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STUDY OF SEED BORNE FUNGI OF DIFFERENT LEGUMES
Narayan M. Ghangaokar 1 and Ayodhya D. Kshirsagar 1*
1 C.T.Bora College, Shirur (Ghodnadi), Tal-Shirur, Dist- Pune Pin- 412210
(*E-mail: drayodhya11@gmail.com nghanu@gmail.com)
ABSTRACT
The study aims at identifying legumes seed borne fungi associated with Pisum sativum L., Macrotyloma uniflorum
(Lam) Verdc., Lens culinaris Medik., Phaseolus vulgaris L., Vigna unguiculata L.Walp., Cajanus cajan L. Millsp.,
Cicer arietinum L. were collected from Shirur tahashil (Pune). The legume seed born fungi was screened by using
blotter plate method from selected untreated and treated seeds. The untreated seeds were found to be associated with
highest number of seed borne fungi. The fungi isolated from these seeds are Alternaria alternata, Chaetomium spp.,
Penicillium citrinum, Aspergillus niger, A. fumigatus, A. flavus, Rhizopus nigricans, Fusarium oxysporum, F.
moniliform , F. solani, Chaetomium sp, Curvularia lunata, Macrophomin sp, Monilia sp., Penicillium sp., Rhizoctonia
sp, Trichoderma etc.
KEY WORDS: blotter plate method, legumes, seed born fungi
INTRODUCTION
Seed is the most important input for crop production. Pathogen free healthy seed is urgently needed for desired plant
populations and good harvest. Many plant pathogens are seed-borne, which can cause enormous crop losses; reduction
in plant growth and productivity of crops (Williams and McDonald, 1983; Kubiak and Korbas, 1999; Dawson and
Bateman, 2001; Islam et al., 2009).
Legume seeds have comparatively higher protein content than non-legume plant. The high protein content makes them
desirable crops in agriculture. The seeds of legumes are second only to cereals as the most important source of food for
humans and animals (National Academy of Sciences, 1979). All varieties of pulses are excellent source of easily
digestible protein. But there are several factors which are responsible for their low production. Among them, diseases
play an important role (Nine, 1986 and Pal, 1996). Numerous examples exist in literature for the international and
national spread of plant diseases as a result of the importation of seeds that were infected or contaminated with
pathogens (Agarwal, 1976). Seed-borne diseases have been found to affect the growth and productivity of crop plants
(Kubiak and Korbas, 1999; Weber et al., 2001; Dawson and Bateman, 2001). Presence or absence of seed borne fungi
on seed surface is one of the important aspects that determine the quality of seed.
The present investigation was undertaken to find out the seed borne fungi associated with the seeds of selected
legumes. Considering the nutritional, agronomical and industrial value of pulses and yield of legumes, the present study
aims to investigate seed-borne fungi associated with Pisum sativum L., Macrotyloma uniflorum (Lam) Verdc., Lens
culinaris Medik., Phaseolus vulgaris L., Vigna unguiculata L. Walp., Cajanus cajan L. Millsp., Cicer arietinum L.
from Shirur Tahsil (Pune, M.H. India).
MATERIALS AND METHOD
Plant Materials:
Pisum sativum L., Macrotyloma uniflorum (Lam) Verdc., Lens culinaris Medik., Phaseolus vulgaris L., Vigna
unguiculata L.Walp., Cajanus cajan L. Millsp., Cicer arietinum L. All legume seed samples were collected from
different localities of Shirur Tahsil (Pune, M.H. India).
Seed Borne fungi analysis:
Detection of seed borne fungi from selected legume seeds was done by blotter paper method as recommended by
International Seed Testing Association (1966), de Tempe (1953), Neergard (1973) and Agrawal (1976). For the
standard blotter method, untreated seeds and seeds after treatment with 0.1% Sodium hypochlorite for 10 minutes were
placed on three layers of moistened blotter, 10 seeds per petri dish. The plates were then incubated in B.O.D incubator
at 25± 2˚C for 8 days under 12 hrs alternating cycles of light and darkness. Thus, the exposed seeds were examined on
the 9 th day under stereo binocular microscope for the presence of seed borne fungi. The fungi found on the seeds were
recorded and percentage of seeds infested and germination of seeds with different fungi was calculated. The isolated
fungi were identified with the help of the keys, monograph and literature provided by Raper and Fennell, (1965);
Booth, (1971); Ellis, (1971); Barnett and Hunter (1972). Untreated seeds were used as controls. All experiments were
carried out in triplet set.
