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Molecular Characterization of Free Radicals Decomposing Genes on Plant Developmental Stages

Authors: R. Haddad, K. Morris, V. Buchanan-Wollaston


Biochemical and molecular analysis of some antioxidant enzyme genes revealed different level of gene expression on oilseed (Brassica napus). For molecular and biochemical analysis, leaf tissues were harvested from plants at eight different developmental stages, from young to senescence. The levels of total protein and chlorophyll were increased during maturity stages of plant, while these were decreased during the last stages of plant growth. Structural analysis (nucleotide and deduced amino acid sequence, and phylogenic tree) of a complementary DNA revealed a high level of similarity for a family of Catalase genes. The expression of the gene encoded by different Catalase isoforms was assessed during different plant growth phase. No significant difference between samples was observed, when Catalase activity was statistically analyzed at different developmental stages. EST analysis exhibited different transcripts levels for a number of other relevant antioxidant genes (different isoforms of SOD and glutathione). The high level of transcription of these genes at senescence stages was indicated that these genes are senescenceinduced genes.

Keywords: Biochemical analysis, Oilseed, Expression pattern, Growth phases

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[1] M. Abler, and J. G. Scandalios. "Isolation and characterization of a genomic sequence encoding the maize Cat3 catalase gene," Plant Molecular Biology 22: (6) 1031-1038, 1993.
[2] A. Ádám, G. S. Bestwick, B. Barna, and J. W. Mansfield. "Enzymes regulating the accumulation of active oxygen species during the hypersensitive reaction of bean to pseudomonas-syringae pv phaseolicola," Planta 197: 240-249, 1995.
[3] H. Aebi. "Catalase in vitro," Methods in Enzymology 105: 121-126, 1984.
[4] C. Ainsworth, J. Beynon, and V. Buchanan-Wollaston. "Techniques in plant molecular biology (The practical mannual)," Wye college, University of London, 1995.
[5] C. Bailly, F. Corbineau, and W. G. van Doorn. "Free radical scavenging and senescence in Iris tepals," Plant Physiology and Biochemistry 39: 649-656, 2001.
[6] M. Bradford. "A rapid and sensitive method for the quantification of micrograms quantities of protein utilising the principle of protein-dye binding," Annals of Biochemistry 72: 248-254, 1976.
[7] V. Buchanan-Wollaston, and C. Ainsworth. "Leaf senescence in Brassica napus: Cloning of senescence related genes by subtractive hybridization," Plant Molecular Biology 33: 821-834, 1997.
[8] V. Buchanan-Wollaston. "Isolation of cDNA clones for genes that are expressed during leaf senescence in Brassica napus," Plant Physiology 105: 839-46, 1994.
[9] V. Buchanan-Wollaston. "The molecular biology of leaf senescence," Journal of Experimental Botany 48: 181-199, 1997.
[10] C. Chevalier, J. Yamaguchi, and P. Mccourt. "Nucleotide sequence of a cDNA for catalase from Arabidopsis thaliana," Plant Physiology 99: 1726-1728, 1992.
[11] M. Dalal; and K. R. Chopra. "Differential response of antioxidant enzymes in leaves of necrotic wheat hybrids and their parents," Physiologia Plantarum 111: 297-304, 2001.
[12] L. De Kok, and I. Stulen. "Role of glutathione in plants under oxidative stress. In LJ De Kok, I Stulen, H Rennenberg, C Brunold, WE Rauser, (eds), Sulfur nutrition and assimilation in higher plants: regulatory agricultural and environmental aspects. SPB Academic Publishing, the Hague, The Netherlands," PP. 125-138, 1993.
[13] T. Demiral, and I. T├╝rkan. "Comparative lipid peroxidation, antioxidant defense systems and proline content in roots of two rice cultivars differing in salt tolerance," Environmental and Experimental Botany 53: 247-257, 2005.
[14] M. Esaka, N. Yamada, M. Kitabayashi, Y. Setoguchi, R. Tsugeki, M. Kondo, and M. Nishimura. "cDNA cloning and differential gene expression of three catalases in pumpkin," Plant Molecular Biology 33: 141-155, 1997.
[15] M. Esaka. "Cat2-Pumpkin, Direct submission to the DDBJ/EMBL/GenBank databases," Hiroshima University, Faculty of Applied Biological Science, Kagamiyama 1-4-4, Higashi-Hiroshima, 739, Japan, 1995a.
[16] M. Esaka. "Cat3-Pumpkin, Direct submission to the DDBJ/EMBL/GenBank databases," Hiroshima University, Faculty of Applied Biological Science, Kagamiyama 1-4-4, Higashi-Hiroshima, 739, Japan, 1995b.
[17] J. Frugoli, M. A. McPeek, T. L. Thomas, and C. R. McClung. "Intron loss and gain during evolution of the catalase gene family in Angiosperms," Genetics 149: (1) 355-365, 1998.
