Article | 08. 2015 Vol. 33, Issue. 4
Antagonistic Mechanisms and Culture Conditions of Isolated Microbes Applied for Controlling Large Patch Disease in Zoysiagrass



Hyosung O&B Co. Ltd.1
Jeollanam-do Agricultural Research & Extension Services,2
Department of Horticulture Science, Chungnam National University3




2015.08. 492:500


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Our previous report demonstrated successful isolation of soil-borne bacteria that suppressed the potential of Rhizoctonia solani AG2-2 (IV) causing turfgrass large patch disease when applied to Korean lawngrass (Zoysia japonica). The current study aimed to uncover the mechanisms of this antagonism of Rhizoctonia solani and to define culture conditions for the isolated microbes. We found that two Bacillus isolates, I-009 and FRIN-001-1 strains, produced cellulase and siderophore, but not chitinase, while the Pseudomonas YPIN-022 strain was found to release only siderophore, implying that three antagonistic bacteria commonly interrupt Fe uptake by the large patch pathogen. The I-009 and FRIN-001-1 isolates grew best at 35 and 30°C in growth medium of pH 5 to 8 for 32 and 28 h, respectively, while optimum growth for the YPIN-022 strain was found at 35°C at pH 5 to 9 for 24 h. Good growth of I-009 and YPIN-022 over 24 h was obtained in M9 minimal medium supplemented with 1% sucrose, 0.5% yeast extract and 0.1% potassium chloride. FRIN-001-1 grew well in M9 medium with 1% mannitol, 0.5% yeast extract and 0.1% potassium phosphate dibasic.



1. Bhat, K.M. and R. Maheshwari. 1987. Sporotrichum thermophile growth, cellulose degradation and cellulase activity. Appl. Environ. Microbiol. 53:2175-2182.  

2. Broque, K., I. Chet, M. Holliday, R. Cressman, P. Biddle, S. Knowlton, C.J. Mauvais, and R. Broglie. 1991. Transgenic plants with enhanced resistance to the fungal pathogen Rhizoctonia solani. Science 254:1194-1197.  

3. Burpee, L.L. and L.G. Goulty. 1984. Suppression of brown patch disease of creeping bentgrass by isolate of nonpathogenic Rhizoctonia spp. Phytopathology 74:692-694.  

4. Chet, I. 1990. Biological control of soil-borne plant pathogens with fungal antagonists in combination with soil treatment, p.15-25. In: D. Hornby(Ed), Biological Control of soil-borne Plant Pathogens. CAB International, Wallingford, Oxon, UK.  

5. Choi, D.H., N.I. Park, S.H. Choi, K.W. Park, J.W. Kim, Y.S. Kwak, and J.J. Lee. 2012. Composition and invading problem of interspecies turfgrass on golf course. Kor. J. Weed Sci. 32:174-179.   

6. Chun, J. Y., I.H. Ryu, S.U. Lee, and K.S. Lee. 2000. Purification and properties of anticaries microbial agent by Bacillus alkalophilshaggy JY-827. Kor. J. Appl. Microbiol. Biotechnol. 28:270-278.  

7. Compant, S., B. Duffy, J. Nowak, C. Clément, and E.A. Barka. 2005. Use of plant growth-promoting bacteria for biocontrol of plant diseases: principles, mechanisms of action, and future prospects. Appl. Environ. Microbiol. 71:4951-4959.  

8. Couch, H. B. 1985. Common names for turfgrass disease. In: Common names for Plant Disease. ed. by J. D. Hansen. Plant Dis. 63: 629-676.  

9. Fridlender, M., J. Inbar, and I. Chet. 1993. Biological control of soilborne plant pathogens by a β-1,3 glucanase-producing Pseudomonas cepacia. Soil Biol. Biochem. 25:1211-1221.  

10. Giesler, L.J. and G,Y. Yuen. 1998. Evaluation of Stenotrophomonas maltophilia strain C3 for biocontrol of brown patch disease. Crop Prot. 17:509-513.  

11. Goodman, D.M. and L.L. Burpee. 1991. Biological control of dollar spot disease of creeping bentgrass. Phytopathology 81:1438-1446.  

12. Hoy, M.A. and D.C. Herzog. 1985. Biological control in agricultural IPM system. Academic Press. Orlando, Florida, USA.  

13. Hsu, S.C. and J.L. Lockwood. 1975. Powdered chitin agar as a selective medium for enumeration of Actinomycetes in water and soil. Appl. Microbiol. 29:422-426.   

14. Kang, S.W., M.J. Chung, I.H. Yeo, and D.H. Kim. 1998. Isolation and characterization of Pseudomonas vesicularis KW-15 for producing chitinase. J. Chitin. Chitosan. 3:303-312.  

15. Kim, H.Y. and T.S. Lee. 2009. Isolation of antifungal substances by Bacillus amyloliquefaciens IUB158-03 and antagonistic activity against pathogenic fungi. Kor. J. Mycol. 37:96-103.  

16. Kim, Y.S., S.K. Ham and H.J. Lim. 2012. Monitoring of soil chemical properties and pond water quality in golf course after application of SCB liquid fertilizer. Asian J. Turfgrass Sci. 26:44-53.  

