A comparative analysis of antioxidant and bioactive constituents in micropropagated and wild Lindernia pusilla medicinal herb

Abstract

Lindernia pusila is one of the important medicinal herbs which is being used to treat various diseases. The present experiment is aimed to develop an efficient protocol for in vitro mass propagation of L. pusila and to study somaclonal variations in the micro propagated plants using RAPD technique. Surface sterilization of collected explants of L. pusila was performed using 0.1% mercuric chloride and cultured in MS media with different concentrations of growth regulators. Explant surface sterilization was most effective at 2 & 3 minutes of treatment with 0.1% mercuric chloride resulting in 75% of the explant survival. The media MS + 1 mg/L BAP + 0.2 mg/L NAA showed maximum shoot proliferation and multiplication per explant. After hardening of tissue cultured plants, the whole genome of wild and micro propagated plants was isolated using the DNeasy Plant Mini Kit and RAPD assay was performed using 14 different RAPD primers. Among the 14 RAPD primers, 7 primers formed polymorphic DNA bands in tissue-cultured Lindernia pusila plants. Comparatively the tissue cultured plant extract showed higher content of phenols, flavonoids, and anti-oxidant properties than the wild extracts of L. pusilla. In Chromatographic study of gallic acid, the tissue cultured extract showed more gallic acid concentration (42.9mg/g) than the wild sample (10.9mg/g) and the quercetin constituent of tissue cultured plant was higher than the wild extract. The volatile bioactive components found in the wild and tissue cultured extracts were studied using GC-MS analysis.