PJB-2022-138
IN VITRO PROPAGATION AND SOMACLONAL VARIATION STUDY OF MEDICINAL HERB LINDERNIA PUSILA
Tikendrajit Baro
Abstract
Lindernia pusila is one of the important medicinal herbs which are being used to treat various diseases. The objectives of present study are 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 treatment with 0.1% mercuric chloride resulting 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 were isolated using DNeasy Plant Mini kit and RAPD assay was performed using 14 different RAPD primers. Among the 14 RAPD primers 7 primers (OPC07, OPC08, OPA04, OPA13, OPA12, OPC01, OPC04) formed monomorphic DNA bands in the wild plants as well as in the tissue cultured plants whereas the other 7 primers (OPC 02, OPC 05, OPC 09, OPA 01, OPA2, OPC 03 and OPC 06) formed polymorphic DNA bands in tissue cultured Lindernia pusila plants.
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