DISPERSAL OF VEGETATIVE MANGO MALFORMATION DISEASE

3398

AMERICAN RESEARCH THOUGHTS ISSN: 2392 – 876X

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Volume 2 │ Issue 3 │ January 2016 Available online at: www.researchthoughts.us

DISPERSAL OF VEGETATIVE MANGO

MALFORMATION DISEASE

Muhammad Ahsan1, Shomaila Iqbal1i, Faisal Sohail Fateh2, Abid Riaz1,

Abdul Sattar1 & Muhammad Waqas1

1 Department of Plant Pathology, PMAS Arid Agriculture University, Rawalpindi, Pakistan

2 Mango Project, Nat-IPM, NARC, Islamabad, Pakistan

Abstract: Vegetative mango malformation is a very severe disease usually appears on young seedlings

at quite early stage and causes the entire death of the plant. Surveys were conducted In order to

monitor and to determine the incidence of MMD in nurseries of traditional and non-traditional areas

of Pakistan. Survey results indicated that higher incidence of 17% and 23.33 was recorded in Mir Pur

Khas Sindh and Hamza nursery Islamabad respectively. Mango variety sunehra was found to be more

susceptible to MMD. Different experiments were conducted in order to study the exact dispersal of

MMD by using Water, Air and Soil. Fusarium mangiferae was found as causal agent of mango

vegetative malformation, easily disperse through air from infected to healthy plant at a distance of 5

feet and due to systemic nature of fungi it can also be dispersed thorough water and soil. Mango bud

mite (Aceria mangiferae) also play an important role in the transmission of Fusarium. Air play a

major role in the dispersal of mango malformation disease. As soon as symptoms appear prune

infected branch and properly dispose off so that it can’t disperse to the healthy ones.

Key Words: Mango, Malformation, Fungus

INTRODUCTION

Mango, Mangifera indica, is a 2nd most important fruit crop in Pakistan (Masood et al.,

2012). It is grown on the area 0.16 million hectares with annual production of 0.175

i Corresponding author: [email protected]

Muhammad Ahsan, Shomaila Iqbal, Faisal Sohail Fateh, Abid Riaz, Abdul Sattar, Muhammad Waqas- DISPERSAL OF VEGETATIVE MANGO MALFORMATION DISEASE

3399 AMERICAN RESEARCH THOUGHTS- Volume 2 │ Issue 3 │ 2016

million tons (Anonymous, 2012). Mango crop is very perishable and is affected by

number of biotic and abiotic agents. Mango diseases especially decline is one of

important disease in reducing production of mango crop. Among the diseases mango

malformation disease (MMD) is very important and severe disease at seedling stage

(Ahmed et al., 2002).

Mango malformation is a very severe and old disease of mango tree. There are

two types of malformation vegetative and floral malformation. Vegetative

malformation is very severe form of disease and mostly appears on very young mango

seedling at nursery stage and causes their death (Rymbai and Rajesh, 2011). Garg 1st

time reported this disease on mature trees in Uttar Pardesh during 1951. He described it

as bunchy top and assumes that it is due to viral infestation (Chakrabarti, 2011). Nirvan

(1953) 1st time reported this disease from hardly 3-4 year old mango seedlings. At first

inflorescence and vegetative malformation were considered as different diseases by

different causes but Tripathi (1954) cleared that these are the two separate

manifestations of same disease by observing high correlation in incidence and severity

between vegetative and inflorescence malformation (Chakrabarti, 2011). Disease

appeared on seedling as small shoot lets with small scaly and brittle leaves with a

bunch type appearance due to hypertrophied growth on shoot apex. From apex of shoot

multi-branches emerge with small scaly brittle leaves and dramatically shortened

internodes referred as “Bunchy Top” (Kanwar and Nijjar, 1979; Ploetz, 2004).

The incidence of vegetative malformation on young plants of 4-8 year age group

was recorded on 90.9% plants and severity of the disease ranges between 2.7-80.4%

(Chakrabarti, 2011). Young plants die due to this disease hence no plants for rootstock

and for grafted purpose. This situation compels researchers to work on it and found

management practices so that young plants can be saved for raising healthy orchards.

The vegetative form of MMD was first reproduced in 1966 with Fusarium

moniliforme (Marasas et al., 2006). Aceria mangiferae was found as vector in dissemination

of fungus from infected nursery plants to healthy plants (Gamliel-Atinsky et al., 2009a).

