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Proline and Betaine Upregulated Glutathione Dependent Detoxification Enzymes in Tolerant Maize Seedlings under Saline Stress
Journal of Plant Sciences
Volume 3, Issue 6, December 2015, Pages: 294-302
Received: Sep. 27, 2015; Accepted: Oct. 26, 2015; Published: Nov. 17, 2015
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Md. Motiar Rohman, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
M. R. Molla, Molecular Biology Lab, Plant Genetic Resources Centre, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
Md. Mahfuzur Rahman, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
Asgar Ahmed, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
Arindam Biswas, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh; RARS, Bangladesh Agricultural Research Institute, Jessore, Bangladesh
Mohammad Amiruzzaman, Molecular Biology Lab, Plant Genetic Resources Centre, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
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Glutathione S-transferases (GSTs, EC: and Glyoxalases [Glyoxalase-I (Gly-I, EC: and glyoxalase-II (Gly-II, EC:] are major glutathione dependent detoxification as well as important antioxidants enzymes in plants. On the other hand, proline and betaine are important osmoprotectants in plants under abiotic stresses including salinity. In this study, roles of GST, Gly-I, Gly-II and glutathione were investigated on cytotoxic metabolites in presence of 15 mM proline and betaine under 16 dSm-1 salinity in leaves of seedlings of a saline tolerant maize inbred CZ-10 to understand the underlying saline tolerant mechanism. The salinity stress increased the contents of H2O2, melondialdehyde (MDA), methylglyoxal (MG) along with decreased reduced glutathione (GSH) and glutathione redox state over control. The activities of GST and Gly-I increased under saline stress. However, activity of Gly-II decreased with stress duration. The application of proline and betaine in saline water reduced the contents of H2O2, MDA and MG. Conversely, proline and betaine increased the activities of GST, Gly-I and Gly-II, and GSH and glutathione-redox state over salinity stress. The western blotting of the soluble protein also suggested the accumulation of maize GST in leaf under salinity stress. The accumulation of GST along with reduced contents of H2O2 and MDA suggested its detoxification roles on organic hydroperoxides under saline stress. The higher activities of Gly-I and Gly-II concurrently with lower content of MG indicated their protective roles from cytotoxic MG. Considering all, this study concluded that both proline and betaine provided protective roles in maize seedlings under salinity stress by maintaining GSH and its related detoxification enzymes.
Maize, Salinity, Glutathione S-transferase, Glyoxalases, Protective Role, Proline, Betaine
To cite this article
Md. Motiar Rohman, M. R. Molla, Md. Mahfuzur Rahman, Asgar Ahmed, Arindam Biswas, Mohammad Amiruzzaman, Proline and Betaine Upregulated Glutathione Dependent Detoxification Enzymes in Tolerant Maize Seedlings under Saline Stress, Journal of Plant Sciences. Vol. 3, No. 6, 2015, pp. 294-302. doi: 10.11648/j.jps.20150306.12
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Demiral T, Türkan I. 2005. Comparative lipid peroxidation, antioxidant defense systems and proline content in roots of two rice cultivars differing in salt tolerance. Environ. Exp. Botany 53 (3): 247-257.
Mandhania S, Madan S, Sawhney V. 2006. Antioxidant defense mechanism under salt stress in wheat seedlings. Biol. Plant 50 (2): 227-231.
Misra N, Gupta AK. 2006. Interactive effects of sodium and calcium on proline metabolism in salt tolerant green gram cultivar. Am. J. Plant Physiol 1 (1): 1-12.
Hasegawa PM, Bressan RA, Zhu JK, Bohnert, HJ. 2000. Plant cellular and molecular responses to high salinity. Annu. Rev. Plant Physiol. Plant Mol. Biol 51: 463-499.
Apel K, Hirt H. 2004. Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu. Rev. Plant. Biol 55: 373-399.
Yadav SK, Singla-Pareek SL, Reddy MK, Sopory SK. 2005a. Transgenic tobacco plants overexpressing glyoxalase enzymes resist an increase in methylglyoxal and maintain higher reduced glutathione levels under salinity stress. FEBS Lett 579 (27): 6265-6271.
