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  • 张增艳.转PvPGIP2基因小麦的获得与纹枯病抗性鉴定[J].植物遗传资源学报,2013,14(1):181-185.    [点击复制]
  • Zhang Zeng yan.Development and Characterization of PvPGIP2 Transgenic wheat Plants with enhanced resistance to Rhizoctonia cerealis[J].植物遗传资源学报,2013,14(1):181-185.   [点击复制]
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转PvPGIP2基因小麦的获得与纹枯病抗性鉴定
张增艳
0
(中国农科院作科所)
摘要:
多聚半乳糖醛酸酶抑制蛋白(PGIP)是植物产生的防卫蛋白,对植物病原真菌具有防御作用。本研究利用基因工程技术构建了PvPGIP2 基因的单子叶植物表达载体pA25- PvPGIP2,其受玉米泛素(ubiquitin)启动子的控制;采用基因枪法将pA25-PvPGIP2 转化小麦推广品种扬麦18 幼胚愈伤组织4000块,获得203 株再生植株,通过PCR检测出阳性植株65 株,转化率为1.625 %。对转PvPGIP2基因小麦T0-T2代植株,进行外源基因的PCR、和 RT-PCR、荧光定量RT-PCR(Q-RT-PCR)分析和小麦纹枯病菌接种及其抗病性鉴定。结果表明,转入的PvPGIP2能够在转基因小麦中遗传、转录与表达。PvPGIP2基因的表达提高了转基因植株对小麦纹枯病的抗性。
关键词:  PvPGIP2基因  转基因小麦  小麦纹枯病菌  抗性
DOI:
投稿时间:2012-03-26修订日期:2012-11-15
基金项目:国家转基因生物新品种培育科技重大专项(2011ZX08002-001,2009ZX08002-006B)
Development and Characterization of PvPGIP2 Transgenic wheat Plants with enhanced resistance to Rhizoctonia cerealis
Abstract:
Polygalacturcuase-inhibing proteins (PGIPs) are defense proteins produced in plants, which can inhibit the growth of pathogenic fungi. In this study, pA25-PvPGIP2, the transformation vector of a bean PGIP gene PvPGIP2, which can be highly expressed in monocot plants, was correctly constructed. In the pA25-PvPGIP2 vector, PvPGIP2 gene was driven by maize ubiquitin promoter. 4000 of embryo callus of Yangmai 18 were bombarded by the particle containing pA25- PvPGIP2. Among the regenerated 203 plants, 65 positive transgenic individuals were identified through PCR assay in the T0 generation with a transformation frequency of 1.625 %. The transgenic wheat plants from T0 to T2 generations were subjected to PCR, RT-PCR, and Q-RT-PCR, and inoculations with Rhizoctonia cerealis as well as the disease resistance tests. Results showed that the alien PvPGIP2 gene was introduced into these transgenic wheat plants, can be inherited from T0 to T2, and expressed in the wheat background. The transgenic wheat plants expressing PvPGIP2 showed enhanced resistance to R. cerealis compared with untransformed Yangmai 18.
Key words:  PvPGIP2 gene  Transgenic wheat  Rhizoctonia cerealis  Resistance

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