in southern
and eastern Australia, and is particularly troublesome in cereal and brassica crops.
It is also thought to be poisonous to livestock. Reproduction is only by seed,
which are spread shorter distances by wind, water, animals and human activities.
Dispersal via contaminated agricultural produce is thought to be the most
significant means of long range spread (Retter and Harden, 2005).
[265]
Several exotic weed species have elderly been introduced to Ethiopia, among
these, R. raphanistrum is now causing a serious problem on crop and rangelands
in Ethiopia. There was no concrete evidence so far regarding the introduction of R.
raphanistrum to Ethiopia. Since its introduction to country the weed is extremely
expanding seriously to highlands ecology and cereal, pulse, oil crops and
rangeland production. It is a serious weed of crop and rangelands, particularly in
West Shewa Welmera and Cheliya South West Shewa (Weliso) and Horo Guduru
Wellega zone areas of Oromiya, regional state, Ethiopia (Personal
communication). According to farmers around Cheliya district (Gedo), R.
raphanistrum was introduced during the war between Ethiopia and Italy (1930-
1935), in such a way that Italy armed forces brought it to Gedo by horse feed
contaminated with R. raphastirum seeds and that it was first observed in Fortino
area; its distribution and spread is more restricted to the normal upland /dry/ type
of grazing area rather than water logged grazing area (Shugute, unpublished,
2015). It is locally known as "Gomana Faranji" (French cabbage) probably
because of its allelopathic potential and strong competitiveness (Personal
communication). The weed has substantial impact on the native biodiversity of
rangelands and arable lands by its canopy cover and is also toxic to animals
causing severe human health problems by making allergic on human hands when
it is hand weeded in n field crops. Therefore, it has become serious problem in
major crops as well as Faba Bean in the study area. Results of field experiment
conducted during 1997 -1998 at Shambu testing site indicated that Raphanus
alone incurred 28.9% grain yield loss in bread wheat compared with the weeded
check and effective control of this test weed was obtained by spraying mecoprop,
2,4-D and the tank mixture of both herbicide and ioxynil + bromoxynil +
mecoprop treated plots (Dhaba et al., 2005).
Cultural, chemical and biological weed control activities can exert a strong
selective influence on the weed populations (Chancellor, 1985). Thus, knowledge
of the weed community structure is an important component of weed
management, and is essential in setting priorities for both weed management and
research. Chemicals nowadays are expensive and are not environmentally friendly.
As a result the “best mix” approach (IPM) is currently advocated for Integrated
Pest Management (IPM) involves the most logical combination of different
compatible tactics for the control of pests (Brader, 1979).
The principal aim of IWM is to improve the health and vigor of crops so that they
may out-compete weeds emerging in the stand. Weed competition is the major
problem reducing crop productivity in study area. Among the major weeds found
in the district wild radish is one. However, there is limited information regarding
its management and effect on the productivity of major crops particularly on faba
bean Therefore, this trial was initiated with the objectives of to enhance faba bean
crop yield by reducing R. raphanistrum weed infestation level using appropriate
[266]
weed management options and to evaluate effective and economical integrated
management package for the control of R. raphanistrum weed in faba bean.

Description of study area
The study was conducted at Weliso district of South West Shewa Zone of
Oromiya Regional State on the farmer’s field during the main cropping season of
2017 and 2018. Testing site was selected after field observation on the availability
of naturally infested heavy and wide population of R. raphanisrtrum weed at
major faba bean growing areas.

The experiment was laid out as a split-plot design with three replications. The
factors include two plowing frequency (two times plowing and three times
plowing) assigned as a main plot and eight weed management practices (S-
metolachlor at 1 litter ha-1 supplemented by one hand weeding at 30-35 days after
sowing; S-metolachlor at 1 litter ha-1 supplemented by twice hand weeding at 30-
35 and 50-55 days after sowing; Pendimethalin at 3 litter ha-1 supplemented by
one hand weeding at 30-35 days after sowing; Pendimethalin at 3 litter ha-1
supplemented by twice hand weeding at 30-35 and 50-55 days after sowing; twice
hand weeding at 30-35 and 50-55 days after sowing; S-Metolachlor at 1 litter ha-1,
Pendimethalin at 3 litter ha-1; Weedy Check) were assigned as sub plot. Details of
treatments are shown in Table 1. The sub plot size was 5m x 4m (20m2) with a
distance of 1m between plots and 1.5 m between blocks were left to avoid spray
drift from adjacent plots. Faba bean variety Walki was sown at 200 kg/ha with a
spacing of 40 cm between rows and 5cm between plants and DAP were applied at
100 kg/ha at sowing time.
[267]
Table 1. Treatment description and corresponding codes
No Treatment description Treatment code

1 Twice plowing+ S-metolachlor 1 liter ha-1 2xP+SM
2 Twice plowing+ weedy check 2xP+WC
3 Thrice plowing + weedy check 3xP+WC
4 Twice plowing+ twice hand weeding at 30-35 and 50-55 DAS 2xP+2Xhwat 30-35 and 50-55 DAS
5 Thrice plowing+ twice hand weeding at 30-35 and 50-55 DAS 3xP+2xhw+at 30-35 and 50-55 DAS
6 Twice plowing+ S-metolachlor 1 liter ha-1+ once hand 2xP+SM+1xhw+ at 30-35 DAS
weeding at 30-35 DAS
7 Thrice plowing+ S-metolachlor 1 liter ha-1+ once hand 3x+SM+1xhw+ at 30-35 DAS
8 Thrice plowing+ S-metolachlor 1 liter ha-1 3x+SM
9 Twice plowing+ S-metolachlor 1 liter ha- + twice weeding at 2xP+SM+2xhw at 30-35 and 50-55 DAS
30-35 and 50-55 DAS
10 Thrice plowing+ S-metolachlor 1 liter ha-1+ twice hand 3xP+SM+2xhw at 30-35 and 50-55 DAS
weeding at 30-35 and 50-55 DAS
11 Twice plowing+ Pendimethalin 3 liter ha-1+ once hand 2xP+P+1xhw at30-35 DAS
12 Twice plowing+ Pendimethalin 3 liter ha-1 2xP+P1xhw30-35 DAS
13 Thrice plowing+ Pendimethalin3 liter ha-1 + once hand 3xP+P+1xhw at 30-35 DAS
14 Thrice plowing+ Pendimethalin3 liter ha-1 3X+P
15 Twice plowing+ Pendimethalin 3 liter ha-1+ twice weeding at 2xP+P+2xhw at 30-35 and 50-55 DAS +
30-35 and 50-55 DAS
16 Thrice plowing+ Pendimethalin 3 liter ha-1 + twice hand 3xP+P+2xhw at 30-35 and 50-55 DAS +
weeding at 30-35 and 50-55 DAS
Land was prepared with oxen plough to fine soil bed which supports easy contact
of the seed with soil. All other agronomic practices required for the crop were
applied according to recommendation given for the crop.
Method of application: The required quantity of the herbicide was calculated and
measured out into manual knapsack sprayer with a pre-determined water volume
for each plot and each herbicide was applied separately one day after sowing
under Culm weather.
Data collection
Data on yield and yield components of faba bean like number of pods per plant,
number of seeds per pod, grain yield, biomass, plant height, and 1000 seed weight
were recorded. Total number of pods in 10 randomly selected plants in each plot
was counted at harvest and expressed as the number of pods plant -1. From these
pods, the seeds were counted to determine the number of seeds pod-1.Thoused
seeds were counted from each plot, and their weight was recorded. Above ground
dry biomass weight was measured at physiological maturity from each plot. This
was converted in to kg ha-1.Grain yield (kg) was recorded from each net plot area.
[268]
In addition, data on R. raphanistrum weed height, density/count after herbicide
application and biomass at harvest was recorded R. raphanistrum weeds present in
the experimental fields of all plots were recorded after application by placing a
quadrant of 0.25mX0.25 m randomly at two spots. Its height was measured from
10 randomly selected weeds before the crop was harvested. Biomass was also
measured during crop harvest from all plots. Partial budget analysis procedure was
employed for economic analysis of herbicide application and it was carried out for
faba bean yield.
Data analysis
The data collected were subjected to analysis of variance by SAS software. Means
that showed a significant difference were separated by least significant difference
at 0.05 probability level by SAS (SAS, 2004).
Economic analysis
The partial budget analysis as described by CIMMYT (1988) was done to
determine the economic advantage of the weed management practice. Economic
analysis was done using the prevailing market prices for inputs at planting and for
the outputs at the time of crop harvest. It was calculated by taking in to account
the additional input and labor cost involved and gross benefits obtained from weed
management practices. Average yield was adjusted downward by 10% to reflect
the difference between the experimental yield and the yield farmers could
obtained from the same weed management practices as described by (CIMMYT,
1988). For economic evaluation, partial budget , value to cost ratio (VCR) and
marginal analysis were used based on the local market price of the faba bean yield
and herbicide cost, the cost of tillage and weeding were also taken from the
farmers own practice.

Plant height
The result of mean separation from the two years data is presented in Table 1.
Plant height was significantly influenced by the interaction effect of plowing
frequency, application of herbicides and hand weeding. The highest mean plant
height (112.91 cm) was obtained from the interaction effect of thrice plowing +
application of Pendimethalin at 3 litter ha-1 followed by thrice plowing +
application of S-metolachlor at 1 litter ha-1. However, he lowest mean plant height
(58.94 cm and 62.23 cm) were obtained from twice plowing + application of S-
metolachlor at 1 litter ha-1 supplemented with twice hand weeding at 30-35 and
50-55 DAS and twice plowing + application of Pendimethalin at 3 litter ha-1
supplemented with twice hand weeding at delete 30-35 and 50-55 DAS ,
respectively. This indicated that thrice plowing was more effective in increasing
[269]
plant height as compared to twice plowing, but plant height was not influenced by
application of herbicide and hand weeding (Table 2).
Number of pods per plant

Analysis of variance revealed that there was a high significant difference among
the treatments in number of pods per plant (Table 1). Combined effect of two
years data indicated that the highest mean number of pods per plant(7.5 and 7.4)
was recorded from thrice plowing + twice hand weeding at 30-35 and 50-55 DAS
and thrice plowing + application of S-metolachlor at 1 litter ha-1 supplemented by
twice hand weeding at 30-35 and 50-55 DAS, respectively. This might be due to
reduced weed competition for nutrient, moisture and light and as result he crop got
higher net assimilation rate thus retaining more flower. The development of more
and vigorous leaves under low weed infestation might have also helped to improve
the photosynthetic efficiency of crop and supported large number of pods
(Hodgson and Blackman, 2005). However, the lowest mean number of pods per
plant (5.3 and 5.6) was recorded from twice plowing + S-metolachlor at 1 litter ha-
1
and twice plowing + Pendimethalin at 3 litter ha-1, respectively (Table 2). This
result indicted that, trice plowing + twice hand weeding at 30-35 and 50-55 DAS
was the most effective followed by thrice plowing+ S-metolachlor 1 liter ha-1+
twice hand weeding at 30-35 and 50-55 DAS in increasing number of pods per
plant in faba bean. These results are in line with Hadi et al. (2006) who observed
an increased number of pods plant-1 where weed population was reduced by
management techniques. Similarly, (Pereira et al., 2015) and (El-Metwally et al.,
2008) reported that the number of pods produced per plant or maintained to final
harvest depends on a number of environmental and management practices.
Number of seeds per pod

The interaction effect of different weed control options showed significant effect
on number of seeds per pods of faba bean (Table 2). The highest mean number of
seeds per pod (3.03) was obtained from the interaction effect of thrice plowing +
application of Pendimethalin 3 litter ha-1supplemented by twice hand weeding at
30-35 and 50-55 DAS followed by thrice plowing + application of Pendimethalin
at 3 litter ha-1supplemented by once hand weeding at 30-35 DAS which having
(2.9) number of seeds per pod. This might be due the reduced interference of
weeds; the vigorous leaves might have helped to improve the photosynthetic
efficiency of crop that support large number of seeds pod-1 which can have
positive contribution for increment of grain yield. However, the lowest mean
number of seeds per pod (2.45) was recorded from twice plowing + twice hand
weeding at 30-35 and 50-55 DAS, but non-significant different existed in twice
plowing + S-metolachlor at 1 litter ha-1, twice plowing + weedy check, thrice
plowing + S-metolachlor at 1 litter ha-1 + once hand weeding at 30-35 DAS, thrice
plowing + twice hand weeding at 30-35 and 50-55 DAS, thrice plowing + S-
[270]
metolachlor at 1 litter ha-1 , thrice plowing + Pendimethalin3 litter ha-1 and trice
plowing + weedy check (Table 3).
Table 2. Effect of integrated management of R. raphanistrum on plant height and yield components of faba bean at
Weliso district, 2017-2018
No of pods per No of seeds

