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An efficient procedure for sprouted seed method of agroinoculation in black gram crop with infectious dimer agro clones of mungbean yellow mosaic virus (MYMV) DNA-A and DNA-B has been developed using black gram susceptible genotype PBG-1. Different parameters like Agrobacterium cell density, concentration of acetosyringone, inoculation buffer, incubation time, incubation temperature and place of pinpricking were standardized for maximum infectivity of MYMV virus in agroinoculated plants. The Agrobacterium cell density at OD₆₀₀ value 0.8 gave high percentage of virus infection followed by OD₆₀₀ value 1 and 0.5. It was observed that high percentage of virus infection gave in 100 µm concentration of acetosyringone followed by 80 µm and 150 µm concentrations of acetosyringone. Agroinoculated blackgram seeds were incubated at three incubation times (1hour, 5hours and overnight) and 80-100% percentage of infection was observed at all three incubation times, similarly incubation temperatures (25^oC, 28^oC and 37^oC were evaluated for maximum percentage of infection and 82% of infection was observed at incubation temperature 28^oC followed by 25^oC and37^oC, even poor germination was noticed at incubation temperature 37^oC. First time we evaluated the place of pinpricking on hypocotyl region for maximum virus infection and found that pinpricking around the hypocotyls gave high percentage of virus infection (79%) in agroinoculated blackgram plants.

Key words: Sprout seed method of agroinoculation, Parameters and MYMV.