DOI QR코드

DOI QR Code

The Biocontrol Activity of Chromobacterium sp. Strain C-61 against Rhizoctonia solani Depends on the Productive Ability of Chitinase

  • Park, Seur-Kee (Department of Agricultural Biology, Suncheon National University) ;
  • Lee, Myung-Chul (Division of Cytogenetics, National Institute of Agricultural Science and technology, RDA) ;
  • Harman, Gary E. (Department of Horticultural Science and Plant Pathology, Cornell University)
  • Published : 2005.09.01

Abstract

A chitinolytic bacterium, Chromobacterium sp. strain C-61, was found strongly antagonistic to Rhizoctonia solani, a causal agent of damping-off of eggplant. In this study, the biocontrol activity and enzymatic characteristics of strain C-61 were compared with its four Tn5 insertion mutants (C61-A, -B, -C, and -D) that had lower chitinolytic ability. The chitinase activity of a 2-day old culture was about $76\%,\;49\%\;and\;6\%$ level in C61-A, C61-B and in C61-C, respectively, compared with that of strain C-61. The $\beta-N-acetylhexosaminidase$(Nahase) activity was little detected in strain C-61 but increased largely in C-61A, C61-B and C61-C. Activities of chitinase and Nahase appeared to be negatively correlated in these strains. Another mutant, C-61D, produced no detectable extracellular chitinase and Nahase. The in vitro and in vivo biocontrol activities of strain C-61 and its mutants were closely related to their ability to produce chitinase but not Nahase. No significant differences in population densities between strain C-61 and its mutants were observed in soil around eggplant roots. The results of SDS-PAGE and isoelectrofocusing showed that a major chitinase of strain C-61 is 54-kDa with pI of approximately 8.5. This study provides evidence that the biocontrol activity of Chromobacterium sp. strain C-61 against Rhizoctonia solani depends on the ability to produce chitinase with molecular weight of 54-kDa and pI of 8.5.

