小麦TaWRKY51基因的克隆及其参与SQ-1诱导小麦花粉败育过程的表达模式研究

Cloning of TaWRKY51 and Analysis of Its Expression Pattern during the Development of Pollen Abortion in Male Sterility Line Induced by SQ-1 in Wheat(Triticum aestivum)

为进一步深入解析化学杂交剂SQ-1诱导小麦花粉败育的机理,本研究从SQ-1诱导的生理型雄性不育系PHYMS-1376及其对照可育系CK-1376各时期花药转录组测序筛选出差异表达基因TraesCS1D02G362900.1,与小麦基因组数据库比对发现,该基因编码的氨基酸与TaWRKY51同源度为97%,说明该差异表达基因为TaWRKY51对其进行生物信息学分析、亚细胞定位、转录激活活性与花药表达模式研究,结果表明,TaWRKY51开放阅读框全长663 bp,可编码220个氨基酸,含有WRKY结构域和锌指结构,分子量约为23.34 kD,等电点为6.42;启动子区包含与光响应、茉莉酸甲酯响应、脱落酸响应等7类顺式作用元件,且TaWRKY51蛋白被定位在细胞核内,具有一定的转录激活活性;与CK-1376相比,PHYMS-1376植株花药中TaWRKY51的表达量在5个发育时期均高于CK-1376,且在四分体时期、单核晚期、二核期和三核期与CK-1376的差异均达极显著水平,表明该基因与PHYMS-1376花粉败育密切相关。

In order to elucidate the molecular mechanism of pollen abortion induced by the chemical hybrid agent SQ-1 in wheat,TraesCS1 D02 G362900.1 was screened out from the significantly different expression genes based on transcriptme sequencing of PHYMS-1376 with pollen abortion in male sterility induced by SQ-1 and its control fertile line CK-1376.Compared with wheat genome database,found that the similarity was 97%between the protein encoded by this gene and TaWRKY51,so the differentially expressed gene TraesCS1 D02 G362900 was TaWRKY51.TaWRKY51 was analysed using bioinformatics technology,transcriptional activation activity,sub-cellular localization,and the anther tissue expression pattern of TaWRKY51 was also performed.The results showed that TaWRKY51 containing an open reading frame with 663 bp and encoding 220 amino acids with a WRKY domain and zinc finger structure.Meanwhile,TaWRKY51 protein was a mono-mer protein with molecular weight of 23.34 kD and isoelectric point of 6.42.Cis-acting elements 7 categeries were found to be distributed in promoter regions,related to light responsiveness,MeJA-responsiveness,abscisic acid responsiveness and so on.TaWRKY51 was located in cell nucleus with certain transcriptional activation.Compared with CK-1376,the expression level of TaWRKY51 was higher at five development stages in anther of PHYMS-1376,with significant difference at tetrad stage,late uninucleate stage,binucleate stage and trinucleate stage,indicating that TaWRKY51 maybe closely associated with pollen abortion in anther of PHYMS-1376.