INTERNSHIP REPORT

AGRICULTURE RESEARCH CENTRE TANDO JAM SINDH PAKISTAN

TITLE:
Survey and identifications of Anthracnose Disease of
mango in the surrounding of Tando Jam
SUBMITTES TO
Research supervisor
Dr. Gul Bahar Poussio
Senior Scientist
Plant Disease Research Institute
Tandojam

SUBMITTED BY
Ahtisham Rasool Khosa
Reg. No. 2K18 -PT-22

DEPARTMENT OF PLANT PROTECTION,

FACULTY OF CROP PROTECTION,
SINDH AGRICULTURE UNIVERSITY TANDO JAM
2022
 LIST OF CONTENTS
S. No PARTICULARS PAGE
1 Acknowledgment 2
2 Plant disease research institute 3
3 Mycology Laboratory 3
4 Nematology Laboratory 3
5 Virology Laboratory 4
6 Tissue culture Laboratory 4
7 Introduction of mango 5
8 Activities carried out 8
9 Survey 8
10 Diseases observed in the field 8
11 Reparations Of Potato Dextrose Agar 9
12 Isolation of diseased samples 9
13 Identifications 10
14 Results 10
15 Conclusion 11
16 References 12-13
 ACKNOWLEDGEMENT

I consider it my duty to offer my humble submission before Almighty Allah who is entire

source of knowledge and wisdom endowed to mankind and who enables me with

potential, ability, health, and strength to complete my research studies successfully. I offer

my humblest thanks from the bottom of my heart to the Holy Prophet (Peace be upon him)

who is always forever a truth of guidance and knowledge for humanity.

In the name of Allah the most beneficial the most merciful whose help we solicit. I have by

acknowledge this humble effort to Honorable Mr. Noor Muhammad Baloch Director

General Agriculture Research Sindh and Dr.Liaqat Ali Bhutto, Director of agriculture

research center for providing such facilities at mycology laboratory. I am' also very much

thankful to my research supervisor Dr. GuI Bahar Poussio, senior scientist for his

continuous guidance cooperation while conducting this internship.

The author is highly thankful to Professor Dr. Manzoor Ali Abro, Professor Department

of Plant Pathology Dr. Sumera Mahessar, Lecturer Department of Plant Pathology, Mr.

Zubair Nizamani, Assistant Professor Faculty of Crop Protection, Sindh Agriculture

University Tandojam, for their moral and scientific support during my one month internship.

And I am also thankful to Mr. Khalid lqbal Rajput, Director Plant Disease Research

Institute Tando Jam and their staff members of this institute for helping me for the positive

cooperation throughout the whole internship Program.

Ahmed Jamal
 Plant Disease Research Institute, Tando jam

Plant disease research institute is working on diagnosis of the plant/crop's diseases

research and their management in-vitro as well as in-vivo conditions. The institute has
different laboratories such as Mycology, Nematology, Bacteriology, Virology, and Tissue
culture laboratory.

Mycology Laboratory

This laboratory has the capacity to diagnose and identify all fungal plant pathogens of
importance to agricultural, horticultural, ornamental, and cereal crops. Samples received
include plant roots, fruits, stems, tubers, and leaves. The lab carries out fungal tests e.g,
by direct isolation and culturing with facilities such as incubation under near-UV to
facilitate fungal sporulation. The results are used for phytosanitary decision making in
support of exports and imports as well as making recommendations on disease
management. This laboratory is fully equipped and working on different aspects of the
fungal diseases and their managements. The scientists are working different chemicals
and bio pesticides for the managements of fungal diseases in the fields as well as in the
laboratory.

Nematology Laboratory

Research of the Laboratory of Nematology is focused on the interactions between

nematodes and (a)biotic environmental factors. The goal is to develop methodologies and
strategies for durable plant parasitic nematode control with a minimal negative impact on
the environment. This laboratory is functional and working on the diagnoses and
management of nematodes diseases in plants.
Virology Laboratory

The plant virology laboratory is working on the diagnosis of viruses as well as viruses
belonging to viral families potyviridac and nanoviridac that infect economically important
plants. The scientists of this institute have worked a lot regarding the survey of viral
diseases of different vegetable and fruit crops in Sindh province. Still the scientists are
working on viral disease of banana, Chilli, Papaya, and Cottoncrops.