Mango malformation disease spread from one place to other places spread from

infected nursery and infected grafted material (Kumar et al., 1993, Haggag, 2010). But

spread with in nurseries is feebly implicit so for studying exact way of dispersal of

MMD in nurseries present study was done. The present study was conducted in order

to find out disease incidence percentage in nurseries of traditional and non-traditional

areas of mango and to study the dispersal of MMD in nursery plants.



MATERIAL AND METHODS

Survey and disease incidence

Surveys were conducted in the nurseries; located in mango growing traditional (Multan

and Muzaffar Garh from Punjab and Tando Allahyar and Mirpurkhas from Sindh

province) and non-traditional areas (Islamabad) in order to estimate the infection

percentage and prevalence of MMD in the mango growing nurseries. Random samples

were collected from each area in order to determine the incidence by using the

following formula of Masood et al, (2010). Selected healthy and infected plants from

nurseries of Islamabad were brought to the green house of NARC, Islamabad.

Number of infected plants

Disease incidence (%) = ----------------------------------------- x 100

Total number of plants

Mode of dispersal

Dispersal of MMD through air, soil and water was studied on nursery plants. Plants

were arranged at a distance of 5ft in such a way that infected plant was placed in center

of healthy plants. Air current was given continuously for 6-8 hours daily in order to

disperse mycelium or spores form infected plant to healthy plants. Dispersal of MMD

through irrigating infected water to healthy plants for this purpose watering the

infecting plant and leach down water were then given to the healthy plants

continuously for two months till the appearance of the symptoms. Dispersal of MMD

through soil, roots were inoculated by the infected (1gm grinded infected tissue in

100ml distilled water) inoculums. (Fig. 1). All plants were kept under controlled

conditions for 8 weeks (Freeman et al.1999) for symptom development. Regularly

checked the plants above ground parts i.e. leaves, buds and stems of plants and observe

under the microscope.



Fig. 1 Experiment for finding mode of Dispersal of vegetative form of MMD 1.A)

Arrangement of plants for finding dispersal through air, infected plant in centre and around it

healthy plants at 5 ft distance 1.B) Close view of infected and a healthy plant in air dispersal

experiment 2) Irrigation of healthy plant with harvested water from infected plant for finding

dispersal through water 3.A) Preparation of inoculum, suspension of MMD infected plant

tissue and water thoroughly mix by vortex mixture 3.B) Application of inoculum to healthy

plant near roots.

Fig. 6 Macro and micro- characteristics of Fusarium mangiferae isolates A) 14 day old Fusarium

colony on PDA B) 7 day old Fusarium purifies colony on PDA C) Micro conidia in false heads

D&E) Mono and poliphyalides F) Macroconidia G) mango bud mites on scotch tape slide from

aerial dispersal experimental plants.



Isolation of pathogen

After 8 weeks of experiments, approximately 5mm pieces were excised from healthy

and infected plants bud junctions and after surface disinfestations with 2% sodium

hypochlorite were placed on PDA Petri plates and incubated at 25±1oC for 7 days. This

isolation was done to confirm fungal pathogen presence.

Identification of pathogen

Fungi were identified on the basis of colony characteristics and spores. Identification of

fungus was done following standard keys (Ellis 1980, Nelson et al. 1983).

RESULTS

Survey and disease incidence

Survey of traditional areas showed the higher disease incidence 17% in Mirpur Khas,

Sindh and lower in Muzaffar Garh, Punjab 5% was observed (Table. 1 and Fig. 2). In

non-traditional areas, nurseries were visited located at Peshawar mor, Margalla town

and Park road Islamabad. Higher disease incidence 23.33% was recorded in Hamza

nursery farm with nursery source from Rabbani Nursery Ghelan Chak # 9 Pattoki while

lowest, 10%, disease incidence was recorded in Lush Green and Fateh Muhammad

nursery farms nursery source from Pattoki (Table. 2 and Fig. 3). 41% prevalence was

reported by visiting total of 12 nurseries out of which 5 have mango malformation

disease infected plants. From varietal point of view Sunehra was recorded with higher

disease incidence of 40% and Dusehri was with the lowest 10% disease incidence (Table.

3 and Fig. 4).



