Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 136, Issue 1, pp 189–196 | Cite as

The effect of explant origin and collection season on stilbene biosynthesis in cell cultures of Vitis amurensis Rupr.

  • A. P. Tyunin
  • A. R. Suprun
  • N. N. Nityagovsky
  • A. Y. Manyakhin
  • Y. A. Karetin
  • A. S. Dubrovina
  • K. V. KiselevEmail author
Research Note


Plant cell and tissue cultures are considered as a source of valuable secondary metabolites but usually produce insufficient level of the compounds, which is the limiting factor for their application in biotechnology. We obtained 18 callus cell cultures from different organs of wild grape Vitis amurensis Rupr. collected at different seasons and analyzed stilbene accumulation in combination with calli growth parameters. This analysis showed that temporal and tissue origin of the calli affected the rate of stilbene biosynthesis. Stem-derived calli accumulated higher stilbene levels and exhibited a higher expression of phenylalanine ammonia-lyase (PAL) and stilbene synthase (STS) genes than calli derived from the leaves and petioles. The highest content of stilbenes was detected in the calli initiated from grapevine stems collected in the autumn. In general, all “autumn” cell cultures contained more than 2 mg g− 1 dry wt (up to 11 mg g− 1 dry wt) and exhibited high PAL and STS genes expression in comparison with the calli initiated in the summer. The content of stilbenes in the “autumn” cell cultures were comparable to the highest stilbene contents detected in other plant sources described in the literature. Thus, selecting the most optimal explant source for cell culture establishment could be an effective approach towards developing plant cell cultures producing high stilbene levels.


Piceid Resveratrol Stilbenoid glucoside Stilbene synthase STS Viniferin 



High-performance liquid chromatography


Stilbene synthase





This work was supported by a Grant from the Russian Science Foundation (17-74-10082).

Author Contributions

ASD and KKV were involved in research design, data analysis, and paper preparation. APT, ARS, and NNN were involved in plant cell cultures management and qRT-PCR. MAY performed HPLC. YAK: analyzed the data.

Compliance with ethical standards

Conflict of interest

We declare that we have no conflict of interest.