Tissue Culture Laboratory

This laboratory is working on banana tissue culture technology of different banana

varieties such as G-9, William hybrid, Pissang and Basrai. The scientists have produced
diseased free nurseries at institutes and have supplied to the growers of Sindh tissue
culture is the growth of tissues or cells separate from the organism. This is typically
facilitated via the use of a liquid, semi-solid, or solid growth medium, such as broth or
agar, in vitro under sterile growing conditions. Banana is typically propagated vegetative
thus, tissue culture as a propagation technique provides a robust means to prepare
disease-free planting materials that can provide the first line of defense in developing an
integrated disease-management program for banana.
 INTRODÚCTION OF MANGO

Mango, (Mangifera indica L.,) the king of all fruits belongs to the family Anacardiaceae
and order Sapindales (Litz et al., 2005). Generally, it is grown in tropical and subtropical
regions of Southeast Asia. The native home of mango is considered as India to Burma
(Myanmar) or maybe from the Malay region. Since the sixteenth century, it traveled to
other parts of the world (Popenoe, 1920). Mango was introduced from India to other
countries of the tropical and subtropical world mainly by the Muslim missionaries, Spanish
voyagers, and Portuguese (Chakrabarti et al., 2011). Mango is a nutritionally rich fruit with
a unique flavor, fragrance, and taste. It is an excellent source of vitamin A. The research
reports revealed that the consumption of natural fruits rich in carotenes is known to
protect against oral cavity and lung cancers. In addition, mango fruit is also a rich source
of vitamins, minerals, fiber, prebiotic dietary, and antioxidant compounds, thus promoting
the benefits for human health. Recent research revealed that the consumption of mango
fruit protects against colon, breast leukemia, and prostate cancer (Anonymous, 2018).

It is grown mostly in sub-tropical and tropical countries and of the world. It is also a
notable cash crop in most of tropical countries (Jahurul et al., 2015). Being 2nd major fruit
after citrus, it is the identity of Pakistan as the demand and market of Pakistani mangoes
are boosting globally because of its marvelous taste and nutritious qualities. The
cultivation of mango is now being distributed in about 85 countries. The important mango
growing countries are. Pakistan, China, Philippines, Sri Lanka, Bangladesh, Indonesia,
Thailand, Burma, Malaysia, Egypt, South Africa, Australia, Brazil, Cuba, USA and India,
(Mukherje and Litz2009).
In Pakistan, over 85,000, tons mango is exported every year and fetches about $36.66
million per annum. The cultivated area of mango is about 4.25 million acres with the
annual yield of l.77 million tons. About 90% production of mango is locally consumed
and province Punjab shares 79% in overall production of mango in the country following
Sindh" province (GoP, 2018). Among the major mango growing areas of Pakistan, the
Bahawalpur, Multan, Muzzatfargarh, Rahim Yar Khan, Hyderabad, Mirpurkhas and
Tando Adam are prominent a (Khan, 2005).

The yield of mango in Pakistan is 27.6 tons/acre is significantly less than its potential yield
of about 61.7 tons/acre and similarly expected exports of mango in Pakistan and it
ismainly associated with grower’s lack of awareness about modern pre-harvest and post-
harvest production methods technology, pests, and diseases (Nafees et al, 2013).
Several pests, diseases, and disorders have because recorded on various mango
varieties, ultimately resulting in severe loses to all parts of the mango around the world.
Approximately 260 pest species including major and minor pests have been recorded
from seedlings to mature trees at harvest and postharvest stages (Veeresh, 1989).
Mango suffers from several infectious diseases Caused by many phytopathogens. More
than 83 different diseases and disorders including 52 fungal, 3 bacterial, and 3 plant-
parasitic nematodes of the mango tree and fruit have been recorded worldwide which
cause losses; however, fortunately no single disease 1s caused by the virus till now in
mango (Pernezny and Simone, 2000).

