Green grams are the most valuable pulse crops in terms of plant-based protein, dietary fiber, and various phytochemicals. Although green gram is found susceptible to the soil-borne fungus Macrophomina phaseolina (Tassi) Goid, it leads to severe root-rot disease and causes a significant reduction in crop yield. Thus, the study aims to determine the bioefficacy of indigenous isolates of biocontrol fungi and bacteria viz., T. asperelloides AMUTASPD-51, T. asperellum AMUTASPM-51, T. asperellum AMUTASPM-52, T. asperellum AMUTASPM-53, T. atroviride AMUTATROV-31, T. harzianum AMUTHZ-71, T. harzianum AMUTHZ-72, T. harzianum AMUTHZ-73, T. harzianum AMUTHZ-74, T. hamatum AMUTHM-31, T. viride AMUTVR-61, T. viride AMUTVR-62, T. virens AMUTVNS-41, T. longibrachiatum AMUTLONB-41, Pochonia chlamydosporia AMUPC-31, Purpureocillium lilacinum AMUPL-31, Aspergillus niger AMUAN-41, Bacillus subtilis AMUBS-80 and Pseudomonas fluorescens AMUPF-80 against Macrophomina phaseolina AMUMP-2 by using dual inoculation technique for seven days incubation at a temperature under in-vitro condition. All species of Trichoderma fungus showed high biocontrol potential to suppress the radial growth of M. phaseolina AMUMP-2 over control. Among the biocontrol fungi and bacteria, T. viride AMUTVR-61 resulted in the highest radial inhibition of M. phaseolina AMUMP-2 by 95.0% over control. The T. harzianum AMUTHZ-72 was second most effective in decreasing the radial growth by 94.2% of the pathogens, followed by T. harzianum AMUTHZ-71 (92.8%), T. asperellum AMUTASPM-53 (86.1%), and T. harzianum AMUTHZ-74 (83.7%) over control. However, B. subtilis AMUBS-80 was found to be the least effective, suppressing radial inhibition of M. phaseolina AMUMP-2 by 21.7% over control. The present study indicates that T. viride AMUTVR-61 and T. harzianum AMUTHZ-72 were the most significant indigenous biocontrol fungi against M. phaseolina AMUMP-2. Furthermore, its application led to a substantial decrease in the soil-borne pathogen population that affects plant health, especially green gram, and adverse environmental and human effects.