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WRKY类转录因子是植物防御反应信号转导的重要组成部分,本研究为了探讨WRKY33基因的功能,以嫩叶为材料,采用RT-PCR和RACE技术相结合的方法,克隆得到WRKY33全长c DNA(命名为Ps WRKY33),对其进行生物信息学分析。以基因组DNA为模板,对该基因的ORF区域进行PCR扩增,获得WRKY33的g DNA全长。采用荧光定量PCR技术检测该基因在水杨酸处理下的表达模式。试验结果表明:Ps WRKY33 c DNA全长为2 113 bp,开放阅读框长为1 770 bp,编码589个氨基酸;Ps WRKY33蛋白含有两个WRKY保守结构域,与Pm WRKY33蛋白同源性最高。Ps WRKY33基因组全长为2 844 bp,存在3个内含子和4个外显子。荧光定量结果表明:Ps WRKY33在1 mmol/L水杨酸处理后,30 h表达量最高,呈先上升后下降的趋势。研究结果为深入研究WRKY33基因与水杨酸诱导植物抗病性的关系提供一定的参考价值。
In order to investigate the function of WRKY33 gene, WRKY class of transcription factors is an important part of plant defense response signal transduction. In this study, RT-PCR and RACE were used to synthesize WRKY33 full-length cDNA (Named Ps WRKY33) for bioinformatics analysis. Using genomic DNA as a template, the ORF region of the gene was amplified by PCR and the full length of g DNA of WRKY33 was obtained. Fluorescent quantitative PCR was used to detect the expression pattern of the gene under salicylic acid treatment. The results showed that the full length of Ps WRKY33 cDNA was 2 113 bp with an open reading frame of 1 770 bp and encoded a protein of 589 amino acids. The Ps WRKY33 protein contained two WRKY conserved domains and had the highest homology with Pm WRKY33. The full-length genome of Ps WRKY33 is 2 844 bp in length with 3 introns and 4 exons. Fluorescence quantification results showed that the expression of Ps WRKY33 was the highest at 30 h after the treatment with 1 mmol / L salicylic acid, and then increased at first and then decreased. The results provide some reference value for further study on the relationship between WRKY33 gene and salicylic acid-induced plant disease resistance.