Glutathione S-transferases (GSTs; EC 2.5.1.18) are abundant proteins encoded by a highly divergent ancient gene family with protective functions through detoxification and non-catalytic roles as carriers of cytotoxins. In this work, we investigated the GST activities in seedlings of 38 crops to obtain a plant with high active GST for further study. In screening of 38 crops, onion seedling showed the highest GST activity (483.54 nmol min-1mg-1 protein) followed by wheat(372.89 nmol min-1mg-1 protein), barley (253.44 nmol min-1mg-1 protein), rice (244.12 nmol min-1mg-1 protein) and proso millet (173.34 nmol min-1mg-1 protein). Carrot seedling showed the lowest activity (3.63 nmol min-1mg-1 protein).In onion plants, both root and leaf showed high GST activity. Onion bulb GSTs were separated DEAE cellulose column chromatography, and purified by affinity chromatography (S-hexyl glutathione-agarose). Three GST peaks were found to elute at 67, 107 and 140mM of KCl gradient solution, and were named as GSTa, GSTb and GSTc. Among the three GSTs, GSTa, GSTb and GSTc contained 9.58, 61.45 and 28.97% of total activity, respectively. In purification, GSTa, GSTb and GSTc had specific activities of 9075, 17259 and 19868 nmol min-1 mg-1 protein, respectively, along with yield of 2.48, 3.17 and 1.28,and purification fold of 15.8, 29.9 and 34.5, respectively. The purity and molecular mass of the fraction was examined by SDS-PAGE. The silver staining of the purified GSTa, GSTb and GSTc indicated that final product of GSTb and GSTc were highly purified and migrated as a single band on SDS-PAGE with an apparent molecular mass of 27 kDa. However, GSTa eluted with Glyoxalase-I (Gly-I) and to purify GSTa, more methodological application are suggested.
| Published in | Journal of Plant Sciences (Volume 3, Issue 3) |
| DOI | 10.11648/j.jps.20150303.17 |
| Page(s) | 162-170 |
| Creative Commons |
This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited. |
| Copyright |
Copyright © The Author(s), 2015. Published by Science Publishing Group |
Comparative GST, Crops, Purification, High Active GST, Onion
| [1] | Edwards R, Dixon PD, Walbat V. 2000. Plant glutathione S-transferases: enzymes with multiple functions in sickness and inhealth. Trends Plant Sci 5: 193-198. |
| [2] | Dean VJ, Devarnne PT, Ik-Soo L, Orlofsky EL. Properties of a maize glutathione S-transferase that conjugates coumaric acid and other phenylpropanoids. 2005. Plant Physiol 108: 985-994. |
| [3] | Bilang K, Widersten M, Engström A, Kozarich JH, Mannervik B. 1993. Detoxification of base propenals and other α,β-unsaturated aldehyde products of radical reactions and lipid peroxidation by human glutathione transferases. Proc. Natl. Acad. Sci. USA 91:1480-1484. |
| [4] | Gonneau J, Mornet R, Laloue AM. 1998. Nicotiana plumbaginifolia protein labeled with an azidocytokininagonist is a glutathione S-transferase. Physiol Plant 103: 114-124. |
| [5] | Loyall L, Uchida Braun KS, Furuya M, Frohnmeyer H. 2000. Glutathione and a UV light-induced glutathione S-transferase are involved in signaling to chalcone synthase in cell cultures. Plant Cell 12: 1939-1950. |
| [6] | Dixon PD, Lapthorn A, Edwards R. 2002. Plant glutathione transferases. Genome Biol 3(3): 1-10. |
| [7] | Mohsenzadeh S, Esmaeili M, Moosavi F, Saffariand B, Mohabatkar H. 2011. Plant glutathione S-transferase classification, structure and evolution. African Journal Biotechnol 10(42): 8160-8165. |
