Expression of resveratrol synthase gene and accumulation of resveratrol in transgenic potatoes (Solanum tuberosum L.)

  • Received : 2009.12.07
  • Published : 20091200

Abstract

A resveratrol synthase (RS) gene was isolated from peanut (Arachis hypogaea, L. cv. Jinpoong) plants. This gene was placed under the control of the cauliflower mosaic virus 35S promoter (CaMV35S) and introduced into two Korean varieties of potato (Solanum tuberosum L. cvs. Jasim and Jowon) plants by Agrobacterium-mediated gene transfer. Putative transformants were screened by PCR with primers designed from CaMV 35S promoter, NOS terminator and RS gene. Most of selected transgenic potato plants showed the amplification of expected fragments by PCR of genomic DNA with gene-specific primers, while they were absent in untransformed control plants. Expression of the resveratrol synthase gene was also examined by northern blot analysis. The transformants showed a band which was lacking in the control plant, confirming that the introduced gene is transcribed into mRNA in the transformants. The strength of the band, which reflected the level of mRNA expression, differed among the individual transformants. Among the transformants obtained, the highest trans-resveratrol content in the transgenic young leaves of purple-fleshed "Jashim" was $2.11{\mu}gg^{-1}$ fresh weight and that in the microtubers in vitro of purple fleshed "Jashim" was $8.31{\mu}gg^{-1}$ fresh weight. This amount of resveratrol may have a positive biological effect on human health.

Keywords

References

  1. An G. 1987. Binary Ti vectors for plant transformation and promoter analysis. Methods Enzymol. 153:292-305 https://doi.org/10.1016/0076-6879(87)53060-9
  2. An G, Ichinose Y, Yamada T, Tanaka Y, Shiraishi T, Oku H. 1993. Organization of the genes encoding chalcone synthase in Pisum sativum. Plant Mol. Biol. 21:789-803 https://doi.org/10.1007/BF00027112
  3. Baek SH, Shin WC, Kim HS, Seo CS, Kang HJ. 2006. Development of transgenic rice producing resveratrol. Korean J of Crop Science 51(sup.2) 141
  4. Bergmann H, Lippmann B, Leinhos V, Tiroke S, Machelett B. 1999. Activation of stress resistance in plants and consequences for product quality. Journal of Applied Botany 73:153-161
  5. Bilodeau P, Udvardi MK, Peacock WJ, Dennis ES. 1999. A prolonged cold treatment-induced cytochrome P450 gene from Arabidopsis thaliana. Plant Cell Environ 22:791-800 https://doi.org/10.1046/j.1365-3040.1999.00444.x
  6. Bokkenheuser VD, Shackleton CH, Winter J. 1987. Hydrolysis of dietary flavonoid glycosides by strains of intestinal Bacteroides from humans. Biochem J 248:953-956
  7. D$\'{e}$stef$\'{a}$no-B$\'{e}$ltran L, Nagpala P, Centiner S, Dodds J, Jaynes J. 1990. Enhancing bacterial and fungal disease resistance in plants. Application to potato. In: Vayda ME, Parks W (eds) The molecular and cellular biology of the potato. CABI. Wallingford, pp. 205-221
  8. Hain R, Reif HJ, Krause E, Langebartels R, Kindl H, Vornam B, Wiese W, Schmelzer E, Schreier PH, Stöcker RH, Stenzel K. 1993. Disease resistance results from foreign phytoalexin expression in a novel plant. Nature 361:153- 156 https://doi.org/10.1038/361153a0
  9. Hart JH. 1981. Role of phytostilbenes in decay and disease resistance. Annu Rev Phytopathol 19:437-458 https://doi.org/10.1146/annurev.py.19.090181.002253
  10. Ingham JL. 1976. 3,5,4’-Trihydroxystilbene as a phytoalexin from groundnuts (Arachis hypogaea). Phytochemistry 15:1791-1793 https://doi.org/10.1016/S0031-9422(00)97494-6
  11. Jang M, Cai L, Udeani GO, Slowing KV, Thomas CF, Beecher CWW, Fong HHS, Farnsworth NR, Kinghorn AD, Mehta RG, Moon RC, Pezzuto JM. 1997. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Scinece 275:218-220 https://doi.org/10.1126/science.275.5297.218
  12. Jeandet P, Breuil AC, Adrian M, Weston LA, Debord S, Meunier P, Maume G, Bessis R. 1999. HPLC analysis of grapevine phytoalexins coupling photodiode array detection and fluorometry. Anal Chem 69:5172-5177 https://doi.org/10.1021/ac970582b
  13. Jeon TW, Cho YS, Lee SH, Cho SM, Cho HM, Chang KS, Park HJ. 2005. Studies on the biological activities and physiochemical characteristics of pigments extracted from Korean purple-fleshed potato. Korean J of Food Sci Technol 37:247-254
  14. Kobayash S, Ding CK, Nakamura Y, Nakajima I, Matsumoto R. 2000. Kiwifruits (Actinidia deliciosa) transformed with a Vitis stilbene synthase gene produce piceid (resveratro-glucoside) Plant Cell Reports 19:904-910 https://doi.org/10.1007/s002990000203
  15. Langcake P, Pryce RJ. 1976. The production of resveratrol by Vitis vinifera and other members of the Vitaceae as a response to infection of injury. Physiol Plant Pathol 9:77- 86 https://doi.org/10.1016/0048-4059(76)90077-1
  16. Lawton MA, Dean SM, Dron M, Kooter, Kragh, Harrison MJ, Yu L, Tanguay L, Dixon RA, Lamb CJ. 1991. Silencer region of a chalcone synthase promoter contains multiple sites for a factor SBF-1, closely related to GT-1 Plant Mol Biol 16:235-249 https://doi.org/10.1007/BF00020555
  17. Leckband G and Lörz H. 1998. Transformation and expression of a stilbene synthase gene of Vitis vinifera L. in barley and wheat for increased fungal reasistance. Theor Appl Genet 96:104-192 https://doi.org/10.1007/s001220050832
  18. Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  19. Ruhmann S, Trutter D, Fritsche S, Briviba K, Szankowski I. 2006. Piceid (resvertrol glucoside) synthesis in stilbene synthase transgenic apple fruit. J Agric food Chem 54: 4633-4640 https://doi.org/10.1021/jf060249l
  20. Stark-Lorenzen P, Nelke B, Hänßler G, Mühlbach HP. Thomzik JE. 1997. Transfer of a grapevine stilbene synthase gene to rice (Oryza sativa L.). Plant Cell Reports 16:668-673 https://doi.org/10.1007/s002990050299
  21. Thomzik JE. 1995. Gene transfer in plants. Pflanzenschutz Nachrichten Bayer 48:5-23
  22. Wu JM, Wang ZR, Hsieh TC, Bruder JL, Zou JG, Huang YZ. 2001. Mechanism of cardioprotection by resveratrol, a phenolic antioxidant present in red wine. Int J Mol Med 8:3-17