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作物学报 ›› 2014, Vol. 40 ›› Issue (03): 550-555.doi: 10.3724/SP.J.1006.2014.00550

• 研究简报 • 上一篇    下一篇

HC-Pro基因片段介导的高抗TuMV

叶艳英1.2,曾钢1,曹鸣庆1,马荣才1,吴才君2,*,姚磊1,*   

  1. 1 北京市农林科学院 / 农业生物技术研究中心,北京 100097; 2 江西农业大学农学院, 江西南昌 330045
  • 收稿日期:2013-07-16 修回日期:2013-12-10 出版日期:2014-03-12 网络出版日期:2014-01-16
  • 通讯作者: 姚磊, E-mail: yaolei@baafs.net.cn; 吴才君, E-mail: wucj12@126.com
  • 基金资助:

    本研究由国家高技术研究发展计划(863计划)项目(2008AA10Z154)和北京市自然科学基金项目(6092010)资助。

HC-Pro Gene Segment Mediated Hyper-Resistance to Turnip mosaic virus

YE Yan-Ying1.2,ZENG Gang1,CAO Ming-Qing1,MA Rong-Cai1,WU Cai-Jun2,*,YAO Lei1,*   

  1. 1 Beijing Ago-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China; 2 College of Agronomy, Jiangxi Agricultural University, Nanchang 330045, China
  • Received:2013-07-16 Revised:2013-12-10 Published:2014-03-12 Published online:2014-01-16
  • Contact: 姚磊, E-mail: yaolei@baafs.net.cn; 吴才君, E-mail: wucj12@126.com

摘要:

芜菁花叶病毒(turnip mosaic viruses,TuMV)是侵染重要经济作物的主要病毒之一,寄主范围十分广泛,尤其是对十字花科作物的危害最为严重。为了获得对TuMV持久稳定的高度抗性,本研究以TuMV HC-Pro基因的453 bp保守序列为靶标,构建了植物表达RNAi载体pBBBTu-HC-Pro,并转化了TuMV的天然宿主拟南芥。对转基因拟南芥用北京地区流行的TuMV强致病株BJ-C4进行了抗病接种鉴定。鉴定的13个单拷贝转基因株系中有4个对TuMVBJ-C4株表现出高度抗性,抗性比对照提高约80%以上,且抗性可以稳定遗传。经半定量和荧光定量PCR方法检测,在高抗转基因植株体内几乎检测不到病毒的累积,抗病效果明显。该载体在利用基因工程抗TuMV育种中具有广阔的应用前景。

关键词: 芜菁花叶病毒, HC-Pro, 基因片段, RNAi, 抗性

Abstract:

 TuMV (turnip mosaic virus) is one of the most major viruses in important economic crops, with broadest known hosts, especially for cruciferous plants. In order to get durable and stable high resistance to TuMV, in this study a 453 bp segment of TuMV HC-Pro gene was chosen as the target to construct pBBBTu-HC-Pro, and transformed into Arabidopsis thaliana, the TuMV nature host. Transgenic plants were inoculated with TuMV strain BJ-C4, which has high pathogenicity and is mainly epidemic in Beijing area. In 13 investigated transgenic lines, four lines showed hyper-resistance to TuMV. The resistance was enhanced 80%, and inherited to progenies. Semi-quantitative RT-PCR and quantitative PCR results revealed that the accumulation of virus was hardly to detect in highly resistant transgenic plants, indicating the resistance is prominent. The plasmid pBBBTu-HC-Pro should have broad applicability in crop breeding of TuMV resistance engineering.

Key words: Turnip mosaic virus, HC-Pro, Gene segment, RNAi, Resistance

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