Eighty three diseases have been reported in the world. Among them the main diseases
are anthracnose, Colletotrichum gloeosporioides, powdery mildew, Oidium mangiferae
(Bert.); malformation, Fusarium spp., bacterial leaf spot, Ewinia mangiferae (Doidge);
crown gall, Agrobacterium fumefaciens; sooty mold, Capnodium mangiferae); fruit rot, C.
gloeosporioides and Aspergillus niger; root rot, Rhizoctonia solani (Kuhn) 0xysporum
(Schl); dieback, Diplodia and F. netalensis and Lasiodiplodia theobromae (Pat.) Griff. and
Maubl. (Syn.) BotryodiplodiaTheobromae Pat.; gummosis, L. theobromae; and mango
sudden decline, a complex Disease (Khaskheli et al., 2008).

Among them Anthracnose is one of the most serious diseases of mangoes in many areas
where the crop is grown. The most devastating effects of anthracnose occur in areas
where it rains during the mango flowering and fruit set stages. Although it appears to be
less of a problem in the NT because flowering and fruit set occur during the dry season,
anthracnose can cause problems here under certain conditions.

Mango anthracnose is caused by the fungus Colletotrichum gloeosporioides var minor

(also known by the name of its perfect stage Glomerella cingulata var minor). Spore
production by this fungus is favored by wet or humid weather. The dispersal of these
spores is particularly favored by rain and wind. This enables spread of the disease over
relatively short distances.

In areas where rain is prevalent during flowering and fruit set, anthracnose can cause
destruction of the inflorescences and infection and drop of young fruit. This can obviously
lead to serious losses.

Keeping in view the importance and occurrence to anthracnose disease in different

countries. In Pakistan, the one-month internship with title studies on survey and
identification of anthracnose disease of mango in the surrounding of Tandojam was
conducted at Plant Disease Research Institute, Agriculture Research Centre Tando jam.
 ACTIVITES CARRIEDOUT

Survey

The survey of the different grower's field of mango orchard was carried out in the
surrounding of Tando jam. The method of survey was zigzag method the diseases such
as mango Anthracnose, mango malformation, Powdery mildew, sooty mold and mango
sudden death was observed in the field.

Diseases observed in the field

Anthracnose Mango malformation

Powdery mildew
 PREPARATION OF POTATO DEXTROSE AGAR

FORMULA AND METHODOLOGY

Potato 200 B, Dextrose 20g, Agar agar 17g, Distill water 1000 ml

1. Boil 200 g of sliced unpeeled potatoes in 1 liter of water for 30minutes.

2. Filter through cloth, saving effluent, which is potato infusion.

3. Add dextrose, agar and boil water in to dissolve completely.

4. Sterilize media by autoclaving at 121°C for 15minutes.

5. Aseptically dispense into sterile Petridishes.

ISOLATION OF DISEASED SAMPLES

The collected diseased leaves showing anthracnose of mango symptoms were processed

by tissue isolation method. The infected leaves were washed with tape water by removing

the dust and mud. These samples were air died and cut into small pieces with sterilized

scissor. The cut samples were surface sterilized with 0.05% sodium hypochlorite NaHCl

for one minute followed by washing with tape water twice for two minutes. These sterilized

samples were dried on sterilized filter paper for removing moisture 5 tissue /samples per

Petri plate were inoculated in sterilized Petri plate containing sterilized PDA. The

inoculated plates were kept at room temperature for the growth of the disease-causing

fungus.
IDENTIFICATIONS

Isolated fungi were identified on basis of morphological characteristics as described by

Simmons (2007) and with the help of the characteristics of the fungus mentioned in
book" isolation and identification of fungi".

RESULTS

SURVEY
The results showed that incidence of anthracnose was recorded higher in mango
orchard followed by sooty mold and bacterial caniker. There were also some sudden
deaths in all fields in the surrounding of Tando jam (Fig. 1). The result on the frequency
of isolated fungi viz., Aspergillus, Penicillium, Fussarium sp., and Colletotrichum
gloeosporioides is mentioned in (Fig.2).

18
16
16

14

12

10

8 7

6 5

4 3

0
Anthracnose Bacterial caniker Sooty mold Sudden death

Fig.1 Disease incidence % in the surrounding of Tandojam

ISOLATION

The maximum fugal frequency was observed by Aspergillus (51.61), Penicillium

(16.12), Fussarium sp., (22.58) and Colletotrichum gloeosporioides (9.69) from the
diseased samples.

