Bt cry1Ia8抗虫基因对结球甘蓝的转化及其表达

园艺学报 ›› 2009, Vol. 36 ›› Issue (8): 1161-1168.

Bt cry1Ia8抗虫基因对结球甘蓝的转化及其表达

崔磊¹;杨丽梅^1*;刘楠²;郎志宏³;刘玉梅¹;庄木¹;张扬勇¹;张友军¹;黄大昉^2,3;方智远¹

(¹中国农业科学院蔬菜花卉研究所, 北京100081; ²中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京100193; ³中国农业科学院生物技术研究所, 北京100081)

收稿日期:2009-04-13 修回日期:2009-06-29 出版日期:2009-08-25 发布日期:2009-08-25
通讯作者: 黄大昉

Transformation and Expressing of Bt Gene cry1Ia8 in Cabbage

CU ILei¹;YANG Li-mei^1*; L IU Nan²;LANG Zhi-hong³;L IU Yu-mei¹;ZHUANG Mu¹;ZHANG Yang-yong¹;ZHANG You-jun¹;HUANG Da-fang^2,3;FANG Zhi-yuan¹

(¹Institute of Vegetables and Flowers, Chinese Academ y of Agricultural Sciences, B eijing 100081, China;² State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing100193, China; ³Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China )

Received:2009-04-13 Revised:2009-06-29 Online:2009-08-25 Published:2009-08-25
Contact: HUANG Da-fang

摘要/Abstract

摘要： 以结球甘蓝(Brassica oleracea L. var. capitata L. ) 的下胚轴和具柄子叶为外植体, 通过根癌
农杆菌(Agrobacterium tum efaciens) 介导法将苏云金芽孢杆菌(Bacillus thuringiensis) 杀虫晶体蛋白基因cry1Ia8导入结球甘蓝中, 共获得38株卡那霉素抗性植株。PCR和Southern blot检测证实cry1Ia8基因已经成功导入并整合到甘蓝的基因组中; RT2PCR和Western blot检测表明, cry1Ia8基因在转录和翻译水平有一定的表达; 转化植株叶片离体饲喂敏感小菜蛾( Plutella xylostella) 和对Cry1Ac蛋白表现抗性的小菜蛾的试验表明, 该基因不仅对敏感小菜蛾有很强的抗性, 对抗性小菜蛾也具有较强的抗性。

关键词: 结球甘蓝, 小菜蛾, Bt基因, cry1Ia8, 转基因, 抗虫性

Abstract: In order to obtain transgenic cabbage lineswith insect2resistance, cry1Ia8 gene was transferred to cabbage (Brassica oleracea L. var. capitata L. ) with the transformation of hypocotyl segments and cotyledon with petiole via Agrobacterium tumefaciens. Thirty-eight transformants with kanamycin-resistance were obtained. PCR and Southern blot analysis showed that cry1Ia8 gene was integrated into the genome of cabbage. Moreover, RT-PCR and Western blot detection demonstrated that cry1Ia8 gene was exp ressed in
RNA and protein levels. The results of bioassay with the suscep tible and resistant diamondback moth (DBM,Plutella xylostella) displayed thatmost of the transgenic plantswere resistant to the larvae of suscep tible DBM and also to the insect strain which is resistant to Cry1Ac toxin.

Key words: cabbage, Plutella xylostella, Bt gene, cry1Ia8, transformation, resistance

中图分类号:

崔磊;杨丽梅;刘楠;郎志宏;刘玉梅;庄木;张扬勇;张友军;黄大昉;方智远. Bt cry1Ia8抗虫基因对结球甘蓝的转化及其表达[J]. 园艺学报, 2009, 36(8): 1161-1168.

CU ILei;YANG Li-mei;L IU Nan;LANG Zhi-hong;L IU Yu-mei;ZHUANG Mu;ZHANG Yang-yong;ZHANG You-jun;HUANG Da-fang;FANG Zhi-yuan. Transformation and Expressing of Bt Gene cry1Ia8 in Cabbage[J]. ACTA HORTICULTURAE SINICA, 2009, 36(8): 1161-1168.

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