Screening of Antagonistic Actinomyces for the Preparation of Bioactive Crude Extract and Development of Starter Culture Powder for Suppression of Rice Pathogen
Abstract
In the present study, several strains of antagonist actinomyces obtained from soils of agricultural fields in Pathum-Thani and Nakhon-Nayok province were screened for antagonistic activity against Xanthomonas oryzae pv. oryzae (Xoo) Pyricularia oryzae and Sclerotium sp., the causative agents of bacterial blight disease, blast disease and stem rot disease of rice, respectively. The isolates were identified as genus Streptomyces based on colony morphology, scanning electron microscope and 16S rRNA gene analysis. Nine Streptomyces isolates produced potential antibacterial or antifungal compounds on screening agars. Among of them, Streptomyces hygroscopicus strain SWU-TH39 showed the highest inhibition zone against both Xoo and rice pathogenic fungi such as Pyricularia oryzae and Sclerotium sp. This strain also produced plant growth promoting compounds including indole-acetic acid (IAA) and siderophores. The antimicrobial compounds produced by strain SWU-TH39 was obtained in modified-antibiotic production medium and extracted by solvent extraction method. n[a1] -Butanol was the best extraction solvent for the recovery of the antimicrobial compounds for inhibition against both Xoo and tested fungal phytopathogen. Furthermore, SWU-TH39 was adapted to the development of starter culture in powder formulation and, remarkably, this was successful as a carrier to maintain bacterial survival with 8x108 CFU/g after 6-month storage at room temperature. Viable cells in the starter retained not only antagonistic potential against rice pathogens, but also capabilities of the production of plant growth promoting substances. Keywords : antagonist actinomyces, rice pathogens, antimicrobial, starter culture powderReferences
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Zaitlin, B., Turkington, K., Parkinson, D., & Clayton, G. (2004). Effects of tillage and inorganic fertilizers on culturable soil actinomycete communities and inhibition of fungi by specific actinomycetes. Applied Soil Ecology, 26, 53-62.
Bressan, W. (2003). Biological control of maize seed pathogenic fungi by use of actinomycetes. BioControl, 48, 233-240.
Compant, S., Duffy, B., Nowak, J., Clément, C., & Barka, E.A. (2005). Use of plant growth-promoting bacteria for biocontrol of plant diseases: principles, mechanisms of action, and future prospects. Applied and Environmental Microbiology, 71, 4951-4959.
Dai, L.Y., Liu, X.L., Xiao, Y.H., & Wang, G.L. (2007). Recent advances in cloning and characterization of disease resistance genes in rice. Journal of Integrative Plant Biology, 49, 112-119.
Harikrishnan, H., Shanmugaiah, V., Balasubramanian, N., Sharma, M. P., & Kotchoni, S.O. (2014). Antagonistic potential of native strain Streptomyces aurantiogriseus VSMGT1014 against sheath blight of rice disease. World Journal of Microbiology and Biotechnology, 30, 3149–3161.
Hastuti, R.D., Lestari, Y., Saraswati, R., Suwanto, A., & Chaerani (2012). Capability of Streptomyces spp. in controlling bacterial leaf blight diseases in rice plants. American Journal of Agricultural and Biological Sciences, 7, 217-223.
Hoa, P. T. P., Quang, N. D., Sakiyama, Y., Hop, D.V., Hang, D. T., Ha, T. H., Van, N. T., Quy, N. T. K., & Dao, N. T. A. (2012). Screening for Actinomyces isolated from soil with the ability to inhibit Xanthomonas oryzae pv.oryzae causing rice bacterial blight disease in Vietnam. African Journal of Biotechnology, 11,
14586–14594.
Ismet, A., Vikineswary, S., Paramaswari, S., Wong, W.H., Ward, A., Seki, T., Fiedler, H. P., & Goodfellow, M. (2004). Production and chemical characterization of antifungal metabolites from Micromonospora sp. M39 isolated from mangrove rhizosphere soil. World Journal of Microbiology and Biotechnology, 20, 523-528.
Kieser, T., Bibb, M.J., Buttner, M.J., Chater, K.F., & Hopwood, D.A. (2000). Practical Streptomyces
Genetics. Norwich, UK: The John Innes Foundation.
Lane, D.J. (1991). 16S/23S rRNA sequencing. In E. Stackebrandt, & M. Goodfellow. (Eds.), Nucleic acid techniques in bacterial systematics. (pp. 115-175). New York: John Wiley & Sons.
Loqman, S., Barka, E. A., Clement, C., & Ouhdouch, Y. (2009). Antagonistic actinomycetes from Moroccan soil to control the grapevine gray mold. World Journal of Microbiology and Biotechnology, 25, 81-91.
Park, S.B., Lee, I.A., Joo, W.S., Jeong, G.K., & Lee, H.W. (2011). Screening and identification of antimicrobial compounds from Streptomyces bottropensis suppressing rice bacterial blight. Journal of Microbiology and Biotechnology, 21, 1236-1242.
Pupakdeepan, W., & Prathuangwong, S. (2010). Formulation development of new bacterial antagonist and control efficacy against bacterial leaf blight of rice. In Proceedings of 48th Kasetsart University Annual Conference: Plants. (pp. 207-214). Bangkok: Kasetsart University.
Schwyn, B., & Neilands, J.B. (1987). Universal chemical assay for the detection and determination of siderophores. Analytical Biochemistry, 160, 47-56.
Souza, R.F., Coelho, R.R.R., Macrae, A., Soares, R.M.A., Nery, D.C.M., Semêdo, L.T.A.S., Alviano, C.S. & Gomes, R.C. (2008). Streptomyces lunalinharesii sp. nov., a chitinolytic streptomycete isolated from cerrado soil in Brazil. International Journal of Systematic and Evolutionary Microbiology, 58, 2774-2778.
Techapun, C., Sinsuwongwat, S., Poosaran, N., Watanabe, M., & Sasaki, K. (2001). Production of a cellulase-free xylanase from agricultural waste materials by a thermotolerant Streptomyces sp. Biotechnology Letter, 23, 1685-1689.
Veronica, Lay, B.W., & Magdalena, S. (2014). Isolation and characterization of new antibiotics from Indonesian coastal marine bacteria. Microbiology Indonesia, 8, 87-93.
Williams, S.T., Goodfellow, M., Alderson, G., Wellington, E.M.H., Sneath, P.H.A. & Sackin, M.J. (1983). Numerical classification of Streptomyces and related genera. Journal of general microbiology, 129, 1743-1813.
Yu, J., Liu, Q., Liu, Q., Liu, X., Sun, Q., Yan, J., Qi, X., & Fan, S. (2008). Effect of liquid culture requirements on antifungal antibiotic production by Streptomyces rimosus MY02. Bioresource Technology, 99,
2087-2091.
Zaitlin, B., Turkington, K., Parkinson, D., & Clayton, G. (2004). Effects of tillage and inorganic fertilizers on culturable soil actinomycete communities and inhibition of fungi by specific actinomycetes. Applied Soil Ecology, 26, 53-62.
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2017-10-20
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