利用农杆菌转化花椰菜组织的研究

供试的四个花椰菜品系对农杆菌T_(37)反应极为敏感,但各品系反应程度有所不同,肿瘤发生率变动于64%-82%之间。利用含有二元载体的农杆菌LBA_(4404)与无菌幼苗的下胚轴切段共培,获得卡拉霉素抗性植株。DNA分子杂交证明,二元载体上的T—DNA确实整合到植物染色体上,其npt嵌合体基因使转化体具有卡拉霉素抗性。由胭脂碱Ti质粒pTiT_(37)诱导的丛生芽,通过组织培养较易分化出不定根,但这些芽状结构很难发育成正常植株。

TRANSFORMATION OF BRASSICA OLERACEA VAR BOTRYTIS MITH AGROBACTERIUM TUMEFACIENS

The sensitivity of several varieties of cauliflower to infection with Agrobac-terium tumefaciens strain T37 was measured at high frequencies, of tumourgenesis raging from 54to 82 per cents on in vivo infected sites after 8 weeks of inoculation. Hypocotyl segmentsfrom germinated sterile-seedlings were CO-cultivated with A. tumefaciens strain LBA4404harbuoring binary vectors, on one of which a achimaeric npt gene codes for the kanamycinresistant determinant as a selection marker in transformants. Resistant plants weresuccessfully obtained under kanamycin selection. Southern hybridization analysis showedthe presence of T-DNA in total DNA isolated from several independant transformants.On the other hand, multiple shoots induced by nopaline Ti plasmid pTiT37 were easy tobe rooted on differentiation mediam but failed to form whole plants.