水曲柳FmWRKY44基因克隆及表达分析

Cloning and Expression Analysis of FmWRKY44 Gene in Fraxinus mandshurica

克隆水曲柳FmWRKY44基因,探究其在非生物胁迫和激素胁迫中的作用。利用水曲柳干旱转录组序列设计特异引物,克隆FmWRKY44基因的完整ORF序列,并对该序列及其编码产物进行生物信息学分析,采用qRT-PCR技术分析FmWRKY44表达模式。克隆了一个水曲柳WKRY基因,编码区长1383bp,编码460氨基酸。对其编码蛋白分析发现其为稳定亲水蛋白,亚细胞定位预测主要在细胞核,进行保守域及同源性分析分析,属于WRKYⅠ类家族,命名为FmWRKY44。qRT-PCR分析发现,FmWRKY44在种子中高度表达,并不同程度响应低温、高温、盐和干旱4种非生物胁迫。同时发现FmWRKY44与NAA、ABA、GA3、JA植物激素响应,水曲柳FmWRKY44基因积极响应低温、高温胁迫。

To clone the FmWRKY44 gene from Fraxinus mandshurica and explore its role in abiotic stress and hormone stress,specific primers were designed using the sequences of the variceal drought-transcriptome,the complete ORF sequence of FmWRKY44 gene was cloned,and bioinformatics analysis was carried out on the sequence and its coded products. The expression pattern of FmWRKY44 was analyzed using qRT-PCR. A WKRY gene of F.mandshurica was cloned,encoding 1 383 bp,encoding 460 amino acid. It was found to be a stable hydrophilic protein by the analysis of its encoded protein. The subcellular localization was mainly predicted in the nucleus,and the conserved domain and homology were analyzed. It belonged to the WRKY class I family and was named FmWRKY44 . By qRT-PCR analysis, FmWRKY44 was highly expressed in the seeds. Different degrees of response to low temperature,high temperature,salt and drought four abiotic stresses. FmWRKY44 and NAA,ABA,GA 3,JA phytohormone responses were also found. FmWRKY44 gene of F.mandshurica positively responds to low temperature and high temperature stress.