식물병연구 (Research in Plant Disease)

한국식물병리학회 (The Korean Society of Plant Pathology)
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Proteomic Analysis of the GacA Response Regulator in Pseudomonas chlororaphis O6

  • Anderson, Anne J. (Department of Biological Engineering, Utah State University) ;
  • Kim, Young Cheol (Department of Applied Biology, Chonnam National University)
  • 투고 : 2018.06.03
  • 심사 : 2018.06.22
  • 발행 : 2018.06.30
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초록

The GacS/GacA system in the root colonizer Pseudomonas chlororaphis O6 is a key regulatory system of many traits relevant to the plant probiotic nature of this bacterium. The work in this paper elucidates proteins using proteomics approach in P. chlororaphis O6 under the control of the cytoplasmic regulatory protein, GacA. A gacA mutant of P. chlororaphis O6 showed loss in production of phenazines, acyl homoserine lactones, hydrogen cyanide, and protease, changes that were associated with reduced in vitro antifungal activity against plant fungal pathogens. Production of iron-chelating siderophore was significantly enhanced in the gacA mutant, also paralleling changes in a gacS mutant. However, proteomic analysis revealed proteins (13 downregulated and 7 upregulated proteins in the mutant compared to parental strain) under GacA control that were not apparent by a proteomic study of a gacS mutant. The putative identity of the downregulated proteins suggested that a gacA mutant would have altered transport potentials. Notable would be a predicted loss of type-VI secretion and PEP-dependent transport. Study of mutants of these GacA-regulated proteins will indicate further the features required for probiotic potential in this rhizobacterium.

