马铃薯StWRKY转录因子的克隆和生物信息学分析

Cloning and bioinformatics analysis of potato WRKY transcription factor

【目的】探明马铃薯StWRKY转录因子的生物信息学特征及其在不同组织、低温和盐胁迫下的表达模式,揭示StWRKY转录因子在逆境下的调控规律,为进一步研究此转录因子的功能奠定基础。【方法】本试验以冀张薯12号马铃薯组培苗为材料,从马铃薯中克隆获得StWRKY基因,并对其进行生物信息学分析和功能的初步研究。【结果】StWRKY基因长度为747 bp,由172个氨基酸编码而成,相对分子量19921.58,理论等电点9.22,带负电残基总数21,带正电残基总数28,不稳定系数40.56。该蛋白质二级结构以无规曲线为主占40.1%,α-螺旋为37.4%、伸展链为15.1%、β-折叠为7.6%,是不稳定蛋白,属于亲水蛋白。通过进化树分析,与番茄同源性最高,为97.1%;利用RT-qPCR分析表明,表达谱分析中StWRKY基因的表达量在叶片中最高;功能初步分析,StWRKY基因的表达量在经过低温胁迫和盐胁迫处理6和7 h表达量显著增加,说明可能参与马铃薯对盐胁迫和低温的响应路径。【结论】StWRKY能够不同程度响应盐胁迫和低温胁迫,说明在马铃薯逆境反应机制中发挥不同作用,初步揭示了马铃薯StWRKY转录因子的功能,为进一步研究其功能提供依据。

[Objective]The present paper aimed to explore the bioinformatics characteristics of StWRKY transcription factor and its expression patterns in different tissues,low temperature and salt stress,and reveal the regulation mechanism of StWRKY transcription factor under stress,so as to provide a foundation for further study.[Method]In the present experiment,StWRKY gene was cloned from jizhangshu No.12,and its bioinformatics analysis and function were studied.[Result]The length of StWRKY gene was 747 bp,which was encoded 172 amino acids.Its molecular weight was 19921.58,theoretical isoelectric point was 9.22,total number of negatively charged residues were 21,total number of positively charged residues were 28,and instability coefficient was 40.56.The secondary structure of the protein was mainly random curve,accounting for 40.1%,ct-helix 37.4%,extended chain 15.1%,0-fold 7.6%.WRKY protein was unstable and belonged to hydrophilic protein.Phylogenetic tree analysis showed that StWRKY had the highest homology with tomato(97.1%);RT-qPCR analysis showed that the expression level of StWRKY gene was the highest in leaves;In the preliminary functional analysis,the expression level of StWRKY gene increased significantly after 6 and 7 hours of low temperature and salt stress treatment,which indicated that StWRKY gene might be related to salt and low temperature tolerance of potato.[Conclusion]StWRKY could respond to salt stress and low temperature stress in different levels,which indicated that it played different roles in the stress response mechanism of potato.These results preliminarily revealed the function of StWRKY transcription factor,which provided a basis for further study of its function.