Han Rong-Chun, Liu Lu-Lu, Liu Jun-Lin, Xie Dong-Mei, Peng Dai-Yin, Yu Nian-Jun
Institute of Traditional Chinese Medicine Resources Protection and Development,Anhui Academy of Chinese Medicine,Anhui University of Chinese Medicine Hefei 230012,China.
Zhongguo Zhong Yao Za Zhi. 2019 Apr;44(8):1552-1557. doi: 10.19540/j.cnki.cjcmm.20190129.013.
In order to understand the function of GDP-mannose pyrophosphorylase(GMPP) function and its regulation in polysaccharide biosynthesis mechanism in Dendrobium. D. huoshanense was used to clone GMPP gene. GMPP gene expression in D. huoshanense,D. officinale and D. moniliforme was also determined by qPCR. The results showed that the length of D. huoshanense GMPP gene c DNA sequence is 1 867 bp,containing 1 245 bp open reading frame(ORF),encoding 415 amino acids. Phylogenetic tree analysis showed that D. huoshanense,D. officinale and D. moniliforme are closely related with GMPP taken into consideration. Bioinformatics analysis demonstrated that GMPP sequence similarity among the three species reached as high as 99%. qPCR results indicated that GMPP genes was highly expressed in stem of D. huoshanense compared with its leaf,flower and root. According to GMPP gene expression profile in D. huoshanense,D. officinale and D. moniliforme grown in Huoshan area,it was clear that GMPP in D. huoshanense showed the highest expression level. Furthermore,our findings of GMPP gene expression profile will facilitate future researches into its polysaccharide biosynthetic mechanism.
为了解GDP-甘露糖焦磷酸化酶（GMPP）在铁皮石斛多糖生物合成机制中的功能及其调控作用，以霍山石斛为材料克隆GMPP基因，并通过实时荧光定量PCR（qPCR）检测霍山石斛、铁皮石斛和细茎石斛中GMPP基因的表达情况。结果显示，霍山石斛GMPP基因cDNA序列长度为1867bp，包含1245bp的开放阅读框（ORF），编码415个氨基酸。系统进化树分析表明，霍山石斛、铁皮石斛和细茎石斛在GMPP方面亲缘关系较近。生物信息学分析显示，这三个物种的GMPP序列相似度高达99%。qPCR结果表明，与叶、花和根相比，霍山石斛茎中GMPP基因表达量较高。根据霍山地区种植的霍山石斛、铁皮石斛和细茎石斛中GMPP基因表达谱可知，霍山石斛中GMPP表达水平最高。此外，我们对GMPP基因表达谱的研究结果将有助于未来对其多糖生物合成机制的研究。