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참고문헌

  1. Anderson, A. J. and Kim, Y. C. 2018. Biopesticides produced by plant-probiotic Pseudomonas chlororaphis isolates. Crop Protect. 105: 62-69. https://doi.org/10.1016/j.cropro.2017.11.009
  2. Anderson, A. J., Kang, B. R. and Kim, Y. C. 2017. The Gac/Rsm signaling pathway of a biocontrol bacterium, Pseudomonas chlororaphis O6. Res. Plant Dis. 23: 212-227. https://doi.org/10.5423/RPD.2017.23.3.212
  3. Bernal, P., Allsopp, L. P., Filloux, A. and Llamas, M. A. 2017. The Pseudomonas putida T6SS is a plant warden against phytopathogens. ISME J. 11: 972-987. https://doi.org/10.1038/ismej.2016.169
  4. Bernal, P., Llamas, M. A. and Filloux, A. 2018. Type VI secretion systems in plant-associated bacteria. Environ. Microbiol. 20: 1-15. https://doi.org/10.1111/1462-2920.13956
  5. Chen, L., Zou, Y., She, P. and Wu, Y. 2015. Composition, function, and regulation of T6SS in Pseudomonas aeruginosa. Microbiol. Res. 172: 19-25. https://doi.org/10.1016/j.micres.2015.01.004
  6. Decoin, V., Barbey, C., Bergeau, D., Latour, X., Feuilloley, M. G. J., Orange, N. et al. 2014. A type VI secretion system is involved in Pseudomonas fluorescens bacterial competition. PLoS One 9: e89411. https://doi.org/10.1371/journal.pone.0089411
  7. Elhai, J. and Wolk, C. P. 1988. A versatile class of positive-selection vectors based on the nonviability of palindrome-containing plasmids that allows cloning into long polylinkers. Gene 68: 119-138. https://doi.org/10.1016/0378-1119(88)90605-1
  8. Gallique, M., Decoin, V., Barbey, C., Rosay, T., Feuilloley, M. G. J., Orange, N. et al. 2017. Contribution of the Pseudomonas fluorescens MFE01 type VI secretion system to biofilm formation. PLoS One 12: e0170770. https://doi.org/10.1371/journal.pone.0170770
  9. Hassan, K. A., Johnson, A., Shaffer, B. T., Ren, Q., Kidarsa, T. A., Elbourne, L. D. H. et al. 2010. Inactivation of the GacA response regulator in Pseudomonas fluorescens Pf-5 has far-reaching transcriptomic consequences. Environ. Microbiol. 12: 899-915. https://doi.org/10.1111/j.1462-2920.2009.02134.x
  10. Heeb, S. and Haas, D. 2001. Regulatory roles of the GacS/GacA two-component system in plant-associated and other gramnegative bacteria. Mol. Plant MIcrobe Interact. 14: 1351-1363. https://doi.org/10.1094/MPMI.2001.14.12.1351
  11. Kang, B. R., Anderson, A. J. and Kim, Y. C. 2018. Hydrogen cyanide produced by Pseudomonas chlororaphis O6 exhibits nematicidal activity against Meloidogyne hapla. Plant Pathol. J. 34: 35-43.
  12. Kang, B. R., Cho, B. H., Anderson, A. J. and Kim, Y. C. 2004. The global regulator GacS of a biocontrol bacterium Pseudomonas chlororaphis O6 regulates transcription from the rpoS gene encoding a stationary-phase sigma factor and affects survival in oxidative stress. Gene 325: 137-143. https://doi.org/10.1016/j.gene.2003.10.004
  13. Kang, B. R., Han, S. H., Zdor, R. E., Anderson, A. J., Spencer, M., Yang, K. Y. et al. 2007. Inhibition of seed germination and induction of systemic disease resistance by Pseudomonas chlororaphis O6 requires phenazine production regulated by the global regulator, gacS. J. Microbiol. Biotech. 17: 586-593.
  14. Kang, B. R., Yang, K. Y., Cho, B. H., Han, T. H., Kim, I. S., Lee, M. C. et al. 2006. Production of indole-3-acetic acid in the plant-beneficial strain Pseudomonas chlororaphis O6 is negatively regulated by the global sensor kinase GacS. Curr. Microbiol. 52: 473-476. https://doi.org/10.1007/s00284-005-0427-x
  15. Kim, C. H., Kim, Y. H., Anderson, A. J. and Kim, Y. C. 2014a. Proteomic analysis of a global regulator GacS sensor kinase in the rhizobacterium, Pseudomonas chlororaphis O6. Plant Pathol. J. 30: 220-227. https://doi.org/10.5423/PPJ.NT.02.2014.0012
  16. Kim, J. S., Kim, Y. H., Anderson, A. J. and Kim, Y. C. 2014b. The sensor kinase GacS negatively regulates flagellar formation and motility in a biocontrol bacterium, Pseudomonas chlororaphis O6. Plant Pathol. J. 30: 215-219. https://doi.org/10.5423/PPJ.NT.11.2013.0109
  17. Kim, J. S., Kim, Y. H., Park, J. Y., Anderson, A. J. and Kim, Y. C. 2014c. The global regulator GacS regulates biofilm formation in Pseudomonas chlororaphis O6 differently with carbon source. Can. J. Microbiol. 60: 133-138. https://doi.org/10.1139/cjm-2013-0736
  18. Kim, Y. C. and Anderson, A. J. 2018. Rhizosphere pseudomonads as probiotics improving plant health. Mol. Plant Pathol. DOI: 10.1111/mpp.12693. (In press)
