Solanum virginianum commonly known as wild eggplant or nightshade plant is a prickly herb that grows throughout Asia including India and Australia. S. virginianum, a member of the Solanaceae family is used by traditional medicinal practitioners to treat different ailments. Several studies have done to scientifically evaluate the potential pharmacological properties of the plant. However, the lack of genetic data on S. virginianum restricts its future research, particularly at the molecular level. The current study aims at transcriptome analysis of the S. virginianum fruit. 18.19 million high-quality reads were obtained. Afterthe de novo transcriptome analysis, 1.4 million unigenes and 60,487 coding sequences were found using Transcoder v5.3.0. 200 maximal length CDS transcripts were translated to protein using the Expasy translate server. Bioactive peptides were identified by different in silico approaches which revealed 58 antimicrobial peptides. All identified peptides were non-toxic. Among the 58 bioactive peptides, 19 are defensins. Four bioactive peptides SVBP1-CITGTTKTFYVN, SVBP-YGKNIVNRGRPRCS, SVBP3-KKCVCGSPRCRGYIGG, and SVBP4-FKIFGCICYAHV have been synthesized, evaluated for hemolytic activity and molecular docking study have been done to evaluate its antimicrobial activity. The identified new bioactive peptides could potentially be used in the next research on antibacterial, anti-inflammatory, and anti-cancer agents.