| [8] | Müller 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: 1561-1570. |
| [9] | Noctor G, Mhamdi A, Chaouch S, Han Y, Neukermans J, Marquez¬ B, Queval G,. Foyer C H. 2012. Glutathione in plants: an integrated overview. Plant Cell Environ 35: 454¬-484. |
| [10] | Blokhina KJM, Van der Zaal BJ, Velterop J, Quint AB, Mennes AM, Hooykaas PJ, Libbenga KR. 2003. Further characterization of expression of auxin-induced genes in tobacco (Nicotiana tabacum) cell-suspension cultures. Plant Physiol 102: 513-520. |
| [11] | Rohman MM, Hossain MD, Suzuki T, Takada G, Fujita M. 2009. Quercetin-4'-glucoside: aphysiological inhibitor of the activities of dominant glutathione S-transferases in onion (Allium cepa L.)bulb.” Acta Physiol. Plant 31: 301-309. |
| [12] | Hossain MA, Fujita M. 2009. Purification of Glyoxalase I from onion bulbs and molecular cloning of its cDNA. Biosci. Biotechnol. Biochem 73 (9): 2007-2013. |
| [13] | 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. |
| [14] | Laemmli UK. 1970. Cleavage of structural proteins during the assembly of head of bacteriophage T4. Nature 227: 680-685. |
| [15] | Dixon PD, Davis GB, Edwards R. 2002. Functional divergence in the glutathione transferase superfamily in plants. J Biol. Chem 277: 30859-30869. |
| [16] | Frova C. 2003. The plant glutathione transferase gene family: genomic structure, functions, expression and evolution.” Physiol.Plantarum 119(4): 469-479. |
| [17] | Cummins I, Cole JD, Edwards R. 1999. A role for glutathione transferases functioning as glutathione peroxidasesin resistance to multiple herbicides in black-grass. Plant Journal 18: 285-292. |
| [18] | Kunieda T, Fujiwara T, Amano T, Sioihi Y. 2005. Molecular cloning and characterization of a senescence-induced tau-class glutathione S-transferase from barley leaves. Plant Cell Physiol 46: 1540-1548. |
| [19] | Levine A, Tenhaken R, Dixon R, Lamb C. 1994. H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell 79: 583-593. |
| [20] | Kim SG, Kim ST, Kang SY, Wang Y, Kim W, Kang KY. 2008. Proteomic analysis of reactive oxygen species (ROS)-related proteins in rice roots. Plant Cell Rep 27: 363-375. |
| [21] | Kiyosue T, Yamaguchi-Shinozaki K, Shinozaki K, 1993. Characterization of two cDNAs (ERD11 and ERD13) for dehydration inducible genes that encode putative glutathione S-transferases in Arabidopsis thaliana L. FEBS Lett 335: 189-192. |
| [22] | Liu X, Deng Z, Gao S, Sun X, Tang K. 2007. Molecular cloning and characterization of a glutathione S-transferase gene from Ginkgo biloba. DNA Seq 18: 371-379. |
| [23] | Anderson JV, Davis DG. 2004. Abiotic stress alters transcript profiles and activity of glutathione S-transferase, glutathione peroxidase, and glutathione reductase in Euphorbia esula. Physiol. Plant 120: 421-433. |
| [24] | Nutricati E, Miceli A, Blando F, Bellis De L. 2006. Characterization of two Arabidopsis thaliana glutathione S-transferases. Plant Cell Rep 25: 997-1005. |
| [25] | Ezaki B, Yamamoto Y, Matsumoto H. 1995. Cloning and sequencing of the cDNAs induced by aluminum treatment and Pi starvation in cultured tobacco cells. Physiol. Plant 93: 11-18. |
| [26] | Sharma KY, Davis RK. 1994. Ozone-induced expression of stress related genes in Arabidopsis thaliana. Plant Physiol 105: 1089-1096. |
| [27] | Ezaki B, Suzuki M, Motoda H, Kawamura M, Nakashima S, Matsumoto H. 2004. Mechanism of gene expression of Arabidopsis glutathione S-transferase, AtGST1, and AtGST11 in response to aluminum stress. Plant Physiol 134: 1672-1682. |