  19. Kim, Y. C., Leveau, J., McSpadden Gardener, B. B., Pierson, E. A., Pierson, L. S., 3rd and Ryu, C. M. 2011. The multifactorial basis for plant health promotion by plant-associated bacteria. Appl. Environ. Microbiol. 77: 1548-1555. https://doi.org/10.1128/AEM.01867-10
  20. Lee, J. H., Ma, K. C., Ko, S. J., Kang, B. R., Kim, I. S. and Kim, Y. C. 2011. Nematicidal activity of a nonpathogenic biocontrol bacterium, Pseudomonas chlororaphis O6. Curr. Microbiol. 62: 746-751. https://doi.org/10.1007/s00284-010-9779-y
  21. Li, J., Yang, Y., Dubern, J. F., Li, H., Halliday, N., Chernin, L., Gao, K. et al. 2015. Regulation of GacA in Pseudomonas chlororaphis strains shows a niche specificity. PLoS One 10: e0137553. https://doi.org/10.1371/journal.pone.0137553
  22. Liu, Y., Wang, Z., Bilal, M., Hu, H., Wang, W., Huang, X. et al. 2018. Enhanced fluorescent siderophore biosynthesis and loss of phenazine-1-carboxamide in phenotypic variant of Pseudomonas chlororaphis HT66. Front. Microbiol. 9: 759. https://doi.org/10.3389/fmicb.2018.00759
  23. Loper, J. E., Hassan, K. A., Mavrodi, D. V., Davis, E. W., 2nd, Lim, C. K., Shaffer, B. T. et al. 2012. Comparative genomics of plant-associated Pseudomonas spp.: insights into diversity and inheritance of traits involved in multitrophic interactions. PLoS Genetics 8: e1002784. https://doi.org/10.1371/journal.pgen.1002784
  24. McClean, K. H., Winson, M. K., Fish, L., Taylor, A., Chhabra, S. R., Camara, M. et al. 1997. Quorum sensing and Chromobacterium violaceum: exploitation of violacein production and inhibition for the detection of N-acylhomoserine lactones. Microbiology 143: 3703-3711. https://doi.org/10.1099/00221287-143-12-3703
  25. Miller, C. D., Kim, Y. C. and Anderson, A. J. 1997. Cloning and mutational analysis of the gene for the stationary-phase inducible catalase (catC) from Pseudomonas putida. J. Bacteriol. 179: 5241-5245. https://doi.org/10.1128/jb.179.16.5241-5245.1997
  26. Natsch, A., Keel, C., Pfirter, H. A., Haas, D. and Defago, G. 1994. Contribution of the global regulator gene gacA to persistence and dissemination of Pseudomonas fluorescens biocontrol strain CHA0 introduced into soil microcosms. Appl. Environ. Microbiol. 60: 2553-2560.
  27. Oh, S. A., Kim, J. S., Han, S. H., Park, J. Y., Dimkpa, C., Edlund, C. et al. 2013a. The GacS-regulated sigma factor RpoS governs production of several factors involved in biocontrol activity of the rhizobacterium Pseudomonas chlororaphis O6. Can. J. Microbiol. 59: 556-562. https://doi.org/10.1139/cjm-2013-0062
  28. Oh, S. A., Kim, J. S., Park, J. Y., Han, S. H., Dimkpa, C., Anderson, A. J. et al. 2013b. The RpoS sigma factor negatively regulates production of IAA and siderophore in a biocontrol rhizobacterium, Pseudomonas chlororaphis O6. Plant Pathol. J. 29: 323-329. https://doi.org/10.5423/PPJ.NT.01.2013.0013
  29. Park, J. Y., Kang, B. R., Ryu, C. M., Anderson, A. J. and Kim, Y. C. 2018. Polyamine is a critical determinant of Pseudomonas chlororaphis O6 for GacS-dependent bacterial cell growth and biocontrol capacity. Mol. Plant Pathol. 19: 1257-1266. https://doi.org/10.1111/mpp.12610
  30. Records, A. R. 2011. The type VI secretion system: a multipurpose delivery system with a phage-like machinery. Mol. Plant Microbe Interact. 24: 751-757. https://doi.org/10.1094/MPMI-11-10-0262
  31. Records, A. R. and Gross, D. C. 2010. Sensor kinases RetS and LadS regulate Pseudomonas syringae type VI secretion and virulence factors. J. Bacteriol. 192: 3584-3596. https://doi.org/10.1128/JB.00114-10
  32. Russell, D. W. and Sambrook, J. 2001. Molecular cloning: a laboratory manual. 3rd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA, 999 pp.
  33. Ryu, C. M., Kang, B. R., Han, S. H., Cho, S. M., Kloepper, J. W., Anderson, A. J. et al. 2007. Tobacco cultivars vary in induction of systemic resistance against Cucumber mosaic virus and growth promotion by Pseudomonas chlororaphis O6 and its gacS mutant. Eur. J. Plant Pathol. 119: 383-390. https://doi.org/10.1007/s10658-007-9168-y
  34. Schwyn, B. and Neilands, J. B. 1987. Universal chemical assay for the detection and determination of siderophores. Anal. Biochem. 160: 47-56. https://doi.org/10.1016/0003-2697(87)90612-9
  35. Seaton, S. C., Silby, M. W. and Levy, S. B. 2013. Pleiotropic effects of GacA on Pseudomonas fluorescens Pf0-1 in vitro and in soil. Appl. Environ. Microbiol. 79: 5405-5410. https://doi.org/10.1128/AEM.00819-13