Articles

Molecular identification of antagonistic bacteria from Tehran soils and evaluation of their inhibitory activities toward pathogenic fungi

Abstract

Background and Objectives: To find antagonistic bacteria with potential antifungal activity against some pathogenic fungi, including Aspergillus niger, A. flavus, Fusarium moniliforme and Penicillium marneffei, a total of 148 agricultural soil samples from different sites of Tehran were examined.
 Materials and Methods: Antagonistic soils were selected by screening against A. niger on glucose-yeast extract (GY) agar using a visual agar plate assay method. All growing bacteria were examined for antifungal activity, and antagonistic bacteria identified based on 16S rRNA sequence analysis. Among a total number of 97 bacteria isolated form inhibitory soils (36 samples), 16 bacteria were reported as strong growth inhibitors in co-cultures on GY agar with all tested fungi at variable degrees. Fungal growth inhibitory bacteria were cultured against all fungi and growth inhibition was measured and analyzed between test and control groups by statistical analysis (ANOVA).
Results: Molecular identification of antagonistic bacteria indicated that most bacterial isolates belonged to the genus Bacillus (81.25%), including B. subtilis (5 isolates), B. amyloliquefaciens (6 isolates) and B. valismortis (2 isolates), followed by one isolate (6.25%) from each Streptomyces sp., Pseudomonas chlororaphis and Acinetobacter baumannii. Based on the visual plate assay results, total fungal growth inhibition of all bacteria was reported in the range of 13.2 to 68.3%. P. chlororaphis S105 was reported as the most potent antagonistic bacterium which inhibited the growth of A. niger by 68.3%, followed by F. moniliforme (66.4%), A. flavus (64.7%) and P. marneffei (57.1%).
Conclusion: P. chlororaphis and some other inhibitory bacteria reported in the present study, they may be considered not only as a rich source of useful metabolites with potential application in antifungal drug discovery, but also as potential candidates for biological control programs.

Samson AR, Hoekstra ES, Frisvad JC, Filtenborg O (2000). Introduction to Food and Airborne Fungi, Sixth edn. The Netherlands: CBS-Utrecht.

Ajello L, Hay RJ (1998). Topley & Wilson´s Microbiology and Microbial Infections, Ninth edn., Volume 4, Medical Mycology, New York: Oxford University Press.

Pitt JI, Hocking AD (1997). Fungi and Food Spoilage.2nd edn., London, Blackie.

Ghisalberti EL. (2000). Bioactive metabolites from soilborne fungi: natural fungicides and biocontrol agents. In: Atta-ur-Rahman (ed.) Studies in Natural Products Chemistry, Volume 21, Part 2: Bioactive Natural Products (Part B). Elsevier Science Publication. pp. 181-250.

Yamaguchi I. (1996). Pesticides of microbial origin and applications of molecular biology. In: Copping LG, eds. Crop protection agents from nature: natural products and analogues. London: The Royal Society of Chemistry. pp. 27-49.

Blondelle SE, Houghten RA. Progress in antimicrobial peptides. Annu Rep Med Chem 1992; 27: 159-168.

Bostian K, Silver L. Screening of natural products for antimicrobial agents. Eur J Clin Microbiol Infect Dis 1990; 9: 455-461.

Lange L, Lopez CS. (1996). Microorganisms as a source of biologically active secondary metabolites. In Copping LG, eds. Crop protection agents from nature: natural products and analogues. London: The Royal Society of Chemistry. pp. 1-26.

Clark AM. Natural products as a resource for new drugs.Pharm Res 1996; 13: 1133-1141.

Hua SS, Baker JL, Flores-Espiritu M. Interactions of saprophytic yeasts with a nor mutant of Aspergillus flavus. Appl Environ Microbiol 1999; 65: 2738-2740.

Lane DJ. (1991). 16S/23S rRNA sequencing. In: E Stackebrandt, M Goodfellow, eds. Nucleic acid techniques in bacterial systematics. New York: John Wiley & Sons, Ltd.; pp. 115-175.

Cole JR, Chai B, Farris RJ, Wang Q, Kulam SA, McGarrell DM, Garrity GM, Tiedje JM. The Ribosomal Database Project (RDP-II): sequences and tools for high-throughput rRNA analysis. Nucleic Acids Res 2005; 33: D294-296.

Gregersen T. Rapid method for distinction of gram- negative from gram-positive bacteria. Eur J Appl Microbiol Biotechnol 1978; 5: 123-127.

Sharma RR, Singh D, Singh R. Biological control of postharvest diseases of fruits and vegetables by microbial antagonists: A review. Biological Control 2009; 50: 205-221.

Ongena M. Jacques P. Bacillus lipopeptides: versatile weapons for plant disease biocontrol. Trends Microbiol 2007; 16: 115-125.16. Raaijmakers JM, Vlami M, de Souza JE. Antibiotic production by bacterial biocontrol agents. Antonie van Leeuwenhoek 2002; 81: 537-547.

Stein T.Bacillus subtilis antibiotics: structures, syntheses and specific functions. Mol Microbiol 2005;56: 845-857.

Arrebola E, Jacobs R, Korsten L. Iturin A is the principal inhibitor in the biocontrol activity of Bacillus amyloliquefaciens PPCB004 against postharvest fungal pathogens. J Appl Microbiol 2010; 108: 386-395.

Akhavan Sepahy A, Selselezakeri SH, Rezapanah MR, Motevaze K. An investigation on activity and compounds of Bacillus subtilis against some pathogenic plant fungi. Iran J Biol Sci 2007; 2: 1-14.

Zhao Z, Wang Q, Wang K, Brian K, Liu C, Gu Y. Study of the antifungal activity of Bacillus valismortis ZZ185 in vitro and identification of its antifungal components. Biores Technol 2010; 101: 292-297.

Shtark OIU, Shaposhnikov AI, Kravchenko LV. The production of antifungal metabolites by Pseudomonas chlororaphis grown on different nutrient sources. Mikrobiologiia 2003; 72: 645-650.

Vandenbergh PA, Gonzalez CF, Wright AM, Kunka BS. Iron-chelating compounds produced by soil pseudomonas: correlation with fungal growth inhibition.Appl Environ Microbiol 1983; 46: 128-132.

Reddy BP, Reddy MS, Kumar KVK. Characterization of antifungal metabolites of Pseudomonas fluorescens and their effect on mycelia growth of Magnaporthe grisea and Rhizoctonia solani. Int J PharmTech Res 2009; 1: 1490-1493.

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IssueVol 3 No 3 (2011) QRcode
SectionArticles
Keywords
Antifungal activity Pathogenic fungi Bacillus 16S rRNA Pseudomonas

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1.
Ranjbariyan A, Shams-Ghahfarokhi M, Kalantari S, Razzaghi-Abyaneh M. Molecular identification of antagonistic bacteria from Tehran soils and evaluation of their inhibitory activities toward pathogenic fungi. Iran J Microbiol. 1;3(3):140-146.