Induced extracellular production of stilbenes in grapevine cell culture medium by elicitation with methyl jasmonate and stevioside

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.

Keywords

Grapevine / Cell cultures / Stilbenes / Viniferin / Resveratrol / Stevioside

References

[1] Aggarwal BB, Bhardwaj A, Aggarwal RS, Seeram NP, Shishodia S, Takada Y. Role of resveratrol in prevention and therapy of cancer: preclinical and clinical studies. Anticancer Res, 2004, 24: 2783-2840.

[2] Almagro L, Carbonell-Bejerano P, Belchí-Navarro S, Bru R, Martínez-Zapater JM, Lijavetzky D, Pedreño MA. Dissecting the transcriptional response to elicitors in Vitis vinifera cells. PLoS ONE, 2014, 9: e109777.

[3] An CH, Lee KW, Lee SH, Jeong YJ, Woo SG, Chun H, Park Y-I, Kwak S-S, Kim CY. Heterologous expression of IbMYB1a by different promoters exhibits different patterns of anthocyanin accumulation in tobacco. Plant Physiol Biochem, 2015, 89: 1-10.

[4] Bavaresco L, Petegolli D, Cantù E, Fregoni M, Chiusa G, Trevisan M. Elicitation and accumulation of stilbene phytoalexins in grapevine berries infected by Botrytis cinerea. Vitis, 1997, 36: 77-83.

[5] Beekwilder J, Wolswinkel R, Jonker H, Hall R, de Vos CHR, Bovy A. Production of resveratrol in recombinant microorganisms. Appl Environ Microbiol, 2006, 72: 5670-5672.

[6] Belchí-Navarro S, Almagro L, Lijavetzky D, Bru R, Pedreño MA. Enhanced extracellular production of trans-resveratrol in Vitis vinifera suspension cultured cells by using cyclodextrins and methyljasmonate. Plant Cell Rep, 2012, 31: 81-89.

[7] Belhadj A, Telef N, Saigne C, Cluzet S, Barrieu F, Hamdi S, Mérillon J-M. Effect of methyl jasmonate in combination with carbohydrates on gene expression of PR proteins, stilbene and anthocyanin accumulation in grapevine cell cultures. Plant Physiol Biochem, 2008, 46: 493-499.

[8] Bhat KPL, Pezzuto JM. Cancer chemopreventive activity of resveratrol. Ann N Y Acad Sci, 2002, 957: 210-229.

[9] Bradamante S, Barenghi L, Villa A. Cardiovascular protective effects of resveratrol. Cardiovasc Drug Rev, 2004, 22: 169-188.

[10] Bru R, Sellés S, Casado-Vela J, Belchí-Navarro S, Pedreño MA. Modified cyclodextrins are chemically defined glucan inducers of defense responses in grapevine cell cultures. J Agric Food Chem, 2006, 54: 65-71.

[11] Calderon AA, Zapata JM, Ros Barcelo A. Peroxidase-mediated formation of resveratrol oxidation products during the hypersensitive-like reaction of grapevine cells to an elicitor from Trichoderma viride. Physiol Mol Plant Pathol, 1994, 44: 289-299.

[12] Chastang T, Pozzobon V, Taidi B, Courot E, Clément C, Pareau D. Resveratrol production by grapevine cells in fed-batch bioreactor: experiments and modelling. Biochem Eng J, 2018, 131: 9-16.

[13] Chialva C, Muñoz C, Miccono M, Eichler E, Calderón L, Prieto H, Lijavetzky D. Differential expression patterns within the grapevine stilbene synthase gene family revealed through their regulatory regions. Plant Mol Biol Rep, 2018, 36: 225-238.

[14] Delaunois B, Cordelier S, Conreux A, Clément C, Jeandet P. Molecular engineering of resveratrol in plants. Plant Biotechnol J, 2009, 7: 2-12.

[15] Donnez D, Jeandet P, Clément C, Courot E. Bioproduction of resveratrol and stilbene derivatives by plant cells and microorganisms. Trends Biotechnol, 2009, 27: 706-713.

[16] Donnez D, Kim K-H, Antoine S, Conreux A, De Luca V, Jeandet P, Clément C, Courot E. Bioproduction of resveratrol and viniferins by an elicited grapevine cell culture in a 2 L stirred bioreactor. Process Biochem, 2011, 46: 1056-1062.

