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Functional identification of phenazine biosynthesis genes in plant pathogenic bacteria Pseudomonas syringae pv. tomato and Xanthomonas oryzae pv. oryzae |
LI Wen1, XU You-ping2, Jean-Pierre Munyampundu1, XU Xin1, QI Xian-fei1, GU Yuan1, CAI Xin-zhong1, 3 |
1 Institute of Biotechnology, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, P.R.China
2 Center of Analysis and Measurement, Zhejiang University, Hangzhou 310058, P.R.China
3 State Key Laboratory of Rice Biology, Zhejiang University, Hangzhou 310058, P.R.China |
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摘要 Phenazines are secondary metabolites with broad spectrum antibiotic activity and thus show high potential in biological control of pathogens. In this study, we identified phenazine biosynthesis (phz) genes in two genome-completed plant pathogenic bacteria Pseudomonas syringae pv. tomato (Pst) DC3000 and Xanthomonas oryzae pv. oryzae (Xoo) PXO99A. Unlike the phz genes in typical phenazine-producing pseudomonads, phz homologs in Pst DC3000 and Xoo PXO99A consisted of phzC/D/E/F/G and phzC/E1/E2/F/G, respectively, and the both were not organized into an operon. Detection experiments demonstrated that phenazine-1-carboxylic acid (PCA) of Pst DC3000 accumulated to 13.4 μg L–1, while that of Xoo PXO99A was almost undetectable. Moreover, Pst DC3000 was resistant to 1 mg mL–1 PCA, while Xoo PXO99A was sensitive to 50 μg mL–1 PCA. Furthermore, mutation of phzF blocked the PCA production and significantly reduced the pathogenicity of Pst DC3000 in tomato, while the complementary strains restored these phenotypes. These results revealed that Pst DC3000 produces low level of and is resistant to phenazines and thus is unable to be biologically controlled by phenazines. Additionally, phz-mediated PCA production is required for full pathogenicity of Pst DC3000. To our knowledge, this is the first report of PCA production and its function in pathogenicity of a plant pathogenic P. syringae strain.
Abstract Phenazines are secondary metabolites with broad spectrum antibiotic activity and thus show high potential in biological control of pathogens. In this study, we identified phenazine biosynthesis (phz) genes in two genome-completed plant pathogenic bacteria Pseudomonas syringae pv. tomato (Pst) DC3000 and Xanthomonas oryzae pv. oryzae (Xoo) PXO99A. Unlike the phz genes in typical phenazine-producing pseudomonads, phz homologs in Pst DC3000 and Xoo PXO99A consisted of phzC/D/E/F/G and phzC/E1/E2/F/G, respectively, and the both were not organized into an operon. Detection experiments demonstrated that phenazine-1-carboxylic acid (PCA) of Pst DC3000 accumulated to 13.4 μg L–1, while that of Xoo PXO99A was almost undetectable. Moreover, Pst DC3000 was resistant to 1 mg mL–1 PCA, while Xoo PXO99A was sensitive to 50 μg mL–1 PCA. Furthermore, mutation of phzF blocked the PCA production and significantly reduced the pathogenicity of Pst DC3000 in tomato, while the complementary strains restored these phenotypes. These results revealed that Pst DC3000 produces low level of and is resistant to phenazines and thus is unable to be biologically controlled by phenazines. Additionally, phz-mediated PCA production is required for full pathogenicity of Pst DC3000. To our knowledge, this is the first report of PCA production and its function in pathogenicity of a plant pathogenic P. syringae strain.
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Received: 02 June 2015
Accepted:
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Fund: This work was financially supported by the grants from the Genetically Modified Organisms Breeding Major Projects, China (2014ZX0800905B), the Fundamental Research Funds for the Central Universities, China, and the Program for New Century 151 Talents of Zhejiang Province, China. |
Corresponding Authors:
CAI Xin-zhong, Tel/Fax: +86-571-88982936, E-mail: xzhcai@zju.edu.cn
E-mail: xzhcai@zju.edu.cn
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About author: LI Wen, E-mail: liwen547249317@126.com |
Cite this article:
LI Wen, XU You-ping, Jean-Pierre Munyampundu, XU Xin, QI Xian-fei, GU Yuan, CAI Xin-zhong.
