Production of Hyoscyamine by ‘Hairy Root’ Cultures of Datura stramonium

Abstract

‘Hairy root’ cultures of Datura stramonium were established following infection of aseptic leaves with Agrobacterium rhizogenes. Transformation was confirmed by Southern blotting using [³² P]-labelled fragments of the T-DNA as probes. The transformed cultures grew in the absence of added phytohormones and cell mass increased 55-fold during 28 days incubation. Hyoscyamine was a major component of the alkaloid fraction and accounted for at least 0.3% of the dry matter, comparable to pot-grown plants from which the cultures were initiated. Production of hyoscyamine followed growth during the first 15 days of incubation, but continued to increase during the early part of stationary phase. The alkaloid was retained almost entirely in the root tissue. The effects of medium composition and pH on growth and hyoscyamine production are reported.