4.6 Article

Expression of PnSS Promotes Squalene and Oleanolic Acid (OA) Accumulation in Aralia elata via Methyl Jasmonate (MeJA) Induction

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GENES
卷 14, 期 6, 页码 -

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MDPI
DOI: 10.3390/genes14061132

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Aralia elata; genetic transformation; heterologous expression; squalene; oleanolic acid; MeJA treatment

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MeJA treatment promoted the accumulation of squalene and OA in transgenic A. elata, with a significant increase in gene expression levels. The maximum content of both products reached 17.34 and 0.70 mg/g after three days of MeJA treatment, showing a significant enhancement compared to the untreated lines. These results suggest that MeJA can activate the biosynthesis pathways of squalene and OA, leading to an increase in yield.
Aralia elata is an important herb due to the abundance of pentacyclic triterpenoid saponins whose important precursors are squalene and OA. Here, we found that MeJA treatment promoted both precursors accumulation, especially the latter, in transgenic A. elata, overexpressing a squalene synthase gene from Panax notoginseng(PnSS). In this study, Rhizobium-mediated transformation was used to express the PnSS gene. Gene expression analysis and high-performance liquid chromatography (HPLC) were used to identify the effect of MeJA on squalene and OA accumulation. The PnSS gene was isolated and expressed in A. elata. Transgenic lines showed a very high expression of the PnSS gene and farnesyl diphosphate synthase gene (AeFPS) and a slightly higher squalene content than the wild-type, but endogenous squalene synthase (AeSS), squalene epoxidase (AeSE), and & beta;-amyrin synthase (Ae & beta;-AS) gene were decreased as well as OA content. Following one day of MeJA treatment, the expression levels of PeSS, AeSS, and AeSE genes increased significantly. On day 3, the maximum content of both products reached 17.34 and 0.70 mg & BULL;g(-1,) which increased 1.39- and 4.90-fold than in the same lines without treatment. Transgenic lines expressing PnSS gene had a limited capability to promote squalene and OA accumulation. MeJA strongly activated their biosynthesis pathways, leading to enhance yield.

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