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目的:研究尿苷二磷酸葡萄糖焦磷酸化酶(UGPase)在铁皮石斛不同组织中的表达情况,探讨其与石斛多糖累积的关系。方法:根据铁皮石斛原球茎转录组数据中的DoUGPase1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不同年份,不同组织中的表达。结果:通过RACE技术得到1个3054 bp基因,命名为DoUGPase1,其ORF长1530 bp,编码510个氨基酸残基,与油棕(Elaeis guineensis)和波斯枣(Phoenix dactylifera)有一定的同源性,且DoUGPase1在3年生的植株中相对表达量整体较高(P<0.05)。结论:我们预测在铁皮石斛的生长发育过程中DoUGPase1基因与多糖累积的活跃程度成正比例。
Objective: To study the expression of uridine diphosphate glucose pyrophosphorylase (UGPase) in different tissues of Dendrobium candidum and to explore its relationship with the accumulation of polysaccharides from Dendrobium. Methods: The primers were designed according to the DoUGPase1 sequence of protoplast transcriptome of D. officinale. The gene was cloned by RACE technique. The protein structure was predicted by bioinformatics software and phylogenetic analysis was carried out. Real-time quantitative PCR (q RT- PCR) technology to study the expression of this gene in different years and different tissues of Dendrobium candidum. Results: A 3054 bp gene was named as DoUGPase1 by RACE. The ORF was 1530 bp in length and encoded 510 amino acid residues. It had some homology with Elaeis guineensis and Phoenix dactylifera, The relative expression level of DoUGPase1 in 3-year-old plants was higher (P <0.05). Conclusion: We predict that the DoUGPase1 gene is directly proportional to the accumulation of polysaccharides during the growth and development of D. officinale.