INTERNATIONAL JOURNAL FOR INNOVATIVE RESEARCH IN MULTIDISCIPLINARY FIELD ISSN – 2455-0620 Volume - 2, Issue - 10, Oct - 2016 Isolation and identification of Seed-Borne Fungi in Green gram Seeds Page 108 Isolation and identification of Seed-Borne Fungi in Green gram Seeds Introduction: Green gram (Vigna radiata (L.) Wilczek) is one of the most important pulse crop of the arid and semiarid tropics (Chen et al., 1987). It is an excellent source of easily digestible protein. Several factors are responsible for low production of Green gram. Among them, diseases play an important role (Nene, 1980; Pal, 1996). 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. However, published reports of seed-borne fungi of Green gram in India are very few. Hence the study was undertaken to isolate and identify the incidence of seed-borne fungi associated with Green gram. Materials and Methods: Green gram varieties, GM-3, GM-4 and K-851 are very popular in south Gujarat region and are grown extensively. The bulk of seed samples of these three varieties grown at Pulse Research Station, Navsari Agricultural University, Navsari were collected at harvest and stored for two months in polythene bags. The agar plate method (ISTA, 1999) was used for the isolation of seed mycoflora of green gram. Ten seeds of each variety were taken randomly and then transferred aseptically in Petri plates containing 20 ml solidified sterilized potato dextrose agar (PDA) medium sterilized in an autoclave at 1.2 kg cm -2 pressure for 20 minutes using laminar air flow system. The seeds were placed at equal distance per plate by keeping one seed at the centre and nine seeds at the periphery. These seeds were incubated under 12/12 hr alternating light and darkened period at 25± 2 0 C for 8 days. Purification of the fungal culture was done from every new growth by hyphal tip culture. Five repetitions were kept for each of each variety and identification of the common fungi occurred in all the three varieties were done by microscopic observations and with the help of literature. These fungi were designated as isolate 1 to 9. Each of the purified isolate was identified on the basis of their cultural and morphological characteristics. The pure culture slants were also sent to ITCC, New Delhi for further confirmation. Results and discussion: Results revealed from the isolation and identification that there is association of nine different fungi. The results obtained from cultural and morphological characteristics were presented hereunder Isolate number 1 The colonies of the fungus grew fast on PDA and attained full plate diameter of about 85mm, profused mycelial growth within 5 to 6 days at 25±20C temperature in incubation with sclerotial formation. The fungus produced initially white mycelial growth later changing to brown black in centre due to the formation of numerous small black sclerotia. The mycelium was hyaline to brown, branched, septate and 1.73 to 6.52 μm in width. The sclerotia formed in culture were black, hard and 67.90 to 195.14 μm in diameter. The pycnidia were not observed in the culture (Plate I-1). The studies on the cultural and morphological characters of isolated fungus showed its close identity with Macrophomina phaseolina as described by Tandel (2004). Moreover, the I.T.C.C., New Delhi also identified the same fungus as M.phaseolina (I.T.C.C. No.7811.10).Thus, the fungus under present study was identified as Macrophomina phaseolina (Tassi) Goid. A.J.Deshmukh 1 , A.N.Sabalpara 2 , V.A. Patil 3 and V.P. Prajapati 4 1 & 4 Asst. Professor, College of Agriculture, NAU, Waghai 2. Director of Research & Dean PG Studies, NAU, Navsari 3. Asst. Research Scientist, NARP, NAU, Navsari Email - amol_deshmukhnau@yahoo.co.in Abstract: An experiment was undertaken to isolate and identify the seed borne fungi of Green gram. Seed samples of three Green gram varieties were collected from Pulse research station, NAU, Navsari. PDA plating method was employed for the isolation of the fungi and cultural, microscopic study was adopted for the identification. Altogether nine fungi were found in association Viz., M. phaseolina, A. alternata, C. capsici, N. sphaerica, Chaetomium sp., Aspergillus sp., R. oryzae, sterile aseptate and sterile septate fungus with all the three variety seed lots. Key Words: Seed-Borne Fungi, Green gram, cultural, morphological study.