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Identification of the protopanaxatriol synthase gene CYP6H for ginsenoside biosynthesis in Panax quinquefolius

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Abstract

Panax quinquefolius is one of perennial herbs and well known for its outstanding pharmacological activity. Ginsenosides are thought to be the main active ingredients in P. quinquefolius and exist in many kinds of plant genus Panax (ginseng). Protopanaxatriol synthase, which is considered cytochrome P450 (CYP450) in ginsenoside biosynthesis pathway can convert protopanaxadiol into protopanaxatriol. However, the protopanaxatriol synthase gene in P. quinquefolius has not been identified. Here, we cloned and identified a protopanaxatriol synthase gene from P. quinquefolius (CYP6H, GenBank accession no. KC190491) at the first time, reverse transcription-PCR (RT-PCR) analysis showed no obvious transcription change of CYP6H in methyl jasmonate (MeJA)-induced hairy roots. Ectopic expression of CYP6H in Saccharomyces cerevisiae resulted in the production of protopanaxatriol with added exogenous protopanaxadiol and confirmed by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC/APCIMS). Moreover, high-performance liquid chromatography (HPLC) analysis shows that RNA interferences of CYP6H in transgenic hairy roots could increase the accumulation of protopanaxadiol-type ginsenosides and decrease the accumulation of protopanaxatriol-type ginsenosides, whereas the effect of overexpression CYP6H in transgenic hairy roots was contrary. Our study indicated that CYP6H is a gene encoding protopanaxadiol 6-hydroxylase which could convert protopanaxadiol into protopanaxatriol in P. quinquefolius ginsenoside biosynthesis, we also have confirmed the function of CYP6H on effect accumulation of ginsenosides.

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Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (863) no. 2013AA102604, projects of the National Science Foundation of China no. 30970259 and 31270337, research fund for the Doctoral Program of Higher Education of China no. 20120061110038, and Scientific and Technological Development Plan Project of Jilin Province no. 20130102041JC.

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Authors and Affiliations

  1. School of Biological and Agricultural Engineering, Jilin University, No. 5988 Renmin Street Nanguan District, Changchun, People’s Republic of China

    Le Wang, Shou-Jing Zhao, Yan-Long Liang & Hao-Jie Cao

  2. The School of Biological and Agricultural Engineering, Changchun University of Technology, No. 2055 Yanan Street, Changchun, People’s Republic of China

    Yao Sun

  3. Jilin Business and Technology College, Changchun, People’s Republic of China

    Ying Han

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  1. Le Wang
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  2. Shou-Jing Zhao
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  3. Yan-Long Liang
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  5. Hao-Jie Cao
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Correspondence to Shou-Jing Zhao.

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Wang, L., Zhao, SJ., Liang, YL. et al. Identification of the protopanaxatriol synthase gene CYP6H for ginsenoside biosynthesis in Panax quinquefolius . Funct Integr Genomics 14, 559–570 (2014). https://doi.org/10.1007/s10142-014-0386-z

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  • DOI: https://doi.org/10.1007/s10142-014-0386-z

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