biologia plantarum

International journal on Plant Life established by Bohumil Němec in 1959

Biologia plantarum 61:169-177, 2017 | DOI: 10.1007/s10535-016-0659-6

Over-expression of CsGSTU promotes tolerance to the herbicide alachlor and resistance to Pseudomonas syringae pv. tabaci in transgenic tobacco

L. Lo Cicero1, V. Catara1, C. P. Strano1, P. Bella1, P. Madesis2, A. R. Lo Piero1,*
1 Department of Agriculture, Food and Environment (Di3A), University of Catania, Catania, Italy
2 Institute of Applied Biosciences, CERTH, Thermi Thessaloniki, Greece

Glutathione transferases (GSTs) mainly catalyze the nucleophilic addition of glutathione to a large variety of hydrophobic molecules participating to the vacuole compartmentalization of many toxic compounds. In this work, the putative tolerance of transgenic tobacco plants over-expressing CsGSTU genes towards the chloroacetanilide herbicide alachlor was investigated. Our results show that the treatment with 0.0075 mg cm-3 of alachlor strongly affects the growth of both wild type and transformed tobacco seedlings with the sole exception of the transgenic lines overexpressing CsGSTU2 isoform that are barely influenced by herbicide treatment. In order to correlate the in planta studies with enzyme properties, recombinant CsGSTs were in vitro expressed and tested for GST activity using alachlor as substrate. The recombinant GSTU2 enzyme was twice more active than GSTU1 in conjugating alachlor to GSH thus indicating that CsGSTU2 might play a crucial role in the plant defense against the herbicide. Moreover, as a consequence of the infiltration with a bacterial suspension of the P. syringae pv. tabaci, transgenic tobacco plants but not wild type plants bestowed the capability to limit toxic metabolite diffusion through plant tissues as indicated by the absence of chlorotic halos formation. Consequently, the transgenic tobacco plants described in the present study might be utilized for phytoremediation of residual xenobiotics in the environment and might represent a model for engineering plants that resist to pathogen attack.

Keywords: biotic stress; glutathione transferase; host-pathogen interaction; phytoremediation
Subjects: CsGSTU; herbicide tolerance; biotic stress; glutathione transferase; host-pathogen interactions; phytoremediation; transgenic plant; tobacco

Received: September 17, 2015; Revised: February 12, 2016; Accepted: March 9, 2016; Published: January 1, 2017  Show citation

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Lo Cicero, L., Catara, V., Strano, C.P., Bella, P., Madesis, P., & Lo Piero, A.R. (2017). Over-expression of CsGSTU promotes tolerance to the herbicide alachlor and resistance to Pseudomonas syringae pv. tabaci in transgenic tobacco. Biologia plantarum61(1), 169-177. doi: 10.1007/s10535-016-0659-6
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References

