Journal of Advances in Biology & Biotechnology

Bacopa monnieri (L.) Pennell, commonly known as “Brahmi,” is a medicinally important plant widely used in Ayurveda for its neuroprotective and memory enhancing properties. The present study aimed to standardize an efficient in vitro micropropagation protocol by optimizing explant type, surface sterilization, hormone concentration, and sucrose level. Nodal and leaf segments were selected as explants. Effective surface sterilization was achieved using 0.1% Bavistin for 30 minutes followed by 0.05% HgCl₂ for 4 minutes, which eliminated contamination while preserving viability. Shoot initiation occurred within 5–8 days, with the highest frequency of multiple shoots observed on MS medium supplemented with 3 ppm BA or 2 ppm BA + 1 ppm NAA. Callus induction was most successful from leaf explants cultured on MS + 2 ppm 2,4-D + 1 ppm kinetin, forming friable and regenerative calli within 30–35 days. For rooting, IBA at 3 ppm promoted direct multiple shoot and root formation, while MS + BAP (5.5 ppm) + NAA (0.2 ppm) supported both elongation and rooting. Optimal growth was obtained at pH 5.8 and 3% sucrose, which enhanced shoot number and explant vigor. The developed protocol is fast, reliable, and reproducible, providing a basis for large scale clonal propagation, conservation of germplasm, and potential metabolite enhancement in B. monnieri.