| [28] | Marrs AK. 1996. The functions and regulation of glutathione S-transferasesinplants. Annu. Rev. Plant Physiol. Plant Mol. Biol 47: 127-158. |
| [29] | Zhou J, Goldsbrough BP. 1993. An Arabidopsis gene with homology to glutathione S-transferase is regulated by ethylene. Plant Mol. Biol 22: 517-523. |
| [30] | Takahashi Y, Nagata T. 1992. parB: an auxin-regulated gene encoding glutathione S-transferase. Proc Natl. Acad. Sci. USA 89: 56-59. |
| [31] | Wagner U, Edwards R, Dixon PD, Mauch F. 2002. Probing the diversity of the Arabidopsis glutathione S-transferase gene family. Plant Mol. Biol 49: 515-532. |
| [32] | Xu F, Lagudah SE, Moose PS, Riechers ED. 2002. Tandemlyduplicated safener-induced glutathione S-transferase genes from Triticum tauschii contribute to genome- and organ-specific expression inhexaploid wheat. Plant Physiol 130: 362-373. |
| [33] | Chen J-H, Jiang H-W, Hsieh E-J, Chen H-Y, Chien C-T, Hsieh H-L, Lin T-P. 2012. Drought and salt stress tolerance of Arabidopsis glutathione S-transferase U17 knockout mutant are attributed to the combined effect of glutathione and abscisic acid. Plant Physiol158(1): 340-351. |
| [34] | Liu D. Liu Y, Rao J, Wang G, Li H, Ge F, Chen C. 2013. Over expression of the glutathione S-transferase gene from Pyrus pyrifolia fruit improves tolerance to abiotic stress in transgenic tobacco.” Plant Mol. Biol 47(4): 515–523. |
| [35] | Kwon S-H, Kwon H-K, Kim W, Noh WE, Kwon M, Choi YI. 2014. Identification of salt and drought inducible glutathione S-transferase genes of hybrid poplar. J. Plant Biotechnol 41: 26-32. |
| [36] | Rohman MM, Begum S, Akhi HA, Ahsan SMFA, Uddin SM, Amiruzzaman M, Banik BR. 2015. Protective role of antioxidants in maize seedlings under saline stress: Exogenous proline provided better tolerance than betaine. Bothalia 45: 17-35. |
| [37] | Rohman MM, Hossain AM, Fujita M. 2010. Levels of a dominant glutathione S-transferase in onion bulbs have a seasonal relationship with physiological inhibitors. Aust. J. Crop Sci 4(1): 55-62. |
| [38] | Rezaei K. M, Shobbar S-Z, Maryam S, Abedini R, Sajjad Z. 2013. Glutathione S-transferase (GST) family in barley: Identification of members, enzyme activity, and gene expression pattern. J Plant Physiol 170: 1277-1284. |
| [39] | Hu T. 2014. A glutathione S-transferase confers herbicide tolerance in rice.” Crop Breed. Applied Biotechnol 14: 76-81. |
| [40] | Sharma R, Sahoo A, Devendran R, Jain M. 2014. Over-expression of a rice Tau class glutathione S-transferase gene improves tolerance to salinity and oxidative stresses in Arabidopsis.”DOI: 10.1371/journal.pone.0092900. |
| [41] | Scarponi L, Quagliarini E, Del DB. 2006. Induction of wheat and maize glutathione S-transferase by some herbicide safeners and their effect on enzyme activity against butachlor and terbuthylazine. Pest Manage Sci 62(10): 927-32. |
| [42] | Galle A, Csiszar J, Seceji M, Tari I, Guoth A. Gyorgyey J. Erdei L. 2008. Monitoring the levels of phi and tau group GST genes in wheat cultivars under osmotic stress.”Acta Biologica Szegediensis 52(1): 95-96. |
| [43] | Pandey B, Sharma P, MPandey D, Varshney J, Sheoran S, Singh M, Singh R, Sharma I, Chatrath R. 2012. Comprehensive computational analysis of different classes of Glutathione S-transferases in Triticum aestivum L. Plant Omics Journal 5(6): 518-531. |
| [44] | 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. |