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RESULTS AND DISSCUSIONS
Seeds play a vital role in the production of healthy crops. Healthy seed is the foundation of healthy plant; a necessary
condition for good yields (Diaz et al., 1998).
Table.1 : Seed borne fungi isolated through blotter method from selected seven sampling legume seeds
Legumes Pisum Macrotyloma Vigna Phaseolus Lens Cajanus Cicer
Fungi
sativum uniforum unguiculata vulgaris culinaris cajan arietinum
Un T Un T Un T Un T Un T Un T Un T
Aspergillus flavus Link + + + + + + + + + + + + +
Aspergillus fumigatus - - - - + - + - - - + - + -
Fresenius
Aspergillus niger van + + + + + + + + + + + + + +
Tieghem
Aspergillus terreus Thom + + - - - - - - + - + + + -
Aspergillus nidulans
- - - - + - + - - - - - - -
(Eidam) Winter
Alternaria longipes Mason - - - - - - - - - - + + - -
Alternaria alternata Keissler + - + - + - - - + - - - + +
Acremonium sp. + - - - - - - - - - + - - -
Botrytis sp. - - - - + + - - - - + - + -
Cladosporium sp - - - - + - - - - - - - + -
Chaetomium globosum + - - - + - - - - - - - - -
kuhze ex steud
Chaetomium sp - - - - - - + - - - + - + -
Curvularia lunata Boedijn - - - - + - + + - - + - + +
Drechslera sp. - - - - + - + - - - + + - -
Fusarium moniliforme - - - - + + - - + - - - + +
Sheldon
Fusarium solani (Mart.) + - - - - - + + + + + + - -
Sacc.
Fusarium oxysporium Schle. - - - - + + + - - - + - + +
ex Fries
Fusarium equiseti (Corda) - - + - - - + - - - - - - -
Sacc.
Helminthosporium sp. - - + - + - - - - - + - - -
Humicola sp. + - + - - - - - + - - - - -
Macrophomina phaseolina + - - - - - - - - - + - + +
(Tassi)
Monilia sitophila Sacc. - - - - + - - - + + - - - -
Mycelia sterilia - - + - - - - - - - + - + +
Mucor hiemalis corti. + - - - + + - - + - - - + -
Mucor racemosus Fresenius - - - + - - + + - - + + - -
Nigrospora oryzae (Berk. & + - - - - + + - - + - - - -
Br.) Petch
Penicillium chrysogenum - - - - - + - - - - - - + -
Thom.
Penicillium citrinum Thom - - - - + + - - + - - - - -
Penicillium sp. + + - - - - + + - + - - + +
Phoma sp - - + - + - - - - - - - + -
Pythium sp - - - - - - + + - - - - - -
Rhizoctonia bataticola + - - - - - - + - - + + + +
(Taub) Butler
Rhizopus nigricans Ehrenb + + - + + + + + + + + + + -
Rhizopus nodosus
- - - - - - + - - + + + + +
Namyslowski
Sclerotium sp - - + - - - - - - - - - + -
Scopularopsis sp. - - + - - - + - - - - - - -
Trichoderma hamatum - - - - + + - - - - - - - -
(Bon.) Bain.
Trichoderma viridae Persoon + - + - - - + - + - + - + +
ex Fries
Verticillium sp - - + + + - - - -- - - - -
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They are carriers of some important seed-borne diseases caused by microorganisms which results in considerable losses
in yields. Many fungal pathogens, some of which are seed transmitted, often reduce the germination ability or kill the
infected plants or substantially reduce the productive capacity. Therefore, control of seed-borne fungi is extremely
important and the damaging effects can be relieved through integrated approaches (Vaidehi, 2002). Altogether 40
species of fungi were isolated from seven samples of legume seeds during present study (Table1). Among the samples
studied the fungi, Aspergillus nidulans, A.niger, A. fumigates, A. flavus, Alternaria alternate, Acremonium sp.,
Chaetomium sp., Rhizopus nigricans Fusarium moniliforme, F. solani, F. oxysporum, Macrophomina phaseolina,
Botrytis sp., Cladosporium, Helminthosporium sp. Curvularia, Drechslera, Humicola,, Penicillium chrysogenum,
Mucor racemosus, Mucor hiemalis Phoma, Monilia sitophila, Pythium, Trichoderma viridae, Trichoderma hamatum,
Nigrospora oryzae, Rhizoctonia bataticola, Sclerotium sp and Verticillium sp etc were reported.
