Evaluation of Chocolate spot (Botrytis fabae) Against Faba bean (Vicia faba L.

)
Varieties in Horro Guduru Wollega District, Western Ethiopia
Fikiru Wakoya1, Tola Abdissa1, Alemayehu Dugasa1

1
Department of Plant Science, College of Agriculture and Natural Resource Shambu Campus,
Wollega University, P. O. Box 38, Shambu, Ethiopia.
ABSTRACT

The quantity and quality of Faba bean (Vicia faba L.) in Ethiopia are reduced due to chocolate
spot (Botrytis fabae) disease. Even though several improved faba bean varieties were released,
more researches’ have not been done especially in Horro Guduru Wollega district, Western
Ethiopia. Therefore, this experiment has been done to evaluate the released varieties against
chocolate spot disease per the objectives to measure the reaction of faba bean varieties to
chocolate spot disease and to select the likely varieties of faba bean for the study areas. The
research was conducted at Guduru and Shambu experimental sites in the 2016 and 2017 main
cropping season, having 14 improved varieties and one local check. From these field
experiments disease severities, PSI, AUDPC, and apparent infection rates were assessed and
these disease parameters were found significantly different (P<0.05) among the varieties in both
locations and seasons. The results revealed that at Shambu the disease host reaction showed
that Mesay and Moti varieties were found moderately susceptible whereas, FB Didia and
Degaga varieties were found moderately resistant in both 2016 and 2017cropping season. At
Guduru location Obse, Hachalu, and Tesfa varieties were found moderately susceptible
whereas, Kasa and Bulga-70 varieties were found moderately resistant in the 2016 cropping
season. However, all varieties were found moderately resistant in the 2017 main cropping
season. The Logistic model (R²=97) was well defined as the epidemiological data than the
Gompertz model (R²=95) for disease development. An estimate of the rate of disease
progression, based on proportional measures of the extent of infection at different times was
varied. The correlation coefficient between the two growing seasons of 2016 and 2017 for all
estimated disease parameters of Guduru was higher than Shambu and the correlation of the
same year was highly significant than between two years (i.e. 2016 with 2017 main growing
seasons). It shows as the faba bean varieties were reacted differently to a disease for these
studies.

Keywords: Chocolate Spot, Infection rate, Vicia faba, Diseases epidemiology

1
1. Introduction

Faba bean (Vicia faba L.) is the third imperative crop in the world which originated in the

Near East and the oldest domesticated legume crops (Torres et al., 2006). Faba bean crop was

introduced to Ethiopia from the Middle East through Egypt about 5000 B.C. and currently,

Ethiopia is one of the centers of secondary diversity for faba bean (Asfaw et al., 1994; Yohannis,

2000 Torres et al., 2006). Faba bean is grown in the highlands of Ethiopia at altitudes of 1800-

3000 m.a.s.l ranges where the requirement of cold temperature is met (Yohannis, 2000). Some

assessments have been done in 2003, the international production of faba bean was 2.6 million

metric tons and over the world, China leads in faba bean production in both area coverage and

production followed by Ethiopia (0.37 million ha, 0.45 Mt), Egypt (0.14 million ha, 0.44 Mt) and

Australia (0.16 million ha, 0.27 Mt). Ethiopia is the second-largest producer of Faba bean crop in

the world next to China, however, the contribution is low about 6.96% of world production and

40.5% within Africa (FOASTAT, 2010). Faba bean crop is full-grown in several regions of the

Ethiopian country; especially Amhara region has contributed to the highest production (47%) in

the country followed by the Oromia region 38% of the area (ICARDA, 2006)

Despite its likely, over the last century, there has been constantly decreasing in the

cultivated areas of faba bean production for several factors, in many countries. According to

Ethiopian circumstances, natural limitations include vulnerability to biotic and abiotic pressures

and integrally low grain yield potential of the home-grown cultivars. Among the diseases,

chocolate Spot disease is a major one characterized by the initial appearance of reddish-brown

spots on leaves, which enlarge and even merge and subsequently lead to severe premature

defoliation. Under favorable conditions, it appears on stems, flowers, and pods, which directly

2
affect seed production. About 94.6% of chocolate spot occurrence is testified in the main faba

bean rising areas of Ethiopia (Hailu et al., 2014), and based on the tolerances and susceptibility

of cultivar, yield loss is reported up to 34% and 61% due to this disease. Faba bean varieties,

respectively (Dereje and Yaynu, 2001). Varietal assessment against diseases might be hopefully

the methods of disease management approaches because of are environmentally eco-friendly,

cost-effective, and free of any toxic substances. Therefore, searching for efficient means for

controlling foliar diseases is an immediate required to increase the Faba bean yield and its

components. For this reason selection of resistant varieties to foliar diseases is the appropriate

means to increase the yield of the crop. Thus, this work focused on evaluating the released

varieties against chocolate spot disease with the objectives to assess the response of Faba bean

varieties to chocolate spot diseases and to select the promising varieties of Faba bean for the

study area.

