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Production of dammarenediol-II triterpene in a cell suspension culture of transgenic tobacco

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Abstract

Key message

Dammarenediol-II is biologically active tetracyclic triterpenoid, which is basic compound of ginsenoside saponin. Here, we established the dammarenediol-II production via a cell suspension culture of transgenic tobacco overexpressing PgDDS.

Abstract

Dammarenediol-II synthase catalyzes the cyclization of 2,3-oxidosqualene to dammarenediol-II, which is the basic triterpene skeleton in dammarene-type saponin (ginsenosides) in Panax ginseng. Dammarenediol-II is a useful candidate both for pharmacologically active triterpenes and as a defense compound in plants. Dammarenediol-II is present in the roots of P. ginseng in trace amounts because it is an intermediate product in triterpene biosynthesis. In this work, we established the production of dammarenediol-II via cell suspension culture of transgenic tobacco. The dammarenediol-II synthase gene (PgDDS) isolated from P. ginseng was introduced into the Nicotiana tobacum genome under the control of 35S promoter by Agrobacterium-mediated transformation. Accumulation of dammarenediol-II in transgenic tobacco plants occurred in an organ-specific manner (roots > stems > leaves > flower buds), and transgenic line 14 (T14) exhibited a high amount (157.8 μg g−1 DW) of dammarenediol-II in the roots. Dammarenediol-II production in transgenic tobacco plants resulted in reduced phytosterol (β-sitosterol, campesterol, and stigmasterol) contents. A cell suspension culture was established as a shake flask culture of a callus derived from root segments of transgenic (T14) plants. The amount of dammarenediol-II production in the cell suspension reached 573 μg g−1 dry weight after 3 weeks of culture, which is equivalent to a culture volume of 5.2 mg dammarenediol-II per liter. Conclusively, the production of dammarenediol-II in a cell suspension culture of transgenic tobacco can be applied to the large-scale production of this compound and utilized as a source of pharmacologically active medicinal materials.

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Abbreviations

HPT:

Hygromycin phosphotransferase

DDS:

Dammarenediol-II synthase

PgDDS:

Panax ginseng dammarenediol-II synthase

BA:

6-Benzylaminopurine

MS:

Murashige and Skoog

MeJA:

Methyl jasmonate

PCR:

Polymerase chain reaction

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Acknowledgments

This work was supported by the Rural Development Administration, Republic of Korea [Next-Generation BioGreen 21 Program (PJ009529)], and Kangwon National University.

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Authors and Affiliations

  1. Department of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chunchon, 200-701, Republic of Korea

    Jung-Yeon Han, Hong-Yan Wang & Yong-Eui Choi

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  1. Jung-Yeon Han
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  2. Hong-Yan Wang
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  3. Yong-Eui Choi
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Correspondence to Yong-Eui Choi.

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Communicated by H. Ebinuma.

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Han, JY., Wang, HY. & Choi, YE. Production of dammarenediol-II triterpene in a cell suspension culture of transgenic tobacco. Plant Cell Rep 33, 225–233 (2014). https://doi.org/10.1007/s00299-013-1523-1

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  • DOI: https://doi.org/10.1007/s00299-013-1523-1

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