转Rs-AFP2基因小麦的分子分析及其纹枯病抗性  

Molecular Analyses on Rs-AFP2 Transgenic Wheat Plants and Their Resistance to Rhizoctonia cerealis

作　　者：路妍[1,2] 张增艳[1] 任丽娟[3] 刘宝业[1] 廖勇[1,4] 徐惠君[1] 杜丽璞[1] 马鸿翔[3] 任正隆[4] 井金学[2] 辛志勇[1] 

机构地区：[1]中国农业科学院作物科学研究所／农作物基因资源与基因改良国家重大科学工程,北京100081 [2]西北农林科技大学植物保护学院,陕西杨凌712100 [3]江苏农业科学院生物技术研究所,江苏南京210014 [4]四川农业大学农学院,四川雅安625000

出　　处：《作物学报》2009年第4期640-646,共7页Acta Agronomica Sinica

基　　金：国家转基因生物新品种培育科技重大专项(2008ZX08002-001)资助

摘　　要：Rs-AFP2属于r-硫堇类抗菌肽,主要通过形成离子通道直接破坏细胞来杀灭病原菌。本研究通过基因枪介导法结合对目标基因的分子检测,证明已将外源Rs-AFP2基因转入小麦推广品种扬麦12中。通过逐株抗纹枯病接种鉴定、PCR、PCR-Southern blot、Southern blot和RT-PCR/荧光定量RT-PCR(Q-RT-PCR)分析,对转Rs-AFP2基因小麦T1至T4代植株跟踪检测。结果表明,Rs-AFP2在转基因小麦中能够稳定遗传,以单拷贝整合到小麦基因组中,遗传方式符合孟德尔遗传规律,并能在转录水平上表达。对转Rs-AFP2基因小麦的抗病性、主要农艺性状以及Rs-AFP2表达活性分析结果表明,与受体扬麦12相比,Rs-AFP2表达活性高的转基因小麦植株对纹枯病抗性有明显提高,其抗病性可以遗传,而主要农艺性状没有明显差异,证明可以利用Rs-AFP2基因和基因工程途径创制抗纹枯病小麦新种质。In this study, the gene encoding Rs-AFP2, a small cyteine-rich antifungal protein from radish, was evidenced to be transformed into a wheat （Triticum aestivum L.） cultivar Yangmai 12 via bombardment of biolistic particle and PCR detection. To evaluate if expression of Rs-AFP2 enhances the transgenic wheat resistance to Rhizoctonia cerealis, a major pathogen of wheat sharp eyespot, the transgenic wheat plants from T1 to T4generations were subjected to R. cerealis inoculation and the disease resistance rating, and PCR, PCR-Southern, Southern blotting, and RT-PCR/Q-RT-PCR analyses for the Rs-AFP2 transgene. Results showed that Rs-AFP2 gene was integrated as a single copy into the susceptible receptor wheat cultivar Yangmai 12, inherited from T1 to T4, and expressed in the wheat background. The transgenic wheat plants expressing Rs-AFP2 showed enhanced resistance to R. cerealis and unchanged agronomic traits compared with nontransgenic Yangmai 12. In the transgenic wheat plants, the express level of Rs-AFP2 was associated with the disease resistance degree. These results suggested that Rs-AFP2 gene can be useful for improving wheat resistance to R. cerealis.

关 键 词：抗菌肽 Rs-AFP2 转基因小麦 基因表达 纹枯病抗性 

分 类 号：S512.1[农业科学—作物学] TQ460.38[化学工程—制药化工]