Yadav SK, Singla-Pareek SL, Ray M, Reddy MK, Sopory SK. 2005b. Methylglyoxal levels in plants under salinity stress are dependent on glyoxalase I and glutathione. Biochem. Biophys. Res. Commun 337 (1): 61-67.
Noctor G, Mhamdi A, Chaouch S, Han Y, Neukermans J, Marquez-Garcia B, Queval G, Foyer CH. 2012. Glutathione in plants: an integrated overview. Plant Cell Environ 35 (2): 454-484.
Noctor G, Foyer CH. 1998. Ascorbate and glutathione: keeping active oxygen under control. Annu. Rev. Plant Physiol. Plant Mol. Biol 49: 249-79.
McNeil SD, Nuccio ML, Hanson AD. 1999. Betaines and related osmoprotectants. Targets for metabolic engineering of stress resistance. Plant Physiol 120 (4): 945-949.
Okuma E, Soeda K, Tada M, Murata Y. 2000. Exogenous proline mitigates the inhibition of growth of Nicotiana tabacum cultured cells under saline conditions. Soil Sci. Plant Nutr 46 (1): 257-263.
Ashraf M and Foolad MR. 2007. Roles of glycinebetaine and proline in improving plant abiotic resistance. Environ. Exp. Bot 59: 206-16.
Okuma E, Murakami Y, Shimoishi Y, Tada M, Murata Y. 2004. Effects of exogenous application of proline and betaine on the growth of tobacco cultured cells under saline conditions. Soil Sci. Plant Nutr 50 (8): 301-1305.
Hasanuzzaman M, Alam MM, Rahman A, Hasanuzzaman M, Nahar K, Fujita M. 2014. Exogenous proline and glycine betaine mediated upregulation of antioxidant defense and glyoxalase systems provides better protection against salt-induced oxidative stress in two rice (Oryza sativa L.) varieties. Bio Med Res Intl. Vol. 2014, Article ID: 757219, doi:10.1155/2014/757219.
Choudhury S, Panda P, Sahoo L, Panda SK. 2013. Reactive oxygen species signaling in plants under abiotic stress. Plant Signal. Behav 8(4): e23681.
Edwards R, Dixon DP. 2005. Plant glutathione transferases. In Glutathione Transferases and Gamma-Glutamyl Transpeptidases (Sies, H. and Packer, L., eds). San Diego, CA: Elsevier Academic Press Inc, pp. 81169-186.
Dixon DP, Steel PG, Edwards R. 2011. Roles for glutathione transferases in antioxidant recycling, Plant Signaling & Behavior 6 (8): 1223-1227.
Chi Y, Cheng Y, Vanitha J, Kumar N, Ramamoorthy R, Ramachandran S, Jiang SY. 2011. Expansion mechanisms and functional divergence of the glutathione S-transferase family in Sorghum and other higher plants. DNA Res 18 (1):1-16.
Chen JH, Jiang HW, Hsieh EJ, Chen HY, Chien CT, Hsieh HL, Lin TP. 2012. Drought and salt stress tolerance of an Arabidopsis glutathione S-transferase U17 knockout mutant are attributed to the combined effect of glutathione and abscisic acid. Plant Physiol 158 (1): 340-351.
Zagorchev L, Seal CE, Kranner I, Odjakova M. 2013. A Central role for thiols in plant tolerance to abiotic stress. Int. J. Mol. Sci 14: 7405-7432.
Marrs KA, Alfenito MR, Lloyd AM, Walbot V. 1995. A glutathione S-transferase involved in vacuolar transfer encoded by the maize gene Bronze-2. Nature 375 (6530): 397-400.
Mueller LA, Goodman CD, Silady RA, Walbot V. 2000. AN9, a petunia glutathione S-transferase required for anthocyanin sequestration, is a flavonoid-binding protein. Plant Physiol 123 (4): 1561-1570.