| [45] | Cetinka H, Seckindinler B, Tascicetinya E, Dinler SB, Tasci E. 2014. Investigation of comparative regulation on antioxidant enzyme system under copper treatment and drought stress in maize (Zea mays L.). Not. Bot. Horti. Agrobot 42(2): 363-371. |
| [46] | Marrs AK, Walbot V. 1997. Expression and RNA splicing of the maize glutathione S-transferase Bronze2 gene is regulated by cadmium and other stresses. Plant Physiol 1997. 113: 93-102. |
| [47] | Hossain MA, Mostofa GM, Fujita M. 2013. Heat-shock positively modulates oxidative protection of salt and drought-stressed mustard (Brassica campestris L.) seedlings. Journal Plant Sci. Mol. Breed http://www.hoajonline.com/journals/pdf/2050-2389-2-2.pdf, doi: 10.7243/2050-2389-2-2.. |
| [48] | Kapoor D, Kaur S, Bhardwaj R. 2014. Physiological and biochemical changes in Brassica juncea plants under Cd-induced stress. BioMed Research International. Article ID 726070, http://dx.doi.org/10.1155/2014/726070. |
| [49] | Shahram N, Shahrokh N, Hossein K. 2012. Induction of phytochelatin and responses of antioxidants under cadmium stress in safflower (Carthamus tinctorius) seedlings. Turkish Journal of Botany 36: 495. |
| [50] | Chan C, Lam M-H. 2014. A putative lambda class glutathione S-transferase enhances plant survival under salinity stress. Plant Cell Physiol 55(3): 570-579. |
| [51] | Dinler SB, Antoniou C, Fotopoulous V. 2014. Interplay between GST and nitric oxide in the early response of soybean (Glycine max L.) plants to salinity stress. Journal Plant Physiol 171(18): 1740-7. |
| [52] | Dalton D, Boniface C, Turner Z, Lindahl A, Kim JH, Jelinek L, Govindarajulu M, Finger ER, Taylor GC. 2009. Physiological roles of glutathione S-transferases in soybean root nodules. Plant Physiol 150: 521-530. |
| [53] | Talukder D. 2012. An induced glutathione-deficient mutant in Grass pea (Lathyrus sativus L.) Modification of plant morphology, Alternations in antioxidants activities, and increased sensitivity to Cadmium. Bioremediation. Biodiversity and Bioavailability 6: 75-86. |
| [54] | Dixit V, Pandey V, Shyam R. 2001. Differential antioxidative responses to cadmium in roots and leaves of pea (Pisum sativum L. cv. Azad). Journal Exp. Bot 52: 1101-1109. |
| [55] | Nahar K, Hasanuzzaman M, Alam MM, Fujita M. 2015. Exogenous glutathione confers high temperature stress tolerance in mung bean (Vigna radiata L.) by modulating antioxidant defense and methylglyoxal detoxification system. Environmental and Experimental Bot 112: 44-54. |
| [56] | Hossain MA, Hasanuzzaman M, Fujita M. 2011. Coordinate induction of antioxidant defense and glyoxalase system by exogenous proline and glycine betaine is correlated with salt tolerance in mung bean. Front. Agric. China 5(1): 1-14. |
| [57] | Molla RM, Ali MR, Hasanuzzaman M, Mamun AHM, Ahmed A, Nazim¬ud¬dowla NAM, Rohman MM. 2014. Exogenous proline and betaine-induced upregulation of glutathione transferase and glyoxalase I in lentil (Lens culinaris ) under drought stress. Not. Bot. Horti. Agrobo 42(1): 73--80. |
| [58] | Gazendam I, Oelofse D. 2007. Isolation of cowpea genes conferring drought tolerance: construction of a cDNA drought expression library. Water SA 33(3): 387-391 |
| [59] | Hossain ZM, Fujita M. 2003. Modulation of pumpkin glutathione S-transferases by aldehydes and related compounds. Plant Cell Physiol 44(5): 481-490. |
| [60] | Hossain ZM, Fujita M, “Purification of a phi-type glutathione S-transferase from pumpkin flowers, and molecular cloning of its cDNA. Biosci. Biotech.Biochem 66(10): 2068-76. |
