AJPS  Vol.3 No.1 , January 2012
Quantitative Analysis of ATP Sulfurylase and Selenocysteine Methyltransferase Gene Expression in Different Organs of Tea Plant (Camellia sinensis)
Abstract: Tea plant (Camellia sinensis) has unique biological features for the study of cellular and molecular mechanisms, an evergreen broad-leaved woody plant which can accumulate selenium in soil abundant of Selenium. Expression of the genes related to Selenium (Se) metabolism is an adaptation to the soil environment for a long period. The purpose of the present study was to explore if there exist differences of expression about these genes in tea plant between growing in Selenium-abundant and normal soil. A quantitative real-time reverse transcription polymerase chain reaction (Q-RT-PCR) assay was done for quantification of ATP sulfurylase (APS) and selenocysteine methyltransferase (SMT) mRNA normalized to Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene in tea plant. Young leaves, mature leaves and tender roots from tea plants growing in soil abundant of Selenium were respectively obtained from Shitai County, Anhui Province, and also the relevant materials of the selenium un-enriched tea plant planted at agricultural garden of Ahui Agriculture University were taken as control for real-time PCR analysis. The results showed that APS1, APS2 and SMT expression levels for either young or mature leaves in selenium-enriched tea plant were lower than that in ordinary (selenium un-enriched) tea plant. In contrast, the APS1, APS2 and SMT expression level of roots in selenium-enriched tea plant were all higher than that in ordinary tea plant. APS1 gene expression level of roots in selenium-enriched tea plant was about 1.6 times higher than that in the ordinary tea plant, APS2 gene expression level was about 4.8-fold higher than that in the ordinary tea plant, SMT gene expression level was about 3.3 times higher than that in the ordinary tea plant. Among various tissues of selenium-enriched tea plant, APS1 gene relative expression level of young leaves was similar to or slightly higher than mature leaves, and the one of roots was the lowest among them; APS2 gene relative expression level of young leaves was similar to or slightly higher than the roots, and the one of mature leaves was the lowest among them; SMT gene relative expression level of young leaves was similar to or slightly higher than mature leaves, and the one of roots was the highest among them. Our results suggest that there existed correlation between selenium and expression levels of these genes.
Cite this paper: S. Tao, J. Li, X. Gu, Y. Wang, Q. Xia, B. Qin and L. Zhu, "Quantitative Analysis of ATP Sulfurylase and Selenocysteine Methyltransferase Gene Expression in Different Organs of Tea Plant (Camellia sinensis)," American Journal of Plant Sciences, Vol. 3 No. 1, 2012, pp. 51-59. doi: 10.4236/ajps.2012.31004.

[1]   N. Terry, A. M. Zayed, M. P. De Souza and A. S. Tarun, “Selenium in Higher Plants,” Annual. Review of Plant Physiology and Plant Molecular Biology, Vol. 51, No. 1, 2000, pp. 401-432. doi:10.1146/annurev.arplant.51.1.401

[2]   B. H. Ng and J. W. Anderson, “Synthesis of Selenocysteine by Cysteine Synthase from Selenium Accumulator and Non-Accumulator Plants,” Phytochemistry, Vol. 17, No. 12, 1978, pp. 2069-2074. doi:10.1016/S0031-9422(00)89282-1

[3]   D. R. Ellis and D. E. Salt, “Plants, Selenium and Human Health,” Current Opinion in Plant Biology, Vol. 6, No. 3, 2003, pp. 273-279. doi:10.1016/S1369-5266(03)00030-X

[4]   D. R. Ellis, T. G. Sors, D. G. Brunk, C. A. lbrecht, C. Orser, B. Lahner, K. V. Wood, H. H. Harris, I. J. Pickering and D. E. Salt, “Production of Se-Methylselenocysteine in Transgenic Plants Expressing Selenocysteine Methyltransferase,” BMC Plant Biology, Vol. 4, 2004, p. 1. doi:10.1186/1471-2229-4-1

[5]   B. Neuhierl, M. Thanbichler, F. Lottspeich and A. B?ck, “A Family of S-Methylmethionine-Dependent Thiol/Selenol Methyltransferases,” Journal of Biological Chemistry, Vol. 274, No. 9, 1999, pp. 5407-5414.

