AGROBACTERIUM-TUMEFACIENS-MEDIATED TRANSFORMATION OF BROCCOLI (BRASSICA-OLERACEA VAR ITALICA) AND CABBAGE (BRASSICA-OLERACEA VAR CAPITATA)

Transgenic broccoli (Brassica oleracea var. italica) was produced by t wo Agrobacterium tumefaciens-mediated transformation methods. One used flowering stalk explants from mature plants; the other used hypocotyl and petiole explants from in vitro-grown seedlings. Several hundred t ransformants containing a Bacillus thuringiensis a-endotoxin gene (Cry IA(c)-type) and the neomycin phosphotransferase gene were recovered, R ooted transformants were obtained in as little as 3 months using seedl ing explants. Transgenic cabbage was also obtained by the seedling exp lant method. Parameters important for high efficiency regeneration and transformation rates included use of a tobacco nurse cell layer, seal ing of petri dishes with a porous surgical tape instead of Parafilm, p reculture of seedling explants and appropriate length of co-cultivatio n with Agrobacterium. Advantages and disadvantages of each transformat ion procedure are discussed.