2. Materials and methods

2.1. Experimental site

The study was conducted in Horro Guduru Wollega district at two sites Shambu and

Guduru during 2016 and 2017 main cropping season; found in western Ethiopia, Oromia

regional state, at a location of longitude 36o39’28.8”-37o40’11.2”E and latitude 9o9’24.6”-10o

20’59.9”N, temperature 8-32oC, annual rainfall 900 mm-2000 mm and altitude 1350-3170 m

a.s.l. where the experiments were conducted (Figure 1).

3
Figure 1. Map of Ethiopia showing the Horro GuduruWollega district and the study area.

2.2. Experimental design and treatments

In both locations 14 improved released varieties and one local check was evaluated with

chocolate spot disease in 2016 and 2017 main cropping season. The varieties were obtained from

Holeta and Kulumsa Agricultural Research Center (Table 1).

4
Table 1. Agronomic characteristics of faba bean varieties used in the study areas (EIAR, 2018)

Varieties Released year Maturity date Production domain(m.a.s.l)a Yield (qt/ha)

CS20DK 1977 145-160 2300-3000 15-30
Bulga-70 1994 143-150 2300-3000 15-35
Holeta- 2 2001 140-150 2200-2800 15-35
Kasa 1980 120-135 1900-2300 15-30
Tesfa 1995 125-135 1900-2300 15-35
Mesay 1995 125-135 1900-2300 30-35
Degaga 2002 116-135 1800-3000 20-37
Moti 2006 116-135 1900-2800 20-35
Obse 2007 103-166 1900-3000 21-35
Dosha 2009 120-130 1900-2800 21-30
Tumsa 2010 120-130 1900-2800 21-35
Gora 2013 126-168 1900-2800 20-40
Hachalu 2010 122-156 1900-2800 20-38
FB Didia 2014 122-145 1900-2800 20-44
a = meter above sea level
The plot size was 2m x 2m having 6-seedling rows 1m spacing between blocks and 0.5m

and 0.3m spacing between plots and rows, respectively. The seeds and fertilizer rates were based

on standard seed rate (30kg/ha) and DAP 75-90kg/ha and UREA 30kg/ha, respectively. The

treatments were arranged in a randomized complete block design (RCBD) with three

replications.

2.3. Data collection

2.3.1. Disease severity

Chocolate spot severity was assessed on 20 randomly selected and pre-tagged plants per

plot at weekly intervals from the time disease first appeared until the crop attained its

physiological maturity. The average severity of the 20 plants per plot was used for analysis.

Disease severity was rated by using 1-9 rating scale (Bernier et al., 1984 and Porta-Puglia et al.,

1993) and classified into five categories (1, 3, 5, 7 and 9 level), where one is a very small spot or

5
no disease indications; three is minor discrete lesions; five is certain merged lesions with

particular defoliation; seven is great merged distinct lesions, and nine is wide-ranging lesions on

leaves, stems and pods and death of plants (Hanounik, 1986) and then transformed into a

proportion for analysis using the following formula.

nxv
Disease severity= ∗100
9N
Where:
n = number of plants in each category
v = Numerical values of symptom category
N = Total number of plants
9 = Maximum numerical value of symptom category

2.3.2. Percent Severity Index

According to Sahile et al., (2008) and Villegas–Fernandez et al., (2012), the disease was

scored in seven days interval starting from the first chocolate spot symptoms presence and

sustained till the ending of podding phase (i.e. when the disease reached a maximum

development stage). Finally, the scores were converted into percent severity index (PSI) for the

analysis (Hanounik, 1988; El-Ghamry et al., 2009; Abo-Hegazy et al., 2012). From the results of

PSI values, the response of tested varieties was categorized into six response clusters following

Abo-Hegazy et al. (2012); where 0% up to 2% is highly resistant indicated by ‘HR’, greater than

2% up to 15% resistant indicated by ‘R’, greater than 15% up to 40% moderately resistant

indicated by ‘MR’, greater than 40% up to 60% is moderately susceptible indicated by ‘MS’ and

greater than 60% up to 80% is susceptible indicated by ‘S’ and greater than 80% is highly

susceptible indicated by ‘HS’.

PSI =
∑ of numericalratings x 100
No . of plants scored x maximum score on scale

6
2.3.3. Area under disease progress curve

The trapezoidal method was used to estimate the AUDPC for the quantitative summary

of average disease strength over time (i.e. the progress of chocolate spot was plotted over time

using mean severity indices for each faba bean variety at each location, and the PSI values were

also used for analyses of the area under disease progressive curve); as the following formula.

x + x )( t
n
AUDPC=∑ [0. 5( i i+1 i+1 −t i )]
i−1

Where: Xi= the average coefficient of infection of the i th note, Xi+1 = the average coefficient of

infection of the i+1th note and, ti+1- ti= the number of days between the ith note and the i+1th note,

n= number of observations.