Keywords

References

  1. Ausubel, F. M., Brent, R., Kingston, R. R., Moore, D. D., Seidman, J. G., Smith, J. A. and Struhl, K. 1992. Short protocols in molecular biology. Second edition. John Wiley and Sons. USA
  2. Chet, I., Ordentlich, A., Shapira, R. and Oppenheim, A. 1990. Mechanisms of biocontrol of soil-borne plant pathogens by rhizobacteria. Plant Soil 129:85-92 https://doi.org/10.1007/BF00011694
  3. Chemin, L. S., Ismailov, Z., Haren, S. and Chet, I. 1995. Chitinolytic Enterobacter agglomerans antagonistic to fungal plant pathogens. Appl. Environ. Microbiol. 57:2426-2428
  4. Chemin, L. S., Fuente, L., Sobolev, V., Haren, S., Vorgias, C., Oppenheim, A. and Chet, I. 1997. Molecular cloning, structural analysis, and expression in Escherichia coli of a chitinase gene from Enterabacter agglomerans. Appl. Environ. Microbiol. 63:834-839
  5. Chemin, L. S., Winson, M. K., Thompson, J. M., Haran, S., Bycroft, B. W., Chet, I., Williams, P. and Stewart, G. S. 1998. Chitinolytic activity in Chromobacterium violaceum: substrate analysis and regulation by quorum sensing. J. Bacteriol. 180:4435-4441
  6. Crosa, J. H. and Falkow, S. 1981. Plasmids, PP. 266-282, In: Manual of methods for general bacteriology, Gerharadt, P., Murray, R. G. E., Costilow, R. N., Nester, E. W., Wood, W. A., Krieg, N. R., Phillips, G. B. (eds), American Society for Microbiology, Washing DC
  7. De Bruijin, F. J. and Lupski, J. R. 1984. The use of transposon Tn5 mutagenesis in the rapid generation of correlated physical and genetic maps of DNA segments cloned into multicopy plasmids-a revew. Gene 27: 131-149 https://doi.org/10.1016/0378-1119(84)90135-5
  8. Dhingra, O. D. and Sinclair, J. B. 1987. Basic plant pathology methods. CRC Press, Inc. Florida
  9. Hames, B. D. and Rickwood, D. 1990. Gel electrophoresis of proteins. 2 nd ed. Oxford University Press, New York
  10. Harman, G. E., Hayes, C. K., Lorito, M., Broadway, R. M., Di Pietro, A., Peterbauer, C. and Tronsmo, A. 1993. Chitinolytic enzymes of Trichoderma harzianum:Purification of chitobiosidase and endochitinase. Phytopathology 83:313-318 https://doi.org/10.1094/Phyto-83-313
  11. Inbar, J. and Chet, I. 1991. Evidence that chitinase produced by Aeromonas caviae is involved in the biological control of soilborne plant pathogens by this bacterium. Soil. Biol. Biochem. 23:973-978 https://doi.org/10.1016/0038-0717(91)90178-M
  12. Jee, H. J., Nam, C. G. and Kim, C. H. 1988. Studies on biological control of Phytophthora blight of red-pepper I. isolation of antagonists and evaluation of antagonistic activity in vitro and in greenhouse. Korean J. Plant Pathol. 4:305-312
  13. Kim, B. S. and Hwang, B. K. 1992. Isolation of antibiotic-producing bacteria antagonistic to Phytophthora capsici from pepper-growing soils and evaluation of their antibiotic activity. Korean J. Plant Pathol. 8:241-248
  14. Laemmli, UK. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227:680-685 https://doi.org/10.1038/227680a0
  15. Lan, X., Ozawa, N., Nishiwaki, N., Kodaira, R., Okazaki, M. and Shim osaka, M. 2004. Purification, cloning, and sequence analysis of beta-N-acetylglucosaminidase from the chitinolytic bacterium Aeromonas hydrophila strain SUWA-9. Biosci. Biotechnol. Biochem. 68: 1082-1090 https://doi.org/10.1271/bbb.68.1082
  16. Lorito, M., Harman, G. E., Hayes, C. K., Broadway, R. M., Tronsmo, A., Woo, S. L. and DiPietro, A. 1993. Chitinolytic enzymes produced by Trichoderma harzianum: antifungal activity of purified endochitinase and chitobiosidase. Phytopathology 83:302-307 https://doi.org/10.1094/Phyto-83-302
  17. Lorito, M., Hayes, C. K., Di Pietro, A., Woo, S. L. and Hannan, G. E. 1994. Purification, characterization, and synergisitic activity of a glucan 1,3-$\beta$-glucosidase and an Nsacetyl-$\beta$-glucosaminidase from Trichoderma harzianum. Phytopathology 84: 398-405 https://doi.org/10.1094/Phyto-84-398
  18. Maniatis, T., Fritsch, E. F. and Sambrook, J. 1982. Molecular cloning.a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
  19. Matsuo, Y., Kurita, M., Park, J. K., Tanaka, K., Nakagawa, T., Kawamukai, M. and Matsuda, H. 1999. Purification, characterization and gene analysis of N-acetylglucosaminidase from Enterobacter sp. G-1. Biosci. Biotechnol. Biochem. 63: 1261-1268 https://doi.org/10.1271/bbb.63.1261
  20. Ordentlich, A., Elad, Y. and Chet, I. 1988. The role of chitinase of Serratia marceacens in biocontrol of Sclerotium rolfsii. Phytopathology 78:84-88
  21. Park, J. H. and Kim, H. K. 1989. Biological control of Phytophthora crown and root rot of greenhouse pepper with Trichoderma harzianum and Enterobacter agglomerans by improved method of application. Korean J. Plant Pathol. 5: 112
  22. Park, S. K., Lee, H. Y. and Kim, K. C. 1995a. Antagonistic effect of chitinolytic bacteria on soilborne plant pathogens. Korean J. Plant Pathol. 11 :47-52
  23. Park, S. K., Lee, H. Y. and Huh, J. W. 1995b. Production and some properties of chitinolytic enzymes by antagonistic bacteria. Korean J. Plant Pathol. 11 :258-264
  24. Park, S. K. and Yoo, J. G. 1995. Isolation and identification of chitinolytic bacteria from soil. J Agric. Sci. Res. Sunchon Nat'l Univ. 9:95-102
  25. Sahai, A. S. and Manocha, M. S. 1993. Chitinases of fungi and plants: their involvement in morphogenesis and host-parasite interaction. FEMS Microbiol. Rev. 11:317-338 https://doi.org/10.1111/j.1574-6976.1993.tb00004.x
  26. Shen, S. S., Choi, O. H., Lee, S. M. and Park, C. S. 2002. In vitro and in vivo activities of a biocontrol agent, Serratia plymuthica A21-4, against Phytophthora capsici. Korean J. Plant Pathol. 18:221-224 https://doi.org/10.5423/PPJ.2002.18.4.221
  27. Singh, P. P., Shin, Y. C. Park, C. S. and Chung, Y. R. 1999. Biological control of Fusarium wilt of cucumber by chitinolytic bacteria. Phytopathology 89:92-99 https://doi.org/10.1094/PHYTO.1999.89.1.92
  28. Sneh, B. 1981. Use of rhizosphere chitinolytic bacteria for biological control of Fusarium oxysporum in carnation. Phytopath. Z. 100:251-256 https://doi.org/10.1111/j.1439-0434.1981.tb03298.x
  29. Tews, I., Vincentelli, R. and Vorgias, C. E. 1996. N-Acetylglu-cosaminidase (chitobiase) from Serratia marcescens: gene sequence, and protein production and purification in Escherichia coli. Gene 170:63-67 https://doi.org/10.1016/0378-1119(95)00848-9
  30. Tronsmo, A. and Harman, G. E. 1993. Detection arid quantification of N-acetyl-$\beta$-D-glucosaminidase, chitobiosidase, and endochitinase in solutions and on gels. Anal. Biochem. 208:74-79 https://doi.org/10.1006/abio.1993.1010
  31. Trudel, K. and Asselin, A. 1989. Detection of chitinase activity after polyacrylcmide gel electrophresis. Anal. Biochem. 178:362-366 https://doi.org/10.1016/0003-2697(89)90653-2
  32. Tsujibo, H., Fujimoto, K., Kimura, Y., Miyamoto, K., Imada, C., Okami, Y. and Inamori, Y. 1995. Purification and characterization of beta-N-acetylglucosaminidase from Alteromonas sp. strain O-7. Biosci. Biotechnol. Biochem. 59:1135-1136 https://doi.org/10.1271/bbb.59.1135
  33. Watanabe, T., Oyanagi, W., Suzuki, K. and Tanaka, H. 1990, Chitinase system of Bacillus circulans W1-12 and importance of chitinase A1 in chitin degradation. J. Bacteriol. 172:4017-4022 https://doi.org/10.1128/jb.172.7.4017-4022.1990
  34. Yabuki, M., Mizushima, K., Amatatsu, T., Ando, A., Fuji, T., Shimada, M. and Yamashita, M. 1986. Purificatioriarid characterization of chitinasee and chitobiase produced by Aeromonas hydrophila subsp. anaerogenesA 52. J. Gen. Appl. Microbiol. 32:25-38 https://doi.org/10.2323/jgam.32.25