Kitamura S, Shikazono N, Tanaka A. 2004. TRANSPARENT TESTA 19 is involved in the accumulation of both anthocyanins and proanthocyanidins in Arabidopsis. Plant J 37 (1): 104-114.
Alfenito MR, Souer E, Goodman CD, Buell R, Mol J, Koes R, Walbot V. 1998. Functional complementation of anthocyanin sequestration in the vacuole by widely divergent glutathione S-transferases. Plant Cell 10 (7): 1135-1149.
Edwards R, Dixon DP, Walbot V. 2000. Plant glutathione S-transferases: enzymes with multiple functions in sickness and in health. Trends Plant Sci 5: 193-198.
Mustafiz A, Sahoo KK, Singla-Pareek SL, Sopory SK. 2010. Metabolic engineering of glyoxalase pathway for enhancing stress tolerance in plants. Methods Mol. Biol 639: 95-118.
Singla-Pareek SL, Yadav SK, Pareek A, Reddy MK, Sopory SK. 2008. Enhancing salt tolerance in a crop plant by overexpression of glyoxalase II. Transgenic Res 17 (2): 171-180.
Saxena M, Bisht R, Roy SD, Sopory SK, Bhalla-Sarin N. 2005. Cloning and characterization of a mitochondrial glyoxalase II from Brassica juncea that is upregulated by NaCl, Zn and ABA. Biochem. Biophys. Res. Commun 336 (3): 813-819.
Saxena M, Deb Roy S, Singla-Pareek S-L, Sopory SK, Bhalla-Sarin N. 2011. Overexpression of the glyoxalase II gene leads to enhanced salinity tolerance in Brassica juncea. The Open Plant Sci. J 5: 23-28.
Hoque MA, Okuma E, Banu MNA, Nakamura Y, Shimoishi Y, Murata Y. 2007a. Exogenous proline mitigates the detrimental effects of salt stress more than exogenous betaine by increasing antioxidant enzyme activities. J. Plant Physiol 164 (5): 553-61.
Hoque MA, Banu MNA, Okuma E, Amako K, Nakamura Y, Shimoishi Y, Murata Y. 2007b. Exogenous proline and glycinebetaine ingresses NaCl-induced ascorbate-glutathione cycle enzyme activities and proline improves salt tolerance more than glycinebetaine in tobacco Bright yellow-2 suspension-cultured cells. J. Plant Physiol 164 (11): 1457-1468.
Hoque MA, Banu MNA, Nakamura Y, Shimoishi Y, Murata Y. 2008. Proline and glycinebetaine enhance antioxidant defense and methylglyoxal detoxification systems and reduce NaCl-induced damage in cultured tobacco cells. J. Plant Physiol 165 (8): 813-824.
Rohman MM, Uddin S, Fujita M. 2010. Up-regulation of onion bulb glutathione S-transferases (GSTs) by abiotic stresses: A comparative study between two differently sensitive GSTs to their physiological inhibitors. Plant Omics 3 (1): 28-34.
Yu CW, Murphy TM, Lin CH. 2003. Hydrogen peroxide-induces chilling tolerance in mung beans mediated through ABA-independent glutathione accumulation. Funct. Plant Biol 30 (9): 955-963.
Heath RL, Packer L. 1968. Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch. Biochem. Biophys 125 (1): 189-198.
Wild R, Ooi L, Srikanth V, Münch G. 2012. A quick, convenient and economical method for the reliable determination of methylglyoxal in millimolar concentrations: the N-acetyl-L-cysteine assay. Anal. Bioanal. Chem 403 (9): 2577-2581.
Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem 72: 248-254.
Laemmli, UK. 1970. Cleavage of structural proteins during the assembly of head of bacteriophage T4. Nature 227 (5259): 680-685.
Edwards R, Dixon DP. 2000. The role of glutathione transferases in herbicide metabolism. In Herbicides and Their Mechanisms of Action (Cobb, A.H. and Kirkwood, R.C., eds). Sheffield: Sheffield Academic Press Ltd, pp. 8138-71.