2.4. Data analysis

The collected data from experimental sites were subjected to analysis of variance with

SAS computer software version 9.1.3 (SAS, 2008). The progress of the disease was transformed

with a logistic model, ln[y/(1 − y)] (Vander plank, 1963). Means of disease parameters were

compared by Least Significance Difference at a 5% probability level.

3. Results

3.1. Disease severity

The mean disease severity in both years of 2016 and 2017 growing seasons was found

statistically significantly different (P<0.05) among the varieties at both Shambu and Guduru

7
experimental sites (Table 2). At Shambu site in 2016 main growing season, the highest mean

disease severity was recorded from Mesay and Moti 48.33% and 43.33%, respectively while, the

lowest disease severity was recorded from FB Didia and Degaga varieties 30.67% and 31.33%,

respectively and about nine varieties were highly infected as compared to local cultivar 35.33%.

In 2017 growing season, the highest mean disease severity was recorded from Obse, Moti and

Gora varieties 46% similar value, while, minimum disease severity was recorded from Dosha

and Kasa varieties 28% and 29.67%, respectively and totally 12 varieties were found highly

diseased as related to local variety was 31.33% (Table 2).

At Guduru experimental site the mean disease severity was significantly different

(P<0.05) in both the 2016 and 2017 growing season (Table 3). In the year 2016, the highest

disease severity was recorded from Obse and Hachalu varieties 43.67% and 41.33%, respectively

while, the minimum mean disease severity was recorded from Kasa and Bulga-70 varieties

33%and 34% respectively (Table 3). In the year 2017, the maximum disease severity was

recorded from Tumsa and Dosha varieties with similar value (40.67%) whereas, the minimum

value was found in both Gora and Mesay varieties (30.67%) (Table 3). The result of the

interaction was not obtainable under these tests because the pooled analysis data of the error

variances were not homogenized and the mean was not significantly different within the varieties

for both locations and years.

3.2. Percent severity index (PSI)

The mean PSI has shown a highly significant difference (P<0.05) in 2017 than in the 2016

main cropping season among the varieties at Shambu experimental site (Table 2). In the 2016

main growing season, the highest PSI was recorded from Mesay and Kasa varieties 65% and

8
61%, respectively while, the lowest PSI was recorded from FB Didia and Degaga varieties with

the value of 34% and 39%, respectively (Table 2). Based on the PSI calculation, the varietal

reaction against diseases on Mesay, Moti, Kasa, Tumsa, and Hachalu varieties were grouped into

moderately susceptible, while the remaining varieties were classified as moderately resistant. In

2017, the highest PSI was recorded from Obse (73.81%) and Moti (60.62%) varieties while the

minimum PSI was recorded from Dosha and Local cultivar with a value of 27.33% and 38.33%

respectively. The reaction of the host on Mesay, Moti, Tumsa, Holeta-02, Obse, and Gora

varieties were grouped into moderately susceptible, while the rest of the varieties were grouped

into moderate resistance (Table 2).

Table 2. Chocolate spot disease (Botrytis fabae) and host reaction of Faba beans (Vicia faba L.) varieties
at Shambu Site in the main growing season

Shambu
2016 2017
Varieties DS a
PSIb HR c
DS PSI HR
Mesay 48.33a 65.00a MS 43.00bac 55.95bac MS
Moti 43.33ba 57.52bdac MS 46.00a 60.62ba MS
Kasa 43.00ba 61.91ba MS 29.67e 34.67ed MR
Tumsa 43.00ba 51.91ebdacf MS 45.67a 58.81ba MS
Hachalu 42.33ba 58.33bac MS 32.67ed 31.67ed MR
Bulga70 40.67bc 51.91ebdacf MR 37.00dc 40.33edc MR
CS20DK 40.67bc 44.14ebdcf MR 31.00ed 34.67ed MR
Dosha 40.33bc 52.38ebdac MR 28.00e 27.33e MR
Holeta02 35.67dc 42.67edcf MR 44.00ba 54.67bc MS
Local 35.33dc 44.00ebdcf MR 31.33ed 30.33ed MR
Tesfa 35.00dc 40.33edcf MR 31.33ed 35.33ed MR
Obse 35.00dc 40.48edcf MR 46.00a 73.81a MS
Gora 33.67d 41.71edcf MR 46.00a 55.95bac MS
Degaga 31.33d 34.00f MR 37.67bdc 46.09bdc MR
FB Didia 30.67d 39.00ef MR 32.33ed 34.67ed MR
CV (%) 10.171 22.257 11.033 23.931
LSD (5%) 6.559 18.000 6.910 18.009
P<value 0.0001 0.0326 <.0001 0.0001
Means with the same letter at the same column are not significantly different at 5% probability level, CV= Coefficient of
Variation, LSD= Least Significant difference, MS= moderately susceptible, MR= Moderately Resistance
a = Disease severity
b = Percent Severity Index
c = Host response to disease

9
 The mean of PSI at Guduru has shown a less significant difference among the varieties

(P<0.05) in both the 2016 and 2017 cropping season (Table 3). In the year 2016, the highest PSI

was calculated on both Obse and Holeta-02 varieties with the value 52.86%, while the minimum

PSI was noticed in Kasa and CS20DK varieties with the values 35.33% and 39% respectively.

