Li H, Wylie S J, Jones M G K
Western Australian State Agricultural Biotechnology Centre (SABC) and Centre for Legumes in Mediterranean Agriculture (CLIMA), School of Biological Sciences and Biotechnology, Division of Science and Engineering, Murdoch University, Perth, W.A. 6150, Australia e-mail:
Plant Cell Rep. 2000 May;19(6):634-637. doi: 10.1007/s002990050785.
Transgenic yellow lupin (Lupinus luteus L.) plants have been generated by meristem co-cultivation with Agrobacterium tumefaciens. The binary plasmid pPZBNIa contains the bar gene under the control of a CaMV 35 S promoter. The transformation method involves inoculation of embryonic axis explants with A. tumefaciens, flooding the meristem with glufosinate, and initial culture on non-selective medium. Shoots were transferred to culture medium containing 20 mg/l glufosinate. Following subculture, shoots were grafted onto non-transgenic narrow-leafed lupin (L. angustifolius L.) seedling rootstocks, or rooted in vitro. The overall transformation efficiency, as determined at the T generation, was 0.05%-0.75%. The transgenic nature of plants grown to the T generation was confirmed by phosphinothricin acetyl transferase, PCR and Southern analyses.
通过与根癌农杆菌共培养分生组织，已培育出转基因黄羽扇豆（Lupinus luteus L.）植株。二元质粒pPZBNIa含有受CaMV 35S启动子控制的bar基因。转化方法包括用根癌农杆菌接种胚轴外植体，用草铵膦淹没分生组织，并在非选择性培养基上进行初始培养。将芽转移到含有20mg/l草铵膦的培养基中。继代培养后，将芽嫁接到非转基因窄叶羽扇豆（L. angustifolius L.）幼苗砧木上，或在体外生根。在T代测定的总体转化效率为0.05%-0.75%。通过膦丝菌素乙酰转移酶、PCR和Southern分析证实了生长到T代的植株的转基因性质。
