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作物学报 ›› 2009, Vol. 35 ›› Issue (4): 640-646.doi: 10.3724/SP.J.1006.2009.00640

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

转Rs-AFP2基因小麦的分子分析及其纹枯病抗性

路妍12，张增艳1*，任丽娟3，刘宝业1，廖勇14，徐惠君1，杜丽璞1，马鸿翔3，任正隆4，井金学2*，辛志勇1   

  1. 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
  • 收稿日期:2008-10-16 修回日期:2008-12-04 出版日期:2009-04-12 网络出版日期:2009-02-13
  • 通讯作者: 张增艳
  • 基金资助:

    本研究由国家转基因生物新品种培育科技重大专项（2008ZX08002-001）资助。

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

LU Yan12,ZHANG Zeng-Yan1*,REN Li-Juan3,LIU Bao-Ye1,LIAO Yong14,XU Hui-Jun1,DU Li-Pu1,MA Hong-Xiang3,REN Zheng-Long4,JING Jin-Xue2*,XIN Zhi-Yong1   

  1. 1National Key Facility for Crop Gene Resources and Genetic Improvement/Institute of Crop Science, chinese Academy of Agricultural Sciences,Beijing 100081,China;2College of Plant Protection,Northwest A&F University,Yangling 712100,China;3Biotechnology Research Institute,Jiangsu Academy of Aaricultural Sciences,Nanjing 210014,China;4College of Agronomy,Sichuan Agricultural University,Ya'an 625000,China
  • Received:2008-10-16 Revised:2008-12-04 Published:2009-04-12 Published online:2009-02-13
  • Contact: ZHANG Zeng-Yan

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2063

被引次数

28

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。

关键词: 抗菌肽, Rs-AFP2, 转基因小麦, 基因表达, 纹枯病抗性

Abstract:

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 T4 generations 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.

Key words: Antimicrobial peptides, Rs-AFP2, Transgenic wheat, Gene expression, Resistance to Rhizoctonia cerealis

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