烟草病程相关蛋白NtPR10基因克隆与表达分析

发布时间：2018-06-03 11:43

本文选题：烟草 + 病程相关蛋白　；参考：《中国烟草科学》2017年03期

【摘要】：为鉴定烟草病程相关蛋白PR10的生物学功能,从普通烟草G28中克隆得到了Nt PR10基因,基因全长483 bp,编码160个氨基酸。基因结构分析表明,该基因编码的蛋白包含病程相关蛋白家族Bet_v_I保守域,具有与核酸酶活性相关的"P-Loop"结构。利用TMHMM、Signal P、Prosite Scan等软件分析发现,Nt PR10不含跨膜区,无信号肽,有胞内定位特征。采用实时荧光定量PCR分别分析了TMV诱导下该基因在抗、感品种枯斑三生和G28中的表达模式。结果表明,在TMV诱导下,Nt PR10基因在感病品种G28中显著上调表达;而在抗病品种枯斑三生中,TMV侵染后6 h,Nt PR10基因显著上调表达,第12小时至第8天显著下调表达,而后至第16天逐渐升高至侵染前水平。综合以上结果,Nt PR10应答了TMV侵染过程,并且在抗、感品种中整体呈现出相反的表达趋势,暗示了Nt PR10在TMV侵染过程中具有重要功能,为深入分析Nt PR10的抗病生物学功能奠定了基础。
[Abstract]:In order to identify the biological function of tobacco pathogenesis-related protein (PR10), the Nt PR10 gene was cloned from common tobacco G28, with a length of 483bpand encoding 160 amino acids. The gene structure analysis showed that the protein encoded by the gene contained the conserved domain of the pathogenesis-related protein family (Bet_v_I) and had a "P-Loop" structure related to the activity of nuclease. It was found that the NT PR10 had no transmembrane region, no signal peptide and intracellular localization characteristics by using TMHMMMM-Signal Pnprosite Scan and other softwares. The results showed that Nt PR10 had no transmembrane region, no signal peptide, and had intracellular localization characteristics. Real-time fluorescence quantitative PCR was used to analyze the expression patterns of the gene induced by TMV in the resistant and susceptible cultivars, Kubansansheng and G28, respectively. The results showed that the expression of Nt PR10 gene was significantly up-regulated in susceptible variety G28 induced by TMV, but significantly up-regulated at 6 h after infection, and significantly down-regulated from 12th hour to 8th day. Then on the 16th day, it gradually increased to the pre-infection level. Combined with the above results, Nt PR10 responded to the process of TMV infection, and showed a contrary expression trend in resistant and susceptible cultivars, suggesting that NT PR10 has an important function in the process of TMV infection. It lays a foundation for further analysis of the biological function of NT PR10 in disease resistance.
【作者单位】：中国农业科学院烟草研究所;中国农业科学院研究生院;
【基金】：国家自然科学基金项目“烟草病程相关蛋白PR10抗病分子机理研究”(31240013) 中国农业科学院烟草研究所青年科学基金项目“对烟草赤星病不同抗性品种蛋白质组差异分析及抗性蛋白挖掘”(2016B04) 中国烟草总公司四川省公司科技项目“优质适产烤烟新品种选育与推广”(SCYC201501)
【分类号】：S435.72

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