The reaction of the host on Obse, Hachalu, Tesfa, Holeta-02, and Dosha varieties were found

statistically on par and graded as moderately susceptible. The remaining ten varieties were

grouped into moderate resistance. In 2017 cropping season the highest PSI was calculated on

Tumsa and Dosha varieties and were found on par statistically with the value of 56.24% and

50.33%, respectively, whereas the minimum PSI was recorded on Degaga variety with the value

of 27.33% 31%, respectively (Table 3). During this year 2017, the results were shown as all

varieties were found moderately resistant to chocolate spot disease (Table 3).

Table 3. Chocolate spot disease (Botrytis fabae) and host reaction of Faba beans (Vicia faba L.) varieties
at Guduru Site in the main growing season

Guduru
2016 2017
Varieties DSa PSIb HRc DS PSI HR
Obse 43.67a 52.86a MS 33.33bdc 31.00de MR
Hachalu 41.33ba 51.09a MS 30.67d 31.00de MR
Tesfa 41b.00ac 51.57a MS 31.33dc 35.33dec MR
Holeta02 41.00bac 52.86a MS 33.00bdc 31.00de MR
Dosha 41.00bac 52.38a MS 40.67a 50.33ba MR
Local 39.67bdc 45.96bac MR 31.33dc 35.33dec MR
Mesay 39.33bdc 45.48bac MR 30.67d 31.67dec MR
FB Didia 38.67bedc 41.62bdc MR 31.00d 31.00de MR
Tumsa 38.33bedc 49.00ba MR 40.67a 56.24a MR
Moti 37.33fedc 46.72bac MR 33.67bdc 31.00de MR
Degaga 36.33fedg 41.62bdc MR 31.33dc 27.33e MR
Gora 35.33feg 40.33bdc MR 30.67d 31.00de MR
CS20DK 35.00feg 39.00dc MR 33.00bdc 36.62bdec MR
Bulga70 34.00fg 40.33bdc MR 38.00bac 44.67bdac MR
Kasa 33.00g 35.33d MR 38.33ba 45.95bac MR
CV (%) 5.962 11.789 12.234 23.783

10
 LSD (5%) 3.822 9.020 6.925 14.572
P<value <.0001 0.0027 0.0253 0.0079
Means with the same letter at the same column are not significantly different at 5% probability level, CV= Coefficient of
Variation, LSD= Least Significant difference, MS= moderately susceptible, MR= Moderately Resistance
a = Disease severity
b = Percent Severity Index
c = Host response to disease

3.3. Area under disease progress curve (AUDPC)

Statistically, a highly significant difference (p<0.05) was noticed for AUDPC among the

varieties at Shambu experimental site in both the 2016 and 2017 main cropping season. In 2016,

the maximum AUDPC was calculated on Mesay and Moti varieties 3125%-days and 2775%-

days respectively, while the minimum AUDPC was recorded on FB Didia and Degaga varieties

with the values of 1937.5%-days and 1962.5 %-days, respectively (Figure2). In the year 2017

main growing season, the highest AUDPC was estimated on Obse, Moti and Gora varieties with

a value of 2962.5%-days, while minimum AUDPC was estimated on Dosha and Kasa varieties

with the values of 1762.5%-days and 1887.5%-days, respectively (Figure 2).

3500
3125
3000 2775 2737.5 2775
2662.5 2612.5
2562.5 2537.5
2500 2300 2275 2250 2200
2100
AUDPC( %days)

1962.5 1937.5
2000

1500

1000

500

0
ay sa lu 0 K 2 l fa se
oti s a
a7 0D sh
a
a0 ca ra ga di
a
es M Ka m cha l g 2 o t Lo Te
s
Ob Go ga Di
M Tu Ha Bu CS D l e
De FB
Ho

11
 3500
2962.5 2925 2962.5 2962.5
3000 2775 2837.5
2400 2387.5
2500
2075 1987.5 1987.5 2012.5 2050
AUDPC( %days)

2000 1887.5
1762.5

1500

1000

500

0
i a sa ha l a e a a ia
ay ot as ha
lu a7
0 K 02 ca sf bs or ag id
es M K m
ac lg 0D os eta Lo Te O G eg
M Tu u 2 D o l D
D
H B CS H FB
Varieties

Figure 2. The area under disease progressive curve (AUDPC) of a Chocolate spot on Faba bean
varieties in 2016 and 2017 cropping season at Shambu

At Guduru, AUDPC was significantly different (p<0.05) among the varieties in both

