Compuestos Antimicrobianos
Páginas: 20 (4893 palabras)
Publicado: 7 de mayo de 2012
J. Microbiol. Biotechnol. (2011), 21(12), 1236–1242 http://dx.doi.org/10.4014/jmb.1106.06047 First published online 11 September 2011
Screening and Identification of Antimicrobial Compounds from Streptomyces bottropensis Suppressing Rice Bacterial Blight
Park, Sait Byul1,2, In Ae Lee3, Joo-Won Suh3, Jeong-Gu Kim2*, and Choong Hwan Lee1*
Division of Bioscience and Biotechnology, KonkukUniversity, Seoul 143-701, Korea Genomics Division, National Academy of Agricultural Science, Rural Development Administration, Suwon 441-857, Korea Division of Bioscience and Bioinformatics, College of Natural Science, Myongji University, Yongin 449-728, Korea.
1 2 3
Received: June 28, 2011 / Revised: August 2, 2011 / Accepted: August 5, 2011
Xanthomonas oryzae pv. oryzae (Xoo) is the mostdevastating pathogen to Oryza sativa and has been shown to cause bacterial blight. Two bioactive compounds showing antimicrobial activities against Xoo strain KACC 10331 were isolated from a Streptomyces bottropensis strain. The ethyl acetate extract was fractionated on a Sephadex LH-20 column, and then purified by preparative HPLC. The purified compounds were identified as bottromycin A2 anddunaimycin D3S by HR/MS and 1H NMR analyses. The MIC value against Xoo and the lowest concentration still capable of suppressing rice bacterial blight were 2 µg/ml and 16 µg/ml for bottromycin A2, and 64 µg/ml and 0.06 µg/ml for dunaimycin D3S, respectively. These two compounds were shown to exert different bioactivities in vitro and in rice leaf explants. Keywords: Bottromycin A2, dunaimycin D3S,Streptomyces bottropensis, Xanthomonas oryzae pv. oryzae, antibacterial activity, rice bacterial blight
fields; however, the world-wide use of artificial pesticides is gradually decreasing because of their excessive toxicity and degradation properties [2]. Thus, it is necessary to identify safe natural bioactive substances that can be used as alternatives to artificial pesticides. Biological controlusing microorganisms for inhibition of plant disease holds promise as a robust and long-lasting alternative to synthetic chemicals [7]. Streptomyces has been historically examined as a microorganism that can produce novel compounds that suppress pathogenic microbes. In recent studies, the antagonistic activities of Streptomyces against several Xanthomonas strains including Xoo were tested andconfirmed [18, 23]. In this study, antimicrobial compounds produced by Streptomyces bottropensis strain MJM3389 against X. oryzae pv. oryzae KACC 10331 were identified. One was a cyclic peptide and the other was a spiroketal macrolide. Their potential bioactivities in regard to inhibiting rice bacterial blight were examined.
Xanthomonas oryzae pv. oryzae (Xoo) causes rice bacterial blight (BB) andhas been reported to result in a severe loss in yield (up to 50%) in tropical Asian countries [1]. Several genomic and genetic studies have been performed to better understand the interaction between pathogens and the host with the aim of developing rice that is broadly resistant against various Xoo strains. For instance, the whole genome map of X. oryzae pv. oryzae str. KACC 10331, which is arepresentatively virulent Korean strain, has been fully defined [12]. In order to control this plant disease, commercial chemicals have been employed in the
*Corresponding author C.H. L. Phone: +82-2-2049-6177; Fax: +82-2-455-4291; E-mail: chlee123@konkuk.ac.kr J.G. K. Phone: +82-31-299-1644; Fax: +82-31-299-1622; E-mail: jkim5aug@korea.kr
MATERIALS AND METHODS
Chemicals and Reagents For thefermentation of Xanthomonas oryzae pv. oryzae KACC 10331 in M210 medium, sucrose and dipotassium phosphate (Junsei Chemical, Japan), NZ Amine (Sigma-Aldrich, MO, USA), and Bacto Yeast Extract (Becton Dickinson, MD, USA) were utilized. Mueller-Hinton broth was purchased from Becton Dickinson (MD, USA) and used to test the in vitro antibacterial activities of different samples. Sephadex LH-20 gel,...
