文章摘要
马柱文,屈玉娇,潘晓英,陈俊标,张振臣,李集勤,黄振瑞,袁清华.广东晒烟大叶密合受青枯病菌侵染后差异表达蛋白分析[J].广东农业科学,2021,48(12):83-90
查看全文    HTML 广东晒烟大叶密合受青枯病菌侵染后差异表达蛋白分析
Analysis on Differentially Expressed Proteins in Tobacco Cultivar Dayemihe after Inoculation with Ralstonia solanacearum
  
DOI:10.16768/j.issn.1004-874X.2021.12.010
中文关键词: 烟草  青枯病  蛋白质组  iTRAQ  烟草凝集素
英文关键词: tobacco  bacterial wilt  proteomics  iTRAQ  nictaba
基金项目:广东省自然科学基金(2016A030313781);广东省烟草专卖局(公司)重大专项(粤烟科项专201701);广东省烟草专卖局(公司)科技项目(粤烟科项 201702,粤烟科项 201908,粤烟科项 201911)
作者单位
马柱文,屈玉娇,潘晓英,陈俊标,张振臣,李集勤,黄振瑞,袁清华  
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中文摘要:
      【目的】大叶密合是广东封开县地方名优晒烟品种,是当前发现烟草青枯病的优良抗源,深入研 究其烟草青枯病抗性机理,寻找其青枯病抗性相关的差异表达蛋白,可为抗青枯病分子机理的深入研究和抗病 育种提供参考。【方法】将烟草品种大叶密合、抗病对照品种D101、感病对照品种长脖黄进行青枯病菌室内接 种鉴定,并利用蛋白质组学(iTRAQ)和实时荧光定量PCR(qRT-PCR)技术分析大叶密合接种前后基因的差 异表达情况。【结果】接种青枯病菌后,大叶密合较长脖黄发病减缓。大叶密合检测到35个接种前后的差异蛋 白,其中21个蛋白下调、14个蛋白上调,共有25个蛋白与植物胁迫相关。上调表达差异倍数最大的蛋白是烟草 凝集素家族(Nictaba)的Hav1。采用实时荧光定量PCR技术研究烟草凝集素基因在接种后12 h内的表达量变化, 发现其在接种后6 h和9 h表达量最高,随后开始下降。【结论】植物响应病菌入侵是一个复杂过程,鉴定到的差 异蛋白中上调幅度最大的是凝集素家族成员之一的Hav1,接种强致病力和无致病力菌株后烟草凝集素呈现类似 的表达模式,表明凝集素蛋白可能对病菌入侵后的上游反应发挥作用。
英文摘要:
      【Objective】Dayemihe, a local famous and excellent tobacco variety in Fengkai County of Guangdong Province, is an excellent source of resistance to bacterial wilt currently. In-depth study of the mechanism of tobacco bacterial wilt resistance and search for differentially expressed proteins related to bacterial wilt resistance can provide a reference for the in-depth study of the molecular mechanism of anti-bacterial wilt and disease resistance breeding. 【Method】With tobacco variety Dayemihe, resistant variety D101 and susceptible variety as materials, inoculation and identification of bacterial wilt were conducted indoor. Differentially expressed genes of Dayemihe before and after inoculation were analyzed by Isobaric Tags for Relative and Absolute Quantitation(iTRAQ)and Quantitative Real-time PCR(qRTPCR).【Result】After inoculation, the development of the bacteria wilt of Dayemihe was slower than that of the susceptible control. Thirty-five proteins were differently expressed in Dayemihe: twenty-one proteins were down-regulated and fourteen proteins were up-regulated. Among them, twenty-five proteins were related to plant stress. The protein with the up-regulated differential expression with largest fold change was Hav1 belonging to the Nicotiana tabacum Agglutinin(Nictaba)family. The nictaba within 12 hours after inoculation was detected by qRT-PCR. It was found that they had the highest expression level at 6 hours and 9 hours after inoculation, and then the level began to decrease.【Conclusion】Plant response to pathogen invasion is a complex process that requires proteins with different functions from different pathways. Plant response to pathogen invasion is a complex process. According to the identification, the protein with the up-regulated differential expression with largest fold change is Hav1, which belongs to the Nicotiana tabacum Agglutinin(Nictaba)family. Tobacco lectins showed similar expression patterns after inoculation with highly pathogenic and non-pathogenic strains, indicating that the lectin protein may play a role in the upstream reaction process after the invasion of pathogens.
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