Treatments Plant height plant /pod
Year 2017 118.265 a 7.6625 a 2.5375 b
Year 2018 52.740 b 4.9854 b 2.8833 a
LSD (0.05) 7.7315 0.5129 0.1184
2xP+SM 76.12 efg 6.3 abcd 2.867abc
2xP+WC 58.94 g 5.92cd 2.55cde
3xP+WC 73.18 efg 6.52 abcd 2.8 abcd
2xP+2Xhwat 30-35 and 50-55 DAS 62.23g 5.98bcd 2.87abc
3xP+2xhw+at 30-35 and 50-55 DAS 77.94 defg 6.07 abcd 2.45 e
2xP+SM+1xhw+ at 30-35 DAS 76.49efg 5.33d 2.533cde
3x+SM+1xhw+ at 30-35 DAS 93.44 abcde 5.6d 2.87 abc
3x+SM 65.16 fg 5.8cd 2.6bcde
2xP+SM+2xhw at 30-35 and 50-55 DAS 80.73defg 5.87 cd 2.533 cde
3xP+SM+2xhw at 30-35 and 50-55 DAS 86.00 cdef 7.4ab 2.8abcd
2xP+P+1xhw at30-35 DAS 104.14abc 7.1 abc 2.90ab
2xP+P1xhw30-35 DAS 89.38bcde 7.23abc 3.03a
3xP+P+1xhw at 30-35 DAS 99.18abcd 7.5a 2.73abcde
3X+P 108.68ab 6.25abcd 2.47de
2xP+P+2xhw at 30-35 and 50-55 DAS + 112.91a 6.3 abcd 2.78abcde
3xP+P+2xhw at 30-35 and 50-55 DAS + 103.53 abc 6.02 bcd 2.58 bcde
LSD (0.05) 21.868 1.4508 0.3348
CV (%) 22.16 19.88 10.70
Note: CV= Coefficient of Variation, LSD=Least significance difference, DAS=Days after sowing, Means followed by the
same letter within a column are not significantly different at 0.05p
Crop biomass
Result for crop biomass yield indicated that significant differences among
treatments were observed (Table 3). The highest mean value of crop biomass
(4241.7 kg/ha) was obtained from the interaction effect of thrice plowing +
application of Pendimethalin3 litter ha-1 supplemented by once hand weeding at
30-35 DAS, but as par with thrice plowing + Pendimethalin at 3 litter ha-
1
supplemented + twice hand weeding at 30-35 and 50-55 DAS, and thrice plowing + S-
metolachlor at 1 litter ha-1 + twice hand weeding at 30-35 DAS. However, the lowest
mean value of crop biomass yield (1883.3 kg/ha) was obtained from twice
plowing + twice hand weeding at 30-35 and 50-55 DAS, but non-significant twice
plowing+ S-metolachlor at 1 litter ha-1,twice plowing+ S-metolachlor at 1 litter ha-
1
+twice hand weeding at 30-35 and 50-55 DAS, trice plowing+ S-metolachlor at 1
litter ha-1. Therefore, thrice plowing + application of pendimethalin at 3 liter ha-1
supplemented by one hand weeding at 30-35 DAS followed by thrice plowing +
application of pendimethalin at 3 liter ha-1 supplemented by twice hand weeding at
30-35 DAS are effective in increasing above ground biomass as compared to
[271]
other treatments. The increased above ground biomass yield in these treatments
might be due to the crop utilized the resources more efficiently leading to initial
crop stand. Similar with present results, (Alfonso et al., 2013) reported good
suppression of weed growth by cultural and herbicidal control measures that lead
to low competition by weeds for light, space and nutrients by which the crop could
utilize both biotic and abiotic resources efficiently, leading to higher dry biomass
production.
Grain yield
Analysis of variance indicated that high significant difference among different
treatments for grain yield (Table 2). The highest mean grain yield (1541.2 kg/ha)
was obtained from the interaction effect of thrice plowing+ application of
pendimethalin at 3 litter ha-1supplemented by one hand weeding at 30-35 DAS
followed by thrice plowing + application of S-metolachlor at 1 litter ha-
1
supplemented by twice hand weeding at 30-35 and 50-55 DAS having 1462.8
kg/ha grain yield. But non-significant trice plowing+ Pendimethalin at 3 litter ha-
1
supplemented + twice hand weeding at 30-35 and 50-55 DAS, trice plowing+ weedy
check, twice plowing + S-metolachlor at 1 litter ha-1suplimented by once hand
weeding at 30-35 DAS and trice plowing+ twice hand weeding at 30-35 and 50-55
DAS. However, the lowest mean grain yield (748.3 kg/ha) was, however, recorded
from the interaction of twice plowing + application of S-metolachlor at 1 litter ha-1
followed by twice plowing + application of pendimethalin at 3 litter ha-1(Table 2).
This result indicated that thrice plowing + application of pendimethalin at 3 litter
ha-1supplemented by one hand weeding at 30-35 DAS and thrice plowing +
application of S-metolachlor at 1 litter ha-1supplemented by twice hand weeding at
30-35 and 50-55 DAS were gave highest grain yield of faba bean as compared to
other treatments. Similarly, research conducted at Holeta and Shamboo showed
that repeated plowings before planting significantly increased seed yield of faba
bean (Endale Asmare and Rezene Fessehaie, 1991; Tolera Abera and Daba Feyisa,
2004). Bellido et al. (2003) also reported that in three rainy years conventional pre
planting productive than no tillage.
Thousand seed weight

High significant difference was observed among treatments on thousand seed
weight (Table 3). The maximum mean thousand seeds weight (696.75 g) and
(693.47g) were obtained from the interaction of thrice plowing + application of
Pendimethalin at 3 litter ha-1 supplemented with twice hand weeding at 30-35
DAS and 50-55 DAS and twice plowing + weedy check, respectively, but most of
means showed statistical parity. Whereas, the lowest value (650.27 g) was
recorded from thrice plowing + application S-metolachlor at 1 litter ha-1
supplemented with twice hand weeding.
[272]
Table 3. Effect of integrated management of R. raphanistrum on yield and yield components of faba bean at Weliso
district, 2017-2018
Treatments Crop Biomass Grain yield 1000 seed weight

(Kg/ha) kg/ha (g)
Year 2017 3595.8 a 1333.27 a 665.919 b
Year 2018 2320.8 b 1025.35 b 682.854 a
LSD (0.05) 275.84 136.83 10.995
2xP+SM 2958.3 cdef 1379.3 abc 684.28 abc
2xP+WC 2425.0 efg 982.2 def 662.03 bcd
3xP+WC 2791.7 def 1150.3 bcde 666.25 abcd
2xP+2Xhwat 30-35 and 50-55 DAS 2775.0 def 1092.7 bcdef 665.83 abcd
3xP+2xhw+at 30-35 and 50-55 DAS 1883.3 g 1108.3 bcdef 678.50 abcd
2xP+SM+1xhw+ at 30-35 DAS 2241.7 fg 748.3 f 656.32 cd
3x+SM+1xhw+ at 30-35 DAS 2725.0 def 795.5 ef 657.52 cd
3x+SM 2750.0 def 1147.2 bcde 693.47a
2xP+SM+2xhw at 30-35 and 50-55 DAS 3066.7 cde 1271.8abcd 673.62 abcd
3xP+SM+2xhw at 30-35 and 50-55 DAS 3625.0 abc 1462.8 ab 650.27 d
2xP+P+1xhw at30-35 DAS 4241.7 a 1541.2 a 683.97 abc
2xP+P1xhw30-35 DAS 3908.3 ab 1384.8 abc 696.75 a
3xP+P+1xhw at 30-35 DAS 2700.0 def 1311.0 abcd 686.38abc
3X+P 2658.3 efg 1098.2 bcdef 675.73 abcd
2xP+P+2xhw at 30-35 and 50-55 DAS + 3366.7 bcd 1064.7cdef 666.43 abcd
3xP+P+2xhw at 30-35 and 50-55 DAS + 3216.7 bcd 1331.0 abcd 692.83 ab
LSD (0.05) 780.19 387.01 31.099
CV (%) 22.85 30.35 3.99
Note: CV= Coefficient of Variation, LSD=Least significance difference, DAS=Days after sowing,
Means followed by the same letter within a column are not significantly different at 0.05p
R. raphanistrum count, height and fresh biomass

There was significant interaction effect of plowing frequency, application of
herbicide and hand weeding on R. raphanistrum count, height and biomass (Table
3). The lowest (18.67 and 21.33 weed/m2) R. raphanistrum count was recorded
from trice plowing + twice hand weeding at 30-35 and 50-55 DAS and trice
plowing + application S-metolachlor at 1 litter ha-1 supplemented by twice hand
weeding at 30-35 and 50-55 DAS, respectively. The highest (62.67 weeds/m2) (R.
raphanistrum ) count was recorded from plowing twice + application of S-
metolachlor at 1 litter ha-1 supplemented by twice hand weeding at 30-35 and 50-
55 DAS followed by plowing twice + application of S-metolachlor at 1 liter ha-1
supplemented by one hand weeding at 30-35 DAS, but most of treatments showed
statistical parity. Therefore, trice plowing + application of S-metolachlor at 1 liter
ha-1 and thrice plowing + application S-metolachlor at 1 liter ha-1 supplemented by
twice hand weeding at 30-35 and 50-55DAS was effective in reducing R.
raphanistrum population as compared to other treatments.
There was significant interaction effect of plowing frequency, application of

herbicide and hand weeding on R. raphanistrum height and weed biomass (Table
4). The shortest (36.36cm) R. Raphanistrum height was recorded from twice
[273]
plowing + application of Pendimethalin at 3 litter ha-1supplemented by twice hand
weeding at 30-35 and 50-55 DAS followed by trice plowing + application of
Pendimethalin at 3 litter ha-1supplemented by twice hand weeding at 30-35 and
50-55 DAS and trice plowing+ application of S-metolachlor at 1 litter ha-1
supplemented by twice hand weeding at 30-35 and 50-55 DAS, respectively, but
as par with twice plowing + S-metolachlor at 1 litter ha-1 supplemented by twice
hand weeding at 30-35 and 50-55 DAS. The tallest (126.1 cm and 124.9 cm) mean
R. raphanistrum height was recorded from plowing thrice + application of
pendimethalin at 3 litter ha-1 and plowing thrice +application of S-metolachlor at 1
litter ha-1, respectively. But non- significant result was obtained from twice
plowing + Pendimethalin at 3 litter ha-1. This indicted that R. raphanistrum height
was highly influenced by application of Pendimethalin at 3 litter ha-1and S-
metolachlor at 1 litter ha-1 when supplemented by twice hand weeding at 30-35
and 50-55 DAS as compared to other treatments. This was because of the fact that
R. raphanistrum can be managed at different germination stage because of the
integration of pre emergence herbicide and twice hand weeding at different time.
The lowest fresh biomass (150.0 kg/ha and 191.7 kg/ha) was recorded from thrice
-1
plowing + application of S-metolachlor at 1 litter ha supplemented by twice hand
weeding at 30-35 and 50-55 DAS and thrice plowing + application of Pendimethalin at 3
litter ha-1supplemented by twice hand weeding at 30-35 and 50-55 DAS, respectively
(Table 3). The highest (9008.5 kg/ha) fresh biomass was recorded from trice
plowing + weedy check. Non-significant difference was observed among all
treatments which where supplemented by twice hand weeding. The higher fresh
biomass weedy check might be due to higher weed density that provided an
opportunity to the weeds to compete vigorously for nutrients, space, light, and
water and carbon dioxide leading to high weed biomass These results are in
agreement with the findings of (Alfonso et al., 2013) and (Das and Yaduraju,
1999) who reported maximum weed dry weight in the weedy check.
[274]
Table 4. Effect of integrated management on R. raphanistrum count, height and fresh biomass of the weed plant at
Weliso, 2017-2018
Fresh biomass
Treatments Count/m2 Height (cm) (kg/ha)
Year 2017 27.833 b 106.835 a 2288.5 b
Year 2018 48.250 a 50.322 b 3311.5 a
LSD (0.05) 10.43 7.0516 571.83
2xP+SM 53.33 ab 65.79 de 2125.0def
2xP+WC 62.67 a 42.29 fgh 266.7gh
3xP+WC 25.33 bc 65.77 de 1875.0 defg
2xP+2Xhwat 30-35 and 50-55 DAS 48.00 abc 36.36 h 391.7 gh
3xP+2xhw+at 30-35 and 50-55 DAS 40.67 abc 58.01 efg 825.0 fgh
2xP+SM+1xhw+ at 30-35 DAS 28.67 bc 102.871 bc 4416.7 c
3x+SM+1xhw+ at 30-35 DAS 28.00 bc 112.024 ab 3225.0 cd
3x+SM 35.33 abc 85.21cd 4458.3 c
2xP+SM+2xhw at 30-35 and 50-55 DAS 36.00 abc 97.98 bc 2458.3 de
3xP+SM+2xhw at 30-35 and 50-55 DAS 21.33 c 39.11 gh 150.0 h
2xP+P+1xhw at30-35 DAS 46.00 abc 87.59 c 1300.0efgh
2xP+P1xhw30-35 DAS 28.67bc 38.89 gh 191.7 h
3xP+P+1xhw at 30-35 DAS 18.67 c 59.09 ef 366.7 gh
3X+P 38.67 abc 124.983 a 7491.7 ab
2xP+P+2xhw at 30-35 and 50-55 DAS + 54.00 ab 126.133 a 6250.0 b
3xP+P+2xhw at 30-35 and 50-55 DAS + 43.33 abc 115.144 ab 9008.5 a
LSD (0.05) 29.502 19.945 1617.4
CV (%) 67.19 21.99 50.05
Note: CV= Coefficient of Variation, LSD=Least significance difference, DAS=Days after sowing, Means followed by the
same letter within a column are not significantly different at 0.05
Economic feasibility of R. raphanistrum management