60
51.61
50

40

30
22.58
20 16.12
9.69
10

0
Aspergillus Penicillium Fusarium sp. Colletotrichum
gloeosporioides

Fig.2 Frequency of fungi isolated

CONCLUSIONS

The Anthracnose disease was found with highest disease intensity. During isolation the
Aspergillus fungus produced highest percentage of fungal colonies in PDA media.
References

1. Ahmed,S.I.2019.Needtotummangoorchardsmorefruitfulforgrowers.Availableonlinec with

updatesathttps://www.thenews.com.pk/print/480307-needto-tum-mango-orchards-more-fr.

2. Anonymous,2018.MangoFruitNutritionFacts.Availablefrom:htps:/www.nutrition-
and:you.com/mango-fruit.html.

3. ChakrabartiDK.MangoMalformation.Vol.1.NY,USA:Springer,2011.p.148.

4. FAOSTAT.FoodandAgricultureOrganizationoftheUnitedNation.ProductionofMangoes,

Mangosteens,andGuavasin2016,Crops/Regions/WorldList/ProductionQuantity(PickLists).

FoodandAgricultureOrganizationoftheUnitedNation,CorporateStatisticalDatabase

(FAOSTAT);2018.

5. HongS.K.,LeeS.Y.,ChoiH.W.,LeeY.K.,JoaJ.H.,ShimH.Occurrenceofstem-endroton

mangofruitscausedbyLasiodiplodiatheobromaeinKorea.PlantPathol.J.2012;28:455.

doi:10.5423/PPJ.DR.09.2011.0174.

6. Jahurul,M.H,I.S.Zaidul,K.Ghafoor,F.Y.Al-Juhaimi,K.L.Nyam,N.A.Norulaini,lF.Sahena

andA.K.M.Omar.2015.Mango(MangiferaindicaL.)byproductsandtheirvaluable

components:Areview.FoodChemistry183:173-180.

7. Khan,S.R.A.2005.Mismanagementinmangoproduction.TheDailyDawn'sEco.Busi.Rev

Karachi,Pakistan.

8. KhaskheliMI,PathanMA,JiskaniMM,WaganKH,SoomroMH,PoussioGB.Firstrecordof

Fusarium nivale (Fr.) Ces. Associated with mango malformation disease (MMD) in Pakistan. Pakistan

Journal of Botany. 2008;40(6):2641-2644.

9. LitzRE,Gomez-LimMA.MangiferaindicaMango.In:LitzRE,editor.BiotechnologyofFruit

andNutCrops.Wallingford,UK:CABInternational;2005.pp.40-6l.

10. MukherjeeSK,LitzRE.Introduction:Botanyandimportance.In:LitzRE,cditor.TheMango:
 Botany,ProductionandUses.Wallingford,UK:CABInternational;2009.pp.01-14.

11. PerneznyK,SimoneGW.DiscasesofMango(MangiferaindicaL.).In:CommonNamesofPlant

Diseases.St.Paul,MN:AmericanPhytopatholoicalSociety;2000.

12. PloetzR.C.Diseasesofmango.In:PloctzR.C.,cditor.DiseasesofTropicalFruitCrops.APSPress;St.
Paul,MN,USA:2003.pp.327-363.

13. PopenoeW.Themango.In:ManualofTropicalandSubtropicalFruits.NewYork:The
MacmillanCo;1920.p.135.
14. Nafees,M.,I.Ahmad,I.Ashraf,M.Jameel,T.HussainandMaryam.2013.Sustainable

productionofmangofruitbymanagingiregularbearingthroughfoliarsprayofurcaatcritical

growthstages.Wor.Appl.Sci.J.24(10):1368-1372.
15. PursegloveJW.MangoeswestofIndia.ActaHorticulturac.1972:(24):107-174.

16. Sakalidis M.L, Ray J.D., Lanoiselet V., Hardy G.E.ST, Burgess Tl. Pathogenic

BotryosphaeriaceaassociatedwithMangjeraindicaintheKimberleyregionofWestern

Australia.Eur.J.PlantPathol.2011;130:379-391.doi:10.1007/s10658.

17. Veeresh GK. Pest problems in mango-world situation. Acta Horticulturae. 1989;(231):551-
565.