[17] Giri CC, Zaheer M. Chemical elicitors versus secondary metabolite production in vitro using plant cell, tissue and organ cultures: recent trends and a sky eye view appraisal. Plant Cell Tiss Org Cult, 2016, 126: 1-18.

[18] Goswami SK, Das DK. Resveratrol and chemoprevention. Cancer Lett, 2009, 284: 1-6.

[19] Grotewold E. The challenges of moving chemicals within and out of cells: insights into the transport of plant natural products. Planta, 2004, 219: 906-909.

[20] Homova V, Weber J, Schulze J, Alipieva K, Bley T, Georgiev M. Devil’s claw hairy root culture in flasks and in a 3-L bioreactor: bioactive metabolite accumulation and flow cytometry. Z Naturforsch C, 2010, 65: 472-478.

[21] Hussain MS, Fareed S, Ansari S, Rahman MA, Ahmad I. Current approaches toward production of secondary plant metabolites. J Pharm Bioallied Sci, 2012, 4: 10-20.

[22] Jaillon O, Aury JM, Noel B, Policriti A, Clepet C, Casagrande A, Choisne N, . The grapevine genome sequence suggests ancestral hexaploidization in major angiosperm phyla. Nature, 2007, 449: 463-467.

[23] Jang M, Cai L, Udeani GO, Slowing KV, Thomas CF, Beecher CWW, Fong HHS, Farnsworth NR, Kinghorn AD, Mehta RG, Moon RC, Pezzuto JM. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science, 1997, 275: 218-220.

[24] Jeandet P, Douillet-Breuil A-C, Bessis R, Debord S, Sbaghi M, Adrian M. Phytoalexins from the Vitaceae: biosynthesis, phytoalexin gene expression in transgenic plants, antifungal activity, and metabolism. J Agric Food Chem, 2002, 50: 2731-2741.

[25] Jeandet P, Clément C, Tisserant L-P, Crouzet J, Courot É. Use of grapevine cell cultures for the production of phytostilbenes of cosmetic interest. C R Chim, 2016, 19: 1062-1070.

[26] Jeandet P, Sobarzo-Sánchez E, Clément C, Nabavi SF, Habtemariam S, Nabavi SM, Cordelier S. Engineering stilbene metabolic pathways in microbial cells. Biotechnol Adv, 2018, 36: 2264-2283.

[27] Jeandet P, Sobarzo-Sánchez E, Silva AS, Clément C, Nabavi SF, Battino M, Rasekhian M, Belwal T, Habtemariam S, Koffas M, Nabavi SM. Whole-cell biocatalytic, enzymatic and green chemistry methods for the production of resveratrol and its derivatives. Biotechnol Adv, 2020, 39: 107461.

[28] Jeong YJ, An CH, Woo SG, Jeong HJ, Kim Y-M, Park S-J, Yoon BD, Kim CY. Production of pinostilbene compounds by the expression of resveratrol O-methyltransferase genes in Escherichia coli. Enzyme Microb Technol, 2014, 54: 8-14.

[29] Jeong YJ, Woo SG, An CH, Jeong HJ, Hong Y-S, Kim Y-M, Ryu YB, Rho M-C, Lee WS, Kim CY. Metabolic engineering for resveratrol derivative biosynthesis in Escherichia coli. Mol Cells, 2015, 38: 318-326.

[30] Jeong YJ, An CH, Woo SG, Park JH, Lee K-W, Lee S-H, Rim Y, Jeong HJ, Ryu YB, Kim CY. Enhanced production of resveratrol derivatives in tobacco plants by improving the metabolic flux of intermediates in the phenylpropanoid pathway. Plant Mol Biol, 2016, 92: 117-129.

[31] Jeong YJ, An CH, Park S-C, Pyun JW, Lee J, Kim SW, Kim H-S, Kim H, Jeong JC, Kim CY. Methyl jasmonate increases isoflavone production in soybean cell cultures by activating structural genes involved in isoflavonoid biosynthesis. J Agric Food Chem, 2018, 66: 4099-4105.