  1. Aleynova OA, Grigorchuk VP, Dubrovina AS, Rybin VG, Kiselev KV (2016) Stilbene accumulation in cell cultures of Vitis amurensis Rupr. overexpressing VaSTS1, VaSTS2, and VaSTS7 genes. Plant Cell Tissue Organ Cult 125:329–339.CrossRefGoogle Scholar
  2. Aleynova OA, Dubrovina AS, Kiselev KV (2017) Activation of stilbene synthesis in cell cultures of Vitis amurensis by calcium-dependent protein kinases VaCPK1 and VaCPK26. Plant Cell Tiss Organ Cult 130:141–152CrossRefGoogle Scholar
  3. Aleynova-Shumakova OA, Dubrovina AS, Manyakhin AY, Karetin YA, Kiselev KV (2014) VaCPK20 gene overexpression significantly increased resveratrol content and expression of stilbene synthase genes in cell cultures of Vitis amurensis Rupr.. Appl Microbiol Biotechnol 98:5541–5549CrossRefGoogle Scholar
  4. Arora J, Goyal S, Ramawat KG (2010) Enhanced stilbene production in cell cultures of Cayratia trifolia through co-treatment with abiotic and biotic elicitors and sucrose. In Vitro Cell Dev Biol Plant 46:430–436CrossRefGoogle Scholar
  5. Chang X, Seo M, Takebayashi Y, Kamiya Y, Riemann M, Nick P (2017) Jasmonates are induced by the PAMP flg22 but not the cell death-inducing elicitor Harpin in Vitis rupestris. Protoplasma 254:271–283CrossRefGoogle Scholar
  6. Chong J, Poutaraud A, Hugueney P (2009) Metabolism and roles of stilbenes in plants. Plant Sci 177:143–155CrossRefGoogle Scholar
  7. Dubrovina AS, Kiselev KV (2016) Age-associated alterations in the somatic mutation and DNA methylation levels in plants. Plant Biol (Stuttg) 18:185–196CrossRefGoogle Scholar
  8. Dubrovina AS, Kiselev KV (2017) Regulation of stilbene biosynthesis in plants. Planta 346:597–623CrossRefGoogle Scholar
  9. Dubrovina AS, Manyakhin AY, Zhuravlev YN, Kiselev KV (2010)) Resveratrol content and expression of phenylalanine ammonia-lyase and stilbene synthase genes in rolC transgenic cell cultures of Vitis amurensis. Appl Microbiol Biotechnol 88:727–736CrossRefGoogle Scholar
  10. Giorcelli A, Sparvoli F, Mattivi F, Tava A, Balestrazzi A, Vrhovsek U, Calligari P, Bollini R, Confalonieri M (2004) Expression of the stilbene synthase (StSy) gene from grapevine in transgenic white poplar results in high accumulation of the antioxidant resveratrol glucosides. Transgenic Res 13:203–214CrossRefGoogle Scholar
  11. Huang KS, Lin M, Cheng GF (2001) Anti-inflammatory tetramers of resveratrol from the roots of Vitis amurensis and the conformations of the seven-membered ring in some oligostilbenes. Phytochemistry 58:357–362CrossRefGoogle Scholar
  12. Kawiak A, Krolicka A, Lojkowska E (2011) In vitro cultures of Drosera aliciae as a source of a cytotoxic naphthoquinone: ramentaceone. Biotechnol Lett 33:2309–2316CrossRefGoogle Scholar
  13. Keskin N, Kunter B (2008) Production of trans-resveratrol in ‘‘Cabernet Sauvignon’’ (Vitis vinifera L.) callus culture in response to ultraviolet-C irradiation. Vitis 47:193–196Google Scholar
  14. Ketel DH, Breteler H, De Groot B (1985) Effect of explant origin on growth and differentiation of calli from Tagetes species. J Plant Physiol 118:327–333CrossRefGoogle Scholar
  15. Kiselev KV (2011) Perspectives for production and application of resveratrol. Appl Microbiol Biotechnol 90:417–425CrossRefGoogle Scholar
  16. Kiselev KV, Dubrovina AS, Veselova MV, Bulgakov VP, Fedoreyev SA, Zhuravlev YN (2007) The rolB gene-induced overproduction of resveratrol in Vitis amurensis transformed cells. J Biotechnol 128:681–692CrossRefGoogle Scholar
  17. Kiselev KV, Dubrovina AS, Isaeva GA, Zhuravlev YN (2010) The effect of salicylic acid on phenylalanine ammonia-lyase and stilbene synthase gene expression in Vitis amurensis cell culture. Russ J Plant Physiol 57:415–448CrossRefGoogle Scholar
  18. Kiselev KV, Tyunin AP, Manyakhin AY, Zhuravlev YN (2011) Resveratrol content and expression patterns of stilbene synthase genes in Vitis amurensis cells treated with 5-azacytidine. Plant Cell Tissue Organ Cult 105:65–72CrossRefGoogle Scholar
  19. Kiselev KV, Dubrovina AS, Shumakova OA, Karetin YA, Manyakhin AY (2013) Structure and expression profiling of a novel calcium-dependent protein kinase gene, CDPK3a, in leaves, stems, grapes, and cell cultures of wild-growing grapevine Vitis amurensis Rupr.. Plant Cell Rep 32:431–442CrossRefGoogle Scholar
  20. Kiselev KV, Aleynova OA, Grigorchuk VP, Dubrovina AS (2017a) Stilbene accumulation and expression of stilbene biosynthesis pathway genes in wild grapevine Vitis amurensis Rupr.. Planta 245:151–159CrossRefGoogle Scholar
  21. Kiselev KV, Aleynova OA, Tyunin AP (2017b) Expression of the R2R3 MYB transcription factors in Vitis amurensis Rupr. plants and cell cultures with different resveratrol content. Russ J Genet 53:465–471CrossRefGoogle Scholar
  22. Langcake P, Pryce RJ (1977) A new class of phytoalexins from grapevines. Experientia 33:151–152CrossRefGoogle Scholar
