AGROBACTERIUM-MEDIATED TRANSFORMATION OF WHITE CLOVER USING DIRECT SHOOT ORGANOGENESIS

White clover (Trifolium repens L.) plants from the cultivars Grassland s Huia and Grasslands Tahora have been transformed using Agrobacterium -mediated T-DNA transfer. Transgenic plants regenerated directly from cells of the cotyledonary axil. To transform white clover, shoot tips from 3 day old seedlings were co-cultivated with A. tumefaciens strain LBA4404 carrying the plasmid vector pPE64. This vector contains the n eomycin phosphotransferase II gene (nptII) and beta-glucuronidase repo rter gene (gus) both under the control of the CaMV 35S promoter. Kanam ycin-resistant plants regenerated within 42 days after transfer onto s elective media. Integration of the nptII and gus genes into the white clover genome was confirmed using Southern blotting, and histochemical analysis indicated that the gus gene was expressed in a variety of ti ssues. In reciprocal crosses between a primary transformant and a non- transformed plant the introduced gus gene segregated as a single domin ant Mendelian trait.