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WRKY转录因子在植物休眠等生长发育过程中具有重要作用。本研究基于芍药芽转录组数据,对芽休眠解除前后显著下调的编号为c22608.graph_c0的序列进行了q PCR验证分析,结果表明其表达模式与转录组一致,说明其与芍药芽休眠解除密切相关。为进一步挖掘和利用该基因,从芍药‘大富贵’芽中克隆了该基因,命名为Pl WRKY40(Gen Bank登录号:KU891820)。其c DNA全长1 347 bp,开放阅读框882 bp,编码293个氨基酸,含有1个典型的WRKY结构域;基因结构分析表明,Pl WRKY40具有4个外显子和3个内含子,其内含子具有特殊结构,可能调控该基因表达;以拟南芥WRKY蛋白的分类为参照,聚类分析结果表明,Pl WRKY40属于Group IIa亚族;系统进化树分析表明,Pl WRKY40与可可Tc WRKY40具有最高的同源性;亚细胞定位观测结果显示,Pl WRKY40转录因子定位在细胞核上;半定量PCR分析显示,Pl WRKY40在芍药的不同组织器官中泛表达,其中芽表达量最高。本研究为深入探索Pl WRKY40的生理学功能奠定了基础。
WRKY transcription factor plays an important role in the growth and development of plant dormancy. Based on the data of Paeonia lactiflora sp. Transcriptome, the sequence of c22608.graph_c0, which was significantly down-regulated before and after bud dormancy release, was verified by qPCR. The results showed that the expression pattern was consistent with that of transcriptome, indicating that it was closely related to the dormancy release of Paeonia lactiflora . In order to further explore and utilize this gene, the gene was cloned from the bud of Paeonia lactiflora and named P1 WRKY40 (Gen Bank accession number: KU891820). The cDNA of c WRKY40 was 1 347 bp in length and 882 bp in open reading frame. It encoded 293 amino acids and contained a typical WRKY domain. Gene structure analysis showed that there are 4 exons and 3 introns in Pl WRKY40 Introns have special structure and may regulate the expression of this gene. Based on the classification of Arabidopsis WRKY protein, cluster analysis showed that P1 WRKY40 belonged to the Group IIa subfamily. Phylogenetic tree analysis showed that Pl WRKY40 and cocoa Tc WRKY40 With the highest homology. The results of subcellular localization showed that Pl WRKY40 transcription factor was located in the nucleus. Semi-quantitative PCR analysis showed that Pl WRKY40 was expressed in different tissues and organs of Paeonia lactiflora with the highest expression of shoots. This study lays the foundation for further exploration of physiological function of Pl WRKY40.