2016.
Functional identification of phenazine biosynthesis genes in plant pathogenic bacteria Pseudomonas syringae pv. tomato and Xanthomonas oryzae pv. oryzae. Journal of Integrative Agriculture, 15(4): 812-821.
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Abken H J, Tietze M, Brodersen J, Bäumer S, Beifuss U, Deppenmeier U. 1998. Isolation and characterization of methanophenazine and function of phenazines in membrane-bound electron transport of Methanosarcina mazei Gö1. Journal of Bacteriology, 180, 2027–2032.Ahuja E G, Janning P, Mentel M, Graebsch A, Breinbauer R, Hiller W, Costisella B, Thomashow L S, Mavrodi D V, Blankenfeldt W. 2008. PhzA/B catalyzes the formation of the tricycle in phenazine biosynthesis. Journal of the American Chemical Society, 130, 17053–17061.Blankenfeldt W, Kuzin A P, Skarina T, Korniyenko Y, Tong L, Bayer P, Janning P, Thomashow L S, Mavrodi D V. 2004. Structure and function of the phenazine biosynthetic protein PhzF from Pseudomonas fluorescens. Proceedings of the National Academy of Sciences of the United States of America, 101, 16431–16436.Camacho C, Coulouris G, Avagyan V, Ma N, Papadopoulos J, Bealer K, Madden T L. 2009. BLAST plus: Architecture and applications. BMC Bioinformatics, 10, 421.Culbertson J E, Toney M D. 2013. Expression and characterization of PhzE from P. aeruginosa PAO1: aminodeoxyisochorismate synthase involved in pyocyanin and phenazine-1-carboxylate production. Biochimica et Biophysica Acta (BBA - Proteins and Proteomics), 1834, 240–246.Dietrich L E, Price W A, Petersen A, Whiteley M, Newman D K. 2006. The phenazine pyocyanin is a terminal signalling factor in the quorum sensing network of Pseudomonas aeruginosa. Molecular Microbiology, 61, 1308–1321.Du X, Li Y, Zhou W, Zhou Q, Liu H, Xu Y. 2013. Phenazine-1-carboxylic acid production in a chromosomally non-scar triple-deleted mutant Pseudomonas aeruginosa using statistical experimental designs to optimize yield. Applied Microbiology and Biotechnology, 97, 7767–7778.Fitzpatrick D A. 2009. Lines of evidence for horizontal gene transfer of a phenazine producing operon into multiple bacterial species. Journal of Molecular Evolution, 68, 171–185.Gebhardt K, Schimana J, Krastel P, Dettner K, Rheinheimer J, Zeeck A, Fiedler H P. 2002. Endophenazines AD, new phenazine antibiotics from the arthropod associated endosymbiont Streptomyces anulatus. I. Taxonomy, fermentation, isolation and biological activities. Journal of Antibiotics, 55, 794–800.Ge Y, Huang X, Wang S, Zhang X, Xu Y. 2004. Phenazine-1-carboxylic acid is negatively regulated and pyoluteorin positively regulated by gacA in Pseudomonas sp. M18. FEMS Microbiology Letters, 237, 41–47.Hu H B, Xu Y Q, Chen F, Zhang X H, Hur B K. 2005. Isolation and characterization of a new fluorescent Pseudomonas strain that produces both phenazine 1-carboxylic acid and pyoluteorin. Journal of Microbiology and Biotechnology, 15, 86–90.Kim K J. 2000. Phenazine 1-carboxylic acid resistance in phenazine 1-carboxylic acid producing Bacillus sp. B-6. Journal of Biochemistry and Molecular Biology, 33, 332–336.Lau G W, Ran H, Kong F, Hassett D J, Mavrodi D. 2004. Pseudomonas aeruginosa