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作物学报 ›› 2012, Vol. 38 ›› Issue (12): 2178-2184.doi: 10.3724/SP.J.1006.2012.02178

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

兼抗全蚀病和白粉病小麦新种质的创制与鉴定

祝秀亮1，李钊1，杜丽璞1，徐惠君1，杨丽华1,2，庄洪涛1，马翎健2，张增艳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 / 农业部作物遗传育种重点开放实验室, 北京 100081; 2西北农林科技大学农学院, 陕西杨凌 712100
  • 收稿日期:2012-04-25 修回日期:2012-07-05 出版日期:2012-12-12 网络出版日期:2012-09-10
  • 通讯作者: 张增艳, E-mail: zhangzy@mail.caas.net.cn, Tel: 010-82108781
  • 基金资助:

    本研究由国家转基因生物新品种培育重大科技专项(2011ZX08002-001和2009ZX08002-006B)资助。

Development and Characterization of Wheat Lines with Resistance to Take-All and Powdery Mildew Diseases

ZHU Xiu-Liang1,LI Zhao1,DU Li-Pu1,XU Hui-Jun1,YANG Li-Hua1,2,ZHUANG Hong-Tao1,MA Ling-Jian2,ZHANG Zeng-Yan1,*   

  1. 1 National Key Facility for Crop Gene Resources and Genetic Improvement / Key Laboratory of Crop Genetic and Breeding of Agriculture Ministry / Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 2 College of Agronomy, Northwest A&F University, Yangling 712100, China
  • Received:2012-04-25 Revised:2012-07-05 Published:2012-12-12 Published online:2012-09-10
  • Contact: 张增艳, E-mail: zhangzy@mail.caas.net.cn, Tel: 010-82108781

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摘要：

TaLTP5是从小麦中分离到的一个脂质转移蛋白编码基因。利用基因枪介导法将TaLTP5表达载体pA25-TaLTP5转入抗白粉病的小麦品种扬麦18 (含抗白粉病基因Pm21)中, 旨在选育兼抗全蚀病和白粉病的小麦新种质。对转基因小麦T0~T3代植株中引入TaLTP5基因进行分子检测和抗病性鉴定。PCR检测、Southern杂交分析结果表明, 外源TaLTP5基因已转入、整合到3个转基因小麦株系的基因组中, 并能稳定遗传; 荧光定量RT-PCR的分析以及全蚀病菌的接种与鉴定结果表明, 与受体小麦扬麦18相比, 这3个转基因小麦株系中TaLTP5表达量显著提高, 其对全蚀病的抗性也明显增强。对3个转基因株系的Pm21分子标记和白粉病抗性鉴定表明, 外源TaLTP5基因的导入没有影响受体小麦对白粉病抗性, 说明这些转基因株系为兼抗全蚀病和白粉病小麦新种质。

关键词: 小麦脂质转移蛋白, TaLTP5, 转基因小麦, 全蚀病, 白粉病, 抗性

Abstract:

“Take-all”, primarily caused by Gaeumannomyces graminis var. tritici (Ggt), and powdery mildew, mainly caused by Blumeria graminis f. sp. tritici (Bgt), are important diseases of wheat (Triticum aestivum L.) worldwide. The wheat cultivar Yangmai 18 carrying a powdery mildewresistance gene Pm21, shows broad-spectrum resistance to powdery mildew. We have isolated a lipid transfer protein gene TaLTP5 from wheat. To study the role of TaLTP5 in wheat defense responses to the major pathogens of take-all, we introduced this gene intowheat cultivar Yangmai 18via bombarding the particle containing the TaLTP5 expressing vector pA25-TaLTP5. The TaLTP5 transgenic wheat plants from T0 to T3 generations were subjected to PCR, Southern blot, RT-PCR, and Q-RT-PCR analyses. We also evaluated the disease resistances of these TaLTP5 transgenic plants against inoculating Ggt and Bgt. The PCR and Southern blotting results showed that the alien TaLTP5 was transferred and integrated into the genomes of three transgenic wheat lines, and inherited stably in the transgenic wheat lines. The RT-PCR and Q-RT-PCR results indicated that the introduced TaLTP5 was over-expressed in transgenic wheat lines, which showed significantly-enhanced resistance to take-all, suggesting that TaLTP5 gene is involved in defense response to Ggt infection. In addition, the resistance of transgenic lines to powdery mildew was not influenced bythe introduced gene TaLTP5. Thus, TaLTP5 transgenic wheat Yangmai 18 exhibits resistance to both “Take-all” and powdery mildew.

Key words: Wheat lipid transfer protein, TaLTP5, Transgenic wheat, Take-all, Powdery mildew, Resistance

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