Abstract

Ginsenosides, the valuable pharmaceutical compounds in Panax ginseng, are triterpene saponins that occur mainly in ginseng plants. It was shown that in vitro treatment with the phytohormone jasmonic acid (JA) is able to increase ginsenoside production in ginseng plants. To understand the molecular link between JA biosynthesis and ginsenoside biosynthesis, we identified a JA biosynthetic 13-lipoxygenase gene (PgLOX6) in P. ginseng that promotes ginsenoside production. The expression of PgLOX6 was high in vascular bundles, which corresponds with expression of ginsenoside biosynthetic genes. Consistent with the role of PgLOX6 in synthesizing JA and promoting ginsenoside synthesis, transgenic plants overexpressing PgLOX6 in Arabidopsis had increased amounts of JA and methyl jasmonate (MJ), increased expression of triterpene biosynthetic genes such as squalene synthase (AtSS1) and squalene epoxidase (AtSE1), and increased squalene content. Moreover, transgenic ginseng roots overexpressing PgLOX6 had around 1.4-fold increased ginsenoside content and upregulation of ginsenoside biosynthesis-related genes including PgSS1, PgSE1, and dammarenediol synthase (PgDDS), which is similar to that of treatment with MJ. However, MJ treatment of transgenic ginseng significantly enhanced JA and MJ, associated with a 2.8-fold increase of ginsenoside content compared with the non-treated, non-transgenic control plant, which was 1.4 times higher than the MJ treatment effect on non-transgenic plants. These results demonstrate that PgLOX6 is responsible for the biosynthesis of JA and promotion of the production of triterpenoid saponin through up-regulating the expression of ginsenoside biosynthetic genes. This work provides insight into the role of JA in biosynthesizing secondary metabolites and provides a molecular tool for increasing ginsenoside production.

Highlights

  • Panax belongs to the family Araliaceae and contains at least as a healing drug and health tonic in China, Japan, and other 17 species (Kim et al, 2015)

  • These results suggest that jasmonic acid (JA) is able to promote the transcription of ginsenoside-synthesizing genes, leading to an increase of JA biosynthesis in ginseng plants

  • Understanding the mechanism of JA synthesis and how to increase ginsenoside accumulation will be helpful for commercial production of ginsenosides

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Summary

Introduction

Panax belongs to the family Araliaceae and contains at least as a healing drug and health tonic in China, Japan, and other 17 species (Kim et al, 2015). There are more than 150 naturally occurring ginsenosides in Panax species, which are classified according to their structure into two types, protopanaxadiols (PPDs) such as ginsenoside Rb1, Rb2, Rb3, Rc, and Rd, and protopanaxatriols (PPTs) such as ginsenoside Re, Rf, Rg1, Rg2, Rh1, and F1. Triterpene ginsenosides are mostly biosynthesized through the mevalonate (MVA) pathway in the cytosol. (S)-2,3-oxidosqualene undergoes additional cyclization, hydroxylation, and glycosylation by the enzymes dammarenediol synthase (DDS), cytochrome P450 and glycosyltransferase (GT), respectively (Kim et al, 2015; Rahimi et al, 2015a)

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