17. Lawton, M.B. and L.L. Burpee. 1990. Effect of rate and frequency of application of Typhulapha corrhiza on biological control of Typhula blight of creeping bentgrass. Phytopathology 80:70-73.  

18. Lee, J. J., H. S. Lim, T. H. Chang, and S. D. Kim. 1999. Isolation of siderophore producing Pseudomonas fluorescens GL7 and its biocontrol activity against root-rot disease. Kor. J. Appl. Microbiol. Biotechnol. 27:427-432.  

19. Limón, M.C., M.R. Chacón, R. Mejías, J. Delgado-Jarana, A.M. Rincón, A.C. Codón. T. Benítez. 2004. Increased antifungal and chitinase specific activities of Trichoderma harzianum CECT 2413 by addition of a cellulose binding domain. App. Microbiol. Biotechnol. 64:675-685.  

20. Lopez, C.S., H. Heras, S.M. Ruzal, C. Sanchez-Rivas, and E. A. Rivas. 1998. Variations of the envelope composition of Bacillus subtilis during growth in hyperosmotic medium. Curr. Microbiol. 36:55-61.  

21. Lounes, A., Lebrihi, A., Benslimane, C., Lefebvre, G. and Germain, P. 1996. Regulation of spiramycin synthesis in Streptomyces ambofaciens: effects of glucose and inorganic phosphate. Appl. Microbiol. Biotechnol. 45:204-211.  

22. Ma, K.Y., S.N. Kwak, and G.J. Lee. 2013. Isolation and selection of antagonistic microbes for biological control of zoysiagrass large patch disease. Kor. J. Hort. Sci. Technol. 31:657-665.  

23. Martínez-Cuesta, C., T. Requena, and C. Peláez. 2002. Effect of bacteriocin-induced cell damage on the branched-chain amino acid transamination by Latococcus lactis. FEMS Microbiol. Lett. 217:109-113.  

24. Melchers, L.S., M.B. Sela-Buuriage, S.A. Vloemans, C.P. Woloshuk, J.S. van Roekel, J. Pen, P.J.M. van den Elzen, and B.J.C. Cornelissen. 1993. Extracellular targeting of the vacuolar tobacco proteins AP24, chitinase and β-1,3-glucanses in transgenic plants. Plant Mol. Biol. 21:583-593.   

25. Mulders, J.W.M., I.J. Boerrigter, H.S. Rollema, R.J. Siezen, and W.M. de Vos. 1991. Identification and characterization of the lantibiotic nisin Z, a natural nisin variant. Eur. J. Biochem. 201:581-584.  

26. Nelson, E.B. and C.M. Craft. 1991. Introduction and establishment of strains of Enterobacter cloacae in golf course turf for the biological control of dollar spot. Plant Dis. 75:510-514.  

27. Payne, S. M. 1994. Detection, isolation, and characterization of siderophore. Methods Enzymol. 235:329-344.  

28. Roberts, W.K., and C.P. Selitrennikoff. 1988. Plant and bacterial chitinases differ in antifungal activity. J. Gen. Microbiol. 134: 169-176.  

29. Sela-Buurlage, M.B., A.S. Ponstein, S.A. Bres-Vloemans, L.S. Melchers, P.J.M. van den Elzen, and B.J.C. Cornelissen. 1993. Only specific tobacco (Nicotiana tabacum) chitinases and β-1,3-glucanases exhibit antifungal activity. Plant Physiol. 101: 857-863.  

30. Shivanna, M.B., M.S. Meera, and M. Hyakumachi. 1994. Sterile fungi from zoysiagrass rhizosphere as plant growth promoters in spring wheat. Can. J. Microbiol. 40:637-644.  

31. Shim, G.Y., J.W. Kim, and H.K. Kim. 1994. Occurrence of Rizoctonia blight of zoysiagrasses in golf courses in Korea. Kor. J. Plant Pathol. 10(1):54-60.   

32. Shim, G.Y. and H.K. Kim. 2000. Control of large patch caused by Rhizoctonia solani AG2-2 by combined application of antagonists and chemicals in golf courses. Kor. Turfgrass Sci. 13:131-138.  

33. Shomura, T., N. Nishizawa, M. Iwata, J. Yoshida, M. Ito, S. Aman, M. Koyama, M. Kojima, and S. Inouye. 1983. Studies on a new nucleoside antibiotic, dapiramicin. I. Producing organism, assay method and fermentation. J. Antibiot. 36:1300-1304.  

34. Smith, J.D. 1980. Is biological control of Marasmius oreades fairy rings possible? Plant Dis. 64:348-354.  

35. Tredway, L.P. and L.L. Burpee. 2001. Rhizoctonia diseases of turfgrass. The Plant Health Instructor. DOI: 10.1094/PHI-I- 2001-1109-01.  

36. Viji, G., W. Uddin, and C.P. Romaine. 2003. Suppression of gray leaf spot (blast) of perennial ryegrass turf by Pseudomonas aeruginosa from spent mushroom substrate. Biol. Control 26:233-243.