[32] Lambert C, Lemaire J, Auger H, Guilleret A, Reynaud R, Clément C, Courot E, Taidi B. Optimize, modulate, and scale-up resveratrol and resveratrol dimers bioproduction in Vitis labrusca L. cell suspension from flasks to 20 L bioreactor. Plants, 2019, 8: 576.

[33] Langcake P, Pryce RJ. A new class of phytoalexins from grapevines. Experientia, 1977, 33: 151-152.

[34] Lijavetzky D, Almagro L, Belchi-Navarro S, Martínez-Zapater JM, Bru R, Pedreño MA. Synergistic effect of methyljasmonate and cyclodextrin on stilbene biosynthesis pathway gene expression and resveratrol production in Monastrell grapevine cell cultures. BMC Res Notes, 2008, 1: 132.

[35] Liu Z (2011) Diterpene glycosides as natural solubilizers. US Patent Application PCT/US2011, 0033525

[36] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCT method. Methods, 2001, 25: 402-408.

[37] Martinez-Esteso MJ, Sellés-Marchart S, Vera-Urbina JC, Pedreño MA, Bru-Martinez R. Changes of defense proteins in the extracellular proteome of grapevine (Vitis vinifera cv. Gamay) cell cultures in response to elicitors. J Proteomics, 2009, 73: 331-341.

[38] Martínez-Márquez A, Morante-Carriel JA, Ramírez-Estrada K, Cusidó RM, Palazon J, Bru-Martínez R. Production of highly bioactive resveratrol analogues pterostilbene and piceatannol in metabolically engineered grapevine cell cultures. Plant Biotechnol J, 2016, 14: 1813-1825.

[39] Maurer M, Kühleitner M, Gasser B, Mattanovich D. Versatile modeling and optimization of fed batch processes for the production of secreted heterologous proteins with Pichia pastoris. Microb Cell Fact, 2006, 5: 37.

[40] Morales M, Bru R, García-Carmona F, Ros Barceló A, Pedreño MA. Effect of dimethyl-β-cyclodextrins on resveratrol metabolism in Gamay grapevine cell cultures before and after inoculation with shape Xylophilus ampelinus. Plant Cell Tiss Org Cult, 1998, 53: 179-187.

[41] Mustafa NR, de Winter W, van Iren F, Verpoorte R. Initiation, growth and cryopreservation of plant cell suspension cultures. Nat Protoc, 2011, 6: 715-742.

[42] Mulabagal V, Tsay H-S. Plant cell cultures an alternative and efficient source for the production of biologically important secondary metabolites. J Appl Sci Eng Tech, 2004, 2: 29-48.

[43] Murthy HN, Lee E-J, Paek K-Y. Production of secondary metabolites from cell and organ cultures: strategies and approaches for biomass improvement and metabolite accumulation. Plant Cell Tiss Org Cult, 2014, 118: 1-16.

[44] Namdeo A. Plant cell elicitation for production of secondary metabolites: a review. Pharmacogn Rev, 2007, 1: 69-79.

[45] Nivelle L, Hubert J, Courot E, Jeandet P, Aziz A, Nuzillard J-M, Renault J-H, Clément C, Martiny L, Delmas D, Tarpin M. Anti-cancer activity of resveratrol and derivatives produced by grapevine cell suspensions in a 14 L stirred bioreactor. Molecules, 2017, 22: 474.

[46] Pan Q-H, Wang L, Li J-M. Amounts and subcellular localization of stilbene synthase in response of grape berries to UV irradiation. Plant Sci, 2009, 176: 360-366.

[47] Pezet R, Perret C, Jean-Denis JB, Tabacchi R, Gindro K, Viret O. δ-Viniferin, a resveratrol dehydrodimer: one of the major stilbenes synthesized by stressed grapevine leaves. J Agric Food Chem, 2003, 51: 5488-5492.

[48] Sako M, Hosokawa H, Ito T, Iinuma M. Regioselective oxidative coupling of 4-hydroxystilbenes: synthesis of resveratrol and ε-viniferin (E)-dehydrodimers. J Org Chem, 2004, 69: 2598-2600.

[49] Shen T, Wang X-N, Lou H-X. Natural stilbenes: an overview. Nat Prod Rep, 2009, 26: 916-935.

