Cloning, Expression Pattern Analysis and Subcellular Localization of Resveratrol Synthase Gene in Peanut (Arachis hypogaea L.)


Resveratrol synthase (RS) is a key enzyme that plays a critical role in the resveratrol synthesis pathway. In this study, six RS genes were isolated and characterized from peanut variety “Zhenzhu Hong” by silico cloning and RT-PCR. Bioinformatics analysis showed that deduced amino acid sequences of the six cloned RS genes were highly conserved with a similarity from 95% to 99% when compared to the RS genes which had been deposited at the GenBank. The results of amino acid sequences analysis showed six RS proteins contained the Chal_Sti_Synt_N and ACP_Syn_III_C domains and can be classified to same family but with different evolutionary distance. Expression pattern analysis by QRT-PCR provided evidence indicating that the mRNA of six RS genes were primarily expressed in the peanut shell at different developmental stages with different expression levels, but only lower levels of them were evident in the peanut kernel. The subcellular localization of RS protein in onion epidermal cell was performed by Agrobacterium tumefaciens-mediated transformation and the green fluorescent was monitored by confocal fluorescence microscopy. The results indicated that, RS1 and RS5 were located in the nucleus and plasma membrane respectively, while the RS2, RS3, RS4 and RS6 were located in both nucleus inner membrane and plasma membrane. The data will provide basic information for elucidating the regulatory mechanisms and enzyme kinetics underlying the RS genes in the resveratrol synthase pathway.

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Zhu, F. , Han, J. , Liu, S. , Chen, X. , Varshney, R. and Liang, X. (2014) Cloning, Expression Pattern Analysis and Subcellular Localization of Resveratrol Synthase Gene in Peanut (Arachis hypogaea L.). American Journal of Plant Sciences, 5, 3619-3631. doi: 10.4236/ajps.2014.524378.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Dao, T.T.H., Linthorst, H.J.M. and Verpoorte, R. (2011) Chalcone Synthase and Its Functions in Plant Resistance. Phytochemistry Reviews, 10, 397-412.
[2] Jeandet, P., Delaunois, B., Conreux, A., et al. (2010) Biosynthesis, Metabolism, Molecular Engineering, and Biological functions of stilbene phytoalexins in plants. BioFactors, 36, 331-341.
[3] Beekwilder, J., Wolswinkel, R., Jonker, H., et al. (2006) Production of Resveratrol in Recombinant Microorganisms. Applied and Environmental Microbiology, 72, 5670-5672.
[4] Horinouchi, S. (2009) Combinatorial Biosynthesis of Plant Medicinal Polyketides by Microorganisms. Current Opinion in Chemical Biology, 13, 197-204.
[5] Csiszar, A., Bagi, Z., Feher, A., et al. (2011) Resveratrol Confers Endothelial Protection via Activation of the Antioxidant Transcription Factor Nrf2. The FASEB Journal, 25, 1093.13.
[6] Lee, H.S., Lee, B.W., Kim, M.R., et al. (2010) Syntheses of Resveratrol and Its Hydroxylated Derivatives as Radical Scavenger and Tyrosinase Inhibitor. Bulletin of the Korean Chemical Society, 31, 971-975.
[7] Elmali, N., Baysal, O., Harma, A., et al. (2007) Effects of Resveratrol in Inflammatory Arthritis. Inflammation, 30, 1-6.
[8] Athar, M., Back, J.H., Kopelovich, L., et al. (2009) Multiple Molecular Targets of Resveratrol: Anti-Carcinogenic Mechanisms. Archives of Biochemistry and Biophysics, 486, 95-102.
[9] Shukla, Y. and Singh, R. (2011) Resveratrol and Cellular Mechanisms of Cancer Prevention. Annals of the New York Academy of Sciences, 1215, 1-8.
[10] Baur, J.A., Pearson, K.J., Price, N.L., et al. (2006) Resveratrol Improves Health and Survival of Mice on a High-Calorie Diet. Nature, 444, 337-342.
[11] Brisdelli, F., D’Andrea, G. and Bozzi, A. (2009) Resveratrol: A Natural Polyphenol with Multiple Chemopreventive Properties (Review). Current Drug Metabolism, 10, 530-546.
