豇豆胰蛋白酶抑制剂转基因棉花的获得  

Obtaining Transgenic Cotton Plants with Cowpea Trypsin Inhibitor Gene

作　　者：李燕娥 朱祯[1,2] 陈志贤 吴霞 王伟 李淑君[1,2] 朱玉 焦改丽 吴家和[1,2] 徐鸿林 范小平 孟晋红 肖桂芳[1,2] 李向辉 

机构地区：[1]山西农科院棉花研究所 [2]中国科学院遗传研究所

出　　处：《棉花学报》1998年第5期237-243,共7页Cotton Science

摘　　要：利用根癌农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍｔｕｍｅｆａｃｉｅｎｓ）介导法将豇豆胰蛋白酶抑制剂（Ｃｏｗ－ｐｅａＴｒｙｐｓｉｎＩｎｈｉｂｉｔｏｒ，ＣｐＴＩ）基因转移进入棉花（ＧｏｓｙｐｉｕｍｈｉｒｓｕｔｕｍＬ．）。棉花幼苗的下胚轴与携带有ＣｐＴＩ基因和Ｎｐｔ－Ⅱ基因的根癌农杆菌共培养，在选择培养基上诱导出了卡那霉素抗性（Ｋｍｒ）愈伤组织。选择Ｎｐｔ－Ⅱ阳性的胚性愈伤组织在胚状体诱导培养基上进行胚状体诱导，继而在分化培养基上进行植株分化，最终获得了再生棉花植株。经Ｎｐｔ－Ⅱ分析、ＰＣＲ及Ｓｏｕｔｈｅｒｎ检测证明，外源ＣｐＴＩ基因和标记基因（Ｎｐｔ－Ⅱ基因）存在于转化棉株及其后代中。抗棉铃虫（Ｈｅｌｉｏｔｈｉｓａｒｍｉｇｅｒａ）生物活性检测表明转基因植株后代具有明显的抗棉铃虫能力。It has been successful to transfer the cowpea trypsin inhibitor(CpTI) gene into cotton(Gossypium hirsutum L.) by A.tumefaciens mediated transformation. Hypocotyl segments from 5 day old seedlings were co-cultured with A.tumefaciens containing foreign CpTI gene and Npt Ⅱ gene. Kanamycin resistant calli were obtained by screening on selective medium. Npt Ⅱ positive calli were subcultured on embryo-inducing medium untile embryos developed. Transgenic cotton plantlets were regenerated on differentiation medium from these embryos. It has been confirmed that transgenic cotton offsprings still carrried foreign CpTI gene and marker gene by PCR, Southern blot and Npt Ⅱ assay. Results of bioassay demonstrated that the transgenic cotton lines were with higher resistance to cotton bollworm(Helicoverpa armigera).

关 键 词：棉花转化 根癌农杆菌 抗虫基因 豇豆 CPTI 

分 类 号：S562.035[农业科学—作物学]