Black gram is one of the most indispensable components of the world food basket and the growth and health of the crop get influenced by biotic and abiotic factors. Beneficial phyto-microbes are one among them that influence the crop growth, more particularly the seed borne microbes are comparatively beneficial, that they pass from generation to generation and are associated with the plants from establishment to development. In the present study, twenty seed-borne yeasts were characterized and tested for growth promotion of black gram and their antagonism against black gram phytopathogens. Two yeasts, Pichia kudriavzevii POY5 and Issatchenkia terricola GRY4, produced indole acetic acid (IAA), siderophore, 1-amino cyclopropane-1-carboxylic acid deaminase (ACCD), and plant defense enzymes. They solubilized phosphate and zinc and fixed atmospheric nitrogen. Inoculation of these two yeast isolates and Rhizobium BMBS1 improved the seed germination, physiological parameters and yield of black gram. Inoculation of Rhizoctonia solani-challenged plants with plant growth-promoting yeasts, resulted in the synthesis of defense-related enzymes such as peroxidases (POD), chitinases, catalase (CAT), and polyphenol oxidases (PPO). Thus, the seed-borne yeasts, Pichia kudriavzevii POY5 and Issatchenkia terricola GRY4, could be used as plant probiotics for black gram.