中国水稻科学

中国水稻科学 ›› 2011, Vol. 25 ›› Issue (6): 599-604.DOI: 10.3969/j.issn.10017216.2011.06.006

• 研究报告 • 上一篇    下一篇

sbk和sck双价抗虫转基因水稻的育成和鉴定

李聪1,#张启军2,#刘少奎1赖东1漆庆明2吕川根2,*   

  1. 1 南京农业大学 农学院, 江苏 南京 210095； 2 江苏省农业科学院 粮食作物研究所/江苏省优质水稻工程技术研究中心,  江苏 南京 210014；
  • 收稿日期:2010-12-08 修回日期:2011-05-17 出版日期:2011-11-10 发布日期:2011-11-10
  • 通讯作者: 吕川根2,*
  • 基金资助:

    江苏省自然科学基金资助项目（BK2008348）；国家转基因生物新品种培育科技重大专项（2008ZX08001004）

Breeding and Identification of InsectResistant Rice with Two Insecticidal Genes, sbk and sck

LI  Cong 1,# , ZHANG Qijun 2,# , LIU Shaokui 1, LAI  Dong 1, QI Qingming 2 , LU Chuangen 2,*   

  1. 1 College of  Agronomy, Nanjing Agricultural University, Nanjing 210095,  China; 2 Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Jiangsu High Quality Rice R ＆ D Center, Nanjing 210014,  China;
  • Received:2010-12-08 Revised:2011-05-17 Online:2011-11-10 Published:2011-11-10
  • Contact: LU Chuangen2,*

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摘要： 将包含2个抗虫基因sbk\[修饰后的Cry1A(c)\]和sck（修饰后的CpTI）的质粒载体pCDMARUBAHyg转入农杆菌EHA105中, 感染南粳45的愈伤组织, 得到再生植株。用sbk和sck基因的引物进行PCR分析, 从97个再生植株中筛选出42个含有2个抗虫基因而没有潮霉素基因的阳性植株。通过Southern杂交发现, 42个阳性植株中具有1~5个外源基因拷贝的分别有23、 11、 5、2和1个单株。用RTPCR检测发现, 4个单拷贝株衍生的28个T3代植株外源基因能正常表达。人工饲喂二化螟幼虫试验也表明, 转基因植株的幼虫死亡率达94%~100%, 抗虫能力比对照显著提高。筛选出3个农艺性状优良的株系。

关键词: 水稻, sbk基因, sck基因, 抗虫, 转基因, 育种

Abstract: The plasmid of pCDMARUBAHyg, which contained two insectresistant genes, sbk\[modified from Cry1A(c)\]  and sck(modified from CpTI), was transformed into an Agrobacterium EHA105 for infection of the calli of   a super japonica rice   Nanjing 45. Plants regenerated from the transformation were subjected to all kinds of gene verifications and selection. Using PCR detection with the primers of sbk and  sck genes, 42 positive transgenic plants that were markerfree and contained the two target genes were selected from 97 regenerated plants. Results of Southernblotting indicated that 23, 11, 5, 2 and 1 plants had one, two, three, four and five copies of the transformed genes, respectively.  Analyses of  RTPCR and  Bt gene testing paper showed that  28 T3 generation plants derived  from four   transgenic plants having a single copy  were insectresistant.  Feeding experiment with rice stem borer revealed  that the insect resistance was greatly increased  with nymph mortality ranging from 94% to 100%. In addition, among the transgenic plants, three T3 transgenic plants  had many desirable characteristics for production. The mechanism of insectresistance of Bt  gene and its application in rice transgenic research were also briefly discussed.

Key words: Oryza sativa, sbk gene, sck gene, insect resistance, transgene, breeding

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