硅酸钠诱导的马铃薯抗黑痣病StWRKY11基因克隆及生物信息学分析

Cloning and Bioinformatics Analysis of Resistant Gene StWRKY11 Induced by Sodium Silicate in Potato by Rhizoctonia solani

为探讨硅酸钠增强马铃薯黑痣病抗性的分子机制,将硅酸钠诱导马铃薯转录组中表达量较高的基因StWRKY11进行克隆和生物信息学分析。从马铃薯中提取总RNA,并利用RT-PCR方法扩增该基因并克隆,通过生物信息学相关软件对其进行结构预测及分析。结果表明,从马铃薯大西洋中克隆了阅读框为1005 bp的StWRKY11基因,编码334个氨基酸,表达蛋白分子式为CHNOS,分子质量为81.86794 ku,理论等电点(pI)为5.09,原子总数为10500。表达蛋白含有一个典型的WRKYGQK保守结构域,锌指结构为CXCXHXH,属于第二类Ⅱd亚家族;表达的为疏水稳定性蛋白,无跨膜区和信号肽结构域,二级结构元件是α-螺旋、延伸链、β-折叠和无规卷曲,其中无规则卷曲比例最高,达61.68%,共有29个磷酸化位点,可能定位于细胞核内。启动子上游具有与抗性胁迫响应相关的顺式调控元件及与生长发育和激素响应顺式作用调控元件。该基因与马铃薯StWRKY5基因的亲缘关系较近,编码蛋白的氨基酸同源性达到95%。

In order to study the molecular mechanism of sodium silicate enhancing risistance of potato to Rhizoctonia solani,a gene StWRKY11 with high expression level in potato transcriptome induced by sodium silicate was cloned and its bioinformatics was analysed.Total RNA was extracted from potato,amplified by RT-PCR and cloned.Through bioinformatics related software,the structure prediction and prediction analysis were carried out.The results showed that StWRKY11 gene with a open reading frame of 1005 bp was cloned from potato Atlantic,encoding 334 amino acids.The molecular formula of the expressed protein was CHNOS,the molecular weight was 81.86794 ku,the theoretical isoelectric point(pI)was 5.09,and the total number of atoms was 10500.The expressed protein contained a typical WRKYGQK conserved domain,and the zinc finger structure was CXCXHXH,belonging to the secondⅡd subfamily.The secondary structural elements wereα-helix,extended chain,β-folding and random coiling,among which the proportion of random curl was the highest,up to 61.68%.There were 29 phosphorylation sites in total,which might be located in the nucleus.There were cis-regulatory elements upstream of the promoter that might related to resistance stress response and cis-regulatory elements related to growth and development and hormone response.The gene was closely related to potato StWRKY5 gene,and the amino acid homology of the coding protein reached 95%.