ISOLATION AND IDENTIFICATION OF BACILLUS SPP. FROM COMPOST MATERIAL, COMPOST AND MUSHROOM CASING SOIL ACTIVE AGAINST TRICHODERMA SPP.
The isolation of bacteria was carried out from samples of straw and chicken manure, compost at various stages of the composting process and casing soil used for growing button mushrooms. A preliminary screening of 108 bacterial isolates for antagonistic activity against Trichoderma aggressivum f. europaeum showed that 23 tested isolates inhibited mycelial growth of the pathogenic fungus. Further screening with four indicator isolates of fungi revealed that all 23 bacterial isolates inhibited the growth of T. aggressivum f. europaeum, T. harzianum and T. koningii, while only 13 isolates inhibited the growth of T. atroviride. T. aggressivum f. europaeum proved to be the most sensitive, with many bacterial isolates generating a high percentage of growth inhibition. Only two bacterial isolates (B-129 and B-268) were successful in inhibiting the growth of all 4 tested pathogens. All 23 bacterial isolates were characterized as Gram-positive and catalase-positive and were subjected to molecular identification based on the partial sequence, the hypervariant region of the 16S rDNA. It was shown that the obtained bacterial strains belong to Bacillus subtilis, B. amyloliquefaciens, B. licheniformis and B. pumilus species.
Key words: Agaricus bisporus; Bacillus; biocontrol; green mold; hypervariant region
Received: November 4, 2015; Revised: December 25, 2015; Accepted: January 21, 2016; Published online: August 9, 2016
How to cite this article: Stanojević O, Milijašević-Marčić S, Potočnik I, Stepanović M, Dimkić I, Stanković S, Berić T. Isolation and identification of Bacillus spp. from compost material, compost and mushroom casing soil active against Trichoderma spp.. Arch Biol Sci. 2016;68(4):845-52.
Chang ST, Wasser SP. The role of culinary-medicinal mushrooms on human welfare with a pyramid model for human health. Int J Med Mushrooms. 2012;14:95-134.
Seaby DA. Investigation of the epidemiology of green mold of mushroom (Agaricus bisporus) compost caused by Trichoderma harzianum. Plant Pathol. 1996;45:913-23.
Kosanović D, Potočnik I, Duduk B, Vukojević J, Stajić M, Rekanović E, Milijašević-Marčić S. Trichoderma species on Agaricus bisporus farms in Serbia and their biocontrol. Ann Appl Biol. 2013;163(2):218-30.
Grogan HM, Gaze RH. Fungicide resistance among Cladobotryum spp. – causal agents of cobweb disease of the edible mushroom Agaricus bisporus. Mycol Res. 2000;104:357-64.
Živković S, Stojanović S, Ivanović Ž, Gavrilović V, Popović T, Balaž J. Screening of antagonistic activity of microorganisms against Colletotrichum acutatum and Colletotrichum gloeosporioides. Arch Biol Sci. 2010;62(3):611-23.
Fravel DR. Commercialization and implementation of biocontrol. Annu Rev Phytopathol. 2005;43:337-59.
Leelasuphakul W, Hemmanee P, Chuenchitt S. Growth inhibitory properties of Bacillus subtilis strains and their metabolites against the green mold pathogen (Penicillium digitatum Sacc.) of citrus fruit. Postharvest Biol Tec. 2008;48:113-21.
Kim P, Chung KC. Production of an antifungal protein for control of Colletotrichum lagenarium by Bacillus amyloliquefaciens MET0908. FEMS Microbiol Lett. 2004;234:177-83.
Leelasuphakul W, Sivanunsakul P, Phongpaichit S. Purification, characterization and synergistic activity of β-1,3-glucanase and antibiotic extract from an antagonistic Bacillus subtilis NSRS 89-24 against rice blast and sheath blight pathogens. Enzyme Microb Tech. 2006;38:990-7.
Food and Drug Administration. Code of Federal Regulations, Title 21: Food and Drugs, Chapter 1: Food and Drug Administration Department of Health and Human Services, Part 184: Direct Food Substances Affirmed as Generally Recognized as Safe. Washington, DC: US Government Printing Office; 1999.
Walker R, Powell AA, Seddon B. Bacillus isolates from the spermosphere of peas and dwarf French beans with antifungal activity against Botrytis cinerea and Pythium species. J Appl Microbiol. 1998;84:791-801.
Suslow TV, Schroth MN, Isaka M. Application of a rapid method for Gram differentiation of plant pathogenic and saprophytic bacteria without staining. Phytopathology. 1982;72:917-8.
Fokkema NJ. Fungal antagonism in the phyllosphere. Ann Appl Biol. 1978;89:115-7.
Korsten L, De Jager ES. Mode of action of Bacillus subtilis for control of avocado postharvest pathogens. SAAGA Yearbook. 1995;18:124-30.
Pastrik KH, Maiss E. Detection of Ralstonia solanacearum in Potato Tubers by Polymerase Chain Reaction. J Phytopathol. 2000;148:619-26.
Goto K, Omura T, Hara Y, Sadaie Y. Application of the partial 16S rDNA sequence as an index for rapid identification of species in the genus Bacillus. J Gen Appl Microbiol. 2000;46:1-8.
Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997;25(17):3389-402.
Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 1997;25(24):4876-82.
Obagwu J, Korsten L. Integrated control of citrus green and blue molds using Bacillus subtilis in combination with sodium bicarbonate or hot water. Postharvest Biol Tec. 2003;28:187-94.
Pusey PL, Wilson CL. Postharvest biological control of stone fruit brown rot by Bacillus subtilis. Plant Dis. 1984;68:753-6.
Lee JP, Lee SW, Kim CS, Son JH, Song JH, Lee KY, Kim H.J, Jung SJ, Moon BJ. Evaluation of formulation of Bacillus licheniformis for the biological control of tomato gray mold caused by Botrytis cinerea. Biol Control. 2006;37(3):329-37.
Yoshida S, Shirata A, Hiradate S. Ecological characteristics and biological control of mulberry anthracnose. Jpn Agr Res Q. 2002;36(2):89-95.
Munimbazi C, Bullerman LB. Isolation and partial characterization of antifungal metabolites of Bacillus pumilus. J Appl Microbiol. 1998;84:959-68.
Chittihunsa T, Bangeekhan E, Wongsamitkul N, Subsomboon T. Screening of Bacillus spp. suppressing the infection of Trichoderma sp. in mushroom cultivation. KMITL Sci Tech J. 2007;7:19-27.
Dimkić I, Živković S, Berić T, Ivanović Ž, Gavrilović V, Stanković S, Fira Đ. Characterization and evaluation of two Bacillus strains, SS-12.6 and SS-13.1, as potential agents for the control of phytopathogenic bacteria and fungi. Biol Control. 2013;65:312-21.
Edwards SG, McKay T, Seddon B. Interaction of Bacillus species with phytopathogenic fungi − Methods of analysis and manipulation for biocontrol purposes. In: Blakeman JP, Williamson B, editors. Ecology of Plant Pathogens. Wallingford, Oxon, UK: CAB International; 1994. p. 101-118.
Stein T. Bacillus subtilis antibiotics: Structures, syntheses and specific functions. Mol Microbiol. 2005;56845-857.
Chen XH, Koumoutsi A, Scholz R, Schneider K, Vater J, Süssmuth R, Piel J, Borriss R. Genome analysis of Bacillus amyloliquefaciens FZB42 reveals its potential for biocontrol of plant pathogens. J Biotechnol. 2009;140(1):27-37.
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