Transformation of White Clover with a Gene Encoding a Lepidopteran-active- endotoxin from Bacillus thuringiensis var. Berliner 1715

During sporulation, Bacillus thuringiensis var. berliner 1715 produces insecticidal proteins (6-endotoxins) that are lethal specifically to Lepidopternn insects. Agrobacterium tumefaciens-mediated T-DNA transfer has been used to introduce a cry/A(b) li-endotoxin gene (the 812 gene) from this B. thuringiensis strain into a genotype of white clover (WR8). The aim is to protect white clover (Trifolium repens L.) from damage caused by Lepidopteran insects, including the pasture pests porina (Wiseana spp.) and sod-web worms (Crombidae), White clover plants were transformed with a binary vector (pGSH 163) containing the bidirectional mannopine synthase promoter which directs simultaneous expression of both the 812 gene, and the neomycin phosphotransfernse II gene (NPTII). Transgenic white clover plants were selected using kanamycin sulphate, and integration of the Bt2 gene was confirmed by polymerase chain reaction (PCR) amplification of the introduced gene from genomic DNA.