on faba bean production
Partial budget analysis for the different treatments is presented (Table 4). The
value to cost ratio for plowing frequency was 6.62 to 42.01 EB per unit
investment. The highest marginal rate of return of 2942% was obtained from trice
plowing with no hand weeding. R. raphanistrum weed management with trice
plowing combined with application of Pendimethalin at 3 litter ha-1 supplemented
with weeding once weeding once at 30-35 DAS gave net profit advantage of
23174 EB with marginal rate of return of 187 % followed by plowing trice +
application of S-metolachlor at 1 litter ha-1 supplemented with twice hand weeding at
30-35 and 50-55 DAS and plowing twice + application of S-metolachlor at 1 litter
ha-1 supplemented by once hand weeding at 30-35 DAS gave net benefit of 22352
and 21208 EB per hectare with marginal rate of return of 527 and 29 %
respectively (Table 5). The economic analysis confirmed that R. raphanistrum
management on faba bean was profitable with plowing trice + application of
Pendimethalin at 3 litter ha-1 herbicide at supplemented with once hand weeding
at 30-35 DAS or plowing trice + application of S-metolachlor at 1 liter ha-1
supplemented by twice hand weeding at 30-35 and 50-55 DAS for Weliso
district.
[275]
Table 5. Effects of integrated management on R. Raphanistrum on economic profitability of faba bean, 2017-2018
Grain yield Adjusted yield Gross field Herbicide Plowing Weeding TCV Net benefit Value to MRR
Treatments kg/ha kg/ha benefit cost cost cost (ETB (ETB/ha) ETB cost ratio (%)
(ETB/ha) ( ETB/ha) (ETB/ha ) ha-1)
2xP+SM 748 673.2 12117.6 0 195 237 432 11686 27.05
2xP+WC 1147 1032.3 18581.4 0 195 237 432 18149 42.01
3xP+WC 1331 1197.9 21562.2 0 293 237 530 21032 39.68 2942
2xP+2Xhw at 30-35 and 50-55 DAS 1108 997.2 17949.6 0 195 356 551 17399 31.58
3xP+2xhw+at 30-35 and 50-55 DAS 1311 1179.9 21238.2 0 293 356 649 20589 31.72
2xP+SM+1xhw+ at 30-35 DAS 1379 1241.1 22339.8 700 195 237 1132 21208 18.73 29
3x+SM+1xhw+ at 30-35 DAS 1272 1144.8 20606.4 700 293 237 1230 19376 15.75
3x+SM 1098 988.2 17787.6 700 293 237 1230 16558 13.46
2xP+SM+2xhw at 30-35 and 50-55 DAS 982 883.8 15908.4 700 195 356 1251 14657 11.72
3xP+SM+2xhw at 30-35 and 50-55 DAS 1463 1316.7 23700.6 700 293 356 1349 22352 16.57 527
2xP+P+1xhw at30-35 DAS 1150 1035 18630 1260 195 237 1692 16938 10.01
2xP+P1xhw30-35 DAS 796 716.4 12895.2 1260 195 237 1692 11203 6.62
3xP+P+1xhw at 30-35 DAS 1541 1386.9 24964.2 1260 293 237 1790 23174 12.95 187
3X+P 1065 958.5 17253 1260 293 237 1790 15463 8.64
2xP+P+2xhw at 30-35 and 50-55 DAS 1093 983.7 17706.6 1260 195 356 1811 15896 8.78
3xP+P+2xhw at 30-35 and 50-55 DAS 1385 1246.5 22437 1260 293 356 1909 20528 10.75
ETB= Ethiopian birr
The result of the present study showed that plowing frequency, application of pre
emergence herbicide supplemented by hand weeding showed significant result on
yield and yield components of faba bean. The highest mean grain yield (1541.2
kg/ha) was obtained from the interaction of three plowings + application of
Pendimethalin at 3 litter ha-1 supplemented by once hand weeding at 30-35 DAS
followed by thrice plowing + application of S-metolachlor at 1 litter ha-1
supplemented with twice hand weeding at 30-35 and 50-55 DAS with mean grain
yields of (1462.8 kg/ha). Similarly, R. raphanistrum population, height and fresh
biomass were significantly different due to the interaction effect of the treatments.
The lowest R. raphanistrum population, height and fresh biomass were obtained
from the interaction of thrice plowing +application of S-metolachlor at 1 litter ha-1
supplemented by twice hand weeding at 30-35 and 50-55 DAS. The economic
analysis also showed that trice plowing + application of Pendimethalin at 3 litter
ha-1 supplemented by once hand weeding at 30-35 DAS gave the highest 23174
EB Net benefit followed by trice plowing + application S-metolachlor at 1 litter
ha-1 supplemented by twice hand weeding at 30-35 and 50-55 DAS that gave
22352 EB net benefit. Therefore, thrice plowing +application of S-metolachlor at
1 litter ha-1 supplemented by twice hand weeding at 30-35 and 50-55 DAS or trice
plowing + application of Pendimethalin at 3 litter ha-1 supplemented by once hand
weeding at 30-35 DAS can be recommended for optimum grain yield, economic
profitability for faba bean production and R. raphanistrum management in Weliso
district of South West Shewa Zone, Ethiopia.
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[278]
Integrated Management of Cotton Weeds at
Middle Awash Rift Valley of Ethiopia
Workishet Taye1, Zemedkun Alemu2, Sharew Abate2 and Bedane Gudeta2
Ethiopian Institute of Agricultural Research, 1Melkessa Agricultural Research Center, 2Werer Agricultural
Research Center, P. O. Box 2003, Addis Ababa, Ethiopia
Abstract
Weed infestation is one of the main problems of cotton production in Ethiopia.
Therefore, an experiment was conducted at Werer Agricultural Research Center and
Gewane cotton commercial farm during the main cropping seasons of 2017 and 2018
to determine better integrated weed management practices for the Middle Awash, Rift
Valley of Ethiopia. A total of eight treatments; hand weeding + Intreculturing, Pre-
irrigation + hand weeding, Pendimethalin + hand weeding, Pre-irrigation +
Intreculturing, Pendimethalin + Intreculturing, Pre-irrigation + Pendimethalin, Pre-
irrigation and Control check were laid out in a Randomized Complete Block Design
(RCBD) with three replications. A total of 40 weed species, belonging to 20 families
were identified in the cotton field. The result showed that abundance of weeds, and ball
number per plant and yield of cotton were significantly affected by the management
practices. A seed cotton yield loss of about 64% was recorded in the weedy control.
This study showed that one hand weeding + Interculturing or Pendamethaline +
Interculturing which resulted better yield and yield components can be used as an
integrated cotton weed management system at Werer and Gewane areas.
Introduction
Yield of cotton greatly reduced due to the naturally-occurring mixed weed
population in which a seed cotton yield loss of 62.43 - 96.21% occurred when
weeding was completely not practiced throughout the crop growing season
(Esayas et al., 2013). An increased higher yield loss on average in the authors
experiment than the former report could be due to increased weed population and
introduction of other new weed species to the area (Esayas et al., 2013). Survey
conducted in Middle Awash during 2000 cropping season indicated that the
infestation level was very high for most of the weed species; broad leaf, grass and
sedges (Esayas and Abraham, 2000). Higher weed density was recorded at
flowering and near harvesting growth stage of cotton resulting in reduction of
yield and harvest efficiency. Weed species such as Xanthium strumarium that
previously was not economical weed had become critical weed in cotton fields.
Similarly, a cotton seed yield loss of 35.03-88.13% and 56.45-94.44% occurred
when weeding was delayed for 60 and 75 Days after crop emergence (DACE),
respectively. This was shown that the major yield loss occurred up to 75 DACE
during the cotton growth period. In Werer State Farm (WSF) and Werer
Agricultural Research Center (WARC) experimental sites, the increase in cotton
seed yield was observed to be consistent with advancement of weed-free period.
On the other hand, the longer the weeds were allowed to grow and compete with
[279]
the crop, the higher the cotton seed yield reduction would be (Esayas et al., 2013).
In agreement with this study, loss of yield with increase in the weed infestation
period was reported in different crops including cotton (Young et al., 1984).
Bishnoi et al., (1993) reported that weed free environment from 20 days after
sowing produced highest seed cotton yields (2798 kg ha-1) compared to unweeded
control (1614 kg ha-1). Weeds can accumulate higher concentration of mineral
nutrients than crops, thereby depleting soil nutrients quickly and reducing the crop
yield (Tomaso, 1995; Gupta, 1998). It is estimated that losses in Arkansas, USA,
due to weeds amount to approximately 34 million dollars annually (Smith, 2000).
Khan and Khan (2003) reported that grassy weeds cause 15 – 40% and broad leaf
weeds 15 – 30% yield losses in cotton crop.
Cotton initial slow growth, wide row spacing, high dose of chemical fertilizers
combined with prostate nature of its growth permit early and severe crop weed
competition resulting in loss of yield to the tune of 45 to 85% (Das, 2008). At
present, manual weeding has become costly due to scarcity of laborers and hence
it has become extremely difficult to keep the crop weed free. Effective and
economical weed control in irrigated cotton is possible through integrating pre-
emergence herbicides along with hand weeding and interculturing. Therefore, the
present study was undertaken to determine better integrated weed management
practices for irrigated cotton production in Middle Awash Rift Valley of Ethiopia.

The experiment was conducted at Werer Agricultural Research Center (WARC)
experimental field and Gewane cotton commercial farm during the May – October
months in main cropping season of 2017 and 2018. Werer is located at 278 km
east of Addis Ababa with an altitude of 740 meters above sea level and at the
coordinates of latitude of 90° 60’22.5’’ N and longitude 40° 09’23’’ E. Whereas
Gewane is located at 350 km east of Addis Ababa, with an altitude of 350 meters
above sea level and at the coordinate of latitude of 90° 59’ 22.5’’ N and 40° 31’
23’’ E longitude.
The dominant soil type of the study areas is described as chromic vertisol; Sand
3.83%, Silt 61.1%, and clay 35.07 % and a bulk density of 1.17. The pH of the soil
is slightly alkaline (7.5 to 8.5). The mean annual rainfall is 540 mm and the mean
maximum and minimum temperatures are 34 0C and 19 0C, respectively (Esayas et
al., 2013).
The popular cotton variety in Ethiopia, Deltapine-90, was planted with a spacing
of 0.20 m x 0.9 m plots each having a size of 31.5 m2. All agronomic practices for
raising a successful crop were employed as per recommendation. The study
[280]
consisted of eight treatments arranged by combining different weed management
practices. The treatments were hand weeding + Intreculturing, Pre-irrigation +
hand weeding, pre-emergence herbicide (Pendimethalin) + hand weeding, Pre-
irrigation + Intreculturing, Pendimethalin +Intreculturing, Pre-irrigation+
Pendimethalin, Pre-irrigation and Control. The study was designed in a
Randomized Complete Block Design (RCBD) with three replications.
A quadrat with a dimension of 1 by 1 m was used in each plot sampling. Weed

type identification and density data collection were done within the quadrat once a
week starting from 20th day to 60th day after emergence of cotton. Data were
collected on yield and yield components of cotton; number of balls, plant
population, and plant height and seed cotton yield. The numbers of balls (open and
unopened) were recorded before the seed cotton picking. Seed cotton yield and
five ball weight was taken at time of picking. Plant height and number of plant
population per quadrat was recorded after picking. The data were recorded weekly
and the collected data were subjected to analysis of variance with SAS statistical
analysis package 9.2. When F-values were significant at P < 0.05 levels, means
were compared by Fisher's least significant difference (LSD) test.

