| 半夏凝集素基因对小麦的遗传转化研究 |
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| 引用本文： | 李淑洁,张正英.半夏凝集素基因对小麦的遗传转化研究[J].麦类作物学报,2012,32(2):191-196. |
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| 作者姓名： | 李淑洁 张正英 |
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| 作者单位： | 甘肃省农业科学院生物技术研究所,甘肃兰州,730070 |
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| 基金项目： | 甘肃省科技攻关项目（GS035A4100103）；甘肃省农业生物技术应用与开发项目（GNSW200406）。 |
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| 摘 要： | 为了拓宽农杆菌介导的小麦遗传转化的受体基因型,同时改良小麦品种的抗蚜特性,以甘肃省主推春小麦品种陇春22和优良品系9614的幼胚为转基因受体材料,以预培养4d的幼胚愈伤组织为受体,以含有半夏凝集素基因的C58c1农杆菌菌株为供体,将重组质粒pBI pta导入小麦,建立了农杆菌介导的小麦遗传转化体系。经G418 25mg.L-1抗性筛选、PCR检测共获得转基因植株7株,其中9614的4株,陇春22的3株;采用荧光定量PCR分析得出转基因植株中单拷贝的有1株,2个拷贝的1株,3个拷贝和4个拷贝的各有2株,1株没有得到扩增。对整合有2个拷贝半夏凝集素基因的陇春22-20转基因小麦的T1代植株进行了PCR检测,其中有50%的植株扩增出目的基因,50%没有扩增出目的基因,初步说明T1代植株中外源基因得到了遗传。
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| 关 键 词： | 小麦(Triticumaestivum L.) 半夏凝集素基因 农杆菌介导 遗传转化 Real-timePCR分析 |
Transformation of pta Gene into Wheat Mediated by Agrobacterium Tumefaciens |
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| LI Shu jie,ZHANG Zheng ying.Transformation of pta Gene into Wheat Mediated by Agrobacterium Tumefaciens[J].Journal of Triticeae Crops,2012,32(2):191-196. |
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| Authors: | LI Shu jie ZHANG Zheng ying |
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| Institution: | (Institute of Biotechnology,Gansu Academy of Agricultural Science,Lanzhou,Gansu 730070,China) |
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| Abstract: | In this study,the genetic transformation system of wheat mediated by Agrobacterium tumefacien was set up by using immature embryos of spring wheat cultivar 9614 and Longchun 22 as explants.The immature embryos after 4-days preculture were transformed with pta gene using Agrobacterium tumefaciens C58c1.Seven transgenic plants(4 come from 9614 and 3 come from Longchun 22) were obtained after screening by G418 resistance and PCR analysis.Real-time PCR analysis showed that the transgenic wheat plant with 1 and 2 copies of pta gene was one for each and with 3 and 4 copies of pta gene was 2 for each,and there is no pta gene amplification in one transgenic plant of 9614-17.50% T1 progeny of transgenic wheat longchun22-20 was integrated with exogenous pta gene by PCR analysis,and the other 50% wasn’t.This was the preliminary evidence that pta gene had been inherited to the T1 progeny of transgenic wheat. |
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| Keywords: | Wheat(Triticum aestivum L ) pta gene Agrobacterium tumefaciens Genetic transformation Real-time PCR analysis |
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