  1. Allocati, N., Federici, L., Masulli, M., Di Ilio, C.: Glutathione transferases in bacteria. - FEBS J. 276: 58-75, 2009. Go to original source...
  2. Alvarez, M.E., Pennell, R.I., Meijer, P.J., Ishikawa, A., Dixon, R.A., Lamb, C.: Reactive oxygen intermediates mediate a systemic signal network in the establishment of plant immunity. - Cell 92: 773-784, 1998. Go to original source...
  3. Bai, H., Lan, J.P., Gan, Q., Wang, X.Y., Hou, M.M., Cao, Y.H., Li, L.Y., Liu, L.J., Hao, Y.J., Yin, C.C., Wu, L., Zhu, L.H., Liu, G.Z.: Identification and expression analysis of components involved in rice Xa21-mediated disease resistance signaling. - Plant Biol. 14: 914-922, 2012. Go to original source...
  4. Bender, C.L.: Chlorosis-inducing phytotoxins produced by Pseudomonas syringae. - Eur. J. Plant Pathol. 105: 1-12, 1999. Go to original source...
  5. Benekos, K., Kissoudis, C., Nianiou-Obeidat, I., Labrou, N., Madesis, P., Kalamaki, M., Makris, A., Tsaftaris, A.: Overexpression of a specific soybean GmGSTU4 isoenzyme improves diphenyl ether and chloroacetanilide herbicide tolerance of transgenic tobacco plants. - J. Biotechnol. 150: 195-201, 2010. Go to original source...
  6. Boparai, H.K., Shea, P.J., Comfort, S.D., Snow, D.D.: Dechlorinating chloroacetanilide herbicides by dithionitetreated aquifer sediment and surface soil. - Environ. Sci. Technol. 40: 3043-3049, 2006. Go to original source...
  7. Bradford, M.M.: 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, 1976. Go to original source...
  8. Choi, I.Y., Noh, E.W., Kim, H.J., Shim, D.: Overexpression of poplar GSTU51 confers selective tolerance to both mercury and methyl viologen but not to CDNB or cadmium in transgenic poplars. - Plant Biotechnol. Rep. 7: 175-184, 2013. Go to original source...
  9. Cintas, N.A., Koike, S.T., Bull, C.T.: A new pathovar, Pseudomonas syringae pv. alisalensis, proposed for the causal agent of bacterial blight of broccoli and broccoli raab. - Plant Dis. 86: 992-998. 2002. Go to original source...
  10. Coleman, J., Blake-Kalff, M., Davies, E.: Detoxification of xenobiotics by plants: chemical modification and vacuolar compartmentation. - Trends Plant Sci. 2: 144-151, 1997. Go to original source...
  11. Cummins, I., Dixon, D.P., Freitag-Pohl, S., Skipsey, M., Edwards, R.: Multiple roles for plant glutathione transferases in xenobiotic detoxification. - Drug Metabol. Rev. 43: 266-280, 2011. Go to original source...
  12. Dean, J.D., Goodwin, P.H., Hsiang, T.: Induction of glutathione S-transferase genes of Nicotiana benthamiana following infection by Colletotrichum destructivum and C. orbiculare and involvement of one in resistance. - J. exp. Bot. 56: 1525-1533, 2005. Go to original source...
  13. Dixit, P., Mukherjee, P.K., Ramachandran, V., Eapen, S.: Glutathione transferase from Trichoderma virens enhances cadmium tolerance without enhancing its accumulation in transgenic Nicotiana tabacum. - PlosOne 6: e16360, 2011. Go to original source...
  14. Dixon, D.P., Lapthorn, A., Edwards, R.: Plant glutathione transferases. - Genome Biol. 3: 3004-1, 2002. Go to original source...
  15. Dudler, R., Hertig, C., Rebmann, G., Bull, J., Mauch, F.: A pathogen-induced wheat gene encodes a protein homologous to glutathione-S-transferases. - Mol. Plant Microbe Interact. 4: 14-18, 1991. Go to original source...
  16. Edwards, R., Dixon, D.P., Walbot, V.: Plant glutathione Stransferases: enzymes with multiple functions in sickness and in health. - Trends Plant Sci. 5: 193-198, 2000. Go to original source...
  17. Goodman, R. N., Novacky, A.J.: The Hypersensitive Reaction in Plants to Pathogens: a Resistance Phenomenon. - American Phytopathological Society Press, St. Paul 1994.