Un: Untreated seeds, T: treated seeds
Figure1: Percentage of seed germination results of seven selected sampling legumes seeds
In this study significant number of fungi was isolated from these seed samples. The untreated seeds were found to be
associated with highest number of seed borne fungi. It was found that Cicer arietinum L. seeds yielded the highest
number of fungal species followed by Cajanus cajan L. Millsp., Vigna unguiculata L.Walp., Phaseolus vulgaris L.,
Pisum sativum L., Lens culinaris Medik. and Macrotyloma uniflorum (Lam) Verdc., seeds showed the least number of
fungal species. That is in order to, Cicer arietinum L.> Cajanus cajan L. Millsp. > Vigna unguiculata L.Walp.>
Phaseolus vulgaris L.> Pisum sativum L.> Lens culinaris Medik.> Macrotyloma uniflorum (Lam) Verdc. during
present study.
The most common fungi were Aspergillus flavus, Aspergillus niger Alternaria alternate, Fusarium moniliforme,
Rhizopus nigricans and Trichoderma viridae were common in all selected seed samples. In Cicer arietinum L. high
incidence of Aspergillus niger Fusarium moniliforme, Rhizopus nigricans and Trichoderma virida was found when
compared to other. Many fungal species viz., Alternaria alternata, Aspergillus terrus, A. flavus, A. fumigatus, A. niger,
Botrytis sp, Cladosporium, Curvularia lunata, Fusarium solani, F. moniliforme, F. oxysporum, Macrophomina
phaseolina, Penicillium notatum, Rhizoctonia sp., and Rhizopus nigricans etc has been reported from Cicer arietinum
L.. Similar observation recorded by Ahmad et al., 1993. The most common seedbrone fungi on Phaselous vulgaris
were Aspergillus niger, A. flavus, Fusarium oxysporum, Botrytis sp, Chaetomium sp, Penicillium notatum., Rhizopus
spp., Cladosporium sp. and Trichothecium.
The fungi isolated from Lens culinaris Medik. treated seeds were Fusarium moniliforme, Alternaria alternata, Mucor
hiemalis, Chaetomium sp, Penicillium citrinum, Aspergillus niger, A. flavus, A .terreus and Nigrospora sp. F.
moniliforme, A. alternata, M. hiemalis, Chaetomium spp., and A. Niger were common in all samples while P. citrinum,
A. flavus, A. terreus and Nigrospora spp., were only isolated from untreated seed. (Muhammad Arshad Hussain et al.,
2007). Richardson, (1979) gave a list of seed-borne diseases of lentil, according to which Botrytis spp., and F.
oxysporum were isolated from lentil seed from Czechoslovakia and Uromyces fabae from debris mixed with seed from
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India. Arun and Mathew (1991) studied mycoflora of pigeon pea seeds and isolated R. nigricans, A. niger, A.
fumigatus, A. alternata, Macrophomina phaseolina, F. pallidoroseum, Chaetomium spp., Drechslera sp. and Phoma sp.
During present study, contaminated seeds result in poor germination and poor seedling vigor, resulting in an unhealthy
crop. Percentage of seed germination results of seven selected sampling legumes seeds were Cicer arietinum L.<
Cajanus cajan L. Millsp. < Vigna unguiculata L.Walp.< Phaseolus vulgaris L.< Pisum sativum L.< Lens culinaris
Medik.L< Macrotyloma uniflorum (Lam) Verdc.(Figure 1). Fungus associated with seeds because deterioration of seed
quality, affect viability and reduces germination (Srivastava and Gupta, 1981). Control of seed-borne fungi will
increase percentage of seed germination and grain yield (DGISP, 1985). Umechuruba and Nwachukwu, (1994);
Umechuruba and Nwachukwu, (1997) also reported significant reduction in seed germination, seedling emergence and
low seed and tuber yield of African yam bean caused by these important seed-borne fungi, as well as reduction in
nutritional qualities of the seeds.
ACKNOWLEDGEMENT
Dr. Narayan M. Ghangaokar and Dr. Ayodhya D. Kshirsagar is grateful to the Principal, Prof. N. S. Nikam, C.T. Bora
College, Shirur. Pune, Pin 412210.
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