Successful cocultivation of Brassica napus microspores and proembryos with Agrobacterium

Brassica napus microspores and microspore-derived proembryos were cocultivated with Agrobacterium tumefaciens harbouring a binary vector. The vector contained selectable genes for kanamycin and hygromycin antibiotic resistance. Microspores and proembryos survived the cocultivation procedure and subsequent antibiotic selection. Thousands of plantlets can be regenerated from a single experiment. Biochemical analysis indicated up to 7.3% of plants exhibited neomycin phosphotransferase II enzyme activity. Success of the cocultivation procedure depended largely on choosing the proper coculture conditions while allowing microspore embryogenesis to proceed.