  23. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408CrossRefGoogle Scholar
  24. Margara J, Piollat MT (1983) New observations on in vitro organogenesis from begonia x elator petals. Comptes Rendus de l’Académie des Sciences - Series III - Sciences de la Vie 297: 161–164Google Scholar
  25. Martini AN, Papafotiou M, Vemmos SN (2013) Season and explant origin affect phenolic content, browning of explants, and micropropagation of X Malosorbus florentina (Zucc.) Browicz. Hortscience 48:102–107Google Scholar
  26. Ochoa-Villarreal M, Howat S, Hong S, Jang MO, Jin YW, Lee EK, Loake GJ (2016) Plant cell culture strategies for the production of natural products. BMB Rep 49:149–158CrossRefGoogle Scholar
  27. Pezet R (1998) Purification and characterization of a 32-kDa laccase-like stilbene oxidase produced by Botrytis cinerea Pers.:Fr. FEMS Microbiol Lett 167:203–208CrossRefGoogle Scholar
  28. Santamaria AR, Mulinacci N, Valletta A, Innocenti M, Pasqua G (2011) Effects of elicitors on the production of resveratrol and viniferins in cell cultures of Vitis vinifera L. cv Italia. J Agric Food Chem 59:9094–9101CrossRefGoogle Scholar
  29. Santamaria AR, Innocenti M, Mulinacci N, Melani F, Valletta A, Sciandra I, Pasqua G (2012) Enhancement of viniferin production in Vitis vinifera L. cv. Alphonse Lavallée cell suspensions by low-energy ultrasound alone and in combination with methyl jasmonate. J Agric Food Chem 60:11135–11142CrossRefGoogle Scholar
  30. Schwekendiek A, Spring O, Heyerick A, Pickel B, Pitsch NT, Peschke F, Keukeleire DD, Weber G (2007) Constitutive expression of a grapevine stilbene synthase gene in transgenic hop (Humulus lupulus L.) yields resveratrol and its derivatives in substantial quantities. J Agric Food Chem 55:7002–7009CrossRefGoogle Scholar
  31. Shankar S, Nall D, Tang SN, Meeker D, Passarini J, Sharma J, Srivastava RK (2011) Resveratrol inhibits pancreatic cancer stem cell characteristics in human and kras(G12D) transgenic mice by inhibiting pluripotency maintaining factors and epithelial-mesenchymal transition. PLoS ONE 6:e16530CrossRefGoogle Scholar
  32. Shumakova OA, Manyakhin AY, Kiselev KV (2011) Resveratrol content and expression of phenylalanine ammonia-lyase and stilbene synthase genes in cell cultures of Vitis amurensis treated with coumaric acid. Appl Biochem Biotechnol 165:1427–1436CrossRefGoogle Scholar
  33. Suwalsky M, Villena F, Gallardo MJ (2015) In vitro protective effects of resveratrol against oxidative damage in human erythrocytes. Biochim Biophys Acta 1848:76–82CrossRefGoogle Scholar
  34. Tassoni A, Fornale S, Franceschetti M, Musiani F, Michael AJ, Perry B, Bagni N (2005) Jasmonates and Na-orthovanadate promote resveratrol production in Vitis vinifera cv. Barbera cell cultures. New Phytol 166:895–905CrossRefGoogle Scholar
  35. Taurino M, Ingrosso I, D’amico L, De Domenico S, Nicoletti I, Corradini D, Santino A, Giovinazzo G (2015) Jasmonates elicit different sets of stilbenes in Vitis vinifera cv. Negramaro cell cultures. Springerplus 4:49CrossRefGoogle Scholar
  36. Tavares S, Vesentini D, Fernandes JC, Ferreira RB, Laureano O, Ricardo-Da-Silva JM, Amâncio S (2013) Vitis vinifera secondary metabolism as affected by sulfate depletion: diagnosis through phenylpropanoid pathway genes and metabolites. Plant Physiol Biochem 66:118–126CrossRefGoogle Scholar
  37. Tyunin AP, Kiselev KV (2016) Alternations in VaSTS gene cytosine methylation and t-resveratrol production in response to UV-C irradiation in Vitis amurensis Rupr. cells. Plant Cell Tissue Organ Cult 124:33–45CrossRefGoogle Scholar
  38. Wilson AG (2008) Epigenetic regulation of gene expression in the inflammatory response and relevance to common diseases. J Periodontol 79:1514–1519CrossRefGoogle Scholar
  39. Xu A, Zhan JC, Huang WD (2015) Effects of ultraviolet C, methyl jasmonate and salicylic acid, alone or in combination, on stilbene biosynthesis in cell suspension cultures of Vitis vinifera L. cv. Cabernet Sauvignon. Plant Cell Tissue Organ Cult 122:197–211CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • A. P. Tyunin
    • 1
  • A. R. Suprun
    • 1
    • 2
  • N. N. Nityagovsky
    • 1
    • 2
  • A. Y. Manyakhin
    • 1
  • Y. A. Karetin
    • 2
    • 3
  • A. S. Dubrovina
    • 1
  • K. V. Kiselev
    • 1
    • 2
    Email author
  1. 1.Laboratory of Biotechnology, Federal Scientific Center of the East Asia Terrestrial BiodiversityFar Eastern Branch of the Russian Academy of SciencesVladivostokRussia
  2. 2.Department of Biodiversity, The School of Natural SciencesFar Eastern Federal UniversityVladivostokRussia
  3. 3.Laboratory of Embryology, National Scientific Center of Marine BiologyFar Eastern Branch of the Russian Academy of SciencesVladivostokRussia

Personalised recommendations