2016 and 2017 years (Figure 3). In 2016, the maximum mean AUDPC was recorded on Obse

and Hachalu varieties with an estimate of 2800%-days and 2650%-days respectively, whereas

the minimum was estimated on Kasa and Bulga-70 varieties with the value of 2087.5%-days and

2162.5%-days, respectively. In 2017 cropping season, the maximum mean AUDPC was

estimated on Tumsa and Dosha varieties with the values of 2625%-days and 2612.5%-days

respectively, while the minimum was estimated on Gora and Hachalu varieties with the value of

1937.5%-days and 1950%-days respectively (Figure 3)

12
 3000

2500

2000
AUDPC( %days)

1500

1000

500

0
se a al ay idia msa i sa
al
u sfa ta02 osh oc ot ga ra K 70
Ob ach Te es M ga Go 20D lga Ka
l e D L M D Tu e
Bu
H Ho FB
D CS

3000

2500

2000
AUDPC( %days)

1500

1000

500

0
se u fa l ay
al s 02 sh
a ca di
a sa oti ag
a ra DK 70 s a
Ob ch Te ta Do Lo es Di m M g Go 20 l ga Ka
a le M
FB Tu De CS
Bu
H Ho
Varieties

Figure 3. The area under disease progressive curve (AUDPC) of a Chocolate spot on Faba bean varieties in
2016 and 2017 cropping season at Guduru

3.4. Disease progress rate

The logistic model (R²=97) was well defined as the epidemiological data than the

Gompertz model (R²=95).for disease development. From the linear regression of the logistic

model the disease progress rate (units day-1) was calculated in both the 2016 and 2017 cropping

season at both Shambu and Guduru experimental sites. At Shambu experimental site in the 2016

13
main cropping season the disease progress rate on Obse and Moti varieties were found faster by

recording 0.0459logits day-1, and 0.0409logits day-1, respectively, whereas the slightly lower rate,

was calculated on Dosha variety with the value of 0.0222logits day -1. The initial percent severity

index was higher on Local, Hachalu, Tumsa, Dosha, CS20DK, and Kasa varieties (>15%), while

(10%) was recorded from others. A minimum final PSI was calculated from Degaga, Bulga-70,

and Gora varieties (45%), and others were accounted for from ranges of (50-65%). In the

Shambu area, in the 2017 main cropping season, the disease progress rate was found faster on

Mesay variety the value of 0.0441logit day-1 whereas, the disease progress rate was found

relatively lower on the variety FB Didia with the value of 0.0271logits day -1 (Table 4). The initial

PSI was recorded higher on FB Didia, Local, Dosha, Kasa, Obse, Tesfa, and Gora varieties

(>15%), while it was recorded low (10%) in other varieties. A minimum final PSI was observed

from FB Didia, Degaga, Bulga-70, and Holeta-02 varieties (45%) and other varieties accounted

for a higher range (50-65%) (Table 4).

Table 4. Infection rates of chocolate spot disease with standard error and mean initial and final PSI on
Faba bean (Vicia faba L.) varieties at Shambu in 2016 and 2017 main growing season

Varieties Shambu
2016 2017

14
 PSI-ia PSI-fb rc Sd R²e PSI-i PSI-f r S R²
FB Didia 0.100 0.500 0.0319 0.0026 0.96 0.150 0.450 0.0271 0.0036 0.91
Hachalu 0.150 0.550 0.0321 0.0041 0.93 0.100 0.500 0.0354 0.0043 0.92
Degaga 0.100 0.450 0.0353 0.0039 0.93 0.100 0.450 0.0294 0.0048 0.86
Tumsa 0.150 0.650 0.0403 0.0042 0.95 0.100 0.550 0.0362 0.0052 0.90
Local 0.200 0.650 0.0284 0.0038 0.93 0.150 0.600 0.0311 0.0038 0.94
Dosha 0.150 0.600 0.0222 0.0044 0.87 0.150 0.600 0.0337 0.0044 0.92
CS20DK 0.150 0.550 0.0268 0.0032 0.94 0.100 0.600 0.0345 0.0046 0.92
Bulga70 0.100 0.450 0.0331 0.0029 0.95 0.100 0.450 0.0353 0.0048 0.89
Kasa 0.150 0.550 0.0251 0.0031 0.94 0.150 0.650 0.0371 0.0064 0.88
Holeta02 0.100 0.500 0.0384 0.0029 0.96 0.100 0.450 0.0326 0.0028 0.95
Gora 0.100 0.450 0.0405 0.0031 0.95 0.150 0.500 0.0301 0.0024 0.97
Mesay 0.100 0.600 0.0366 0.0044 0.93 0.100 0.650 0.0441 0.0044 0.95
Obse 0.100 0.600 0.0459 0.0036 0.97 0.150 0.500 0.0304 0.0027 0.96
Moti 0.100 0.550 0.0409 0.0048 0.92 0.100 0.550 0.0371 0.0042 0.93
Tesfa 0.150 0.550 0.0316 0.0035 0.95 0.150 0.600 0.0305 0.0051 0.90
a = Percent severity index initial
b = Percent severity index final
c = Disease progress rate from logistic line
d = Standard error of the rate
e = Coefficient of determination for the logistic model ln(y/1-y))

