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Purpose The secondary metabolites in plants are the basis of defense and stress balance, which is an important aspect in plant growth. The UV-B treatment (a biotic stress) and salt stress on bitter melon ( Momordica charantia L.) were studied, and the impact of pre-sowing seed treatment was evaluated on the basis of biochemical and enzymatic biomarkers, antioxidants, and wound-healing potential during early growth stages. Methods The UV-B treatment for 5 and 10 min and salt stress 250 mM and 500 mM treatments were applied, and 21-day seedling tissue were collected for total phenolic contents (TPC), total flavonoid contents (TFC), antioxidant, chlorophyll contents, hydrogen peroxide, total soluble sugar, enzymes activities, and wound-healing potential studies. Results The TPC, TFC, diphenyl picrylhydrazyl (DPPH), chlorophyll contents, and total soluble sugar were recorded higher at 5 min treatment with UV-B and salt stress at 250 mM concentration. Antioxidant enzymes activities were recorded higher for 10 min UV-B treatment and 500 mM salt treatment. Wound-healing potential was found significant at 5 min treatment with UV-B radiation, which was studied in vivo in rabbits. The LC-MS analysis revealed a variety of phenolic compounds in the seedlings. Conclusion The study concluded that treatments significantly affect the biological activities of bitter melon seeds at the seedling stage, and the seeds contain important phenolic compounds responsible for its antioxidant potential and enzymatic activities. Future studies could be focused on the later stages of growth, development, and yield characteristics subjected to salt stress along with UV-B radiation treatment.