Abstract
We report the high production of stilbenes, including resveratrol and viniferin, in grapevine (Vitis labruscana L.) cell cultures through elicitation with methyl jasmonate (MeJA) and stevioside (STE). Methyl-β-cyclodextrin (MeβCD) is widely used as a solubilizer for resveratrol production. For the first time, we used STE as a solubilizer for stilbene production in plant cell cultures. MeJA was most effective elicitor in activating VvSTS expression and stimulating stilbene biosynthesis in grapevine cell cultures. The maximum concentration of δ-viniferin (892.2 mg/L) production with a small amount of trans-resveratrol (12.2 mg/L) was observed in the culture medium by co-treatment of cells with MeJA and STE, whereas the highest level of trans-resveratrol (371.9 mg/L) with a slight amount of δ-viniferin (11.5 mg/L) was accumulated in the culture medium of cells treated with MeJA and MeβCD. However, neither trans-resveratrol nor δ-viniferin were significantly elevated within the cells by the applications. Notably, predominant production of δ-viniferin and trans-resveratrol was observed in shake and static flask culture medium, respectively, by co-treatment of MeJA and STE. Furthermore, stilbene compounds of resveratrol, ε-viniferin, and δ-viniferin were mainly produced in a 3-L bioreactor culture following elicitation of cells with MeJA and STE. These results provide new strategies for conditional, high-level production of resveratrol and viniferin in cell cultures by utilizing the solubilizing properties of STE or MeβCD.
Highlights
Plants synthesize a wide range of secondary metabolites in response to various environmental stresses (Langcake and Pryce 1977; Zamboni et al 2006)
Establishment and optimization of grapevine cell cultures The grapevine cell suspension cultures were established using calli derived from anthers and maintained by sub-culturing homogeneous cells in MS1D liquid medium several times at 2-week intervals
To obtain the best conditions for resveratrol production in grapevine cell suspension cultures, the expression patterns of the grapevine VvSTS genes (VvSTS6, VvSTS7, VvSTS36, and VvSTS42) were analyzed using specific primers by Reverse transcription-polymerase chain reaction (RT-PCR) and qRTPCR using flask culture samples treated with methyl jasmonate (MeJA), salicylic acid (SA), abscisic acid (ABA), and ET as elicitors (Fig. 2b; Additional file 1: Fig. S1)
Summary
Plants synthesize a wide range of secondary metabolites in response to various environmental stresses (Langcake and Pryce 1977; Zamboni et al 2006). Grapevines produce stilbenes derived from the phenylpropanoid pathway. (2020) 7:38 infection stimulates biosynthesis of δ- and ε-viniferin in grapevine leaves (Bavaresco et al 1997; Pezet et al 2003). In most plants producing stilbenes, STS genes exist as a family of closely related genes. Genome-wide analysis of the STS gene family based on the grapevine PN40024 genome revealed the identification of 48 putative STS genes, designated VvSTS1 to VvSTS48, with at least 33 full-length coding genes (Jaillon et al 2007; Vannozzi et al 2012). Expression of VvSTS genes is differentially regulated by biotic and abiotic stresses, pathogen infection, mechanical wounding or hormones (Almagro et al 2014; Chialva et al 2018; Lijavetzky et al 2008; Vannozzi et al 2012)
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