[6]   T. Leustek, M. Murillo and M. Cervantes, “Cloning of a cDNA Encoding ATP Sulfurylase from Arabidopsis Thaliana by Functional Expression in Sac-Charomyces Cerevisiae,” Plant Physiology, Vol. 105, No. 3, 1994, pp. 897-902.

[7]   B. Neuhierl and A. B?ck, “On the Mechanism of Selenium Tolerance in Selenium-Accumulating Plants: Puri?cation and Characterization of a Speci?c Selenocysteine Methyltransferase from Cultured Cells of Astragalus Bisulcatus,” European Journal of Biochemistry, Vol. 239, No. 1, 1996, pp. 235-238. doi:10.1111/j.1432-1033.1996.0235u.x

[8]   L. Zhu, W. W. Deng, A. H. Ye, M. Yu, Z. X. Wang and C. J. Jiang, “Cloning of Two cDNAs Encoding a Family of ATP Sulfurylasefrom Camellia Sinensis Related to Selenium or Sulfur Metabolism and Functional Expression in Escherichia Coli,” Plant Physiology and Biochemistry, Vol. 46, No. 8-9, 2008, pp.731-738. doi:10.1016/j.plaphy.2007.03.029

[9]   L. Zhu, C. J. Jiang, W. W. Deng, X. Gao, R. J. Wang and X. C. Wan, “Cloning and Expression of Selenocysteine Methyltransferase cDNA from Camellia Sinensis,” Acta Physiologiae Plantarum, Vol. 30, No. 2, 2008, pp. 167-174. doi:10.1007/s11738-007-0105-7

[10]   T. G. Sors, D. R. Ellis, G. N. Na, B. Lahner, S. Lee, T. Leustek, I. J. Pickerin and D. E. Salt, “Analysis of Sulfur and Selenium Assimilation in Astragalus Plants with Varying Capacities to Accumulate Selenium,” The Plant Journal, Vol. 42, No. 6, 2005, pp. 785-797. doi:10.1111/j.1365-313X.2005.02413.x

[11]   J. N. Qi, S. C. Yu, F. L. Zhang, X. Q. Shen, X. Y. Zhao, Y. J. Yu and D. S. Zhang, “Reference Gene Selection for Real-Time Quantitative Polymerase Chain Reaction of mRNA Transcript Levels in Chinese Cabbage (Brassica rapa L. ssp. pekinensis),” Plant Molecular Biology Reporter, Vol. 28, No. 4, 2010, pp. 597-604. doi:10.1007/s11105-010-0185-1

[12]   S. J. Vandecasteele, W. E. Peetermans, R. Merckx and J. Van Eldere, “Quantification of Expression of Staphylococcus Epidermidishousekeeping Genes with Taqman Quantitative PCR during in Vitro Growth and under Different Conditions,” Journal of Bacteriology, Vol. 183, No. 24, 2001, pp. 7094-7101. doi:10.1128/JB.183.24.7094-7101.2001

[13]   J. D. Dean, P. H. Goodwin and T. Hsiang, “Comparison of Relative RT-PCR and Northern Blot Analyses to Measure Expression of β-1,3-Glucanase in Nicotiana benthamiana Infected with Colletotrichum destructivum,” Plant Molecular Biology Reporter, Vol. 20, No. 4, 2002, pp. 347-356. doi:10.1007/BF02772122

[14]   S. A. Bustin, “Absolute Quantification of mRNA Using Real-Time Reverse Transcription Polymerase Chain Reaction Assays,” Journal of Molecular Endocrinology, Vol. 25, No. 2, 2000, pp. 169-193. doi:10.1677/jme.0.0250169

[15]   M. Orsel, A. Krapp and F. Daniel-Vedele, “Analysis of the NRT2 Nitrate Transporter Family in Arabidopsis. Structure and Gene Expression,” Plant Physiology, Vol. 129, No. 2, 2002, pp. 886-888. doi:10.1104/pp.005280