Species composition
The data showed that a total of 40 weed species within 20 families recorded at
Werer and Gewane (Table 1). The most dominant families according to the
numbers of represented weed species were Euphorbiaceous, Poaceae,
Amaranthaceae, Convolvulaceae, Asteraceae, Fabaceae, Malvaceae and
Zygophyllaceae. Most of the weed species (77.5%) were erect annual broad leave
herbs, (12.5%) grasses, (2.5%) sedges and the rest (7.5%) were annual or
perennial climbers or perennial shrubs.
[281]
Table 1. Weed species life form, number and proportion (%) within the twenty plant families at study area
No Family Life form No. of species Percent flora

1 Euphorbiaceae Herb 6 15
2 Poacea Grass 5 12.5
3 Amaranthaceae Herb 5 12.5
4 Convolvulaceae Herb 3 7.5
5 Asteraceae Herb 3 7.5
6 Fabaceae Shrub 2 5
7 Malvaceae Herb 2 5
8 Zygophyllaceae Herb 2 5
9 Cucurbitaceae Shrub 1 2.5
10 Solanaceae Herb 1 2.5
11 Acanthaceae Herb 1 2.5
12 Tiliaceae Herb 1 2.5
13 Portulacaceae Herb 1 2.5
14 Nyctaginaceae Herb 1 2.5
15 Commelinaceae Herb 1 2.5
16 Chenopodiaceae Herb 1 2.5
17 Capparideae Herb 1 2.5
18 Polygonaceae Shrub 1 2.5
19 Papaveraceae Herb 1 2.5
20 Cyperaceae Sedge 1 2.5
Effect of weed management on weed abundance

The least weed abundance was recorded from pre-irrigation + Pendimethalin
application (Table 2) but the highest weed abundance recorded by pre-irrigation +
Interculturing and followed by weedy check at 20 days after cotton emergence
(DACE). Weed abundance at 35 DACE was significantly affected by the different
weed management practices. Hand weeding + Interculturing and Pendimethalin +
Interculturing were out performed the other treatments in reducing the weed
abundance. Weed management practices at 50 DACE was significantly different
that hand weeding + Interculturing, Pendimethalin herbicide + Interculturing and
Pre-irrigation outperformed the weedy control.
Management practices had a significant effect on the population of weed. Overall

hand weeding + Interculturing and Pendimethalin + Interculturing were recorded
the smallest weed abundance among the management practices. Tillage systems
and management practices determines the composition and abundance of weed
species in crop fields and can be helpful in understanding how particular weed
species increase or decrease, in terms of numbers and diversity, and how crop
management can contribute to the suppression of weeds (Travlos et al., 2018).
Takim and Uddin, (2010) had reported a significantly higher weed density in un-
weeded plots, which is in agreement with this finding. On the contrary, weed
density in growing seasons decreased with increasing duration of the weed-free
period (Dragica et al., 2008). As it was reported in results of weed surveys on
different crops in other places, field pea, faba bean, barley, wheat (Kedir et al.,
[282]
1999 a,b) and teff (Taye and Yohannes, 1998); there was a positive and significant
relationship among the weed species density, dominance and frequency. It was
recognized that the dominance level of individual weed species varied across the
crop growth stage.
Table 2. Effect of weed management on weed abundance at Werer and Gewane, 2017 and 2018
Treatment Weed abundance at Weed abundance at Weed abundance at

20 DACE 35 DACE 50 DACE
Hand weeding + Interculturing 126.7a 56.5b 57.08b
Pre-irrigation + hand weeding 143.4a 162.4ab 122.4ab
Pendamethaline + hand weeding 163.3a 79.63ab 84.88b
Pre- irrigation + Interculturing 199.5a 152.2ab 141.8ab
Pendamethaline + Interculturing 111.1a 56.5b 55.38b
Pre- irrigation + Pendamethaline 77.58a 73.83ab 94.33ab
Pre- irrigation 107.5a 191.9a 98.46ab
Control 181.3a 192.4a 205a
Mean 138.7813 110.7552 105.0439
C.V. (%) 26.1945 24.473 29.6279
LSD (0.05) 131.6 112.5 111.12
Effect of weed management on yield and

yield component of cotton
Different yield attributes, such as number of balls per plant and plant height were
affected significantly by different weed removal practices (Table 3). Hand
weeding + Interculturing, Pendimethalin + Interculturing and Pendimethalin +
hand weeding was found considerably increasing plant height unlike weedy
control and pre-irrigation. The decrease in plant height due to weed competition
was also observed by different authors (Singh, 1983 and Rushing et al., 1984).
Cotton stem diameter and height also decreased with increasing weed competition
(Keeley and Thullen, 1991b). Plant height and stem diameter reduced by weed
competition was also reported by (Snipes and Byrd, 1994).
Data of ball numbers per plant showed that there was significant difference among
the management practices. Hand weeding + Interculturing and Pendimethalin +
Interculturing were outperformed in decreasing weed abundance in the field and
then increasing the number of cotton balls per plant. Velayutham et al., (2002) and
Srinivasan (2003) reported that un-weeded check reduced the ball number per
plant and ball weight of cotton. Mohamed and Bhanumurthy (1985) reported
significant reduction in fruiting points due to uncontrolled weed growth in the
field.
The study showed that seed cotton yield was significantly affected by the weed
management practices. Hand weeding + Interculturing and Pendimethalin +
Interculturing were recorded higher yield than pre-irrigation and weedy check
(Table 3). A yield loss of 64% in seed cotton was recorded when weeding was
[283]
completely denied throughout the crop growing season. The hand weeding +
Interculturing and Pendimethalin herbicide + Interculturing management practices
were resulted high seed cotton yield advantages of 60.74 and 64 percent as
compared with weedy check. This result agrees with Esayas, et al. (2012) who
reported seed cotton yield loss of 62.43 - 96.21% when weeding was completely
denied throughout the crop growing season. Bishnoi et al. (1993) weed free
environment from 20 days after sowing produced highest seed cotton yields (2798
kg ha-1) compared to unweeded control (1614 kg ha-1).
Table 3. Effect of weed management on yield and yield component of cotton at Werer and Gewane during 2017 and 2018
crop season
Treatment Plant population Plant height Ball number Yield

Hand weeding + Interculturing 82a 71.917ab 16.15ab 23.978a
Pre-irrigation + hand weeding 88.67a 60.233bc 12.608abc 13.228bc
Pendimethalin + hand weeding 86a 62.95ab 15.25abc 19.272ab
Pre- irrigation + Interculturing 90.42a 54.533bc 10.95bc 12bc
Pendimethalin + Interculturing 91.17a 81.083a 19.992a 26.15a
Pre- irrigation + Pendimethalin 86.92a 54.533bc 9.008bc 10.44bc
Pre- irrigation 78.25a 51.4c 11.15bc 9.603c
Control 76.83a 50.967c 8.175c 9.414c
Mean 89.4063 61.41875 12.91042 15.51125
C.V. (%) 35.5869 29.35771 24.47796 27.50684
LSD(0.05) 25.965 19.727 7.8469 8.925
Effect of weed management practices on cotton lint quality

There was no significant difference among the weed management practices on
their effect on quality parameters (Micronior, Upper half mean length, Uniformity
index and Strength) of cotton lint (Table 4). Similarly, it had been reported that the
quality of cotton (Rushing et al., 1984) and ginning percentage of cotton (Singh
and Nagwekar, 1989) were not affected by weed infestation.
Table 4. Effect of weed management on quality of cotton at Werer and Gewane, 2017 and 2018
Treatment Micronior UHML (mm) UI (%) Strength (g/tex) Eig (%)

