Chemical fertilization in agriculture is threatening to the ecosystem. Therefore, the use of eco-friendly stimulant for crop revolution is highly desirable. This study investigates the effects of plasma activated water (PAW) created by treating de-ionized water with high voltage discharge on black gram. Results showed significant improvements in different agronomic traits. Significant changes in H2O2, a reactive-oxygen-species (ROS) were observed in seeds, leaves, and roots. Increase in catalase (ROS scavenger) was also observed in roots of plants grown from 3 and 6 min PAW treated seeds which was consistent with the upregulation of VmCAT gene. This reveals that PAW is associated with elevated H2O2 in black gram in a tightly regulated manner by the upregulation of CAT activity resulted in increased germination, growth, and development. Non-covalent binding pattern of H2O2 with CAT through docking strongly supports this phenomenon. It does suggest that the increase of H2O2 tightly regulated by the upregulation of CAT provides optimum conditions for improved seed germination and growth in black gram. These findings reveal the benefits of PAW for enhanced crop production.