[50] Shin S-Y, Han NS, Park Y-C, Kim M-D, Seo J-H. Production of resveratrol from p-coumaric acid in recombinant Saccharomyces cerevisiae expressing 4-coumarate:coenzyme A ligase and stilbene synthase genes. Enzyme Microb Technol, 2011, 48: 48-53.

[51] Sotheeswaran S, Pasupathy V. Distribution of resveratrol oligomers in plants. Phytochemistry, 1993, 32: 1083-1092.

[52] Tassoni A, Fornalè S, Franceschetti M, Musiani F, Michael AJ, Perry B, Bagni N. Jasmonates and Na-orthovanadate promote resveratrol production in Vitis vinifera cv. Barbera cell cultures. New Phytol, 2005, 166: 895-905.

[53] Uchiyama H, Tozuka Y, Nishikawa M, Takeuchi H. Nanocomposite formation between alpha-glucosyl stevia and surfactant improves the dissolution profile of poorly water-soluble drug. Int J Pharm, 2012, 428: 183-186.

[54] Valenzano DR, Terzibasi E, Genade T, Cattaneo A, Domenici L, Cellerino A. Resveratrol prolongs lifespan and retards the onset of age-related markers in a short-lived vertebrate. Curr Biol, 2006, 16: 296-300.

[55] Vannozzi A, Dry IB, Fasoli M, Zenoni S, Lucchin M. Genome-wide analysis of the grapevine stilbene synthase multigenic family: genomic organization and expression profiles upon biotic and abiotic stresses. BMC Plant Biol, 2012, 12: 130.

[56] Vezzulli S, Civardi S, Ferrari F, Bavaresco L. Methyl jasmonate treatment as a trigger of resveratrol synthesis in cultivated grapevine. Am J Enology Vitic, 2007, 58: 530-533.

[57] Vitrac X, Bornet A, Vanderlinde R, Valls J, Richard T, Delaunay JC, Mérillon JM, Teissédre PL. Determination of stilbenes (ε-viniferin, trans-astringin, trans-piceid, cis- and trans-resveratrol, ε-viniferin) in Brazilian wines. J Agric Food Chem, 2005, 53: 5664-5669.

[58] Wan Z-L, Wang J-M, Wang L-Y, Yang X-Q, Yuan Y. Enhanced physical and oxidative stabilities of soy protein-based emulsions by incorporation of a water-soluble stevioside–resveratrol complex. J Agric Food Chem, 2013, 61: 4433-4440.

[59] Wang Y, Yu O. Synthetic scaffolds increased resveratrol biosynthesis in engineered yeast cells. J Biotechnol, 2012, 157: 258-260.

[60] Wu J, Liu P, Fan Y, Bao H, Du G, Zhou J, Chen J. Multivariate modular metabolic engineering of Escherichia coli to produce resveratrol from L-tyrosine. J Biotechnol, 2013, 167: 404-411.

[61] Xue Y-Q, Di J-M, Luo Y, Cheng K-J, Wei X, Shi Z. Resveratrol oligomers for the prevention and treatment of cancers. Oxid Med Cell Longev, 2014, 2014: 9.

[62] Yazaki K. Transporters of secondary metabolites. Curr Opin Plant Biol, 2005, 8: 301-307.

[63] Yu CKY, Lam CNW, Springob K, Schmidt J, Chu IK, Lo C. Constitutive accumulation of cis -piceid in transgenic Arabidopsis overexpressing a sorghum stilbene synthase gene. Plant Cell Physiol, 2006, 47: 1017-1021.

[64] Zamboni A, Vrhovsek U, Kassemeyer H-H, Mattivi F, Velasco R. Elicitor-induced resveratrol production in cell cultures of different grape genotypes (Vitis spp.). Vitis, 2006, 45: 63-68.

[65] Zhang F, Koh GY, Jeansonne DP, Hollingsworth J, Russo PS, Vicente G, Stout RW, Liu Z. A novel solubility-enhanced curcumin formulation showing stability and maintenance of anticancer activity. J Pharm Sci, 2011, 100: 2778-2789.

[66] Zhao J, Davis LC, Verpoorte R. Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnol Adv, 2005, 23: 283-333.

Funding

Korea Research Institute of Bioscience and Biotechnology(The KRIBB Initiative Program)

The Next-Generation BioGreen 21 Program, Rural Development Administration (KR)(Grant #: PJ01318604)