人参转录因子ERF基因家族的表达分析

Expression analysis of transcription factor ERF gene family of Panax ginseng

ERF转录因子是植物最大的转录因子家族之一,对药用植物次生代谢具有重要的调控意义。该研究运用转录组热图方法从空间和时间2个层面分析了ERF家族基因的表达,筛选出6个可能参与人参皂苷调控的ERF基因。采用UPLC-MS/MS测定了不同浓度MeJA处理的人参不定根中人参皂苷Rg1,Re,Rb1的含量。运用实时荧光定量PCR测定MeJA处理的人参不定根中人参皂苷生物合成关键基因(DDS,CYP716A47,CYP716A53v2)和ERF家族基因的表达;采用Pearson相关对DDS,CYP716A47,CYP716A53v2基因与ERF家族基因表达特异性进行分析。结果表明,不同浓度MeJA处理的人参不定根中原人参二醇型人参皂苷Rb1含量上升,原人参三醇型人参皂苷Rg1和Re含量有所下降,与DDS,CYP716A47的表达量上升以及CYP716A53v2基因表达量下降相一致;ERF003,ERF118,ERF012的表达与CYP716A53v2呈显著正相关,而与DDS呈显著负相关,说明ERF003,ERF118,ERF012很可能通过抑制DDS表达和促进CYP716A53v2表达来调控人参皂苷合成;ERF1B表达则与CYP716A47呈显著负相关,说明ERF1B很可能抑制CYP716A47的表达从而参与调控人参皂苷合成。该文为后期深入研究ERF转录因子对于人参皂苷生物合成调控的功能验证奠定基础。

Ethylene responsive factor(ERF),one of the largest families of transcriptional factors in plants,plays a key role in se-condary metabolism of herbal plants.To analyze the expression of ERF family genes,the heat map clustering method was used by analyzing the ginseng transcriptomes of different parts and different growth years.The contents of ginsenosides Rg1,Re and Rb1 in various concentrations of MeJA-treated ginseng adventitious roots were determined by UPLC-MS/MS method.The expression of key genes of ginsenoside biosynthesis(DDS,CYP716A47,CYP716A53v2)and ERF family genes in MeJA-treated ginseng adventitious roots were determined by using real-time quantitative PCR.Pearson correlation was adopted to analyze the gene expression pattern of DDS,CYP716A47,CYP716A53v2 gene and ERF family.The results showed that the content of ginseng diol ginsenoside Rb1 in ginseng adventitious roots treated with different concentrations of MeJA increased,and the content of ginseng triol ginsenoside Rg1 and Re decreased.It is consistent with the increase of DDS and CYP716A47 expression and the decrease of CYP716A53v2 gene expression.The expression of ERF003,ERF118 and ERF012 genes was significantly positively correlated with CYP716A53v2,but negatively correlated with DDS.While the expression of ERF1B was significantly negatively correlated with CYP716A47.It is proved that ERF003,ERF118 and ERF012 were likely to inhibit the expression of DDS and promote the expression of CYP716A53v2,and ERF1B was likely to inhibit CYP716A47.This work could provide theoretical basis of ERF functional verification of regulating the biosynthesis of ginsenosides.