Abstract
Primers for the PCR amplification of homologous genes encoding polyketide coronafacic acid and coronafacic ligase in the cells of Pectobacterium atrosepticum SCRI1043 (BX950851) were developed to study the presence of these genes in the genome of Pectobacterium sp. and Dickeya sp. Coronafacic ligase catalyses the formation of coronatine from polyketide coronafacic acid and coronamic acid. Coronatine is a toxin produced by Pseudomonas syringae and is one of the major virulence factors in this bacterium. This study using several strains of P. atrosepticum, P. carotovorum subsp. carotovorum and Dickeya sp. isolated in different countries, indicated that all strains of P. atrosepticum possess genes coding coronafacic acid (cfa gene cluster) and coronafacic ligase (cfl). However, these genes were present only in the genome of five out of 50 tested P. carotovorum subsp. carotovorum strains and two out of 34 strains of Dickeya sp. tested. The PCR products homologous to the sequence of cfa7 and cfl gene fragments were sequenced in order to check the level of homology between genes of P. atrosepticum, P. carotovorum subsp. carotovorum and Dickeya sp. The sequences of the gene fragments amplified from all P. atrosepticum strains were almost identical (100% and 99.97%, respectively). The homology of the sequences obtained for P. atrosepticum and sequences of five P. carotovorum subsp. carotovorum and two Dickeya sp. was lower, between 89.69% to 95.00% for the cfl gene fragment, and about 94% for the cfa7 gene fragment.
References
Bell, K. S., Sebaihia, M., Pritchard, L., Holden, M. T. G., Hyman, L. J., Holeva, M. C., et al. (2004). Genome sequence of the enterobacterial phytopathogen Erwinia carotovora subsp. atroseptica and characterization of virulence factors. Proceedings of the National Academy of Sciences of the United States of America, 101, 11105–11110. doi:10.1073/pnas.0402424101.
Bender, C. L., Young, S. A., & Mitchell, R. E. (1991). Conservation of plasmid DNA sequences in coronatine-producing pathovars of Pseudomonas syringae. Applied and Environmental Microbiology, 57, 993–999.
Bender, C. L., Alarcon-Chaidez, F., & Gross, D. C. (1999). Pseudomonas syringae phytotoxins: mode of action, regulation, and biosynthesis by peptide and polyketide synthetases. Microbiology and Molecular Biology Reviews, 63, 266–292.
Bereswill, S., Bugert, P., Völksch, B., Ullrich, M., Bender, C. L., & Geider, K. (1994). Identification and relatedness of coronatine-producing Pseudomonas syringae pathovars by PCR analysis and sequence determination of the amplification products. Applied and Environmental Microbiology, 60, 2924–2930.
Darrasse, A., Priou, S., Kotoujansky, A., & Bertheau, Y. (1994). PCR and restriction fragment length polymorphism of a pel gene as a tool to identify Erwinia carotovora in relation to potato diseases. Applied and Environmental Microbiology, 60, 1437–1443.
da Silva, A. C. R., Ferro, J. A., Reinach, F. C., Farah, C. S., Furlan, L. R., Quaggio, R. B., et al. (2002). Comparison of the genomes of two Xanthomonas pathogens with differing host specificities. Nature, 417, 459–463. doi:10.1038/417459a.
DeBoer, S. H., & Ward, L. J. (1995). PCR detection of Erwinia carotovora subsp. atroseptica associated with potato tissue. Phytopathology, 85, 854–858. doi:10.1094/Phyto-85-854.
Dickey, R. S. (1979). Erwinia chrysanthemi: a comparative study of phenotypic properties of strains from several hosts and other Erwinia species. Phytopathology, 69, 324–329. doi:10.1094/Phyto-69-324.
Hugouvieux-Cotte-Pattat, N., Condemine, G., Nasser, W., & Reverchon, S. (1996). Regulation of pectinolysis in Erwinia chrysanthemi. Annual Review of Microbiology, 50, 213–257. doi:10.1146/annurev.micro.50.1.213.