Hand weeding + Interculturing 4.9683a 27.708ab 82.25abc 23.9167a 8.6167a
Pre-irrigation + hand weeding 5.2217a 27.35ab 81.7833abc 22.4333b 8.5a
Pendimethalin + hand weeding 5.2833a 27.538ab 82.3833abc 22.4b 8.9a
Pre- irrigation + Interculturing 5.145a 27.315ab 81.25c 23.1667ab 8.7333a
Pendimethalin + Interculturing 5.38a 27.51ab 81.5333bc 23.0167ab 8.3167a
Pre- irrigation + Pendimethalin 5.1817a 27.197ab 82.3167abc 22.85ab 8.8667a
Pre- irrigation 4.945a 28.072a 82.8333a 23.65ab 8.8a
Control 5.4067a 23.48b 82.5333ab 23.45ab 8.4333a
Mean 5.191458 27.02125 82.1104 23.1104 8.645833
C.V. (%) 7.91985 13.5473 1.195396 5.19369 6.617975
LSD (0.05) 0.4848 4.3163 1.1573 1.4153 0.6747
[284]
The degree of damage of cotton from weed competition is related to the weed
species composition (type of weeds), weed abundance and the duration of weed-
cotton competition as related to the lifecycle of the cotton plants. Different weed
management practices affect the abundance of weed species in the cotton field. As
the abundance and duration of weeds in the cotton field increases the damage on
yield and yield component also increase. The current study showed that hand
weeding + interculturing or Pendimethalin + Interculturing shall be used as an integrated
weed management practice in cotton at Werer and Gewane areas.
References
Bishnoi LK, Panwar RS, Malik RK and Rathi SS. 1993. Effect of varieties and weed free
maintenance period on weed competition in cotton. In: Integrated weed management for
sustainable agriculture. Proc. of the Indian Society of Weed Science, International
symposium, held on the 18-20 November. Hisar, Haryana, India; 3:182-183.
Das TK. 2008. Weed Science: Basic and Applications. Jain Brothers, New Delhi, India
Dragica Spasova S, Mitrev S, Spasov D and Biljana A. 2008. Critical Period of weed Competition
in Cotton. Union Scientist Stara Zagora International Scientific Conference Held on Jun 5-6,
2008 GoceDelcev University - Stip, R. Of. Macedonia.
Esayas T and Abraham GH. 2000. Quantitative and qualitative survey of weeds growing in
association with cotton in the Middle and the Lower Awash. Progress report for the period of
2000/2001: Werer Agricultural Research Center.
Esayas T, Abraham GH and Mashila D. 2013. Ethiopian Journal Applied Science Technology 4:
41-49.
Esayas T, Abraham GH and Mashila D. 2012. Ethiopian Journal of Applied Science Technology 3
(1): 57- 69.
Gupta OP. 1998. Modern Weed Management, Agro Botanica Publishers, Bikaner, India, 488 PP.
Kedir N, Feyissa T and Tilahun G. 1999a. Results of weed survey in the major barley- and wheat
growing areas of the Bale highlands. Arem 5: 85-95.
Kedir N, Feyissa T and Tilahun G. 1999b. Results of weed survey in the major field pea and faba
bean growing-areas of the Bale highlands. Arem. 5: 109-121.
Keeley PE and Thullen RJ. 1991a. Growth and Interaction of Bermuda grass (Cynodon dactylon)
with cotton (Gossypium hirsutumL.). Weed Sci., 39(4): 570-574.
Keeley PE and Thullen RJ. 1991b. Growth and interaction of barnyard grass (Echinochloa
crusgalli) with cotton (Gossypium hirsutum). Weed Sci. 39:369–375.
Khan NU and Khan SU. 2003. Integrated weed management in upland cotton. Pak. J. Weed Sci.
Res., 9(3 and 4):185-192.
Mofett JE and Mcclosky WB. 1998. Effects of soil moisture and yellow nut sedge (Cyperuse
sculentus) density on cotton (Gossypium hirsutum L.). Weed Sci., 46(2): 231-237.
Mohamed Ali A and Bhanumurthy VB. 1985. Trianthema portulacastrum L. under irrigated
conditions. Trop. Pest Mgt., 31(3): 232-234
Rushing DW, Murray DS and Varhalen LM. 1984. Tumple pigweed interference with cotton.
Weed Abstr.,35: 179.
Singh JN. 1983. Mechanical and chemical weed control in cotton. Indian J. Weed Sci. 15: 69-71.
Singh V and Nagwekar SN. 1989. Effect of weed control and nitrogen levels on quality characters
in cotton. J. Indian Soc. Cott. Improv., 14(1): 60-64.
[285]
Smith KL. 2000. Controlling weeds in cotton. In: Proceedings of the 2000 Cotton Research
Meeting. AAES: Special report, 33-37.
Snipes CE and Byrd JRJD. 1994. The influence of fluometuron and MSMA on cotton yield and
fruiting characteristics. Weed Sci., 42(2): 210-215.
Srinivasan G. 2003. Boiefficacy of prometryn for weed control in summer irrigated cotton. Madras
Agric. J., 90 (4-6): 243-246.
Srinivasulu G and Rao AS. 2000. Effect of sequential application of herbicides on weed
management in cotton. In: proc. Of symposium on challenges in Agronomic crop
management in early 21st century May, 24-25 organized by society of Agronomists,
Hyderabad,71-74 PP.
Tadesse E and Ahmed S. 1985. A review of weed research in Ethiopia. In Proc. of the first crop
protection symposium, 4-7 February, Institute of Agricultural Research, Addis Ababa,
Ethiopia. 233-244 PP.
Takim FO and Uddin RO. 2010. Effect of weed removal on insect populations and yield of
Cowpea [Vignaun guiculata (L) Walp] Australian Journal of Agricultural Engineering,
1(5):194-199.
Taye Tand Yohannes L. 1998. Qualitative and quantitative determination of weeds in teff in west
Shewa Zone, Arem, 4: 46-60.
Tomaso JM. 1995. Approaches for improving crop competitiveness through the manipulation of
fertilization strategies. Weed Sci., 43(3): 491-497.
Travlos IS, Cheimona N, Roussis I and Bilalis DJ. 2018. Weed-Species Abundance and Diversity
Indices in Relation to Tillage Systems and Fertilization. Front. Environ. Sci. 6:11. doi:
10.3389/fenvs.2018.00011
Velayutham A, Ali AM and Veerabadran V. 2002. Influence of intercropping systems and weed
management practices on the growth and yield of irrigated cotton. Madras Agric. J., 89 (1-3):
59-62.
[286]
PART III
Agricultural Entomology
[287]
[288]
Studies on Geographic Distribution, Seasonal
Abundance and Insecticidal Control of White
Mango Scale (Aulacaspis tubercularis)
(Hemiptera: Diaspididae)
Birhanu Sisay, Abiy Fekadu, and Gashawbeza Ayalew
Ethiopian Institute of Agricultural Research, Melkassa Center, PO Box 436, Adama
Abstract
Series of studies on geographic distribution and severity of white mango scale (WMS)
infestation in mango fields of northern, central and southern Ethiopia; seasonal
abundance of the pest in the Central Rift valley region and screening of insecticides
against white mango scale were made between 2016 and 2019. In 2017, a total of 61
mango fields were surveyed. The pest was detected only in central Ethiopia (Adama,
Melkawoba and Degaga) with 60 to 100% infestation. In 2018 a total of 78 mango
fields were surveyed and the pest was detected only in Bahar dar and Sodo towns with
infestation level ranging from 0 to 100%. Population of the pest increased during
hotter period (February through May) and decreased during the rainy period and
cooler months (July to December). The systemic insecticide, Movento OD with
adjuvant significantly reduced the WMS population. The performance of the registered
insecticide, spark WG (Thiametoxam), for the control of white mango scale on mango
was on par with Movento OD. The detections of the pest in scattered mango trees of
bigger towns and its absence or low infestation level in close and distant areas of
northern western and southern regions suggest that bigger towns are gateways for the
WMS invasion. Hence, a containment activity is critical to eradicate or slow the
spread of WMS. Chemical control of WMS should target the breeding period of WMS
which was observed to peak in hotter months in CRV. Integration of cultural control
such as removing of heavily infested branches with more effective soil and foliarly
applied systemic insecticides along with biological control should form the basis of
Integrated WMS management.
Introduction
A range of tropical and sub-tropical fruit crops are cultivated in Ethiopia, among
which banana, mango, papaya, citrus, avocado, grape, guava and pineapple are the
most important. The area under fruit crops is estimated at about 94.6 thousand
hectares of land, most of which is owned by smallholder farmers, who produced
over 0.8 million tons of fruits in the meher (main production) season of 2014/15
with average productivity of 7.8 tons/-ha (CSA, 2015). Mango crop is an
important trading commodity worldwide and currently, it is the second most
internationally traded tropical fruit (FAO, 2013). In Ethiopia, mango is produced
on about 8809 ha with a total annual production of about 697, 507 quintals
(Tesfaye et al., 2014).
[289]
Cultivation of mango is currently constrained by emerging pests and diseases,
poor and inadequate supply of improved quality and certified seedlings. Yield
losses due to pests and diseases and poor post-harvest handling of produce are
enormous. Insect pests, notably the White mango scale (WMS) (Aulacaspis
tubercularis) are among the largest threats to mango production in Ethiopia
(Mohammed et al., 2012, Gashawbeza et al., 2015; Djirata et al., 2018). Besides
mango, several plant species including citrus, papaya, avocado, ginger, cinnamon,
pumpkin etc., are hosts to A. tubercularis (http://www.
plantwise.org/KnowledgeBank /Datasheet.aspx). The pest is Asian origin and
distributed all over the world. CABI distribution map of pests show that A.
tubercularis occurs in several countries of West Africa and several Sub-Saharan
African countries (http://www.plantwise.org/ KnowledgeBank/Datasheet.aspx).
Threats by invasive pests result from loose quarantine system, globalization,
increased human mobility, and climate change (Eshetu and Binyam, 2012).
Occurrence of WMS in Ethiopia was first confirmed following its outbreak in
Weke site of Green Focus Ethiopia in Guto Gida district of East Wollega zone of
Oromia region (Mohammed et al., 2012). Efforts in the management of the pest
has so far been limited to advising growers to prune and burn heavily infested
branches, use of insecticides registered for the control of related armored scale
and use of seedlings free from the pest for planting. Information on seasonal
fluctuation of the pest with the aim of timing the application period was not
available. The current studies were therefore initiated to determine the distribution
of the insect in selected mango producing regions of Ethiopia, identify effective
insecticides for use as an integral component of IPM and to determine seasonal
variability in the population of the insect.

Survey on geographical distribution and damage of White Mango Scale
Mango orchards were identified by communicating with the agricultural bureaus
of the regions and districts. Orchards located along the main roads of South,
North-west and Central Ethiopia were considered. The level of leaf infestation was
determined by visual estimation of proportion of leaf infested. Estimation was
made on whole tree (canopy) basis. Location details of the areas where the survey
conducted were recorded using GPS.
Seasonal Abundance of white mango scale

Mango orchards that have never been treated with insecticides were selected for
this study. A total of six mango trees were selected randomly at Melkassa
Agricultural Research Center from May 2016 to November 2018 (Fig. 3). Five
mango leaves per tree were randomly picked in different direction (North, east,
south and west) from lower branches. Sampled leaves were kept in a plastic bag
[290]
and transported to the entomology laboratory of Melkassa Agricultural research
center (MARC) for counting female, male and crawlers of white mango scale. The
life stages of the insect were examined under a microscope by lifting or breaking
the scale cover with a thin needle. Data were collected at ten days intervals and
summarized in to monthly basis.
Screening of insecticide against white mango scale

Five different insecticides, namely Confidence 350 SC, Spark 250 WG, Best 5
EC, Movento 150 OD and Movento energy SC 240 were tested at manufacturers’
rate against white mango scale. Except Spark 250 WG which was applied by
drenching, the foliar applied insecticides were tested with and without adjuvant.
A total of eleven treatments were arranged in RCBD with three replications. The
experiment was conducted in a mango orchard located at Doni. Mango trees with
similar level of white mango scale infestation were selected for treatment
application. Thirty-three mango trees were selected for the experiment where a
single mango tree forms a plot. Prior to commencement of treatment application,
data on the initial white mango scale population was collected. Five mango leaves
per tree were sampled and examined under microscope to determine white mango
scale population. Data on white mango scale population were collected prior to
each treatment application. A motorized backpack sprayer of 15lt capacity was
used to apply treatments (i.e. for foliar applied treatments). Treatments were
applied for a total of six times at biweekly starting from May 2018. Collected data
was subjected to analysis of variance using SAS statistical package (SAS system
9.1version). Students Newman Keuls test at 5% Significance level was used to
separate treatment means of significant difference.

Survey on geographical distribution and damage of white mango scale
In 2016 the pest was detected only in central Ethiopia (Adama, Melkawoba, and
Degaga) (Fig 1). The survey was repeated in December 2017(Fig. 2) covering the
same area as in 2016. During this time (December 2017), mango trees in Bahir
Dar town (Kebele 14, 15, 16, and 17) showed up to 100 % infestation. Similarly,
up to 80% trees were infested in mango fields located in the outskirt of Sodo in
Southern Ethiopia. This suggests that due to absence of domestic quarantine,
mango orchards in big towns are being invaded by the pest probably due to
transportation of infested fruits.
[291]
Fig 1. Distribution and severity of WMS in different parts of Ethiopia: Yellow (60- 100% Infestation; green (no infestation)
Fig 2. Distribution of white mango scale in northwest, CRV and southern Ethiopia, 2017
[292]
Seasonal abundance of white mango scale
Population of white mango scale increased during hotter period (February through
May) and lowered rainy cooler months (July to December) (Fig. 3). This finding
agrees with Djirata et al. (2018) who recorded higher population density of white
mango scale during months of high maximum temperature.
33.0
Temperature (Max.)
31.0
29.0
27.0
25.0
400
WMS populaton
300
200 mea…
100
0 Sep-18
Sep-16
Sep-17
Nov-16
Nov-17
May-16
Mar-17
May-17
Mar-18
May-18
Jul-16
Jan-17
Jul-17
Jan-18
Jul-18
Nov-18
Sampling months
Fig 3. Fluctuation in the population of white mango scale on mango (May 2016 to Nov 2018) (bottom) and maximum
temperature during the study period (top)
Screening of insecticide against white mango scale

Significant differences in the number of white mango scale between treatments
were observed after the second, third, fifth and sixth sprays (Fig. 4). The mean
number of white mango scale per leaf during these periods ranged from 6-124, 17-
188, 5-51 and 27-439, respectively after the second, third, fifth and sixth sprays.
Differences in the rest of the sampling occasions were not significant including the
overall mean (Figs. 4 and Fig. 5). Trees treated with Movento energy SC 240 with
adjuvant resulted in significantly lower mean number of white mango scale than
the treatments Movento energy SC 240 without adjuvant, Best 5 EC and Movento
150 OD and Confidence 350 SC respectively after the third, fifth and sixth sprays
(Fig. 4). However, treatments did not differ significantly for overall mean
although population of WMS appeared to be lower in trees treated with Movento
energy SC 240 with adjuvant (Fig. 5).
[293]
Number of white mango scale
180 A
160
140
120
100
80
60
40
20
Treatments
160 B
140
120
100
80
ab
60
40
20
Treatments
[294]
250

C
200
ab
150
ab
100 ab
ab ab
50 ab ab
ab
Treatments
100
D
90
80
70
60
50
40
30
20
10
Treatments
[295]
60 E

a a
50 ab
40 ab ab
ab
30 ab ab
ab
20 ab
10
Treatments
500
F
450
400
350
300
250
200
ab
150
ab ab ab
100
50
Treatments
Fig 4. A,B,C,D,E and F shows mean number of white mango scale after first, second, third, fourth, fifth and sixth sprays
respectively
[296]
120
100
80
60
40
20
Treatments
Fig. 5. Mean number of white mango scale for six after spray counts (overall spray effect)

