关键词: 颠茄, 托品烷类生物碱, 发根, 外源基因, 代谢工程

Abstract: The disarmed Agrobacteriumtum efaciens strain C58C1 harboring the plasmid pRiA4 was engineered by introducing the plant expressing vector pCAMBIA1304⁺ . The bacteria-free well-growing true leaves of A. belladonna were infected with the engineered recombined C58C1. The hairy roots were induced from the wounded sites of the leaves at the frequency of 100%. Then, genomic PCR was applied to detect rolB , rolC, and antihygromycin gene in different monoclonal hairy root lines. All the three geneswere simultaneously detected in 30% hairy root clones. The results demonstrated that exogenous genes could be exp ressed in hairy roots of A. belladonna at a relatively high frequency by the method in the present studies. These were fundamental works for tropane alkaloidsmetabolic engineering in hairy roots and molecular breeding of A. belladonna.

Key words: Atropa belladonna, Tropane alkaloids, Hairy root, Exogenous gene, Metabolic engineering Regeneration