Hyman, L. J., Toth, I. K., & Pérombelon, M. C. M. (1998). Isolation and identification. In M. C. M. Pérombelon, & J. M. van der Wolf (Eds.), Methods for the detection and quantification of Erwinia carotovora subsp. atroseptica in potatoes (pp. 60–65). Invergowrie: Laboratory Manual Scottish Crop Research Institute Occasional Publication No. 10In pages 60–65.
Jafra, S., Bielawski, K., & Lojkowska, E. (1998). Application of the pelL gene based primers for PCR diagnostics of Erwinia chrysanthemi. Abstracts of the 13th symposium of the Polish genetics society. Journal of Applied Genetics, 39A, 167.
Nassar, A., Darrasse, A., Lemattre, M., Kotoujansky, A., Dervin, C., Vedel, R., et al. (1996). Characterization of Erwinia chrysanthemi by pectinolytic isozyme polymorphism and restriction fragment length polymorphism analysis of PCR-amplified fragments of pel genes. Applied and Environmental Microbiology, 62, 2228–2235.
Pérombelon, M. C. M. (2002). Potato diseases caused by soft rot erwinias: an overview of pathogenesis. Plant Pathology, 51, 1–12. doi:10.1046/j.0032-0862.2001.Short title.doc.x.
Pérombelon, M. C. M., & Kelman, A. (1980). Ecology of the soft rot erwinias. Annual Review of Phytopathology, 18, 361–387. doi:10.1146/annurev.py.18.090180.002045.
Pitman, A. R., Wright, P. J., Galbraith, M. D., & Harrow, S. A. (2008). Biochemical and genetic diversity of pectolytic enterobacteria causing soft rot disease of potatoes in New Zealand. Australasian Plant Pathology, 37, 559–568. doi:10.1071/AP08056.
Py, B., Barras, F., Harris, S., Robson, N., & Salmond, G. P. C. (1998). Extracellular enzymes and their role in Erwinia virulence. In P. Williams, J. Ketley, & G. Salmond (Eds.), Bacterial Pathogenesis: Methods in Microbiology (vol. 27, (pp. 157–168)). London, UK: Academic Press Ltd.
Salanoubat, M., Genin, S., Artiguenave, F., Gouzy, J., Mangenot, S., Arlat, M., et al. (2002). Genome sequence of the plant pathogen Ralstonia solanacearum. Nature, 415, 497–502. doi:10.1038/415497a.
Sledz, W., Jafra, S., Waleron, M., & Lojkowska, E. (2000). Genetic diversity of Erwinia carotovora strains isolated from infected plants growing in Poland. Bulletin OEPP/EPPO Bulletin, 30, 403–407.
Toth, I. K., Bell, K. S., Holeva, M. C., & Birch, P. R. J. (2003). Soft rot erwiniae: from genes to genomes. Molecular Plant Pathology, 4, 17–30. doi:10.1046/j.1364-3703.2003.00149.x.
Toth, I. K., Pritchard, L., & Birch, P. R. J. (2006). Comparative genomics reveals what makes an enterobacterial plant pathogen. Annual Review of Phytopathology, 44, 305–336. doi:10.1146/annurev.phyto.44.070505.143444.
Waleron, M., Waleron, K., Podhajska, A. J., & Lojkowska, E. (2002). Genotyping of bacteria belonging to the former Erwinia genus by PCR-RFLP analysis of a recA gene fragment. Microbiology, 148, 583–595.
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Slawiak, M., Lojkowska, E. Genes responsible for coronatine synthesis in Pseudomonas syringae present in the genome of soft rot bacteria. Eur J Plant Pathol 124, 353–361 (2009). https://doi.org/10.1007/s10658-008-9418-7
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DOI: https://doi.org/10.1007/s10658-008-9418-7