Population of the pest increased during hotter period (February through May) and
decreased during the rainy period and cooler months (July to December).
Chemical control of white mango scale should target periods of population
buildup during hotter months in the CRV. The detections of the pest in scattered
mango trees of bigger towns and its absence or low infestation level in close and
distant areas of northern western and southern regions suggest that bigger towns
are gateways for the WMS invasion. Hence, a containment activity is critical to
eradicate or slow the spread of WMS. The results on chemical control study
suggest that, trees treated with Movento energy and adjuvant suffered less from
white mango scale infestation and damage. Hence, chemical control of WMS may
consider the use of Movento energy OD as foliar application and the registered
soil applied insecticide Spark WG250. However, the emphasis on the integrated
management of WMS should be on biological control using effective and proven
natural enemy along with cultural methods such as removal of heavily infested
branches.
Acknowledgment
The study was financed by the Ethiopian Institute of Agricultural Research. We
are grateful to the technical and field assistants of entomology section of MARC
for their assistance in execution of the study and data collection.
[297]
References
CSA (Central Statistical Agency). 2015. Statistical bulletin of agricultural sample survey. Report
on area and production of crops. Addis Ababa.
Djirata O, Getu E, Ruth K. 2018. Population dynamics of white mango scale, Aulacaspis
tubercularis Newstead (Hemiptera: Diaspididae) in Western Ethiopia. Afr. J. Agric. Res.
http://www.academicjournals.org/AJAR ,Vol. 13(31), pp. 1598-1605, 2.
Eshetu Derso and Binyam Yalemtesfa. 2012. Citrus canker (Xanthomonas axonopodis pv. citri)
and Huanglongbing (greening) (Candidatus liberibacter) Invasive Citrus Diseases in Ethiopia
FAO. 2013. Rome, United Nations Food and Agriculture Organization.
(http://faostat3.fao.org/faostat-gateway/go/to/home/E., Accessed on March 28, 2015).
Gashawbeza Ayalew, Abiy Fekadu and Birhanu Sisay. 2015. Appearance and chemical control of
White mango scale (Aulacaspis tubercularis) in Central Rift Valley. Sci,Technol.Arts Res.J.
4(2):59-63.
Mohammed Dawd, Belay H/Gebriel, Lemma Ayele, KonjitFeleke, Seyoum Hailemariam and
Teshome Burka. 2012. White Mango Scale: A new insect pest of mango in western Ethiopia.
pp 257-267.
Tesfaye Hailu, Solomon Tsegaye, and Tadele Wakuma. 2014. White mango scale insect
infestation and its implication in GutoGida and Diga districts of East Wollega Zone. ABC
research Alert 2 (2): pp 33.
[298]
Screening of Common bean Genotypes for their
Resistance to the Mexican Bean Weevil
(Zabrotes subfasciatus)
Mulatwa Wondimu and Berhanu Amsalu
Ethiopian Institute of Agricultural Research, Melkassa Centre, P.O.Box 436, Adama
Abstract
Seed beetles (Bruchids) cause great damage to stored beans. They are important on
common bean (Phaseolus vulgaris L.) and include Zabrotes subfasciatus
(Boheman) Acanthoscelides obtectus (Say) and Callosobruchus maculatus (Fab.).
A variety of methods including host plant resistance are used to minimize damage by
these pests. Bean materials introduced by the lowland pulse breeding program which
consisted of 64-MAZ lines, 14-released varieties and 23 candidate genotypes were
screened for their resistance to Z. subfasciatus using artificially reared adults in the
entomology laboratory of Melkassa Agricultural Research Center. A completely
randomized design with three replications was used. Data on number of eggs laid,
number of hole per seed, F1 progeny emerged, percent damaged seeds, median
developmental period, Dobi susceptible index and percent weight loss were collected
to measure resistance variability. Fewer number of F1 progeny emerged from eggs
laid on six Maz-lines: Maz-2, Maz-7, Maz-17, Maz-21, Maz-26, Maz-68 and six
varieties, Hawassa Dume, Glp2, MLRB, NLRB, SW (N1) and LRB. Percent seed
damage was lesser on nine Maz-lines: Maz-5, Maz-50, Maz-64, Maz-67, Maz-74,
Maz-84, Maz-76, Maz-77, Maz- and Maz-99. Lower percent weight loss was
recorded on seven genotypes, Median development period ranged from 28-37, and
most Maz-lines prolonged the developmental time indicating the resistance exhibited
against Z. subfasciatus. Based on Dobie index of susceptibility seventeen genotypes
were found resistant and twenty five as moderate resistant. The study enabled to
identify substantial number of resistant and moderately resistant genotypes and these
genotypes will be used for bruchid resistance breeding.
Introduction
Common bean (Phaseolus vulgaris L.) is one of the principal food and cash crop
legume grown in the tropical world (Abate and Ampofo, 1996; Songa and Rono,
1998; Schmale et al., 2002). A wide range of seed beetles attack the grain of
common bean varieties (Mulungu et al., 2007). However, the predominant
damaging pests of stored grain legumes mainly in the tropics are Callosobruchus
maculatus (Fab.), C. chinensis (L.), Caryedon serratus (Oliver), Zabrotes
subfasciatus (Boheman) and Acanthoscelides obtectus (Say) (Nichimbi-Msolla
and Misangu, 2002; Emana et al., 2003). In Ethiopia, Z. subfasciatus and A.
obtectus are the major pests of stored beans. These storage pests have been found
to cause average grain losses of 60% within 3 to 6 months of storage period
(Emana et al., 2003). To offset and/or minimize the losses, farmers use different
control measures which range from synthetic insecticides to cultural control
[299]
methods. Synthetic insecticides may play a significant role in reducing storage
losses due to insect pests (Tapondjou et al., 2002). However, application of these
pesticides for the control of storage insect pests is limited because of resistance
development by the pest, consumers concern, widespread environmental hazards
and increasing costs of application (Bekele et al., 1995; Bekele, 2002).
Additionally, other control measures also have their own side effects. Hence,
increasing attention is being given to the development of resistance genotypes.
Thus, this study was carried out to identify bean genotypes resistance to the most
important damaging species, Z. subfasciatus, in the Central Rift Valley region of
Ethiopia.
Material and methods

This experiment was conducted in the entomology laboratory of Melkassa
Agricultural Research Center (MARC) from February to May of 2017 & 2018.
MARC is located in the Central Rift Valley of Ethiopia at 8o 24’N latitude and 39o
12’E longitude and at an elevation of 1550 meter above sea level (masl). The
maximum and minimum annual mean temperatures are 28oC and 14oC,
respectively (MARC, 1996).
Bean Seed preparation

A total of 101 (64 –Maz lines, 15 released varieties, 22 candidate genotypes)
common bean materials obtained from CIAT and national lowland pulse research
program were screened using laboratory reared adult Z. subfasciatus in 2017 and
2018 at MARC. Before the commencement of the screening, the seeds of each line
or genotype were multiplied to get sufficient seeds for the experiment at MARC.
Harvested seeds were disinfected by storing them in deep freezer following
storage in cold room to avoid contamination. The characteristics of the bean
genotypes evaluated are described in Table 1.
Table 1. Characteristics of the bean genotypes tested for resistance to Z. subfasciatus

Characteristics
Bean genotypes Seed size (Large/ Seed color and shape Growth Maturity
Medium/Small habit (days)
Maz - lines Large Red Bush 85-92
Zebra medium Cream bush Bush 88-92
Dimtu small Red Elong Bush 91-93
Argene small White Bush 90-95
Goberasha large Red mottled Elong Bush 92-100
Red wolayta small Red Elong Bush 85-90
Morka large Red mottled Elong/kidney Bush 84-115
Awash-2 small White round Bush 85-90
GLP-2 large Red mottled Elong/kideny Bush 85-90
Ada medium Yellow Round Bush 75-79
MelkaDima medium Red Elong Bush 79-102
Nasir small Red Bush 86-88
Awash-1 Small White Round Bush 90
[300]
Rearing and inoculation of Z. subfasciatus
To obtain the same age group and required numbers of adult bruchids for the
experiment, Z. subfasciatus was reared in the entomology laboratory of MARC.
The materials used for rearing were collected from MARC stores and reared in
three small bags having 5 kg capacity each. About 100 adults of Z.
subfasciatus were added to each small bag by keeping the proportion of male and
female equal (1:1). Rearing was done at room temperature. Inspection of the
culture for emergence of the progeny was made daily starting from twenty two
days after parent bruchids introduction (Bhardwaj and Verma, 2012). The newly
emerged one-day old adult F1 progenies were used for the experiment (Bhardwaj
and Verma, 2012). Accordingly, 5 male and 5 female F1 progeny adults were
transferred to each transparent 200 ml capacity jar. Fifty common bean seeds were
added to each jar for the test. The jars were covered with screw caps having
muslin cloth to prevent bruchids from escaping. The beetles were allowed to mate
for seven days at 27 ± 1°C and 60 to 90% relative humidity for egg laying and
then removed.
Experimental design and data collection

All the jars were laid out on laboratory shelves at room temperature in a
completely randomized design with three replications. The bruchids were allowed
to oviposit for seven days and then discarded. The following data were collected:
initial and final weight of seed, number of emerged bruchids, median development
period, number of damaged and undamaged seeds, number of holes per seed,
weight of damaged and undamaged seeds.
Resistance classification
Resistance was measured by the number of holes per seed, seed damage
percentage, seed weight loss incurred due to damage caused by bruchids, the
number of emerged F1 bruchids, and the period required for 50% of the insects to
develop within the bean seeds.
Seed weight loss was calculated after 90 days using the following formula
(Kamanulaa, et al., 2010)
(WuXNd )  (WdXNu )
Weight loss (%) = x 100
WuX ( Nd  Nu )
Weight loss (%) = Where;
Wu= weight of undamaged seed and Wd= weight of damaged seed

Nd= number of damaged seed and Nu= of undamaged seed
The number of holes per seed (NHPS) was calculated as follows:

NH
Nd
[301]
NHPS = Number of Holes per Seed. NH = Total
number of holes
Nd = Total number of damaged seeds
The median bruchid development period (MDP) on each genotype was calculated at
mid-period of oviposition (5 days from initial infestation time) to the emergence of
50% of the first generation out of total adults (Derera et al., 2010),.
The number of emerged adult bruchid and the median development period were
combined to calculate the susceptibility index (Dobie, 1974) for each variety using
the equation:
LogeY
x 100
MDT
DSI =
Where; Y = total number of adult bruchid emerged MDT= median development period
Based on Dobie susceptibility Index, bean genotypes were grouped into four
resistance classes (Table 2).
Table 2. Classification of resistance based on Dobie Susceptibility Index (DSI)
DSI Resistance class

0-3 Resistant
4-7 Moderate resistant
8-10 Susceptible
≥11 Highly Susceptible
Data were analyzed using the Proc GLM procedure of SAS (SAS, 9.0). Significant means
were separated using the Student – New man-Keuls test.

Significant differences were observed between genotypes on a number of
parameters considered. Smaller number of F1 progeny weevils emerged from six
Maz lines, Maz-2, Maz-7, Maz-17, Maz-21, Maz-26, Maz-68 and six varieties,
Hawassa Dume , Glp2, MLRB,NLRB,SW(N1) and LRB from high number of
eggs laid (Fig, 1).
A lower percent damaged seed was recorded on nine Maz-lines: Maz-5, Maz-50,
Maz-64, Maz-67, Maz-74, Maz-84, Maz-76, Maz-77, Maz-and Maz-99 as
compared to susceptible check (Table 3). Lower percent weight loss was similarly
recorded from seven genotypes, Maz-1, Maz-5, Maz-34, Maz-74, Maz-80, Maz-
84 and Maz-90, (Table 3).
Mean of Median Development period also ranged 28 up to 37 days, greater than

