低温胁迫对川续断中川续断皂苷Ⅵ积累的影响研究

Effect of Low Temperature Stress on the Accumulatim of Asperosaponin Ⅵ of Dipsacus asper

目的:探究低温胁迫对川续断皂苷Ⅵ积累的影响,为揭示海拔影响川续断质量形成的分子机制提供研究基础。方法:以二叶期川续断植株于15℃条件下进行低温处理,分别处理10、20、50 d后取样,通过HPLC检测川续断皂苷Ⅵ的含量,并以DaACT103为内参基因,实时荧光定量PCR法测定6个参与川续断皂苷Ⅵ合成的相关酶基因(DaAACT、DaMVD、DaHMGS、DaHMGR、DaSS、Daβ-AS)的相对表达量。结果:低温胁迫下川续断皂苷Ⅵ含量极显著升高,且各基因的相对表达量均较对照组极显著或显著升高;DaAACT、DaMVD、DaHMGR、DaSS、Daβ-AS基因的相对表达量与川续断皂苷Ⅵ含量达极显著正相关。结论:低温胁迫有利于川续断皂苷Ⅵ的积累,且DaAACT、DaMVD、DaHMGR、DaSS、Daβ-AS基因对川续断皂苷Ⅵ的积累作用显著。

Objective:To explore the effect of low temperature stress on the accumulation of asperosaponin Ⅵ,so as to provide research basis to reveal the formation law of altitude affecting the quality of Dipsacus asper.Methods:The low temperature treatment was performed by selecting second leaf of Dipsacus asper at 15℃,and harvested after 10,20 and 50 days treatment for further analysis.The content of asperosaponin Ⅵ was determined by HPLC.DaACT103 was used as a reference gene,the relative expression levels of six enzyme genes(DaAACT,DaMVD,DaHMGS,DaHMGR,DaSS,Daβ-AS)involved in the synthesis of asperosaponin Ⅵ im Dipsacus asper were determined by real-time fluorescence quantitative PCR.Results:The content of asperosaponin Ⅵ was highly significant increased under low temperature stress and the relative expression levels of each genes were highly significant or significant increased than control treatment.The correlation between the relative expression levels of DaAACT,DaMVD,DaHMGR,DaSS,Daβ-AS genes and the content of asperosaponin Ⅵ is highly significant positive.Conclusion:The low temperature stress is beneficial to the accumulation of asperosaponin Ⅵ.The accumulation of asperosaponin Ⅵ by DaAACT,DaMVD,DaHMGR,DaSS and Daβ-AS genes of the biosynthesis pathway is significant.