Mode of dispersal

After six week of inoculation plant start to develop symptoms as apical and lateral buds

of plants start blackening and no shoots was developed and plant start drying from tips

and shows stunted growth (Fig. 5). Spores of Alternaria, Aspergillus and micro-conidia of

Fusarium was found under the light microscope. Mango bud mite, Aceria mangiferae, was

also observed during microscopic study of the infected plant (Fig. 6), which is known as

the vector of fungus and transmit spores from infected buds to healthy ones (Gamliel-

Atinsky et al., 2009a). Presence of Aceria mangiferae clear that it also perform its role in

dissemination of fungus spores along with air current. Same symptoms of bud drying

and stunting was also observed from soil inoculated with infected inoculum and

infested water through irrigating healthy plants. But they did not dry completely (Fig.

5).

From isolation of pathogens, colonies of Fusarium, Aspergillus and Alternaria was

obtained from above ground parts of plants of aerial dissemination while from roots

and above ground parts of other plants Fusarium and Aspergillus was isolated. Fusarium

was identified by observing its characteristics macro and micro conidia (Fig. 6)

S. No

Varieties Number Infected

Total Number

D.I %age

1 Chaunsa 0 40 0

2 Desi 5 30 16.67

3 Sunsaishun 1 5 20

4 Sunehra 2 5 40

5 Anwar rataul 0 5 0

6 Langra 0 25 0

7 Dusehri 2 20 10

Table. 3 Disease incidence %age in different varieties Fig. 4 Disease incidence %age in different varieties



Table.1 Disease incidence %age in traditional areas Fig. 2 Disease incidence %age in traditional areas

Table. 2 Disease incidence in nurseries of non-traditional areas Fig. 3 Disease incidence in nurseries of non-traditional areas

DISCUSSION

Present study was conducted in order to find the exact dispersal way of the causal agent

of mango malformation disease i.e. Fusarium mangiferae .Many scientists all over the

world works on MMD but not confirmed the exact way of dispersal till date. Fusarium

mangiferae being dominant fungi in causation of mango malformation disease was also

confirmed by Iqbal et al., 2006. Macro and micro conidia were disperse through air as

observed by the Gamliel-Atinsky et al., 2009b. Conidia disperse through air current and

cause disease in other healthy plants. Aceria mangiferae act as vector for the transmission

of MMD (Gamliel-Atinsky et al., 2009a). Haggag et al., 2010 studied the systematic

transmission of MMD as our results shows that Fusarium spores inoculated in soil first

colonize the roots of mango plant and then through vascular bundle moves towards

above ground parts and produced symptoms of MMD which lead to the complete

Province Location No. of Nurseries

Average %age of MMD

Punjab Multan 10 14

Muzaffar Garh 5 5

Sindh Mirpur Khas 7 17

Tando Allahyar 5 10

S. No

Nursery Name Name of Nursery source Farm

Infected Plants

Total plants

D.I %age

1 Saleem Brothers

Saleem Farms Mardan 1 5 20

2 Fateh M. Nursery Farms

Dr. Nursery Farm GhelanChak#9 Pattoki

2 20 10

3 Makkah Nursery

Multan 4 20 20

4 Hamza Nursery Farm

Rabbani Nursery Ghelan Chak#9 Pattoki

7 30 23.33

5 Lash Green Nursery

Rabbani Nursery Ghelan Chak#9 Pattoki

1 10 10



death of plants. Haggag et al., 2010 confirmed this by isolating fungus on artificial

media from root and apical buds as well as through scanning electron microscopy in In

vitro conditions. About dispersal through water, very minimum or no literature is

available. This is the first report confirming that through water conidia also enter in

roots from lesion points and then became systematically, move towards above ground

parts. It is assume that when plant roots were colonized by fungus, conidia collected at

the bottom of pod with drainage water. When this drainage water was applied to new

healthy plant, at particular concentration fungus start colonizing roots and became

systemic. Due to systemic nature of fungi, it produced disease on above ground parts as

confirmed by Haggag et al., 2010. This present study will help researcher in future for

verification of dispersal way and for adopting control measures. Our results confirm the

dispersal way of causal agent of MMD i.e. it can be disperse through air soil as well as

through water. Air born inoculum is more than any other way and may serve as

important source of disease development. Therefore, it may be probable to develop

control measure in such a way that inoculum load may reduce i.e. by pruning infected

branches as early as they develop and by adopting sanitation practices i.e. removal of

plant disease debris from the orchards. Don’t raise nurseries under or around the

mature infected trees because spores from infected floral heads can infect the newly

developed buds of young plants.

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DISPERSAL OF VEGETATIVE MANGO MALFORMATION DISEASE