作为植物转录因子家族最大的成员之一,WRKY蛋白在植物的生长发育、防御反应以及胁迫响应中都发挥着极为关键的作用.本研究以玉米自交系 B73 为材料,克隆了一个玉米 WRKY 转录因子 Zm-WRKY36,并对其进行生物信息学与功能分析.结果表明,ZmWRKY36 基因的开放阅读框为1 002 bp,编码333个氨基酸.ZmWRKY36 蛋白分子量为35.8 kDa,理论等电点为5.66,属于酸性蛋白;不稳定系数为67.16,为极不稳定蛋白;亲水指数为-0.526,为亲水性蛋白;具有 1 个WRKY结构域和 1 个C2 HC锌指结构基序,属于WRKY转录因子Ⅲ类亚家族成员;与高粱SbWRKY64、稷PmWRKY4 和狗尾草SvWRKY4 有着较近的亲缘关系.进一步分析发现,ZmWRKY36 蛋白定位于细胞核内,并且有着较强的转录激活活性.qRT-PCR结果显示,ZmWRKY36 基因在玉米根和叶中表达量较高,具有组织特异性;并且受到干旱胁迫和盐胁迫的显著诱导,表明转录因子ZmWRKY36 可能在玉米抵御外界干旱胁迫和盐胁迫的过程中发挥着重要作用.
As one of the largest families of transcription factors in the plant,the WRKY proteins play crucial roles in plant growth and development,defense regulation and stress response.In this research,we cloned a maize WRKY transcription factor gene ZmWRKY36,and conducted the bioinformatics and functional analysis.The results showed that the open reading frame of ZmWRKY36 gene was 1 002 bp,and it encoded 333 amino acids.The molecular weight of ZmWRKY36 was 35.8 kDa,and its isoelectric point was 5.66,so it belonged to an acidic protein;its instability coefficient was 67.16,indicating that it was a highly unstable pro-tein;its hydrophilicity index was-0.526,indicating that it was a hydrophilic protein;it contained one WRKY domain and one C2 HC zinc-finger motif,so it was a member of the subgroupⅢfamily of WRKY transcription factor;it showed close relationships with SbWRKY64,PmWRKY4 and SvWRKY4.Further analysis results showed that ZmWRKY36 protein was localized in the nucleus and had strong transcriptional activation activity.The results of qRT-PCR indicated that the expression of ZmWRKY36 gene was tissue-specific with higher level in root and leaves,and was significantly induced by drought stress and salt stress,which suggested that the transcription factor ZmWRKY36 might play a key role in the process of maize resisting environmental drought and salt stresses.