Ding L-C, Hu C, Yeh K-W, Wang P-J
Graduate Institute of Agricultural Biotechnology, National Chung Hsing University, Taichung, Taiwan, , , , , , TW.
Center for Applied Science, William Paterson University, Wayne, NJ 07470, USA Fax no.: +1-973-7202338 e-mail:
Plant Cell Rep. 1998 Aug;17(11):854-860. doi: 10.1007/s002990050497.
Agrobacterium-mediated transformation was used to introduce a trypsin inhibitor gene into Taiwan cauliflower (Brassica oleracea var. botrytis L.) cultivars. The TI gene was isolated from a well-adapted Taiwan sweet potato cultivar and was expected to be especially effective in combating local pests. In vitro regeneration studies indicated that 4-day-old cauliflower seedling hypocotyl segments, pretreated with 2,4-dichlorophenoxyacetic acid for 3 days and incubated on a silver-ion-containing shoot induction medium, gave regeneration rates greater than 95%. Optimum transformation conditions were determined. G418 selection at 15 mg/l was initiated 1 week after cocultivation, and the dose was doubled 1 week later. Over 100 putative transgenic plants were produced. Transgenic status was confirmed by in vitro TI activity, and Southern and Western hybridization assays. The transgenic plants demonstrated in planta resistance to local insects to which the control plants were vulnerable.
利用农杆菌介导的转化方法将一种胰蛋白酶抑制剂基因导入台湾花椰菜（Brassica oleracea var. botrytis L.）品种。该TI基因从一个适应性良好的台湾甘薯品种中分离出来，预计在对抗当地害虫方面特别有效。体外再生研究表明，用2,4-二氯苯氧乙酸预处理3天的4日龄花椰菜幼苗下胚轴切段，在含银离子的芽诱导培养基上培养，再生率超过95%。确定了最佳转化条件。共培养1周后开始用15 mg/l的G418进行选择，1周后剂量加倍。产生了100多株推定的转基因植株。通过体外TI活性以及Southern和Western杂交分析确认了转基因状态。转基因植株在植株内对对照植株易受侵害的当地昆虫表现出抗性。
