Production of ÿ-ecdysone in in vitro-cultured Brazilian ginseng, Pfaffiaglomerata(Spreng.) Pedersen

Abstract

SummaryIn Brazil, several pharmaceutical industries produce phytomedicines from the roots of Pfaffia glomerata (Spreng.) Pedersen. In this work, callus cultures of P. glomerata were established in order to study whether in vitro-cultured tissue could produce _-Ecdysone. To induce calli, nodal segments were cultured on Murashige and Skoog (MS) medium with various levels of 6-benzylaminopurine (BAP; 1.0, 5.0, or 10.0 ?M), _-naphthaleneacetic acid (NAA; 0.5, 1.0, 5.0, or 10.0 ?M) or 2,4-dichlorophenoxyacetic (2,4-D; 0.5, 1.0, 5.0, or 10.0 ?M). _-Ecdysone concentrations [on a (w/w) dry weight (DW) basis] in in vitro-cultured and field-grown tissues were measured by high performance liquid chromatography.The level of _-ecdysone was related to the friability and shoot regeneration capacity of the different calli. Organogenic, friable calli grown on media supplemented with 10.0 ?M BAP, plus either 5.0 ?M NAA or 0.5 ?M 2,4-D, resulted in the highest _-ecdysone concentrations [0.282% (w/w) and 0.211% (w/w), respectively]. In contrast, the _-ecdysone concentrations in field-grown plants were 0.194% (w/w) in roots and 0.333% (w/w) in the aerial parts. Low levels of _-ecdysone [0.007% (w/w)] were detected in plantlets cultured on hormone-free MS medium, indicating the importance of culture conditions for the in vitro production of _-ecdysone.This is the first report on the presence of ecdysteroids in calli of P. glomerata.