张杰;刘娟;蒋超;南铁贵;康利平;周利;袁媛;黄璐琦;
ZHANG Jie;LIU Juan;JIANG Chao;NAN Tie-gui;KANG Li-ping;ZHOU Li;YUAN Yuan;HUANG Lu-qi;School of Pharmacy, Jiangsu University;National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences;

• 1:江苏大学药学院
• 2:中国中医科学院中药资源中心

摘要(Abstract)：

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Rg_1,Re,Rb_1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Rb_1含量上升,原人参三醇型人参皂苷Rg_1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 Rg_1, Re and Rb_1 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 Rb_1 in ginseng adventitious roots treated with different concentrations of MeJA increased, and the content of ginseng triol ginsenoside Rg_1 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.

关键词(KeyWords)： 人参;人参皂苷生物合成;转录因子;ERF
Panax ginseng;ginsenoside biosynthesis;transcriptional factor;ERF

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81974516,81703653,81891013);; 中央级公益性科研院所基本科研业务费专项(ZZ10-008,ZZ13-YQ-093);; 广西科技重大专项(桂科AA18242040)

作者(Author): 张杰;刘娟;蒋超;南铁贵;康利平;周利;袁媛;黄璐琦;
ZHANG Jie;LIU Juan;JIANG Chao;NAN Tie-gui;KANG Li-ping;ZHOU Li;YUAN Yuan;HUANG Lu-qi;School of Pharmacy, Jiangsu University;National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences;

Email:

DOI: 10.19540/j.cnki.cjcmm.20200329.101

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