[16]   B. R. Kim, H. Y. Nam, S. U. Kim, S. I. Kim and Y. J. Chang, “Normalization of Reverse Transcription Quantitative-PCR with Housekeeping Genes in Rice,” Biotechnology Letters, Vol. 25, No. 3, 2003, pp. 1869-1872. doi:10.1023/A:1026298032009

[17]   A. Bézier, B. Lambert and F. Baillieul, “Study of Defense-Related Gene Expression in Grapevine Leaves and Berries Infected with Botrytis Cinerea,” European Journal of Plant Pathology, Vol. 108, No. 2, 2002, pp. 111-120. doi:10.1023/A:1015061108045

[18]   C. Thomas, D. Meyer, M. Wolff, C. Himber, M. Alioua and A. Steinmetz, “Molecular Characterization and Spatial Expression of the Sunflower ABP1 Gene,” Plant Molecular Biology, Vol. 52, No. 5, 2003, pp. 1025-1036. doi:10.1023/A:1025482432486

[19]   J. K. Livak and D. T. Schmittgen, “Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2–??CT Method,” Methods, Vol. 25, No. 4, 2001, pp. 402-408. doi:10.1006/meth.2001.1262

[20]   J. Logemann, J. Schell and L. Willmitzer, “Improved Method for the Isolation of RNA from Plant Tissues,” Analytical Biochemistry, Vol. 163, No. 1, 1987, pp. 16-20. doi:10.1016/0003-2697(87)90086-8

[21]   E. A. H. Pilon-Smits, S. Hwang, C. M. Lytle., Y. L. Zhu, J. C. Tai, R. C. Bravo, Y. C. Chen, T. Leustek and N. Terry, “Overexpression of ATP Sul-Furylase in Indian Mustard Leads to Increased Selenate Uptake, Reduction,and Tolerance,” Plant Physiology, Vol. 119, No. 1, 1999, pp. 123-132. doi:10.1104/pp.119.1.123

[22]   D. L. LeDuc, A. S. Tarun, M. Montes-Bayon, J. Meija, M. F. Malit, C. P. Wu, et al., “Overexpression of Selenocysteine Methyltransferase in Arabidopsis and Indian Mustard Increases Selenium Tolerance and Accumulation,” Plant Physiology, Vol. 135, No. 1, 2004, pp. 377-383. doi:10.1104/pp.103.026989

[23]   C. S. Wang and L. O. Vodkin, “Extraction of RNA from Tissues Containing High Levels of Procyanidins That Bind RNA,” Plant Molecular Biology Reporter, Vol. 12, No. 2, 1994, pp. 132-145.

[24]   S. L. Dellaporta, J. Wood and J. B. Hicks, “A Plant DNA Minipreparation,” Plant Molecular Biology Reporter, Vol. 1, No. 4,1983, pp. 19-21. doi:10.1007/BF02712670

[25]   M. Mejjad, F. Vedel and G. Ducreux, “Improvement of DNA Preparation and of PCR Cycling in RAPD Analysis of Marine Microalgae,” Plant Molecular Biology Reporter, Vol. 12, No. 2, 1994, pp. 101-105. doi:10.1007/BF02668370

[26]   J. J. Doyle and J. L. Doyle, “Isolation of Plant DNA from Fresh Tissue,” Focus, Vol. 12, No. 12, 1990, pp. 13-15

[27]   G. Fang, S. Hammar and R. Rebecca, “A Quick and Inexpensive Method for Removing Polysaccharides from Plant Genomic DNA,” Bio Techniques, Vol. 13, No. 1, 1992, pp. 52-56

[28]   M. A. Lodhi, G. N. Ye, N. F. Weeden and B. I. Reisch, “A Simple and Ef?cient Method for DNA Extraction from Grapevine Cultivars and vitis Species,” Plant Molecular Biology Reporter, Vol. 12, No. 1, 1994, pp. 6-13. doi:10.1007/BF02668658

[29]   A. B. Iandolino, F. Goes da Silva, H. Lim, H. Choi, L. E. Williams and D. R. Cook, “High-Quality RNA, cDNA, and Derived EST Libraries from Grapevine (Vitis vinifera L.),” Plant Molecular Biology Reporter, Vol. 22, No. 3, 2004, pp. 269-278. doi:10.1007/BF02773137

