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作物学报 ›› 2010, Vol. 36 ›› Issue (06): 911-917.doi: 10.3724/SP.J.1006.2010.00911

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦TaPIM1基因的克隆及其转基因烟草的抗病性分析

周贤尧1,2,董 娜1,3,**,刘红霞1,张怀渝2,张增艳1,*   

  1. 1中国农业科学院作物科学研究所/农作物基因资源与教育改良国家重大科学
  • 收稿日期:2009-12-29 修回日期:2010-03-29 出版日期:2010-06-12 网络出版日期:2010-04-14
  • 通讯作者: 张增艳, E-mail: zhangzy@mail.caas.net.cn, Tel: 010-82108781
  • 基金资助:

    本研究由国家自然科学基金项目(30871523)和国家转基因植物研究与产业化专项(2008ZX08002-001)资助。

Cloning of Wheat TaPIM1 Gene and Analysis of Disease Resistance in TaPIM1 Transgenic Tobacco

ZHOU Xian-Yao1,2,DONG Na1,3**,LIU Gong-Xia1,ZHANG Fu-Yu2,ZHANG Ceng-Yan1*   

  1. 1National Key Facility for Crop Gene Resources and Genetic Improvement/Key Laboratory of Crop Genetic and Breeding,Ministry of Agriculture/Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 100081,China;2State Key Laboratory of Plant Breeding and Genetics,Sichuan Agricultural University,Ya'an Sichuan 625014,China;3Henan Institute of Science and Technology,Xinxiang 453003,China
  • Received:2009-12-29 Revised:2010-03-29 Published:2010-06-12 Published online:2010-04-14
  • Contact: ZHANG Zeng-yan,E-mail: zhangzy@mail.caas.net.cn, Tel: 010-82108781

摘要:

TaPIM1从小麦中克隆获得的1个病原诱导的小麦MYB基因,编码由323个氨基酸残基组成的蛋白TaPIM1TaPIM1具有R2R3MYB转录因子的典型结构,即2个保守的MYB DNA结合域(R2R3)、核定位位点和酸性激活区。TaPIM1的全长氨基酸序列与已克隆MYB蛋白的一致性仅为43.69%以下,为植物MYB转录因子家族R2R3亚群的一个新成员。小麦纹枯病菌(Rhizoctonia cerealis)、根腐病菌(Bipolaris sorokiniana)侵染可快速诱导抗病小麦中TaPIM1基因的上调表达,说明TaPIM1可能参与小麦对纹枯病菌、根腐病菌的防御反应。将TaPIM1基因构建到由组成型强启动子CaMV35S控制的双子叶转化载体pBI121通过农杆菌介导法将其转入烟草W38品系。通过卡那霉素抗性筛选和PCR检测,鉴定出转TaPIM1基因烟草T0代株系M66M102M11012个株系。对转基因烟草T1代株系进行PCRRT-PCR分析和青枯病菌抗性分析,结果表明,TaPIM1超量表达的转基因烟草株系M66M102M110对青枯病菌(Ralstonia solanacearum)的抗性显著高于未转基因烟草对照TaPIM1正向调控烟草对某些病原菌的防御反应。

关键词: 小麦, MYB转录因子, 转基因烟草, 青枯病抗性

Abstract:

In plants,MYB transcription factors play various roles in developmental processes and in responses to biotic and abiotic stresses. In this paper, a pathogen-induced MYB gene of wheat, TaPIM1, protein TaPIM1 consisting of 323 amino acids. TaPIM1 protein contains two MYB DNA binding domains (R2, R3), two nuclear localization sites and two transcription activation domains. The entire sequence of TaPIM1 protein shares a low identity of below 43.69% with other plant MYB proteins. TaPIM1 is a new member of R2R3 type of MYB transcription factors of transcription families. The transcript of TaPIM1 was obviously upregulated after challenge with fungal pathogens Rhizoctonia cerealis or Bipolaris sorokiniana, respectively. The gene transformation vector pBI-35S:TaPIM1 was constructed, in which the TaPIM1 was driven by the CaMV 35S promoter. The vector pBI-35S:TaPIM1 wastransferred into tobacco cultivar W38 by Agrobacterium-mediated transformation method. Twelve TaPIM1 transgenic tobacco lines, such as M66, M102 and M110, were obtained through screening. The T1 plants of these transgenic lines were subjected to PCR, RT-PCR analyses and evaluated for Ralstonia solanacearum resistance. The results showed that three transgenic lines, including M66, M102, and M110, overexpressed TaPIM1 and showed significantly enhanced resistance to R.  solanacearum compared with the host tobacco W38 plants.The resistance degree was correlated with the transcript level of the TaPIM1 inthese transgenic lines. These results suggested that TaPIM1 may positively modulate defense response to certain pathogens.was isolated from cDNAs of wheat leaves inoculated with Rhizoctonia cerealis. TaPIM1 gene encodes a

Key words: Wheat, MYB transcription factor, Transgenic tobacco, Resistance to Ralstonia solanacearum

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