and equal to thirty MDT were found (MDT) on nine genotypes, Maz-7, Maz-9,
[302]
Maz-13, Maz-84, Maz-85, Maz-55, Maz-57, Maz-89, and MICPY. Giga and
Chinwada (1993) indicated that the development periods of those adults that
emerged from the RAZ lines were significantly longer (39.6-69.0 days) than those
from the susceptible check (29.4 days).
Number of holes per seed showed similar trend to weight loss of genotypes (Fig,
2). Mean of two year results (2017 & 2018) revealed that among 101 genotypes,
only sixteen Maz lines Maz-1, Maz-5, Maz-7, Maz-9, Maz-13, Maz-31, Maz-34,
Maz- 35, Maz-57, Maz-55, Maz-72, Maz-85, Maz-89, Maz-99, Maz-76, Maz-77,
and one variety Morka were identified as resistance genotypes. The other eighteen
Maz- lines were classified as moderate resistance material like, Maz-17, Maz-21,
Maz- 26, Maz-32, Maz-33, Maz-50, Maz-61, Maz-62, Maz-64, Maz-67, Maz-73,
Maz-74, Maz-79, Maz-80, Maz-82, Maz-84, Maz-90, Maz-75, and seven released
varieties Tabor, LRMB, SW(N1), LRB, MICPY, Red kidney and Awash-1
susceptible, based on Dobi susceptible index (DSI) (Table 3).
Total eggs laid and F1 progeny emerged
20
15
10
Total egg laid
F1 progeny emerged
Genotypes
Figure 1. Total eggs laid and the corresponding F1 progeny emerged
[303]
10.00
12.00
14.00
0.00
2.00
4.00
6.00
8.00
MAZ -2
MAZ -10
MAZ -23
MAZ -28
MAZ -42
MAZ -56
MAZ -96
MAZ -111
MAZ -139
MAZ -142
MAZ -145
MAZ -148
MAZ -151
MAZ -154
MAZ -157
MAZ -160
MAZ -163
MAZ -166
Genotypes
MAZ -169
MAZ -172
MAZ -175
MAZ -178
Argene
Morka
GLP-2
Figure 2. Number of holes per seed and percent weight loss in bean genotypes screened for resistance to Z. subfasciatus
Chore
MIRB
SW(N1)
SW(N1)
Number of holes per seed (NHPS) and percent weight loss%
NLRB
MLCPY
LS-sugar
Red kideny
F8 drought line
NHPS
Table 3. Resistance class, damage level, weight loss and reproduction of Z. subfasciatus in bean genotypes tested for resistance
No Genotypes Total F1 progeny Damaged seed Median Dobi Susceptibility Weight Resistance class
emerged (%) Development period loss index (DSI) %
1 MAZ- 1 2b 1.44ab 19 ns 1.731bcd 1.97bc Resistant
2 MAZ - 5 1.67b 0.88b 29 0d 1.95bc Resistant
3 MAZ - 7 3.67b 3.17b 35 1.185bcd 2.159bc Resistant
4 MAZ- 9 31.67ab 13.83ab 31 0.00d 2.42bc Resistant
5 MAZ -13 6.67b 4.10b 37 2.741bcd 2.32bc Resistant
6 MAZ - 17 2b 1.26b 23 4.308abcd 2.302bc Moderate Resistant
7 MAZ- 21 8.67b 3.66b 32 6.724abcd 2.253bc Moderate Resistant
8 MAZ- 26 14b 5.86b 30 5.742abcd 2.155bc Moderate Resistant
9 MAZ - 31 1b 2.11b 29 1.850bcd 2.24bc Resistant
11 Maz- 33 2.33b 1.96b 13 5.320abcd 2.04bc Moderate Resistant
12 Maz-34 38.33ab 25.39ab 29 1.588bcd 1.74bc Resistant
13 Maz- 35 30ab 17.95ab 30 1.308bcd 2.04bc Resistant
15 MAZ - 57 13,67b 3.69b 30 1.962bcd 2.163bc Resistant
16 MAZ - 55 14.33b 8.20b 30 2.524bcd 2.18bc Resistant
18 MAZ - 62 8.33b 4.56b 20 6.307abcd 2.22bc Moderate Resistant
20 MAZ- 67 1b 0.449b 22 3.797abcd 2.96bc Moderate Resistant
21 MAZ - 72 2,67b 5.10b 14 1.68bc 3.062bcd Resistant
22 MAZ - 73 7.33b 3.14b 19 4.943abc 2.177bcd Moderate Resistant
23 MAZ - 74 1.67b 0.772b 25 3.779abc 1.815bcd Moderate Resistant
28 MAZ - 85 8b 2.31b 33 0d 2.26bc Resistant
29 MAZ- 89 32.33ab 14.88ab 30 0.797abc 2.134bc Resistant
30 MAZ- 90 18b 10.86ab 29 5.476abcd 1.859bc Moderate Resistant
31 MAZ- 99 0.67b 0.265b 20 1.650bcd 2.18bc Resistant
[305]
Cont’d Table 3
32 Maz- 75 10.67b 5.77b 30 6.172abcd 2.32bc Moderate Resistant
33 Maz- 76 3b 0.749b 29 2.635bcd 2.26bc Resistant
34 Maz- 77 0.67b 0.238b 10 0.745cd 2.36bc Resistant
35 Tabore 53.33ab 14.13ab 19 5.594abc 2.856bcd Moderate Resistant
36 LRMB 35.33ab 23.12ab 29 6.77abc 3.721abcd Moderate Resistant
37 SW(N1) 28.33ab 5.93b 19 5.84abc 2.726bcd Moderate Resistant
38 LRB 80.33a 25.33ab 29 6.627abc 4.774abcd Moderate Resistant
39 MLCPY 51.67ab 14.71ab 31 6.942abc 3.673abcd Moderate Resistant
40 Red kidney 19.33ab 2.32b 22 4.479abc 2.364bcd Moderate Resistant
41 Chore 20ab 4.59b 28 9.37a 6.402abcd Susceptible
42 Morka 2.33b 1.96b 13 1.663bcdd 2.04bc Resistant
43 Awash1 19.33ab 2.32b 22 3.916abcd 3.03bc Moderate Resistant
CV% 16.2 14.87 10.99 15.13 12.34
ns=non-significant
[306]
Twelve genotypes, 6 - MAZ lines, 4-candidate genotypes and 2 released varieties
have been found to be resistant to bean bruchid according to Dobi’s susceptibility
index. Most of the resistant MAZ lines are known to have arceline gene that inhibits
growth of larvae. Nine Maz lines prolonged median development period indicating
the level of resistance against to Z. subfacicatus. Similarly, lower percent weight
loss was recorded in Maz lines. Adult emerging holes showed similar trend to
weight loss. In summary, this study identified 17 Z. subfasciatus resistant sources or
genotypes, 16 Maz lines and one released variety, and 25 moderately resistant
sources, eighteen Maz-lines and 7 varieties based on Dobi’s susceptible index. We
therefore recommend the identified resistant and moderately resistant sources for
further bruchid resistance breeding program.
Acknowledgement
The authors are very much grateful for the germplasm, technical and financial
support of Tropical legume III (TL III) through International Centre of Tropical
Agricultural (CIAT) Cali, Colombia and Pan Africa Bean Research Alliance
(PABRA), Uganda for execution of this experiment.
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[308]
Evaluation of Botanical and Synthetic Insecticides
against the Cabbage Aphid Brevicoryne brassicae
(Linnaeus) on Ethiopian Kale
at Debre Zeit, Ethiopia
Ermias Shonga, Fasil Tadesse and Getachew Tabor
Debre Zeit Agricultural Research Center, POBox 32, Debre Zeit
Abstract
Cabbage aphid, Brevicoryne brassicae L. is one of the most important pests on cabbage
worldwide and in Ethiopia. Field experiment was carried out at Debre Zeit Agricultural
Research Center during 2017 and 2018 cropping seasons by using a kale variety locally
called ‘Gurage gomen’ to evaluate the efficacy of selected plant extracts and insecticides
for the control of the pest. Complete Randomized Block Design with three replications
was used. The effect of botanical treatments and synthetic insecticides alone or in
combination against B. brassicae and effect on associated beneficial insects were
assessed. Treatments showed significant differences on cabbage aphid infestation and
beneficial insects population reduction. Effects were non- significant for fresh leaf yield
and plant canopy spread. The results obtained proved that garlic+onion+chilli extract
and Neem oil were the best indigenous products for aphids control on kale with
comparable performance to the synthetic insecticide Dimethoate 40% EC. The use of
natural and easily biodegradable crop protection products like Azadirachtin can be a
useful component of an IPM program because of minimal harm to beneficial insects.
Therefore, these products can form integral components in the IPM program of cabbage
aphid in the Central Rift Valley of Ethiopia.
Introduction
Ethiopian Kale, (Brassica carinata L) commonly known as ‘Gurage Gommen’ is a
cool season crop that belongs to the Brassica family. It is an important smallholder
subsistence crop in Kenya, Ethiopia, Zimbabwe and Mozambique (Lo’hr and Kfir,
2004). Kale is grown for its leaves which are rich in vitamin A, thiamine, ascorbic
acid and essential mineral elements. It is widely utilized alone or mixed with other
vegetables, pulses or meat. It is an all-year crop and important for food security.
Kale is grown in a wide range of climatic conditions provided water is available. In
Ethiopia Kale is grown at an altitude ranging from 800 – 3200 masl. According to
the CSA (2015/16) report, Ethiopia produces 3.3 million MT of Cabbage in the
main season of 2015/16 from 33,942.01 ha area with an average productivity of
about 9.71 t/ha.
Among the yield limiting factors, insect pests are major ones and sometimes cause
complete failure of the crop. Cabbage Aphid, Brevicoryne brassicae (L) is an
[309]
important pest of cabbage in all growing areas of the world (Bhatia and Verma,
1994; Dattu and Dattu, 1995). Adults and nymphs feed by sucking plant juices
causing yellowing, curling of leaves, wilting and stunting of plants. In seasons of
severe infestation, yield may decrease by up to 80% (Atwal, 1976). Cabbage aphids
also transmit a number of viruses of brassica crops like cauliflower and turnip
mosaic viruses which can be managed by effective control of aphid (Chowfla and
Baruah, 1990).
According to Tsedeke and Gashawbeza (1994), the cabbage aphid is a major pest
causing economic damage to Brassica crops production in Ethiopia. Under
favorable conditions, feeding damage from large number of aphids can kill
seedlings and young transplants. On larger plants, its feeding results in curling and
yellowing of leaves, stunting plant growth and damaging of flowers (Ahmad and
Akhtar, 2013).
The current insect pest management strategy largely depends on the use of synthetic
chemical insecticides. Indiscriminate use of insecticides pose some hazardous
effects such as development of insecticide resistance in insect pests, adverse effects
on friendly organisms, environmental pollution and accumulation of toxic elements
in food and ultimately pesticide residue-induced diseases in human beings
(Ambethger, 2009). Use of bio-pesticides is progressively becoming recognized as
one option to address yield losses due to pest attacks as well as reducing
environmental degradation associated with the use of synthetic pesticides
(Kopondo, 2004).
Meanwhile, the use of new generation insecticides or botanical insecticide could be

considered as possible alternative for suppressing aphids because these are claimed
to be relatively safer or have little impact on beneficial organisms compared to
other conventional insecticides, and they can be included in the future IPM
programs. The use of natural and easily biodegradable crop protection products like
Azadirachtin can be useful components of an IPM program because of their low
toxicity to beneficial insects (Pavela, 2009).
On the other hand, aphids are attacked by a wide range of natural enemies including
ladybird beetle adults and larvae, lacewing larvae, syrphid fly larvae, damsel bugs
and minute pirate bugs. Parasitic wasps are also important in long- term control of
aphids. Lady birds (Coccinellid beetles), Ccocinella septempunctata are the
important entomophagous predators against many species of aphids and observed as
an efficient predator of cabbage aphid in field conditions (Singh and Singh 2013).
Hence, exploring potential of biological pesticides that are relatively cheap,
accessible, safer and environmentally friendly is timely. These biological pesticides
have been shown to have little impact on natural enemies of pests (Schmutterer,
1997) and are easily biodegradable (Devlin
[310]
and Zettel, 1999). However, there is very little information about their effective
active ingredients, preparations and application rates.
The objective of the current study was to evaluate the efficacy of the biopesticides
Nimbecidine 0.03% EC, aqueous extract of Neem seed and botanical mixture
against Brevicoryne brassicae L. on kale under field conditions, and assess their
relative toxicity to the natural enemy of the pest.

The experiment was conducted at Debre Zeit Agricultural Research Center
experimental field during 2017 and 2018 cropping seasons. Ethiopian Kale,
Brassica carinata L. commonly known as ‘Gurage Gommen’ which is the most
common cultivar in central rift valley region of Ethiopia was planted on 15 August
2017 and on 21 August 2018. Agronomic practices were followed as per the culture
of kale production in the area. The experimental field was divided into three blocks
and each block into five plots.
Plant preparation
Kale seeds were drilled on the seed bed at a depth of about 2 cm and covered with
light soil and dry grass (mulch), which was removed after seedling emergence.
Water was applied daily to ensure the soil remained wet. Seedlings emerged within
5 – 8 days. After a week, the covers were removed and the seedlings were hardened
for 3 to 4 days. After germination, seedlings were thinned to maintain a plant
spacing of 2-3cm to reduce competition for space and nutrients, as well as
preventing the development of damping off disease. Then, seedlings were watered
regularly. The seedlings became ready for transplanting after 4 weeks, after
attaining 3-4 leaves. The 30-days old seedlings were transplanted into 3 X 2m area
plots. Spacing between rows and plants were 60 cm and 30 cm, respectively. Each
plot had 30 plants.
Preparation of botanicals
Single bulb of garlic and onion was peeled and grinded in a mortar; mixed with one
teaspoonful dry pepper powder. The product was soaked in one liter of water for
overnight and filtered using muslin cloth. A 0.01% (v/v) of liquid soap was added
to the mixture prior to spraying as a sticking agent. Then, the solution was sprayed
at a volume of 500 liter per hectare using a manually operated knapsack sprayer.
Preparation of neem seed kernel extract (NSKE)

Neem seed kernel extract was prepared by gently grinding a one kilo of well dried
neem seed kernels into a fine powder using mortar and pestle. Then, the resulting
[311]
powder was soaked for overnight in two liters of water. In the morning, the solution
was stirred with wooden plank till it formed milky white solution. Then the solution
was filtered through a double layer of fine muslin cloth with repeated washing to
bring the volume level to 10 liter. The volume was maintained by adding the
required quantity of water to get 10 percent solution. One percent dish washing
liquid detergent soap was added at one ml per liter of spray solution.
Dimethoate 40% EC, a synthetic insecticide was used as a standard check and
applied at the rate of 1.5liter /ha as per manufacturer’s recommendation. Similarly,
Nimbicidin 0.03% EC (the neem-oil-based botanical insecticide) commercial
formulation was used at a rate of 3 liters per hectare.
Application of treatments
Knapsack sprayer was used for applying the treatments. Care was taken to prevent
the drift effects of treatments by putting polythene sheet screen around each plot
during spraying. A total of three spray applications were made biweekly.
Application volume was maintained at 500 liters per ha.
Cabbage aphid count

The experiment was conducted using natural aphid infestation. Data were collected
weekly from the first occurrence of aphid till crop harvest (first week of September
to last week of November). Aphid counts were recorded from each of the 10
randomly selected plants from each plot, by examining the whole plant (all leaves,
flower shoots and the main stem). Percentage of aphid population reduction (%
efficacy) was estimated according to the equation of Henderson and Tilton (1955):
% population reduction (efficacy) = [1-(Ta/Ca × Cb/Tb] × 100.

