Isolation and characterization of a novel PDR-type ABC transporter gene PgPDR3 from Panax ginseng C.A. Meyer induced by methyl jasmonate
Isolation and characterization of a novel PDR-type ABC transporter gene PgPDR3 from Panax ginseng C.A. Meyer induced by methyl jasmonate
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DOI：
10.1007/s11033-013-2731-z
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发表时间：
2013-09
影响因子：
2.8
通讯作者：
Ru Zhang;Jingjia Huang;Jie Zhu;Xiaolei Xie;Q. Tang;Xianghui Chen;Jun Luo;Zhiyong Luo
中科院分区：
文献类型：
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作者：
Ru Zhang;Jingjia Huang;Jie Zhu;Xiaolei Xie;Q. Tang;Xianghui Chen;Jun Luo;Zhiyong Luo
为了筛选人参皂苷生物合成积累的新基因，研究人参皂苷积累的分子机制，采用dna扩增片段长度多态性（cDNA-AFLP）方法对茉莉酸甲酯（MeJA）处理的人参悬浮细胞和未处理的人参悬浮细胞进行差异表达分析。共获得25个转录衍生片段（transcript-derived-fragments, TDFs），其中14个参与了次生代谢物的生物合成、转运、信号转导和应激反应。在meja处理的人参细胞中检测到PgTDF3(JK974248)，并分离其全长cDNA。pgpdr3基因（GenBank登录号KC013238）全长4515 bp，包含4137 bp的开放阅读框（ORF）。推断的PgPDR3氨基酸序列与其他植物多效性耐药（PDR）转运蛋白具有高度相似性，在N端和c端分别具有植物PDR转运蛋白Walker A、Walker B和ABC特征域。RT-PCR和定量实时PCR （qRT-PCR）分析表明，pgpdr3在根和不定根中的表达水平高于叶、种子和芽，且MeJA对pgpdr3的表达具有强烈的诱导作用，且具有时间依赖性，因为MeJA是介导次生代谢物生物合成和积累的信号分子。这些结果证明了pgpdr3基因在meja诱导人参次生代谢产物积累中的潜在作用。
In order to screen novel genes involved in the biosynthesis and accumulation of ginsenosides in ginseng for investigating the molecular mechanism of ginsenosides accumulation, the differential expression analysis was performed by cDNA-amplified fragment length polymorphism (cDNA-AFLP) in methyl jasmonate (MeJA) treated ginseng suspension cells and untreated controls. Twenty-five transcript-derived-fragments (TDFs) were obtained and 14 of them was involved in secondary metabolite biosynthesis, transportation, signal transduction and stress response.PgTDF3(JK974248) was detected in MeJA-treated ginseng cells and its full-length cDNA was subsequently isolated. The gene designatedPgPDR3(GenBank accession number KC013238) has 4515 bp in length containing a 4137 bp open reading frame (ORF). The deduced amino acid sequence of PgPDR3 shares high similarity to other plant pleiotropic drug resistance (PDR) transporters and have the characteristic domain of plant PDR transporter Walker A, Walker B and ABC signature both at the N- and C-terminal ends respectively. RT-PCR and quantitative real-time PCR (qRT-PCR) analysis indicated thatPgPDR3was expressed at a high level in the roots and adventitious roots compared to leaves, seeds and buds, and the expression was induced strongly by MeJA in a time-dependent manner, as it is well known MeJA is a signaling molecule that mediates the biosynthesis and accumulation of secondary metabolites. These results demonstrate a potential role of thePgPDR3gene in the accumulation of secondary metabolites in MeJA-induced ginseng.