Double transformation: Producing transgenic phosphinothricin-resistant plants of commercial pea lines

The worked-out protocol for the double transformation of the garden pea (Pisum sativum L.) comprises two steps: (1) obtaining the regenerable lines in an in vitro culture by transforming plants with the shooty mutant of Agrobacterium tumefaciens pGV2206; and (2) producing transgenic plants containing the foreign gene in question from the regenerable lines. The protocol was employed to produce transgenic pea plants containing the gene for resistance to herbicide L-phosphinothricin (BASTA) and the neomycin phosphotransferase gene (nptII), The synthetic bar gene for resistance to BASTA was transferred to pea plants using a binary vector pMP90/pK3 driven by cauliflower mosaic virus 35S promoter. The molecular biological analysis confirmed bar and nptII integration into the plant genome. A protocol was developed for seed production in in vitro culture. The transferred bar gene was inherited in the T-2 generation.