TRAITS OF TRANSGENIC ATROPA-BELLADONNA DOUBLY TRANSFORMED WITH DIFFERENT AGROBACTERIUM-RHIZOGENES STRAINS

Hairy root cultures of Atropa belladonna L. were established by infect ion either with Agrobacterium rhizogenes ATCC 15834 or MAFF 03-01724, and transgenic plants were obtained from both hairy root cultures. Dou bly transformed roots were induced by re-infection of the leaf segment s of transgenic Atropa belladonna plants (A. rhizogenes 15834) with MA FF 03-01724. Shoots and viviparous leaves were regenerated from the do ubly transformed roots. The genetic transformation was determined by t he opine assay (agropine, mannopine and/or mikimopine) and polymerase chain reaction. Physiological changes and tropane alkaloid biosynthesi s in the hairy roots (singly and doubly transformed) were investigated . The alkaloid content in the doubly transformed root strain was inter mediate as compared to the root strains which were singly transformed. On the other hand endogenous IAA levels in doubly transformed roots w ere significantly decreased compared to both singly transformed roots.