Fini A, Brunetti C, Ferdinando MD, Ferrini F, Tattini. 2011. Stress-induced flavonoid biosynthesis and the antioxidant machinery of plants. Plant Signal. Behav 6 (5): 709-711.
Rohman MM, Hossain MD, Suzuki T, Takada G, Fujita M. 2009. Quercetin-4'-glucoside: a physiological inhibitor of the activities of dominant glutathione S-transferases in onion (Allium cepa L.) bulb. Acta. Physiol. Plant 31 (2): 301-309.
Seppanen MM, Cardi T, Hyokki MB, Pehu E. 2000. Characterisation and expression of cold-induced glutathione S-transferase in freezing tolerant Solanum commersonii, sensitive S. tuberosum and their interspecific somatic hybrids. Plant Sci 153 (2): 125-133.
Moons, A. 2003. Osgstu3 and osgtu4, encoding tau class glutathione S-transferases, are heavy metal- and hypoxic stress-induced and differentially salt stress-responsive in rice roots. FEBS Lett 553 (3): 427-432.
Kiyosue T, Yamaguchi-Shinozaki K, Shinozaki K. 1993. Characterization of two cDNAs (ERD11 and ERD13) for hydration-inducible genes that encode putative glutathione S-transferases in Arabidopsis thaliana L. FEBS Lett 335 (2): 189-192.
Bianchi MW, Roux C, Vartanian N. 2002. Drought regulation of GST8, encoding the Arabidopsis homologue of ParC/Nt107 glutathione transferase/peroxidase. Physiol. Plant. 116 (1): 96-105.
Vollenweider S, Weber H, Stolz S, Chetelat A, Farmer EE. 2000. Fatty acid ketodienes and fatty acid ketotrienes: Michael addition acceptors that accumulate in wounded and diseased Arabidopsis leaves. Plant J 24 (4): 467-476.
Mauch F, Dudler R. 1993. Differential induction of distinct glutathione-S-transferases of wheat by xenobiotics and by pathogen attack. Plant Physiol 102 (4): 1193-1201.
Zhou J, Goldsbrough PB. 1993. An Arabidopsis gene with homology to glutathione S-transferases is regulated by ethylene. Plant Mol. Biol 22: 517-523.
Chen W, Chao G, Singh KB. 1996. The promoter of a H2O2 inducible, Arabidopsis glutathione S-transferase gene contains closely linked OBF- and OBP1-binding sites. Plant J 10 (6): 955-966.
Gronwald JW, Plaisance KL. 1998. Isolation and characterization of glutathione S-transferase isozymes from sorghum. Plant Physiol. 117 (3): 877-892.
Marrs KA. 1996. The functions and regulation of glutathione S-transferases in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol 47: 127-158.
Chattopadhayay MK, Tiwari BS, Chattopadhyay G, Bose A, Sengupta DN, Ghosh B. 2002. Protective role of exogenous polyamines on salinity-stressed rice (Oryza sativa) plants. Physiol. Plant 116 (2): 192-199.
Bor M, Ozdemir F, Turkan I. 2003. The effect of salt stress on lipid peroxidation and antioxidants in leaves of suger beet Beta vulgaris L. and Beta maritime L. Plant Sci 164 (1): 77-84.
Banu MNA, Hoque MA, Watanble-Sugimoto M, Masuoka K, Nakamura Y and Shimoishi Y. 2009. Proline and glycinebetaine induce antioxidant defense gene expression and suppress cell death in cultured tobacco cells under salt stress. J. Plant Physiol 166 (2): 146-156.
Park EJ, Jeknic Z, Chen THH. 2006. Exogenous application of glycinebetaine increases chilling tolerance in tomato plants. Plant Cell Physiol 47 (6): 706-714.
Smirnoff N. 2005. Antioxidants and reactive oxygen species in plants. 1st ed. Oxford: Blackwell Publishing Ltd. 302 p.
May MJ, Leaver CJ.1993. Oxidative stimulation of glutathione synthesis in Arabidopsis thaliana suspension cultures. Plant Physiol 103: 621-7.
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