At Guduru, in 2016 the disease progress rate was higher on Tumsa and Dosha varieties with the

estimate of 0.0354logits day-1 and 0.0351logits day-1, respectively while slower disease progress rate was

calculated on Gora and Hachalu varieties with the value of 0.0283logits day -1 and 0.0287logits day-1

respectively (Table 5). An initial PSI was found high on Local, Dosha, Kasa, CS20DK, Mesay and Moti

varieties with the value of 15%, while the remaining varieties were 10% of initial PSI. A minimum final

PSI was observed from Hachalu, Degaga and Gora varieties with the value of 45% while a high final PSI

was found high in the remaining varieties with the value of ≥50% of the disease indices. In year 2017

cropping season the faster disease progress rate was observed on Obse and Hachalu varieties with the

value of 0.0421logits day -1 and 0.0391logits day-1 respectively, whereas on Kasa variety was found slow

with the value of 0.0284logits day-1. The initial PSI was found higher on Local, Tumsa, CS20DK, Kasa,

Mesay and Tesfa varieties with the value of 15% index whereas, others recorded low with the value of

15
10% index. A minimum final PSI was observed from Degaga, Bulga-70 and Gora varieties with the value

of 45%, while it was a higher range for other varieties with the values of 50-65% (Table 5).

Table 5. Infection rates of chocolate spot disease with standard error and mean initial and final PSI on
Faba bean (Vicia faba L.) varieties at Guduru in 2016 and 2017 main growing season

Guduru
varieties 2016 2017
PSI-ia PSI-fb ra Sb R²c PSI-i PSI-f r S R²
FB Didia 0.100 0.500 0.0294 0.0049 0.88 0.100 0.500 0.0348 0.0031 0.96
Hachalu 0.100 0.450 0.0287 0.0038 0.93 0.100 0.600 0.0391 0.0035 0.96
Degaga 0.100 0.450 0.0298 0.0038 0.95 0.100 0.450 0.0311 0.0022 0.97
Tumsa 0.100 0.550 0.0354 0.0047 0.97 0.150 0.600 0.0335 0.0036 0.95
Local 0.150 0.650 0.0309 0.0048 0.96 0.150 0.650 0.0369 0.0046 0.94
Dosha 0.150 0.650 0.0351 0.0051 0.95 0.100 0.550 0.0372 0.0032 0.96
CS20DK 0.150 0.600 0.0312 0.0048 0.95 0.150 0.550 0.0298 0.0029 0.96
Bulga 70 0.100 0.500 0.0338 0.0045 0.95 0.100 0.450 0.0296 0.0033 0.93
Kasa 0.150 0.650 0.0341 0.0051 0.92 0.150 0.550 0.0284 0.0026 0.96
Holeta 02 0.100 0.500 0.0321 0.0042 0.96 0.100 0.500 0.0376 0.0035 0.95
Gora 0.100 0.450 0.0283 0.0039 0.83 0.100 0.450 0.0301 0.0024 0.96
Mesay 0.150 0.600 0.0289 0.0043 0.96 0.150 0.650 0.0351 0.0032 0.97
Obse 0.100 0.500 0.0321 0.0043 0.92 0.100 0.600 0.0421 0.0031 0.97
Moti 0.150 0.550 0.0326 0.0046 0.96 0.100 0.500 0.0315 0.0051 0.88
Tesfa 0.100 0.550 0.0309 0.0043 0.95 0.150 0.650 0.0381 0.0028 0.98
a = Percent severity index initial
b = Percent severity index final
c = Disease progress rate from logistic line
d = Standard error of the rate
e = Coefficient of determination for the logistic model ln(y/1-y))

The appearance and amount of inoculums were found varied or the epidemic dynamics over time

among varieties during the growing period might be due to the prevalence of conducive

environments where the field experiment was conducted and susceptible hosts tested in the

experiments. The tradeline of the logistic regression line shown as Mesay variety was found

lined over the progress of other varieties but Gora variety was found relatively under (Figure 4).

16
 0.7 0.7
0.6 0.6

Proportion of PSI
Proportion of PSI
0.5 0.5
0.4 0.4
0.3 0.3
0.2 0.2
0.1 0.1
0 0
40 55 70 85 100 115 40 55 70 85 100 115
Days After Planting
0.7
0.7
0.6
0.6

Proportion of PSI
0.5
Proportion of PSI

0.5
0.4
0.4
0.3
0.3
0.2
0.2
0.1
0.1
0
0 40 55 70 85 100 115
40 55 70 85 100 115
Days After Planting

Figure 4. Spread of Chocolate spot (Botrytis fabae) in Faba bean (Vicia faba L.) varieties at Shambu
and Guduru in 2016 and 2017 growing seasons

3.5. Disease correlations

The correlation between all pairs of disease severity, PSI, and AUDPC within two

locations at Shambu and Guduru sites and two main cropping seasons during the year 2016 and

2017 were found positively correlated. At Shambu experimental site the estimated disease

parameters of the same year have shown a highly significant difference (p<0.01) and high

positive correlation (r = >0.85). However, the correlation between different years or the

estimated disease parameters of 2016 with 2017 was not a significant difference and weakly

correlated (p<0.01) (Table 6).

