Extract of Allium sativum L. as a sustainable biopesticide candidate: Phytochemical, toxicity, and molecular characterization

The excessive use of synthetic pesticides has raised growing concerns about environmental safety, human health risks, and the emergence of pest resistance. In addressing these issues, this study evaluates the biopesticidal potential of Allium sativum L. (garlic) extract through a multidisciplinary approach involving phytochemical screening, toxicity assessment, and molecular identification. Two hypotheses were proposed: (1) that garlic ethanol extract containing alkaloids, saponins, and steroids would exhibit significant bioactivity against Artemia salina with an LC₅₀ below 100 ppm; and (2) that molecular validation using the chloroplast-encoded matK gene would confirm species identity with ≥ 99% sequence similarity to known A. sativum cultivars. Garlic was extracted via maceration using 70 % ethanol, yielding 19.94% crude extract. Phytochemical tests confirmed the presence of alkaloids, saponins, and steroids. Acute toxicity analysis using the Brine Shrimp Lethality Test (BSLT) yielded an LC₅₀ of 93.54 ppm, classifying the extract as moderately toxic by EPA guidelines. Sequencing of matK gene (854 bp) revealed 99.74% similarity to A. sativum cv. Cang ShanPuKe (GenBank: MK335928.1), supported by a 98% bootstrap in phylogenetic analysis. These findings validate both hypotheses and establish garlic extract as a bioactive, molecularly verified candidate for eco-friendly pest control within sustainable agricultural frameworks.