In vitro regeneration and chromatographic fingerprint analysis of Aphanamixis polystachya (Wall.) Parker by HPTLC technique

Abstract

• In vitro regeneration on nodes of Aphanamixis polystachya tree showed breakage of recalcitrance. • Biomass content was higher on cytokinins 2-ip (N 6 -(2-Isopentyl) Adenine) supplemented Murashige and Skoog's medium. • HPTLC analysis of showed higher diversity of phytochemicals in the in vitro shoots without any supplementation of elicitors. • Comparison between the in vitro shoots and mature plants shown the presence of majority of phytochemicals in both groups. Aphanamixis polystachya is an evergreen tree belonging to the Meliaceae family used for centuries in traditional medicines. Various metabolites that were isolated from the barks showed high therapeutic value. Isolation of pharmaceutically active compounds imposes pressure on harvesting the plants from the wild, resulting in the endangerment of the tree. The development of an alternate method for the isolation of phytochemicals is the need of the hour. The current study evaluated the phytochemical screening of bioactive compounds from the tree parts and in vitro regenerated shoots of A. polystachya . Due to its recalcitrancy, tissue culture data remains inaccessible, and the present investigation was first attempt on developing a protocol for generating plantlets using in vitro culture. Murashige and Skoog's medium fortified with cytokinins like BAP (6-Benzylaminopurine), Kinetin, 2-ip (N 6 -(2-Isopentyl) Adenine), and TDZ (Thidiazuron) were used, and maximum biomass was produced from 3 mg/L of 2-ip. Meanwhile, HPTLC fingerprinting analysis on the tree parts (bark and leaves) and in vitro shoots showed similar diversity among metabolites. The secondary metabolites produced in the in vitro plants were the same with mature trees for standards like Rutin, Stigmasterol, β-Sitosterol, Piscidinol A, Bourjotinolone B, and Niloticin.