17
Table 6. Correlation analysis among Chocolate spot disease parameters at Shambu for two years of 2016
and 2017 main cropping seasons

Shambu
Disease Parameter DS16a DS17b PSI16c PSI17d AUDPC16e AUDPC17f
Disease severity of 2016 - .227 ns .869** .027ns .992** .232 ns
Disease severity of 2017 .227ns - .016 ns .915** .224ns .997**
PSI of 2016 .869** .016ns - .180 ns .852** .021 ns
PSI of 2017 .027ns .915** .180ns - .022 ns .914**
AUDPC of 2016 .992** .224ns .852** .022ns - .228 ns
AUDPC of 2017 .232ns .997** .021ns .914** .228ns -
**. Two way tabulation of the relation is significant at the 0.01 level and ns = non-significant
a= Disease severity recorded in 2016 main cropping season
b=Disease severity recorded in 2017 main cropping season
c= Percent severity index in 7 days interval calculated from disease severity in 2016 main cropping season
d= Percent severity index in 7 days interval calculated from disease severity in 2017 main cropping season
e= Area under disease progress curve from PSI in 2016 main cropping season
f= Area under disease progress curve from PSI in 2017 main cropping season

At Guduru experimental site the estimated disease parameters within the year (2016;

2017) and between the years have shown significant difference positively (p<0.01) and a high

correlation was noticed in the same year (r = >0.85); then between two years (2016 with 2017 of

growing seasons) at r = <0.665 (Table 7).

Table 7. Correlation analysis among Chocolate spot disease parameters at Guduru for two years of 2016
and 2017 main cropping seasons

Guduru
Disease Parameter DS16 a
DS17 b
PSI16c PSI17d AUDPC16e AUDPC17f
Disease severity of 2016 - .656** .896** .588** .994** .659**
Disease severity of 2017 .656** - .515** .935** .663** .998**
PSI of 2016 .896** .515** - .416** .884** .519**
PSI of 2017 .588** .935** .416** - .586** .935**
AUDPC of 2016 .994** .663** .884** .586** - .665**
AUDPC of 2017 .659** .998** .519** .935** .665** -
**. Two way tabulation of the relation is significant at the 0.01 level and ns = non-significant
a= Disease severity recorded in 2016 main cropping season
b=Disease severity recorded in 2017 main cropping season
c= Percent severity index in 7 days interval calculated from disease severity in 2016 main cropping season
d= Percent severity index in 7 days interval calculated from disease severity in 2017 main cropping season
e= Area under disease progress curve from PSI in 2016 main cropping season
f= Area under disease progress curve from PSI in 2017 main cropping season

4. Discussion

18
 Chocolate spot disease (Botrytis fabae) is a faba bean disease having the ability to

producing several infections cycle in a season and hence the polycyclic epidemics are potentially

concerned. This disease apparent on itself as small red-brown spots on leaves, stems, and flowers

of faba bean plants and spores form on the dead tissues that spread the infection to other plants.

In this field experiment, the resistances of Faba bean varieties were evaluated against disease and

disease parameters because it is crucial for quantifying disease intensity or to assess the average

disease intensity over time and location (Laurence et al., 2007). In these two field experiments

the disease progress rate, disease severity, PSI, and AUDPC influences greatly varied against

Faba bean varieties at Shambu and Guduru sites in both 2016 and 2017 main cropping seasons.

The logistic model was well shown as the normal epidemics in both the main growing seasons

and location than the Gompertz model because the logistic model has been described as the most

suitable for time-related investigation of disease development. The proportion of the measured

chocolate spot disease was related to the logarithm of the proportion of the quantity of diseased

and health plant parts (Madden and Campbell, 1990). Error mean squares from each environment

and time were tested to ensure the collective analysis of variance across environments and times

and the variances were not homogenized.

The results of the present work revealed that at Shambu high disease severity were

recorded on Mesay variety followed by Moti, Kasa, and Tumsa varieties in 2016 cropping season

and 2017 cropping season Moti, Obse and Gora varieties were moderately susceptible to disease.

This result is aligned with the reports of Mesele et al (2016). At Guduru similar results were

reported by Mulualem et al (2012) who reported that Moti and Degaga have a moderate

resistance to chocolate spot severity in both 2016 and 2017 main cropping seasons. Villegas-

Fernandez et al (2012) reported that the varieties used represented a wide range of responses to

19
chocolate spot infection; further reported by referring Basandrai et al (2007) that there are known

cases of resistance varieties become susceptible.