| [61] | Hahn K, Strittmatter G. 1994. Pathogen-defence gene prpl-1 from potato encodes an auxin-responsive glutathione S-transferase. Eur. J. Biochem 226: 619-626. |
| [62] | Peil D, Jinping D, Zilei D, Meng L, Yiyang F, Chengwei L. 2012. Cloning and expression of a tomato glutathione S-transferase (GST) in Escherichia coli. African Journal Biotechnol 11(23): 6402-6408. |
| [63] | Csiszar J, Horvath E, Vary Z, Galle A, Bela K, Burnner S, Tari I. 2014. Glutathione transferase supergene family in tomato: Salt stress-regulated expression of representative genes from distinct GST classes in plants primed with salicylic acid. Plant Physiol and Biochem 78: 15-26. |
| [64] | Kumchail J, Huang J-Z, Lee C-Y, Chen F-C, Chin S-W. 2012. Proline partially overcomes excess molybdenum toxicity in cabbage seedlings grown in vitro. Genet. Mol. Res 12: 5589-5601. |
| [65] | Huang S-S, Hung J-H, Chiu S-C, Chen J-H, Lin S-S, Lin C-Y, Yi C-H, Huang J-G, Iy H-L. 2009. Sweet potato trypsin inhibitor with thioltransferase-like and glutathione S-transferase-like activities. Botanical Studies 50: 443-450. |
| [66] | Gong H, Jiao Y, Hu WW, Pua CE 2005. Expression of glutathione S-transferas and its role in plant growth and development in vivo and shoot morphogenesis in vitro. Plant Mol. Biol 57: 53-66. |
APA Style
Md. Raisul Islam, Abul Kashem Chowdhury, Md. Mahfuzur Rahman, Md. Motiar Rohman. (2015). Comparative Investigation of Glutathione S-transferase (GST) in Different Crops and Purification of High Active GSTs from Onion (Allium cepa L.). Journal of Plant Sciences, 3(3), 162-170. https://doi.org/10.11648/j.jps.20150303.17
ACS Style
Md. Raisul Islam; Abul Kashem Chowdhury; Md. Mahfuzur Rahman; Md. Motiar Rohman. Comparative Investigation of Glutathione S-transferase (GST) in Different Crops and Purification of High Active GSTs from Onion (Allium cepa L.). J. Plant Sci. 2015, 3(3), 162-170. doi: 10.11648/j.jps.20150303.17
AMA Style
Md. Raisul Islam, Abul Kashem Chowdhury, Md. Mahfuzur Rahman, Md. Motiar Rohman. Comparative Investigation of Glutathione S-transferase (GST) in Different Crops and Purification of High Active GSTs from Onion (Allium cepa L.). J Plant Sci. 2015;3(3):162-170. doi: 10.11648/j.jps.20150303.17
Share This Article
Twitter
Linked In
Facebook
Copy
Download@article{10.11648/j.jps.20150303.17,
author = {Md. Raisul Islam and Abul Kashem Chowdhury and Md. Mahfuzur Rahman and Md. Motiar Rohman},
title = {Comparative Investigation of Glutathione S-transferase (GST) in Different Crops and Purification of High Active GSTs from Onion (Allium cepa L.)},
journal = {Journal of Plant Sciences},
volume = {3},
number = {3},
pages = {162-170},
doi = {10.11648/j.jps.20150303.17},
url = {https://doi.org/10.11648/j.jps.20150303.17},
eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jps.20150303.17},
abstract = {Glutathione S-transferases (GSTs; EC 2.5.1.18) are abundant proteins encoded by a highly divergent ancient gene family with protective functions through detoxification and non-catalytic roles as carriers of cytotoxins. In this work, we investigated the GST activities in seedlings of 38 crops to obtain a plant with high active GST for further study. In screening of 38 crops, onion seedling showed the highest GST activity (483.54 nmol min-1mg-1 protein) followed by wheat(372.89 nmol min-1mg-1 protein), barley (253.44 nmol min-1mg-1 protein), rice (244.12 nmol min-1mg-1 protein) and proso millet (173.34 nmol min-1mg-1 protein). Carrot