INTERGENERIC COCULTURE OF GENETICALLY TRANSFORMED ORGANS FOR THE PRODUCTION OF SCOPOLAMINE

This work demonstrates the effectiveness of interspecies and intergenu s organ co-culture for the production of plant secondary metabolites. Go-cultures of Atropa belladonna transformed roots and Duboisia hybrid shooty teratomas produced significant levels of scopolamine, whereas no scopolamine could be detected in separate root and shoot cultures. Maximum scopolamine levels found in co-cultured shoots were 4.8 mg g(- 1) dry weight in a 2 x 2.5-1 dual bioreactor system, and 3.7 +/- 0.9 m g g(-1) in dual 1-1 shake flasks. Periodic crushing of the roots in sh ake flasks was effective in releasing hyoscyamine, but impaired growth and overall alkaloid synthesis. Less severe root damaging in the bior eactor was more successful in improving scopolamine accumulation. Upta ke of hyoscyamine by shooty teratomas was inhibited at high hyoscyamin e concentrations, was reduced in the presence of ATPase inhibitors, an d could be significantly enhanced by pre-incubating the shoots on medi um containing hyoscyamine. (C) 1997 Elsevier Science Ltd.