AVNI DAHIYA, ADHISHA DAHIYA, SUBHASH KAJLA AND NAMITA SINGH*
Department of Biotechnology, Guru Jambeshwar University of Science & Technology, Hisar-125 001 (Haryana),
India
*(e-mail: namitasingh71@gmail.com; Mobile: 94169 28883)
(Received: July 1, 2025; Accepted: August 15, 2025)
ABSTRACT
Bacopa monnieri (L.) Wettst., a medicinally important and endangered herb rich in the nootropic compound
bacoside-A, faces severe constraints in conventional propagation due to poor seed viability and overharvesting of wild populations. This study developed an optimized in vitro protocol to enhance callus induction, biomass production and bacoside-A accumulation. Leaf explants from in vitro grown plants were cultured on Murashige and Skoog (MS) medium with varying plant growth regulators (PGRs), where a combination of 1.5 mg/l BAP and 2.5 mg/l NAA produced the highest callus induction (90%) within 12 days. Callus proliferation and secondary metabolite synthesis were further improved by supplementing the medium with adenine sulphate (ADS) and casein hydrolysate (CH), individually and in combination. The synergistic treatment of 50 mg/l ADS and 100 mg/l CH yielded the maximum biomass (0.294 g dry weight) and bacoside-A content (15.75 mg/g DW in ethanol extract), significantly surpassing control and single-additive treatments. Ethanol consistently proved superior to aqueous extraction for bacoside-A recovery. These findings demonstrated that the strategic use of optimized PGRs with ADS and CH can greatly enhance in vitro propagation and metabolite production in B. monnieri, offering a scalable approach for conservation and commercial exploitation of this high-value medicinal plant.
Key words: Bacopa monnieri, callus induction, bacoside-A production, adenine sulphate (ADS), casein hydrolysate (CH)