In vitro Callus Induction and LC-MS-based Phytochemical Analysis of Salacia macrosperma Wight

Abstract

The present study aimed to develop a standardized tissue culture protocol for callus induction in Salacia macrosperma using juvenile leaf explants. Fifteen treatments were evaluated in a completely randomized design to optimize in vitro callus induction. A comprehensive protocol for efficient callus induction was established. Plant Preservative Mixture (PPM) was identified as the most effective agent for minimizing microbial contamination. Callus induction was achieved from the midrib region of leaf explants cultured on Murashige and Skoog (MS) medium supplemented with B5 vitamins, 2,4-dichlorophenoxyacetic acid (2,4-D), and kinetin (Kn). The highest callus response (181.33 ± 4.10) was recorded in the medium containing 7 mg L⁻¹ 2,4-D and 3 mg L⁻¹ Kn, whereas the lowest response (14.33 ± 2.40) was observed at lower concentrations of the growth regulators. The study also investigated the phytochemical composition of Salacia macrosperma roots using liquid chromatography–mass spectrometry (LC–MS). LC-MS analysis revealed a total of 1,775 metabolite features, comprising 849 identified compounds and 926 unidentified features. These compounds may play important roles in the management of metabolic disorders such as diabetes and chronic inflammation, highlighting the therapeutic potential of Salacia macrosperma.