PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY

The present study was conducted to develop a protocol for the conservation of P. kurroa through in vitro culture of leaf segments of plant. MS+2, 4-D (8μM) was found to be extremely effective for the development of callus from leaf segments. The medicinal plant has been utilized in traditional as well as a modern system of medicine since ages. However, overexploitation has rendered several medicinal plants endangered. Picrorhiza kurroa is one such crucial medicinal plant which has become endangered primarily due to unrestricted collection from wild stands for traditional as well as commercial medicinal purposes. Regeneration of shoot from the subculture of callus onto MS+2,4-D (2-10μM) + Kn (4-10μM) and MS+2,4- D (2-10 μM) +BAP (4-10 μM) was achieved. Medium supplemented with 2,4-D and Kn was found to be superior as compared to medium containing 2,4-D and BAP. Callus obtained was fragile and green. MS + 2,4-D (10μM) + Kn (8μM) resulted in regeneration of about 6.2±0.4 shoots per culture with a maximum of 8 shots within 7-10 days of subculture of callus. Regenerated shoots were rooted in MS+IBA (10-20 μM). A maximum of 82.4% explants developed in vitro roots on ½MS+12μM IBA. About 62.4% regenerated plants were successfully transferred to natural conditions during the process of acclimatization. The GC-MS analysis of a methanolic extract of leaves of wild and in vitro regenerated plants revealed the presence of 26 and 46 phytocompounds respectively. Phytocompounds phytol; vitamin-E; glycidyl palmitate; linalyl cinnamate; eicosanol; cis-caryophyllene were specifically present in the extract of in vitro raised plant whereas benzestrol; cedrene 13 ol; thione; thujone; bisalolol; curcumene; nerodilol were present in extract of mother plant.