Screening and Identification of Antimicrobial Compounds from Streptomyces bottropensis Suppressing Rice Bacterial Blight
Park, Sait Byul1,2, In Ae Lee3, Joo-Won Suh3, Jeong-Gu Kim2*, and Choong Hwan Lee1*
Division of Bioscience and Biotechnology, KonkukUniversity, Seoul 143-701, Korea Genomics Division, National Academy of Agricultural Science, Rural Development Administration, Suwon 441-857, Korea Division of Bioscience and Bioinformatics, College of Natural Science, Myongji University, Yongin 449-728, Korea.
1 2 3
Received: June 28, 2011 / Revised: August 2, 2011 / Accepted: August 5, 2011
Xanthomonas oryzae pv. oryzae (Xoo) is the mostdevastating pathogen to Oryza sativa and has been shown to cause bacterial blight. Two bioactive compounds showing antimicrobial activities against Xoo strain KACC 10331 were isolated from a Streptomyces bottropensis strain. The ethyl acetate extract was fractionated on a Sephadex LH-20 column, and then purified by preparative HPLC. The purified compounds were identified as bottromycin A2 anddunaimycin D3S by HR/MS and 1H NMR analyses. The MIC value against Xoo and the lowest concentration still capable of suppressing rice bacterial blight were 2 µg/ml and 16 µg/ml for bottromycin A2, and 64 µg/ml and 0.06 µg/ml for dunaimycin D3S, respectively. These two compounds were shown to exert different bioactivities in vitro and in rice leaf explants. Keywords: Bottromycin A2, dunaimycin D3S,Streptomyces bottropensis, Xanthomonas oryzae pv. oryzae, antibacterial activity, rice bacterial blight
fields; however, the world-wide use of artificial pesticides is gradually decreasing because of their excessive toxicity and degradation properties [2]. Thus, it is necessary to identify safe natural bioactive substances that can be used as alternatives to artificial pesticides. Biological controlusing microorganisms for inhibition of plant disease holds promise as a robust and long-lasting alternative to synthetic chemicals [7]. Streptomyces has been historically examined as a microorganism that can produce novel compounds that suppress pathogenic microbes. In recent studies, the antagonistic activities of Streptomyces against several Xanthomonas strains including Xoo were tested andconfirmed [18, 23]. In this study, antimicrobial compounds produced by Streptomyces bottropensis strain MJM3389 against X. oryzae pv. oryzae KACC 10331 were identified. One was a cyclic peptide and the other was a spiroketal macrolide. Their potential bioactivities in regard to inhibiting rice bacterial blight were examined.
Xanthomonas oryzae pv. oryzae (Xoo) causes rice bacterial blight (BB) andhas been reported to result in a severe loss in yield (up to 50%) in tropical Asian countries [1]. Several genomic and genetic studies have been performed to better understand the interaction between pathogens and the host with the aim of developing rice that is broadly resistant against various Xoo strains. For instance, the whole genome map of X. oryzae pv. oryzae str. KACC 10331, which is arepresentatively virulent Korean strain, has been fully defined [12]. In order to control this plant disease, commercial chemicals have been employed in the
*Corresponding author C.H. L. Phone: +82-2-2049-6177; Fax: +82-2-455-4291; E-mail: chlee123@konkuk.ac.kr J.G. K. Phone: +82-31-299-1644; Fax: +82-31-299-1622; E-mail: jkim5aug@korea.kr
MATERIALS AND METHODS
Chemicals and Reagents For thefermentation of Xanthomonas oryzae pv. oryzae KACC 10331 in M210 medium, sucrose and dipotassium phosphate (Junsei Chemical, Japan), NZ Amine (Sigma-Aldrich, MO, USA), and Bacto Yeast Extract (Becton Dickinson, MD, USA) were utilized. Mueller-Hinton broth was purchased from Becton Dickinson (MD, USA) and used to test the in vitro antibacterial activities of different samples. Sephadex LH-20 gel,...
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