Upregulation of ginsenoside and gene expression related to triterpene biosynthesis in ginseng hairy root cultures elicited by methyl jasmonate
Upregulation of ginsenoside and gene expression related to triterpene biosynthesis in ginseng hairy root cultures elicited by methyl jasmonate
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DOI：
10.1007/s11240-009-9535-9
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发表时间：
2009-07-01
影响因子：
3
通讯作者：
Cha, Seon Woo
中科院分区：
文献类型：
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作者：
Kim, Ok Tae;Bang, Kyong Hwan;Cha, Seon Woo
本研究分析了茉莉酸甲酯（methyl jasmonate， MJ）诱导人参毛状根中三萜皂苷含量的变化及相关应答基因的表达谱。MJ诱导毛状根培养体中角鲨烯合成酶（PgSS）、角鲨烯环氧化酶（PgSE）和达马烯二醇合成酶（PNA）基因的转录量较对照增加，而环蒿烯合成酶（PNX）基因的转录量略有下降。为了选择与三萜生物合成相关的细胞色素p450依赖性羟化酶或葡萄糖基转移酶的候选基因，在MJ激发后进行RT-PCR分析。与对照组相比，与细胞色素P450家族相关的5个基因的表达没有差异。对于葡萄糖基转移酶候选基因，EST id PG07020C06、PG07025D04和PG07029G02的表达上调。为了研究MJ激发对三萜皂苷生物合成的影响，采用高效液相色谱法测定了毛状根中原人参二醇皂苷（Rb组）和原人参三醇皂苷（Rg组）的含量。MJ诱导7 d后，两组人参皂苷含量均显著高于对照组。其中，原人参二醇型皂苷含量比对照提高了5.5 ~ 9.7倍，原人参三醇型皂苷含量比对照提高了1.85 ~ 3.82倍。MJ诱导后，人参毛状根中Rg1人参皂苷含量呈负相关。
In this study, methyl jasmonate (MJ)-induced changes of triterpene saponins in ginseng (Panax ginseng C.A. Meyer) hairy roots and expression profiling of relevant responsive genes were analyzed. The transcription of PgSS (squalene synthase), PgSE (squalene epoxidase), and PNA (dammarenediol synthase-II) genes in hairy root cultures elicited by MJ treatment increased as compared with the controls, whereas that of PNX (cycloartenol synthase) decreased slightly. In order to select candidate genes encoding for cytochrome P450-dependent hydroxylase or glucosyltransferase associated with triterpene biosynthesis, RT-PCR analysis was conducted following MJ elicitation. No differences were observed in any expression among the five genes associated with the cytochrome P450 family, when compared to that of control. For candidates of the glucosyltransferase gene,expression of EST IDs PG07020C06, PG07025D04, and PG07029G02 was upregulated. In an effort to assess the effects of MJ elicitation on the biosynthesis of triterpene saponin, protopanaxadiol saponin (Rb group) and protopanaxatriol saponin (Rg group) contents in hairy roots were evaluated by HPLC analysis. With 7 days of MJ elicitation, levels of all ginseonsides of the two-groups increased much higher than that observed in the control. In particular, protopanaxadiol-type saponin contents increased by 5.5-9.7 times that of the control, whereas protopanaxatriol-type saponin contents were increased by 1.85-3.82-fold. In the case of Rg1 ginsenoside after MJ elicitation, the content was affected negatively in ginseng hairy root cultures.