  18. Greenberg, J.T., Guo, A., Klessig, D.F., Ausubel, F.M.: Programmed cell death in plants: a pathogen-triggered response activated coordinately with multiple defense functions. - Cell 77: 551-563, 1994. Go to original source...
  19. Heath, M.C.: Hypersensitive response-related death. - Plant mol. Biol. 44: 321-334, 2000. Go to original source...
  20. Hirano, S.S., Upper, C.D.: Population biology and epidemiology of Pseudomonas syringae. - Annu. Rev. Phytopathol. 28: 155-177, 1990. Go to original source...
  21. Hu, T., He, S., Yang, G., Zeng, H., Wang, G., Chen, Z., Huang, X.: Isolation and characterization of a rice glutathione Stransferase gene promoter regulated by herbicides and hormones. - Plant Cell Rep. 30: 539-549, 2011. Go to original source...
  22. Iwasaki, I., Utsumi, S., Ozawa, T.: New colorimetric determination of chloride using mercuric thiocyanate and ferric ion. - Bull. Chem. Soc. Japan 25: 226, 1952. Go to original source...
  23. Jha, B., Sharma, A., Mishra, A.: Expression of SbGSTU (tau class glutathione S-transferase) gene isolated from Salicornia brachiata in tobacco for salt tolerance. - Mol. Biol. Rep. 38: 4823-4832, 2011. Go to original source...
  24. Ji, W., Zhu, Y., Li, Y., Yang, L., Zhao, X., Cai, H., Bai, X.: Over-expression of a glutathione S-transferase gene, GsGST, from wild soybean (Glycine soja) enhances drought and salt tolerance in transgenic tobacco. - Biotechnol. Lett. 32: 1173-1179, 2010. Go to original source...
  25. Jo, H.J., Lee, J.J., Kong, K.H.: A plant-specific tau class glutathione S-transferase from Oryza sativa with very high activity against 1-chloro-2,4-dinitrobenzene and chloroacetanilide herbicides. - Pestic. Biochem. Phys. 101: 265-269, 2011. Go to original source...
  26. Karavangeli, M., Labrou, N.E., Clonis, Y.D., Tsaftaris, A.: Development of transgenic tobacco plants overexpressing maize glutathione S-transferase I for chloroacetanilide herbicides phytoremediation. - Biomol. Eng. 22: 121-128, 2005. Go to original source...
  27. King, E.O., Ward, M.K., Raney, D.E.: Two simple media for the demonstration of pyocyanin and fluorescin. - J. Lab. clin. Med. 44: 301-307, 1954.
  28. Klement, Z.: Rapid detection of the pathogenicity of phytopathogenic Pseudomonads. - Nature 199: 299-300, 1963. Go to original source...
  29. Labrou, N.E., Papageorgiou, A.C, Pavli, O., Flemetakis, E.: Plant GSTome: structure and functional role in genome network and plant stress response. - Curr. Opin. Biotechnol. 32: 186-194, 2015. Go to original source...
  30. Lee, S., Yang, D.S., Uppalapati, S.R., Sumner, L.W., Mysore, K.S.: Suppression of plant defense responses by extracellular metabolites from Pseudomonas syringae pv. tabaci in Nicotiana benthamiana. - BMC Plant Biol. 13: 65, 2013. Go to original source...
  31. Levine, A., Tenhaken, R., Dixon, R., Lamb, C.: H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. - Cell 79: 583-593, 1994. Go to original source...
  32. Liu, D., Liu, Y., Rao, J., Wang, G., Li, H., Ge, F., Chen, C.: Overexpression of the glutathione S-transferase gene from Pyrus pyrifolia fruit improves tolerance to abiotic stress in transgenic tobacco plants. - Mol. Biol. 47: 515-523, 2013. Go to original source...
  33. Lo Cicero, L., Madesis, P., Tsaftaris, A., Lo Piero, A.R.: Tobacco plants over-expressing the sweet orange tau glutathione transferases (CsGSTUs) acquire tolerance to the diphenyl ether herbicide fluorodifen and to salt and drought stresses. - Phytochemistry 116: 69-77, 2015. Go to original source...
  34. Lo Piero, A.R., Mercurio, V., Puglisi, I., Petrone, G.: Gene isolation and expression analysis of two distinct sweet orange [Citrus sinensis L.(Osbeck)] tau-type glutathione transferases. - Gene 443: 143-150, 2009. Go to original source...