[18] J. Frugoli, H. H. Zhong, M. L. Nuccio, P. McCourt, M. A. McPeek, T. L. Thomas, and C. R. McClung. "Catalase is encoded by a multigene family in Arabidopsis thaliana (L) Heynh," Plant Physiology 112: (1) 327-336, 1996.
[19] L. Guan, S. Ruzsa, R. W. Skadsen, and J. G. Scandalios. "Comparison of the Cat2 complementary -DNA sequences of a normal catalase activity line (w64a) and a high catalase activity line (R6- 67) of maize," Plant Physiology 96: (4) 1379-1381, 1991.
[20] R. Haddad, K. Morris K and V. Buchanan-Wollaston. "Expression analysis of genes related to oxidative protection during senescence in Brassica napus," Iranian Journal of biotechnology 2(4): 269-278, 2004.
[21] R. Haddad, K. Morris, and V. Buchanan-Wollaston. "Characterisation and functional analysis of senescence-enhanced genes in Brassica napus and Arabidopsis thaliana," PhD Thesis, Wye College, University of London, 1999.
[22] C. Hill, S. A. Pearson, A. J. Smith, and L. J. Rogers. "Inhibition of chlorophyll synthesis in Hordeum vulgare by 2,3 dihydrobenzoic acid (gabaculin)," Bioscience Reports 5: 775-781,
[23] S. Jebara, M. Jebara, F. Limam, and M. E. Aouani. "Changes in ascorbate peroxidase, catalase, guaiacol peroxidase and superoxide dismutase activities in common bean (Phaseolus vulgaris) nodules under salt stress," Journal of Plant Physiology 162: (8) 929-936, 2005.
[24] S. Jung. "Effect of chlorophyll reduction in Arabidopsis thaliana by methyl jasmonate or norflurazon on antioxidant systems," Plant Physiology and Biochemistry 42: (3) 225-231, 2004.
[25] J. S. Kim, B. W. Yun, J. S. Choi, T. J. Kim, S. S. Kwak, and K. Y. Cho. "Death mechanisms caused by carotenoid biosynthesis inhibitors in green and in undeveloped plant tissues," Pesticide Biochemistry and Physiology 78: 127-139, 2004.
[26] B. Lewin. "Gene VI," Oxford University Press and Cell Press, 1997.
[27] O. Loew. "Catalase: A new enzyme of general occurrence," Government printing office, Washington, DC, 1901.
[28] S. Lortz, and M. Tiedge. "Sequential inactivation of reactive oxygen species by combined overexpression of SOD isoforms and catalase in insulin-producing cells," Free Radical Biology and Medicine 34: 683- 688, 2003.
[29] H. Luck. "Catalase. In: Bergmeyer HU (ed.): Methods for enzymatic analysis," Academic Press, NY, PP. 885-894, 1965.
[30] H. Nam. "The molecular genetic analysis of leaf senescence," Current Opinion in Biotechnology 8: (2) 200-207, 1997.
[31] A. S. Nandwal, S. Kukreja, N. Kumar, P. K. Sharma, M. Jain, A. Mann, and S. Singh. "Plant water status, ethylene evolution, N2-fixing efficiency, antioxidant activity and lipid peroxidation in Cicer arietinum L. nodules as affected by short-term salinization and desalinization," Journal of Plant Physiology 164: (9) 1161-1169, 2007.
[32] K. Ogawa, S. Kanematsu, and K. Asada. "Generation of superoxide anion and localization of Cu,Zn- superoxide dismutase in the vascular tissue of spinach hypocotyls: Their association with lignification," Plant and Cell Physiology 38: 1118-1126, 1997.
[33] A. K. Parida, A. B. Das, and P. Mohanty. "Defense potentials to NaCl in a mangrove, Bruguiera parviflora: Differential changes of isoforms of some antioxidative enzymes," Journal of Plant Physiology 161: 531-542, 2004.
[34] M. Redinbaugh, G. J. Wadsworth, and J. G. Scandalios. "Characterization of catalase transcripts and their differential expression in maize," Biochimica et Biophysica Acta 951: (1) 104-116, 1988.
[35] S. Reid S, and G. S. Ross. "Up-regulation of two cDNA clones encoding metallothionein-like proteins in apple fruit during cool storage," Physiologia Plantarum 100: (1) 183-189, 1997.
[36] J. Scandalios. "The antioxidant enzyme genes Cat and SOD of maize - regulation, functional-significance, and molecular-biology," Isozymes- Current Topics in Biological and Medical Research 14: 19-44, 1987.
[37] J. Sumner, and A. L. Dounce. "Crystalline catalase," Journal of Biological Chemistry 121: 417-424, 1937.
[38] R. Trelease. "Cotton Catalase, Direct submission to the DDBJ/EMBL/GenBank databases," Arizona State University, Department of Botany, Temp. Arizona 85287-1601, USA, 1990.
[39] H. Willekens, R. Villarroel, M. Vanmontagu, D. Inze, and W. Vancamp. "Molecular-identification of catalases from Nicotiana- plumbaginifolia (L)," Febs Letters 352: 79-83, 1994.
[40] H. Willekens. Direct submission to the DDBJ/EMBL/GenBank databases, Universiteit Gent, Laboratoratorium Genetika, K.L. Ledeganckstraat 35, Gent, Belgium, B 9000, 1994.
[41] M. Zenk. "Heavy metal detoxification in higher plants - A review," Gene 179: 21-30, 1996.
[42] Y. Zhang, W. Guo, S. Chen, L. Han, and Z. Li. "The role of Nlauroylethanolamine in the regulation of senescence of cut carnations (Dianthus caryophyllus)," Journal of Plant Physiology 164: (8) 993- 1001, 2007.