[12] Fulda, S. (2010) Resveratrol and Derivatives for the Prevention and Treatment of Cancer. Drug Discovery Today, 15, 757-765.
[13] Hsieh, T. and Wu, J.M. (2010) Resveratrol: Biological and Pharmaceutical Properties as Anticancer Molecule. BioFactors, 36, 360-369.
[14] Vang, O., Ahmad, N., Baile, C.A., Baur, J.A., Brown, K., Csiszar, A., et al. (2011) What Is New for an Old Molecule? Systematic Review and Recommendations on the Use of Resveratrol. PLoS ONE, 6, e19881.
[15] Kasiotis, K.M., Pratsinis, H., Kletsas, D. and Haroutounian, S.A. (2013) Resveratrol and Related Stilbenes: Their Anti-Aging and Anti-Angiogenic Properties. Food and Chemical Toxicology, 61, 112-120.
[16] Chong, J., Poutaraud, A. and Hugueney, P. (2009) Metabolism and Roles of Stilbenes in Plants. Plant Science, 177, 143-155.
[17] Guerrero, R.F., Puertas, B., Fernández, M.I., Palma, M. and Cantos-Villar, E. (2010) Induction of Stilbenes in Grapes by UV-C: Comparison of Different Subspecies of Vitis. Innovative Food Science & Emerging Technologies, 11, 231-238.
[18] Tang, K., Zhan, J.C., Yang, H.R. and Huang, W.D. (2010) Changes of Resveratrol and Antioxidant Enzymes during UV-Induced Plant Defense Response in Peanut Seedlings. Journal of Plant Physiology, 167, 95-102.
[19] Hammerbacher, A., Ralph, S.G., Bohlmann, J., Fenning, T.M., Gershenzon, J. and Schmidt, A. (2011) Biosynthesis of the Major Tetrahydroxystilbenes in Spruce, Astringin and Isorhapontin, Proceeds via Resveratrol and Is Enhanced by Fungal Infection. Plant Physiology, 157, 876-890.
[20] Dubrovina, A.S. and Kiselev, K.V. (2012) Effect of Long-Term Cultivation on Resveratrol Accumulation in a High-Producing Cell Culture of Vitis amurensis. Acta Physiologiae Plantarum, 34, 1101-1106.
[21] Lu, D., Zhao, W., Zhu, K. and Zhao, S. (2012) Relevant Enzymes, Genes and Regulation Mechanisms in Biosynthesis Pathway of Stilbenes. Open Journal of Medicinal Chemistry, 2, 15-23.
[22] Schröder, G., Brown, J.W. and Schröder, J. (1988) Molecular Analysis of Resveratrol Synthase. European Journal of Biochemistry, 172, 161-169.
[23] Lanz, T., Schroder, G. and Schroder, J. (1990) Differential Regulation of Genes for Resveratrol Synthase in Cell Cultures of Arachis hypogaea L. Planta, 181, 169-175.
[24] Wang, B.Y., Pan, H.F., Ye, B.Y., et al. (2005) Cloning and Sequence Analysis of Resveratrol Synthase DNA from Peanut (Arachis hypogaea L.). Journal of Fujian Normal University (Natural Science), 21, 56-60.
[25] Huang, X.Q., Guo, L.Q., Li, X.M., Lin, J.F., Yuan, Z.H. and Tan, M.C. (2012) Cloning and Sequence Analysis of Resveratrol Synthase Gene from Peanut(Arachis hypogaea L.). Biotechnology Bulletin, 3, 69-74.
[26] Delaunois, B., Cordelier, S., Conreux, A., Clément, C. and Jeandet, P. (2009) Molecular Engineering of Resveratrol in Plants. Plant Biotechnology Journal, 7, 2-12.
[27] Trantas, E., Panopoulos, N. and Ververidis, F. (2009) Metabolic Engineering of the Complete Pathway Leading to Heterologous Biosynthesis of Various Flavonoids and Stilbenoids in Saccharomyces cerevisiae. Metabolic Engineering, 11, 355-366.
[28] Jeandet, P., Delaunois, B., Aziz, A., Donnez, D., Vasserot, Y., Cordelier, S. and Courot, E. (2012) Metabolic Engineering of Yeast and Plants for the Production of the Biologically Active Hydroxystilbene, Resveratrol. Journal of Biomedicine and Biotechnology, 2012, 1-14.