The calculated disease parameters of PSI and AUDPC were found varying among the

varieties in both the 2016 and 2017 cropping years. For instance, in 2016 cropping season results

at Shambu location and other research results reported a high disease level on Kasa and Degaga

varieties (Mesele et al., 2016 and Yitayih and Azmeraw, 2018); however, controversial results

were reported on Moti and Degaga varieties (Degife and Kiya, 2018 and Wolde & Mitiku, 2018).

In 2017 cropping season at the same location it had a severe infection on Moti, Obse, Gora,

Holeta-2 and Tumsa varieties, while low on Dosha, Kasa, CS20DK, Local and Tesfa varieties

which is inlined again with reports of work done in southwest Ethiopia (Mesele et al., 2016).

Similar results were also reported on CS20DK and Degaga varieties by Degife and Kiya (2018)

and Yitayih and Azmeraw (2018). In 2017 cropping season study from the same location PSI and

AUDPC were high on Gora and Moti varieties, whereas low on Dosha and CS20DK varieties,

which is supported by reports of Degife and Kiya (2018) and Mesele et al (2016). However,

opposite results were reported by Wolde & Mitiku (2018) and Yitayih and Azmeraw (2018).

Sahile et al (2011) report supported that high PSI 52.29% in average was recorded from the non-

sprayed treatment of CS20DK variety. Research results pointed out by Mesele et al (2016) and

Wolde & Mitiku (2018) as well as the result of this work in 2016 main cropping season at

Guduru location, the disease severity was found high on Obse, Hachalu and Tesfa varieties while

it was low on Kasa, Bulga-70, CS20DK and Gora varieties.

High PSI and AUDPC were calculated in 2016 cropping season at Shambu on Hachalu

variety, whereas a low estimate is obtained from Dosha and Bulga-70 varieties, and the same

results also supported by the reports of Mesele et al (2016), Wolde & Mitiku (2018) and Degife

20
and Kiya (2018). At the same area in 2017 main cropping season, the disease severity was higher

on Dosha, Tumsa and Kasa varieties, whereas low on Gora, Mesay, FB Didia and Holeta-2

varieties, (Degife and Kiya, 2018; Mesele et al., 2016 and Yitayih and Azmeraw, 2018),

however, Tumsa variety has not coincided with reports of Yitayih and Azmeraw (2018). In the

2017 cropping season, PSI and AUDPC high values were obtained from Tumsa and Kasa

varieties, which inlined with Mesele et al (2016) and Yitayih and Azmeraw (2018) reports,

whereas low on Hachalu variety which supported by Degife and Kiya (2018) reports. However,

on Dosha, Tumsa, and Gora varieties, opposite results were reported by Wolde & Mitiku (2018).

The degree of host reaction against chocolate spot disease across the environments is fluctuated

in between moderately resistant to moderately susceptible.

The mean disease frequency per unit time of fungal pathogenic chocolate spot varied

significantly different at (P < 0.05) among all varieties at the two sites. Low infection rates of

disease logits per day were estimated on Degaga, CS20DK, and Tesfa (Mesele et al., 2016).The

presented results supported by Kora et al (2016) report, the symptoms appeared and performed

through the times vary among varieties, even with the local check. Similarly, the work of

Bouhassan et al (2004) reports that the disease progress curves of the resistant and susceptible

varieties with checks were similar scorings. The occurrence and intensity of recorded and

calculated disease severity; PSI and AUDPC were higher in Shambu than Guduru. Generally,

disease severity and AUDPC were higher in 2016 than the 2017 cropping season in both

locations, but PSI was higher in 2017 than the 2016 cropping season. Although a slight

difference in disease onset might have passed during inspection before the disease was first

observed. The experiments revealed that the figures of disease growth curves were almost similar

for all during the cropping season. The appearance and amount of inoculum changes in varieties

21
during the growing period because the epidemic dynamics over time among varieties might be

environmentally conducive where the field experiment was conducted (Agrios, 2005). The data

obtained under these conditions provides empirical evidence that the disease influenced by

seasonal condition shows varying in progress and host reaction among the varieties, for all

experiments the disease epidemic was increased over the time increased (Dereje et al., 1994;

Laurence et al., 2007 and Villegas-Fernández et al., 2012). Finally, improving the resistance

crops and crop protection are priorities in research should be given to chocolate spot disease, the

number one yield-limiting factor in Faba bean production across the nation. Stress may be given

to screening an enormous number of exotic and indigenous materials to identify genetic

resistance and to control the disease.

22
Acknowledgment

The authors, we would like to thank Wollega University Research and community

service Directorate for their financial support and Shambu Campus College of Agriculture and

natural resources coordinator and all Plant Science staff for their active participation throughout

the experiment was conducted.

23
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