  35. Lo Piero, A.R., Mercurio, V., Puglisi, I., Petrone, G.: Different roles of functional residues in the hydrophobic binding site of two sweet orange tau glutathione S-transferases. - FEBS J. 277: 255-262, 2010. Go to original source...
  36. Lo Piero, A.R., Puglisi, I., Mercurio, V., Petrone, G.: Engineering the xenobiotic substrate specificity of sweet orange tau glutathione S-transferase. - Acta Hort. 892: 143-147, 2011. Go to original source...
  37. Lo Piero, A.R., Puglisi, I., Petrone, G.: Gene isolation, analysis of expression, and in vitro synthesis of glutathione Stransferase from orange fruit [Citrus sinensis L. (Osbeck)]. - J. Agr. Food Chem. 54: 9227-9233, 2006. Go to original source...
  38. Marrs, K.A.: The functions and regulation of glutathione Stransferases in plants. - Annu. Rev. Plant Biol. 47: 127-158, 1996. Go to original source...
  39. Mauch, F., Dudler, R.: Differential induction of distinct glutathione-S-transferases of wheat by xenobiotics and by pathogen attack. - Plant Physiol. 102: 1193-1201, 1993. Go to original source...
  40. Melotto, M., Underwood, W., Koczan, J., Nomura, K., He, S.Y.: Plant stomata function in innate immunity against bacterial invasion. - Cell 126: 969-980, 2006. Go to original source...
  41. Melotto, M., Underwood, W., He, S.Y.: Role of stomata in plant innate immunity and foliar bacterial diseases. - Annu. Rev. Phytopathol. 46: 101-122, 2008. Go to original source...
  42. Mohsenzadeh, S., Esmaeili, M., Moosavi, F., Shahrtash, M., Saffari, B., Mohabatkar, H.: Plant glutathione S-transferase classification, structure and evolution. - Afr. J. Biotechnol. 10: 8160-8165, 2011. Go to original source...
  43. Murashige, T., Skoog, F.: A revised medium for rapid growth and bio assays with tobacco tissue cultures. - Physiol. Plant. 15: 473-497, 1962. Go to original source...
  44. Öztetik, E.: A tale of plant glutathione S-transferases: since 1970. - Bot. Rev. 74: 419-437, 2008. Go to original source...
  45. Puglisi, I., Lo Cicero, L., Lo Piero, A.R.: The glutathione Stransferase gene superfamily: an in silico approach to study the post translational regulation. - Biodegradation 24: 471-485, 2013a. Go to original source...
  46. Puglisi, I., Lo Cicero, L., Lo Piero, A.R.: Fingerprint analysis of selected glutathione S-transferase superfamily genes from rape (Brassica napus). - Acta Hort. 1005: 261-267, 2013b. Go to original source...
  47. Sambrook, J., Fritsch, E.F., Maniatis, T.: Molecular Cloning: a Laboratory Manual. - Cold Spring Harbor Laboratory Press, New York 1989.
  48. Skopelitou, K., Labrou, N.E.: A new colorimetric assay for glutathione transferase-catalyzed halogen ion release for high-throughput screening. - Anal. Biochem. 405: 201-206, 2010. Go to original source...
  49. Soranzo, N., Gorla, M.S., Mizzi, L., De Toma, G., Frova, C.: Organization and structural evolution of the rice glutathione S-transferase gene family. - Mol. Genet. Genom. 271: 511-521, 2004. Go to original source...
  50. Szatmari, A., Ott, P.G., Varga, G.J., Besenyei, E., Czelleng, A., Klement, Z., Bozsó, Z.: Characterisation of basal resistance (BR) by expression patterns of newly isolated representative genes in tobacco. - Plant Cell Rep. 25: 728-740, 2006. Go to original source...
  51. Tremblay, R., Wang, D., Jevnikar, A.M., Ma, S.: Tobacco, a highly efficient green bioreactor for production of therapeutic proteins. - Biotechnol. Adv. 28: 214-221, 2010. Go to original source...
  52. Woolley, D.W., Pringle, R.B., Braun, A.C.: Isolation of the phytopathogenic toxin of Pseudomonas tabaci, an antagonist of methionine. - J. biol. Chem. 197: 409-417, 1952. Go to original source...
  53. Yen, J.H., Chang, S.L., Wu, C.C., Wang, Y.S.: Behavior of acetanilide herbicides in soils and the possibility of groundwater contamination. - Bull. Environ. Contam. Toxicol. 75: 1086-1093, 2005. Go to original source...