A stable transformation system for the ornamental plant, Datura meteloidesDC

A transformation system is described for Datura meteloides using the superv irulent Agrobacterium tumefaciens strain 1065, carrying both the beta-glucu ronidase (gusA) and neomycin phosphotransferase II (nptII) genes between th e T-DNA border sequences of the binary vector. The importance of conditions such as the preculture period of the plant tissues, wounding, bacterial di lution and incubation time were evaluated in terms of transgenic plant prod uction. A preculture period of 2-3 days, using a 1 : 20 or 1 : 10 (vol:vol) dilution of an overnight bacterial culture, resulted in optimum shoot rege neration, with 48% from a total of 576 explants regenerating transformed sh oots. Expression of the gusA and nptII genes was confirmed by a GUS fluorom etric assay and by NPTII ELISA. Southern analysis revealed the integration of both transgenes, which segregated as dominant Mendelian traits in seed p rogeny.