seedling showed the lowest activity (3.63 nmol min-1mg-1 protein).In onion plants, both root and leaf showed high GST activity. Onion bulb GSTs were separated DEAE cellulose column chromatography, and purified by affinity chromatography (S-hexyl glutathione-agarose). Three GST peaks were found to elute at 67, 107 and 140mM of KCl gradient solution, and were named as GSTa, GSTb and GSTc. Among the three GSTs, GSTa, GSTb and GSTc contained 9.58, 61.45 and 28.97% of total activity, respectively. In purification, GSTa, GSTb and GSTc had specific activities of 9075, 17259 and 19868 nmol min-1 mg-1 protein, respectively, along with yield of 2.48, 3.17 and 1.28,and purification fold of 15.8, 29.9 and 34.5, respectively. The purity and molecular mass of the fraction was examined by SDS-PAGE. The silver staining of the purified GSTa, GSTb and GSTc indicated that final product of GSTb and GSTc were highly purified and migrated as a single band on SDS-PAGE with an apparent molecular mass of 27 kDa. However, GSTa eluted with Glyoxalase-I (Gly-I) and to purify GSTa, more methodological application are suggested.},
year = {2015}
}
TY - JOUR T1 - Comparative Investigation of Glutathione S-transferase (GST) in Different Crops and Purification of High Active GSTs from Onion (Allium cepa L.) AU - Md. Raisul Islam AU - Abul Kashem Chowdhury AU - Md. Mahfuzur Rahman AU - Md. Motiar Rohman Y1 - 2015/06/13 PY - 2015 N1 - https://doi.org/10.11648/j.jps.20150303.17 DO - 10.11648/j.jps.20150303.17 T2 - Journal of Plant Sciences JF - Journal of Plant Sciences JO - Journal of Plant Sciences SP - 162 EP - 170 PB - Science Publishing Group SN - 2331-0731 UR - https://doi.org/10.11648/j.jps.20150303.17 AB - Glutathione S-transferases (GSTs; EC 2.5.1.18) are abundant proteins encoded by a highly divergent ancient gene family with protective functions through detoxification and non-catalytic roles as carriers of cytotoxins. In this work, we investigated the GST activities in seedlings of 38 crops to obtain a plant with high active GST for further study. In screening of 38 crops, onion seedling showed the highest GST activity (483.54 nmol min-1mg-1 protein) followed by wheat(372.89 nmol min-1mg-1 protein), barley (253.44 nmol min-1mg-1 protein), rice (244.12 nmol min-1mg-1 protein) and proso millet (173.34 nmol min-1mg-1 protein). Carrot seedling showed the lowest activity (3.63 nmol min-1mg-1 protein).In onion plants, both root and leaf showed high GST activity. Onion bulb GSTs were separated DEAE cellulose column chromatography, and purified by affinity chromatography (S-hexyl glutathione-agarose). Three GST peaks were found to elute at 67, 107 and 140mM of KCl gradient solution, and were named as GSTa, GSTb and GSTc. Among the three GSTs, GSTa, GSTb and GSTc contained 9.58, 61.45 and 28.97% of total activity, respectively. In purification, GSTa, GSTb and GSTc had specific activities of 9075, 17259 and 19868 nmol min-1 mg-1 protein, respectively, along with yield of 2.48, 3.17 and 1.28,and purification fold of 15.8, 29.9 and 34.5, respectively. The purity and molecular mass of the fraction was examined by SDS-PAGE. The silver staining of the purified GSTa, GSTb and GSTc indicated that final product of GSTb and GSTc were highly purified and migrated as a single band on SDS-PAGE with an apparent molecular mass of 27 kDa. However, GSTa eluted with Glyoxalase-I (Gly-I) and to purify GSTa, more methodological application are suggested. VL - 3 IS - 3 ER -
Email: