【摘要】 目的:植物生长发育由多种转录因子调控,探索WRKY转录因子调控铁皮石斛组织生长发育机理。方法:根据铁皮石斛原球茎转录组数据中的WRKY6序列设计引物,并利用RACE技术克隆该基因;用生信分析软件预测其蛋白结构,并进行系统发育分析;利用实时荧光定量PCR（q RT-PCR）技术研究该基因在铁皮石斛幼苗根、茎、叶等组织中的表达。结果:通过RACE技术得到1个1,238bp的Ⅱ类WRKY转录因子基因,命名为DoWRKY6,其ORF长1,017 bp,编码339个氨基酸残基,与梅[PrmuWRKY27（XM₀08236601.1）]及大豆[GlmaWRKY27（XM₀03555347.3）]有较近的亲缘关系,且DoWRKY6在茎中的相对表达量最大。结论:我们预测在铁皮石斛的生长发育过程中DoWRKY6基因对根、叶、茎的调控作用依次增强,表明DoWRKY6与已发表的铁皮石斛WRKY基因在组织的差异性表达上有较大差别,这对丰富WRKY家族对铁皮石斛的生长发育调控机制提供更多依据。更多还原

【Abstract】 Objective: Many transcription factors regulate the growth and development of plant, it is very important to explore WRKY transcription regulation function about the tissue of Dendrobium officinale. Methods: Primers were designed according to the transcriptomic WRKY6 unigene sequence of protocorm in D. officinale, then I cloned DoWRKY6 transcription factors by RACE techniques. Through bioinformatics analysis and phylogenesis analysis we predicted its protein structure, and got its affinity. I analyzed the relative expression of D.officinale seedling’s root, stem and leaf through Roche Light circle 96 real-time fluorescent quantitative PCR（q RT-PCR） experiment. Results: By RACE technique, I obtain a new 1,238 bp long D. officinale Ⅱ kind of WRKY transcription factors named DoWRKY6, its ORF（Open reading frame） is 1,017 bp encoding 339 amino acids. The DoWRKY6 has the close relatives with Prunus mume [Prmu WRKY27（XM₀08236601.1）] and Glycine max [Glma WRKY27（XM₀03555347.3）]. The DoWRKY6 expression in stem is the biggest. Conclusions: We predicted that DoWRKY6 in D. officinale’s growth and development regulate the root, leaf and stem enhanced in turn, and DoWRKY6 had great different expression on the different organizations with the published D.officinale WRKY, which enriched WRKY family to provide more evidence on D. officinale growth regulatory mechanism.更多还原