【摘要】 为探讨多糖合成基因对铁皮石斛生长发育的调控作用及低温胁迫对多糖合成基因的影响,利用实时荧光定量PCR(qRT-PCR)技术对铁皮石斛多糖合成途径中UAM、UGP、RHM、SPS、CSLA和PGM基因在不同组织(根、茎、叶、花)及0℃胁迫24 h的表达水平进行定量分析。结果显示:GAPDH基因在5个候选基因中稳定性最高,可作为本研究的内参基因;7个多糖合成相关基因在铁皮石斛根、茎、叶、花中均有表达,且在花和根中的表达量高于成熟茎和叶;低温胁迫下,PGM基因呈持续下调表达,RHM和SPS基因呈先下调后上调至正常表达水平,UGP、UAM和CSLA基因则呈现先下调后上调再下调表达的波动。研究结果提示,GAPDH基因可作为内参基因用于铁皮石斛不同组织及低温胁迫基因表达水平研究中;7个与多糖合成相关的基因在铁皮石斛生长发育中也起到一定的调控作用;夜间0℃低温对铁皮石斛多糖合成相关基因的表达有显著的影响。更多还原

【Abstract】 The aim of this work was to estimate the regulation of polysaccharide synthesis genes on the growth and development and low temperature of Dendrobium officinale.In this study, real-time quantitative PCR(qRT-PCR) were used for determination of expression levels of UAM,UGP,RHM,SPS,CSLA and PGM genes in different tissues and 24 h treatment at 0 ℃ of D.officinale.The results showed that the GAPDH gene was used as the internal standard on account of its highest stability.The results showed that seven genes involved in polysaccharide synthesis were expressed in the roots, stems, leaves and flowers of D.officinale,and the expression levels in flowers and roots were higher than that in mature stems and leaves.Under low temperature stress, PGM gene was continuously down-regulated, RHM and SPS genes showed down-regulated firstly and then up-regulated to the normal expression level, while UGP,UAM and CSLA genes showed down-regulated first, and then up and down regulated alternatively.The results showed that GAPDH gene could be used as an internal standard gene in this study.Seven genes involved in polysaccharide synthesis play an important role in the growth and development of D.officinale.Low temperature treatment at 0 ℃ during night had significant effects on the expression of genes related to polysaccharide synthesis in D.officinale.更多还原