Hairy root culture of Przewalskia tangutica for enhanced production of pharmaceutical tropane alkaloids

The bacteria-free seedlings of Przewalskia tangutica were germinated from seeds. The leaves were used as initial explants for genetic transformation mediated byAgrobacterium rhizogenes strain A4, with the induction of Acetosyringone. The hairy roots were induced from the wounded sites of the leaves at the frequency of 100%. The independently transformed and fast growing hairy roots were subcultured in MS medium to establish the hairy root culture lines. Genomic DNA PCR analysis was applied to detect the presence of the rooting genes, includingrolB and rolC that had integrated into the genome of Przewalskia tangutica. After 5-week culture in MS liquid medium, the hairy roots produced scopolamine and hyoscyamine at higher levels by comparison to the wild-type roots. The best scopolamine-production line produced 0.68 mg/g DW scopolamine, and the best hyoscyamine-production line produced 1.13 mg/g DW hyoscyamine. It was the first time that hairy root cultures of P. tangutica were established to produce tropane alkaloids.   Key words: Przewalskia tangutica, hairy root, scopolamine, hyoscyamine.