[30]   S. E. Denman and C. S. Mcsweeney, “Quantitative (Real-Time) PCR,” In: H. P. S. Makkar and C. S. McSweeney, Eds., Methods in Gut Microbial Ecology for Ruminants, CSIRO Livestock Industries, Queensland, 2005, pp. 105-115. doi:10.1007/1-4020-3791-0_8

[31]   M. W. Pfaf?, “A New Mathematical Model for Relative Quanti?cation in Real-Time RT-PCR,” Nucleic Acids Research, Vol. 29, No. 9, 2001, pp. 2002-2007. doi:10.1093/nar/29.9.e45

[32]   M. P. Arvy, “Selenate and Selenite Uptake and Translocation in Bean Plants (Phaseolus vulgaris),” Journal of Experimental Botany, Vol. 44, No. 6, 1993, pp. 1083-1087. doi:10.1093/jxb/44.6.1083

[33]   M. Montes-Bayón, D. L. LeDuc, N. Terry and J. A. Caruso, “Selenium Speciation in Wild-Type and Genetically Modified Se Accumulating Plants with HPLC Separation and ICP-MS/ES-MS Detection,” Journal of Analytical Atomic Spectrometry, Vol. 17, No. 8, 2002, pp. 872-879.

[34]   P. Cartes, L. Gianfreda and M. L. Mora, “Uptake of Selenium and Its Antioxidant Activity in Ryegrass When Applied as Selenate and Selenite Forms,” Plant and Soil, Vol. 276, No. 1-2, 2005, pp. 359-367. doi:10.1007/s11104-005-5691-9

[35]   T. Leustek, M. N. Martin, J. A. Bick and J. P. Davies, “Pathways and Regulation of Sulfur Metabolism Revealed through Molecular and Genetic Studies,” Annual Review of Plant Physiology and Plant Molecular Biology, Vol. 51, 2000, pp. 141-165. doi:10.1146/annurev.arplant.51.1.141

[36]   J. W. Anderson and A. R. Scarf, “Selenium and Plant Metabolism,” In: D. A. Robb and W. S. Pierpoint, Eds., Metals and Micro Nutrients: Uptake and Utilization by Plants, Academic, London, 1983, pp. 241-275.

[37]   G. L. Dilworth and R. S. Bandurski, “Activation of Selenate by Adenosine 5’-Triphosphate Sulfurylase from Saccharmoyces Cereviseae,” Biochemical Journal, Vol. 163, No. 3, 1977, pp. 521-529.

[38]   T. J. McCluskey, A. R. Scarf and W. Anderson, “Enzyme-Catalyzed αβ-Elimination of Selenocystathionine and Selenocystine and Their Sulfur Isologues by Plant Extracts,” Phytochemistry, Vol. 25, No. 9, 1986, pp. 2063-2068. doi:10.1016/0031-9422(86)80067-X

[39]   Y. Gavel and G. V. Heijne, “A Conserved Cleavage-Site Motif in Chloroplast Transit Peptides,” FEBS Letters, Vol. 261, No. 2, 1990, pp. 455-458. doi:10.1016/0014-5793(90)80614-O

[40]   F. Renosto, H. C. Patel, R. L. Martin, C. Thomassian, G. Zimmerman and I. H. Segel, “ATP Sulfurylase from Higher Plants: Kinetic and Structural Characterization of the Chloroplast and Cytosol Enzymes from Spinach Leaf,” Archives of Biochemistry, Biophysics, Vol. 307, No. 2, 1993, pp. 272-285. doi:10.1006/abbi.1993.1590

[41]   A. Zayed, C. M. Lytle and N. Terry, “Accumulation and Volatilization of Different Chemical Species of Selenium by Plants,” Planta, Vol. 206, No. 2, 1998, pp. 284-292. doi:10.1007/s004250050402

[42]   D. C. Eustice, I. Foster and F. J. Kull, “In Vitro Incorporation of Selenomethionine into Protein by Vigna Radiata Polysomes,” Plant Physiology, Vol. 66, No. 1, 1980, pp. 182-186. doi:10.1104/pp.66.1.182