Where;
Tb is infestation in treated plot prior to application; Ta is infestation in treated plot
after application; Cb is infestation in control plot prior to application; Ca is
infestation in control plot after treatment application.
Statistical analysis
Data were subjected to one way analysis of variance (ANOVA) in a randomized
complete block design (RCBD). In case of significant F-values, treatment means
were separated using Tukey's Studentized Range (HSD) test at 5% level of
significance (SAS Institute, 2009).
[312]
Plant morphological parameters
Treatments differed significantly for plant height and leaf canopy spread (Table 1).
The mean plant height ranged from 67.28 to 80.78 cm (Table 1). The highest (80.78
cm) and the lowest (67.28 cm) mean plant height were recorded from botanical
mixture and Nimbicidine treated plots, respectively (Table 1). Differences between
the rest of the treatments were insignificant.
Effects on aphid population

Treatments effect on mortality of cabbage aphid is shown in Table 1. All the
botanical and insecticidal treatments resulted in significantly lower aphid
population than the untreated control.
The mean post treatment data showed that at 7 DAS, the control plots had
significantly higher numbers of aphids (72.69 /plant) than plots treated with
botanicals and synthetic insecticides. Whereas, the lowest aphid population (9.065
aphids/plant) was recorded on Neem seed extract treated plot followed by botanical
mixture and Nimbicidine oil extract with 19.67 and 20.27 aphids/plant, respectively
(Table 1).
Differences between Dimethoate, Neem seed extract and botanical mixture were
significant for aphid population. However, the botanical mix did not differ
significantly from Nimbicidine in both seasons. All tested botanical insecticides
resulted in higher mortality of cabbage aphid. Neem seed extract and Dimethoate
40% EC resulted in higher percent aphid population reduction (84.07 and 83.4%)
than botanical mix and Nimbicidene (78.62 and 78.21 %,) (Table 1). The results
agree with previous findings (Pavela, 2009; Shiberu and Negeri, 2016; Desale and
Getnet, 2018) who reported that Neem seed extract was effective for the control of
aphid. Similar results of botanicals effects were reported by Bhathal et al., (1994),
Pandey et al., (1987) and Singh and Singh (1995).
Effects on fresh leaf yield

Differences were non-significant between treatments although yield in the untreated
control (19.78 t/ha) was lower than the rest of the treatments (Table 2). Highest
yield of 23.22 t/ha was recorded from botanical mix followed by Nimbicidine,
Dimethoate and Neem with yield of 22.75, 21.02, 20.62 t/ha, respectively (Table 2)
[313]
Table 1. Mean aphids population density, canopy spread, plant height of kale and percent efficacy
(mean of 2017 and 2018)
Treatments Aphid population Canopy spread Plant height at Aphid population reduction
1DBS 7DAS at maturity (cm) maturity (cm) (Efficacy %) at 7 DAS
Untreated control 30.64 C 72.69 A 43.90 B 71..88AB -
Dimethoate 40% EC 87.56 A 34.55 B 48.33 AB 75.44AB 83.37
Nimbicidine 0.03% EC 39.22 B 20.27 C 48.87 AB 67.28C 78.21
Neem seed extract 23.98 C 9.065 D 51.07 A 76.68 AB 84.07
Botanical mixture** 38.78 B 19.67 C 51.17 A 80.78 A 78.62
Mean 44.03 31.25 48.79 74.41
SE 2.497 2.043 1.79 2.402
CV 5.671 6.54 3.66 3.23
LSD 7.0433 5.764 5.035 6.76
*Means followed in the same column by the same letter are not significantly different (P ≤ 0.05; Tukey’s HSD test)
**1Garlic bulb + 1 Onion bulb + 1TSF dry pepper + 2ml of liquid soap/ lt water DBS=
Day Before spray; DAS= Days After Spray
Relative toxicity to Lady beetles (Coccinella septempunctata)

The results presented in Table 3 indicate significant variations in lady beetle
population between treatments. The highest population of Lady beetle was observed
in plots treated with botanical mixture (10.93, 11.27 and 11.17 beetles/ 5plants) at
3, 5 and 7 DAS, respectively. Plots treated with Dimethoate resulted in 71.6, 70.6
and 75.1 % reduction in the lady beetle population at 3, 5 and 7 DAS, respectively
(Table 3). In Neem seed extract and botanical mixture treated plots, population
reduction was lower with 24.44 and 7.95%, respectively. Differences in beetle
population between Dimethoate, Nimbicidene and Neem seed extract treated plots
were insignificant at 3, 5 and at 7 DAS (Table 3). In addition to lady beetles, other
beneficial insects in biocontrol of aphids such as parasitic wasp (mummified aphids
with Diaeretiella rapae), syrphid fly larvae and lace wing larvae were observed.
Table 2. Fresh leaf yield (t/ha) of kale treated with neem based products and botanical mixture against the
cabbage aphid in 2017/18 and 2018/19
Treatments 2017/18 2018/19 Mean

Fresh wt. yield (t/ha) Fresh wt. yield (t/ha) Yield t/ha
Untreated control (water spray only) 19.53 A* 20.033A 19.783 A
Dimethoate 40% EC 20.767 A 21.267A 21.017 A
Nimbicidine 0.03% EC 22.50 A 23.00A 22.75A
Neem seed extract 20.367 A 20.867A 20.617 A
Botanical mixture** 22.967 A 23.467A 23.217 A
Mean 21.7 21.73 21.477
SE 2.295 2.295 2.295
CV 10.56 10.56 10.69
LSD NS NS NS
*Means followed in the same column by the same letter are not significantly different (P ≤ 0.05; Tukey’s HSD test)
**1Garlic bulb + 1 Onion bulb + 1TSF dry pepper + 2ml of liquid soap/ lt water
[314]
Table 3. Combined mean relative toxicity of different treatments on the population of Natural enemies (Lady beetles) in kale crop
Treatments Mean Lady beetle population/5 plants Percent reduction of Natural Enemies population
at
1 DBS 3 DAS 5DAS 7 DAS 3 DAS 5DAS 7 DAS
Untreated control 8.10b* 10.77a 6.033b 5.533b - - -
Dimethoate 40% EC 13.7a 5.167b 3.00b 2.33b 71.63 70.60 75.10
Nimbicidine 0.03% EC 9.43b 8.10ab 6.30b 5.80ab 35.40 10.30 9.96
Neem seed extract 7.83b 7.867ab 5.967b 5.467b 24.44 -2.32 -2.21
Botanical mixture** 8.933b 10.933a 11.267a 11.167a 7.95 -69.34 -83.01
Mean 9.60 8.57 6.51 6.06
SE 0.583 1.35 1.65 1.94
CV 6.074 15.81 25.304 32.016
LSD 1.6448 3.8208 4.649 5.4729
*Means having same letter(s) in a column are not significantly different at P< 0.05 followed by Tukey's. DBS= Day Before spray; DAS= Days after Spray
**1Garlic bulb + 1 Onion bulb + 1TSF dry pepper + 2ml of liquid soap/ lt water
Findings of the study clearly indicated that seed extract of Neem was the most
effective against cabbage aphid followed by Dimethoate 40% EC. On the other
hand, Dimethoate 40% EC was found to be more toxic to natural enemies of
cabbage aphid. On the average 78-84% mortality of aphids was observed from
treatment plots. A single application of the treatments appeared to be sufficient to
provide a significant reduction in the number of aphids although further reduction
could be achieved with repeated applications. Botanicals proved to be the most
effective in controlling aphids with minimal harm to the natural enemy, the lady
beetle. They also gave comparable leaf yield with the synthetic insecticide, In
general, because of their effect on aphids and minimal harm to the natural enemies,
botanicals can be used as a component of aphid IPM on kale.
Acknowledgement
This study was fully financed by the Ethiopian Institute of agricultural Research
(EIAR). We are grateful to the Debre Zeit Agricultural Research Center (DZARC)
for providing assistance in the form of farm machinery, labor and resources for
conducting this research. Special thanks to the research and technical staff of the
Horticultural Research (DZARC), program for their unreserved technical support.
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Screening of Linseed (Linum Usitatissimum) Accessions For Resistance To Fusarium Oxysporum F. Sp. Lini

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Screening of Linseed (Linum Usitatissimum) Accessions For Resistance To Fusarium Oxysporum F. Sp. Lini

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Results of

የኢትዮጵያ የግብርና ምርምር ኢንስቲትዩት

Proceedings of the Completed Plant

Copy Editing and Design: Elizabeth Baslyos

PART I: Plant Pathology

Evaluation of Trichoderma Species as Biocontrol Agents against Fusarium Wilt of

Collection, Isolation and Identification of Entomopathogenic Fungi from

Chitinolytic Activity of Entomopathogenic Fungi Collections from Ethiopia

Evaluation of Faba Bean Genotypes for Faba bean Gall Resistance

Evaluation of Sweetpotato Varieties against Major Sweetpotato Viruses in Ethiopia

Screening of Rice Germplasms for Resistance against Sheath Rot

Development of Integrated Management of Faba bean Gall

Evaluation of Fungicides Efficacy against Rice sheath Rot (Sarocladium oryzae)

PART II: Weed Science

Critical Weed Competition Period on Roselle (Hibiscus sabdariffa L.)

Critical Weed Competition Period on Basil (Ocimum basilicum L.)

Critical Weed Competition Period on Chamomile (Matricaria chamomilla L.)

Critical Weed Competition Period on Roselle (Hibiscus sabdariffa L.)

Herbicidal Control of Chrysanthemum segetum in Wheat Field at Cheliya District

Herbicidal Control of Weeds in Turmeric at Tepi, South western Ethiopia

Management of Striga (Striga hermonthica) in Sorghum (Sorghum bicolor) at Jeldu District

Integrated Management of Wild Radish (Raphanus raphanistrum)

Integrated Management of Cotton Weeds at Middle Awash Rift Valley of Ethiopia

Screening of Common bean Genotypes for their Resistance to the

Evaluation of Botanical and Synthetic Insecticides against the Cabbage Aphid

The productivity of crops is severely threatened by various factors in which pests

The information presented in this proceeding is aimed at availing information and

Diriba Geleti (PhD)

Materials and Methods

Disease incidence: DI (%) = Number of diseased plants in a quadrant X 100

Disease severity: DS (%) = Area of a plant tissue affected X 100

Prevalence (%) = No of fields affected X 100

In addition, data on geographical information (latitude, longitude and elevation) of

Results and Discussion

Growing resistant cultivars is a mostly preferred approach for control of rusts

No of Yellow rust Stem rust Leaf rust

SNNP Sidamo 20 17 85.0 0 0.0 0 0.0

Region Zone Yellow rust Stem rust Leaf rust

Yellow rust Stem rust

Generally, most recently released varieties in the country were succumbed to

Conclusion and Recommendation

Materials and Methods

Isolation and multiplication of single-pustule isolates

Inoculation of wheat stem rust differential lines

Infection types produced on near-isogenic Sr lines

Results and Discussion

This first confirmation of TTKTT in Ethiopia is important because of its

Races Virulence Avirulence

Conclusion and Recommendation

Materials and Methods

Results and Discussion

Zone District Willt Blight Powdery mildew

Conclusion and Recommendation

Disease Incidence (%)

Isolation and identificationof the pathogen

Results and Discussion

S/no Zone Districts Disease incidence (%)

Morphological view of the cultures

Conclusion and Recommendation

Rosemary is native to Mediterranean and Asia. However, it is still reasonably

Materials and Methods

Antagonistic activity of Trichoderma species against

Successful control of Fusarium wilt in many crops by application of different