[29] Hain, R., Bieseler, B., Kindl, H., Schröder, G. and Stöcker, R. (1990) Expression of a Stilbene Synthase Gene in Nicotiana tabacum Results in Synthesis of the Phytoalexin Resveratrol. Plant Molecular Biology, 15, 325-335.
[30] Yu, C.K.Y., Springob, K., Schmidt, J., Nicholson, R.L., Chu, I.K., Yip, W.K. and Lo, C. (2005) A Stilbene Synthase Gene (SbSTS1) Is Involved in Host and Nonhost Defense Responses in Sorghum. Plant Physiology, 138, 393-401.
[31] Christine, K.Y., Lam, C.N.W., Springob, K., Schmidt, J., Chu, I.K. and Lo, C. (2006) Constitutive Accumulation of cis-Piceid in Transgenic Arabidopsis Overexpressing a Sorghum Stilbene Synthase Gene. Plant and Cell Physiology, 47, 1017-1021.
[32] Halls, C. and Yu, O. (2008) Potential for Metabolic Engineering of Resveratrol Biosynthesis. Trends in Biotechnology, 26, 77-81.
[33] Pan, L.P., Yu, S.L., Chen, C.J., Li, H., Wu, Y.L. and Li, H.H. (2012) Cloning a Peanut Resveratrol Synthase Gene and Its Expression in Purple Sweet Potato. Plant Cell Reports, 31, 121-131.
[34] He, X., Wang, L., Szklarz, G., Bi, Y.Y. and Ma, Q. (2012) Resveratrol Inhibits Paraquat-Induced Oxidative Stress and Fibrogenic Response by Activating the Nuclear Factor Erythroid 2-Related Factor 2 Pathway. Journal of Pharmacology and Experimental Therapeutics, 342, 81-90.
[35] Preisig-Müller, R., Schwekendiek, A., Brehm, I., Reif, H.J. and Kindl, H. (1999) Characterization of a Pine Multigene Family Containing Elicitor-Responsive Stilbene Synthase Genes. Plant Molecular Biology, 39, 221-229.
[36] Zhou, Y., Yang, Y., Huang, J., et al. (2008) Cloning and Analysis of Resveratrol Synthase Gene Family. Chinese Journal of Oil Crop Sciences, 2, 8.
[37] Chen, R.S., Wu, P.L. and Chiou, R.Y.Y. (2002) Peanut Roots as a Source of Resveratrol. Journal of Agricultural and Food Chemistry, 50, 1665-1667.
[38] Chung, I.M., Park, M.R., Rehman, S. and Yun, S.J. (2001) Tissue Specific and Inducible Expression of Resveratrol Synthase Gene in Peanut Plants. Molecules and Cells, 12, 353-359.
[39] Chung, I.M., Park, M.R., Chun, J.C. and Yun, S.J. (2003) Resveratrol Accumulation and Resveratrol Synthase Gene Expression in Response to Abiotic Stresses and Hormones in Peanut Plants. Plant Science, 164, 103-109.
[40] Galgut, J.M. and Ali, S.A. (2011) Effect and Mechanism of Action of Resveratrol: A Novel Melanolytic Compound from the Peanut Skin of Arachis hypogaea. Journal of Receptors and Signal Transduction, 31, 374-380.
[41] Adrian, M., Jeandet, P., Bessis, R. and Joubert, J.M. (1996) Induction of Phytoalexin (Resveratrol) Synthesis in Grapevine Leaves Treated with Aluminum Chloride (AlCl3). Journal of Agricultural and Food Chemistry, 44, 1979-1981.
[42] Rudolf, J.R. and Resurreccion, A.V.A. (2005) Elicitation of Resveratrol in Peanut Kernels by Application of Abiotic Stresses. Journal of Agricultural and Food Chemistry, 53, 10186-10192.
[43] Hrazdina, G. and Jensen, R.A. (1992) Spatial Organization of Enzymes in Plant Metabolic Pathways. Annual Review of Plant Physiology and Plant Molecular Biology, 43, 241-267.
[44] Pan, Q.H., Wang, L. and Li, J.M. (2009) Amounts and Subcellular Localization of Stilbene Synthase in Response of Grape Berries to UV Irradiation. Plant Science, 176, 360-366.

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