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作物学报 ›› 2016, Vol. 42 ›› Issue (10): 1429-1436.

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

抗根腐病的TaMYB86过表达转基因小麦的创制与分子功能鉴定

单天雷,洪彦涛,杜丽璞,徐惠君,魏学宁,张增艳*   

  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 / 农业部麦类生物学与遗传育种重点实验室, 北京 100081
  • 收稿日期:2016-03-10 修回日期:2016-07-11 出版日期:2016-10-12 网络出版日期:2016-08-01
  • 通讯作者: E-mail: zhangzengyan@caas.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专项(2016ZX08002-001-004)和国家自然科学基金项目(31471494)资助。

Development and Characterization of TaMYB86-Overexpressing Transgenic Wheat Lines with Resistance to Common Root Rot

SHAN Tian-Lei,HONG Yan-Tao,DU Li-Pu,XU Hui-Jun,WEI Xue-Ning,ZHANG Zeng-Yan*   

  1. National Key Facility for Crop Gene Resources and Genetic Improvement / Key Laboratory of Biology and Genetic Improvement of Triticeae Crops, Ministry of Agriculture / Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2016-03-10 Revised:2016-07-11 Published:2016-10-12 Published online:2016-08-01
  • Contact: E-mail: zhangzengyan@caas.cn, Tel: 010-82108781
  • Supported by:

    The study was supported by the National Major Project for Developing New GM Crops (2016ZX08002-001-004) and the National Natural Science Foundation of China (31471494).

摘要:

小麦根腐病是一种难以防治的小麦土传病害。TaMYB86是一个小麦中受根腐病菌诱导表达的MYB编码基因。本文构建了TaMYB86的过表达转基因载体pUbi:MYC-TaMYB86,利用基因枪介导法将其转入推广小麦品种扬麦16。对转TaMYB86基因小麦T0-T3代植株进行分子特征分析和抗病鉴定。PCR检测结果表明,外源TaMYB86已转入3个转基因小麦株系中; qRT-PCR结果显示,TaMYB86在3个转基因小麦株系中的表达量显著高于在未转基因扬麦16中,约为未转基因扬麦16中的5~6倍,表明TaMYB86可在转基因小麦中过量转录; Western杂交结果表明,引入的TaMYB86可在上述3个转基因小麦株系中翻译表达。对转TaMYB86基因小麦与未转基因扬麦16进行根腐病菌接种与抗病鉴定表明,3个转TaMYB86基因小麦株系在T1-T3代的根腐病病情指数分别为31.75、50.00、45.00; 37.75、37.50、38.50; 41.75、31.25、37.50; 在3次鉴定中未转基因扬麦16的根腐病病情指数分别为75.04、54.17、65.38,转TaMYB86基因小麦T1~T3代的根腐病抗性均显著高于未转基因扬麦16 (P < 0.01)。与未转基因扬麦16相比,转TaMYB86基因小麦中3个下游防卫基因(PR10、PR17c和Chit1)的转录水平也显著上调。以上结果说明,TaMYB86过表达可显著增强转基因小麦的根腐病抗性,在小麦防御根腐病过程中起正向调控作用。

关键词: 小麦, MYB转录因子, 防御反应, 小麦根腐病, 转基因

Abstract:

Wheat common root rot, mainly caused by Bipolaris sorokiniana, is a difficultly prevent soil-borne disease of wheat (Triticum aestivum L.) worldwide. TaMYB86, a B. sorokiniana -induced wheat MYB gene, encodes a MYB transcription factor. We constructed the TaMYB86 overexpression vector pUbi:MYC-TaMYB86 and introduced TaMYB86 into Yangmai 16 via the particle bombardment. The TaMYB86 transgenic wheat lines on generations of T0–T3 were underwent by molecular characteristics analysis and disease resistance evaluation. The PCR and quantitative RT-PCR results showed that the alien TaMYB86 was introduced into three transgenic wheat lines, and the relative transcriptional level of TaMYB86 was apparently higher in transgenic wheat lines than in non-transformed Yangmai 16. As Western blot results presented, the introduced MYC-TaMYB86 gene was translated into the MYC-TaMYB86 protein in the three overexpressing transgenic lines, but not in non-transformed Yangmai 16. The infection types and disease indexes of three TaMYB86 transgenic wheat lines were significantly lower than those of non-transformation Yangmai 16 (t-test, P < 0.01). The transcript levels of 3 wheat defense genes (PR10, PR17c, and Chit1) were significantly elevated in three transgenic wheat lines than in the non-transformed Yangmai 16. These results indicate that overexpression of TaMYB86 enhances significantly resistance to B. sorokiniana in transgenic wheat lines and TaMYB86 plays a positive role in defense response to B. sorokiniana.

Key words: Triticum aestivum, MYB transcription factor, Defense response, Common root rot, Transgene

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