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农杆菌介导的半夏凝集素基因(Pinellia Ternate Agglutinin Gene,pta)对小麦的遗传转化及鉴定
引用本文:李淑洁,李静雯,张正英.农杆菌介导的半夏凝集素基因(Pinellia Ternate Agglutinin Gene,pta)对小麦的遗传转化及鉴定[J].中国生物工程杂志,2012,32(2):50-56.
作者姓名:李淑洁  李静雯  张正英
作者单位:甘肃省农业科学院生物技术研究所 兰州 730070
基金项目:甘肃省科技攻关项目(GS035-A41-001-03);甘肃省农业生物技术应用与开发项目(GNSW-2004-06)资助项目
摘    要:以甘肃主要推广春小麦品种陇春22幼胚为转基因受体材料,建立了农杆菌介导的小麦遗传转化体系。以预培养4天的幼胚愈伤组织为受体,C58c1农杆菌菌株为供体,将含有半夏凝集素基因的重组质粒pBIpta转入了小麦,经G418 25 mg/L抗性筛选、PCR检测和荧光定量PCR检测共获得转基因植株3株,外源基因的插入拷贝数分别为2、1、3。同时对转基因小麦的T1代植株进行了PCR检测和抗虫性分析,表明半夏凝集素基因在转基因植株的后代中得到了遗传并有一定的抗蚜虫作用。

关 键 词:半夏凝集素基因  陇春22(Triticum  aestivum  L.)  农杆菌介导法  Real-time  PCR  分析  
收稿时间:2011-11-15
修稿时间:2011-12-01

Transformation and Identification of pta Gene into Wheat Cultivar Longchun22 Mediated by Agrobacterium tumefaciens
LI Shu-jie , LI Jin-wen , ZHANG Zheng-ying.Transformation and Identification of pta Gene into Wheat Cultivar Longchun22 Mediated by Agrobacterium tumefaciens[J].China Biotechnology,2012,32(2):50-56.
Authors:LI Shu-jie  LI Jin-wen  ZHANG Zheng-ying
Institution:(Institute of Biotechnology,Gansu Academy of Agricultural Science,Lanzhou 730070,China)
Abstract:Agrobacterium tumefaciens-mediated wheat genetic transformation system was set up by using immature embryos of spring wheat cultivar Longchun 22 as explant.4-day-preculture immature embryos were transformed with pta gene using Agrobacterium tumefaciens C58c1.Three transgenic plants were gained after G418 resistant screening and PCR analysis.Real-time PCR analysis showed that pta gene copy number in transgenic wheat plant is 2,1,3,respectively.The exogenous pta gene have integrated into three plants out of six T1 progeny plants confirmed by PCR analysis,and the transgenic wheat showed good resistance to aphids in greenhouse.
Keywords:pta gene Longchun 22(Triticum aestivum L  ) Agrobacterium tumefaciens Real-time PCR analysis
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