Cited by

  1. Insight Into Genes Involved in the Production of Extracellular Chitinase in a Biocontrol Bacterium Lysobacter enzymogenes C-3 vol.28, pp.4, 2012, https://doi.org/10.5423/PPJ.NT.07.2012.0115
  2. Cloning and high-level production of a chitinase from Chromobacterium sp. and the role of conserved or nonconserved residues on its catalytic activity vol.74, pp.4, 2007, https://doi.org/10.1007/s00253-006-0614-0
  3. An effective biocontrol bioformulation against Phytophthora blight of pepper using growth mixtures of combined chitinolytic bacteria under different field conditions vol.120, pp.4, 2008, https://doi.org/10.1007/s10658-007-9227-4
  4. Both extracellular chitinase and a new cyclic lipopeptide, chromobactomycin, contribute to the biocontrol activity ofChromobacteriumsp. C61 vol.15, pp.2, 2014, https://doi.org/10.1111/mpp.12070
  5. Isolation and characterization of chitinolytic rhizobacteria for the management of Fusarium wilt in tomato vol.30, pp.12, 2011, https://doi.org/10.1016/j.cropro.2011.02.032
  6. Antifungal Action of NewTrichoderma Spp.Romanian Isolates on Different Plant Pathogens vol.23, pp.sup1, 2009, https://doi.org/10.1080/13102818.2009.10818536
  7. Identification and characterization of a chitinase of Stenotrophomonas maltophilia, a bacterium that is antagonistic towards fungal phytopathogens vol.113, pp.1, 2012, https://doi.org/10.1016/j.jbiosc.2011.08.023
  8. Draft Genome Sequence of the Biocontrol Bacterium Chromobacterium sp. Strain C-61 vol.193, pp.23, 2011, https://doi.org/10.1128/JB.06191-11