pyocyanin is critical for lung infection in mice. Infection and Immunity, 72, 4275–4278.Li W, Xu Y P, Zhang Z X, Cao W Y, Li F, Zhou X, Chen G Y, Cai X Z. 2012. Identification of genes required for nonhost resistance to Xanthomonas oryzae pv. oryzae reveals novel signaling components. PLoS One, 7, e42796.Mahajan-Miklos S, Tan M W, Rahme L G, Ausubel F M. 1999. Molecular mechanisms of bacterial virulence elucidated using a Pseudomonas aeruginosa-Caenorhabditis elegans pathogenesis model. Cell, 96, 47–56.Mavrodi D V, Blankenfeldt W, Thomashow L S. 2006. Phenazine compounds in fluorescent Pseudomonas spp. biosynthesis and regulation. Annual Review of Phytopathology, 44, 417–445.Mavrodi D V, Ksenzenko V N, Bonsall R F, Cook R J, Boronin A M, Thomashow L S. 1998. A seven-gene locus for synthesis of phenazine-1-carboxylic acid by Pseudomonas fluorescens 2-79. Journal of Bacteriology, 180, 2541–2548.Mavrodi D V, Peever T L, Mavrodi O V, Parejko J A, Raaijmakers J M, Lemanceau P, Mazurier S, Heide L, Blankenfeldt W, Weller D M. 2010. Diversity and evolution of the phenazine biosynthesis pathway. Applied and Environmental Microbiology, 76, 866–879.McDonald M, Mavrodi D V, Thomashow L S, Floss H G. 2001. Phenazine biosynthesis in Pseudomonas fluorescens: Branchpoint from the primary shikimate biosynthetic pathway and role of phenazine-1,6-dicarboxylic acid. Journal of the American Chemical Society, 123, 9459–9460.Penfold R J, Pemberton J M. 1992. An improved suicide vector for construction of chromosomal insertion mutations in bacteria. Gene, 118, 145–146.Podojil M, Gerber N N. 1967. The biosynthesis of 1,6-phenazinediol 5,10-dioxide (iodinin) by Brevibacterium iodinum. Biochemistry, 6, 2701–2705.Rahme L G, Ausubel F M, Cao H, Drenkard E, Goumnerov B C, Lau G W, Mahajan-Miklos S, Plotnikova J, Tan M W, Tsongalis J. 2000. Plants and animals share functionally common bacterial virulence factors. Proceedings of the National Academy of Sciences of the United States of America, 97, 8815–8821.Saand M A, Xu Y P, Li W, Wang J P, Cai X Z. 2015. Cyclic nucleotide gated channel gene family in tomato: genome-wide identification and functional analyses in disease resistance. Frontiers in Plant Science, 6, 303.Saleh O, Gust B, Boll B, Fiedler H P, Heide L. 2009. Aromatic prenylation in phenazine biosynthesis dihydrophenazine-1-carboxylate dimethylallyltransferase from Streptomyces anulatus. Journal of Biological Chemistry, 284, 4439–14447.Thomashow L S, Weller D M, Bonsall R F, Pierson L S. 1990. Production of the antibiotic phenazine-1-carboxylic acid by fluorescent Pseudomonas species in the rhizosphere of wheat. Applied and Environmental Microbiology, 56, 908–912.Webby C J, Baker H M, Lott J S, Baker E N, Parker E J. 2005. The structure of 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase from Mycobacterium tuberculosis reveals a common catalytic scaffold and ancestry for type I and type II enzymes. Journal of Molecular Biology, 354, 927–939.Xu X Q, Pan S Q. 2000. An Agrobacterium catalase is a virulence factor involved in tumorigenesis. Molecular Microbiology, 35, 407–414. |
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