Plant Protect. Sci., 2002, 38(11):288-290 | DOI: 10.17221/10470-PPS
Genotypic characterisation of the Erwinia genus by PCR-RFLP analysis of rpoS gene
- Laboratory of Plant Protection & Biotechnology, Department of Biotechnology, Faculty of Biotechnology UG & AMG, 80-822 Gdañsk, Poland
Genotypic characterisation of the members of the genus Erwinia, based on the PCR-RFLP analysis of a fragment of the rpoS gene was done. PCR primers deduced from described rpoS gene sequences of E. carotovora allowed the amplification of about 880 bp DNA fragments from all tested Erwinia species. The rpoS fragments, amplified from 20 species of the studied Erwinia genus, were compared by RFLP analysis with 4 enzymes (AluI, Hin6I, HinfI, and Tru1I). Restriction analysis allowed drawing 63 common profiles of RFLP products for all tested Erwinia. From 1 to 3 specific RFLP profiles were identified among most of the species tested. However, in two cases: E. chrysanthemi and E. c. subsp. carotovora 15 and 20 specific RFLP groups were detected, respectively. High variability of genetic profiles of the E. chrysanthemi and E. c. subsp. carotovora can be explained by the wide spectrum of plants, which they infect. The results indicated that rpoS PCR-RFLP analysis is a useful tool for identification of species and subspecies belonging to the former Erwinia genus, as well as for differentiation of strains within E. c. subsp. carotovora and E. chrysanthemi.
Keywords: Erwinia; fingerprinting; PCR-RFLP; rpoS
Published: December 31, 2002 Show citation
References
- AUSUBEL F.M., BRENT R., KINGSTON R.E., MOORE D.D., SEIDMAN J.G., SMITH J.A., STRUHL K. (eds) (1992): Short Protocols in Molecular Biology. 2nd ed. Green Publ. Ass. and John Wiley & Sons, New York.
- BRENNER D.J., FANNING G.R., STEIGERWALT A.G. (1974): Deoxyribonucleic acid relatedness among Erwiniae and other Enterobacteriaceae: the gall, wilt, and drynecrosis organisms (Genus Erwiniae Winslow et al., sensu stricto). Int. J. Syst. Bacteriol., 24: 197-204.
Go to original source... - BROWN E.W., DAVIS R.M., GOUK C., VAN DER ZWET T. (2000): Phylogenetic relationships of necrogenic Erwinia and Brenneria species as revealed by glyceraldehyde-3phosphate dehydrogenase gene sequences. Int. J. Syst. Evol. Microbiol., 50: 2057-2068. Go to original source...
Go to PubMed... - 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. Appl. Environ. Microbiol., 60: 1437-1443. Go to original source... Go to PubMed...
- DYE D.W. (1968): A taxonomic study of the genus Erwinia. I. The"Amylovora" group. N.Z. J. Sci., 11: 590-607.
- DYE D.W. (1981): A taxonomic study of the genus Erwinia. N.Z. J. Agric. Res., 24: 223-231. Go to original source...
- FESSEHAIE A., DE BOER S.H., LÉVESQUE C.A. (2002): Molecular characterization of DNA encoding 16S-23S rRNA intergenic spacer regions and 16S rRNA of pectolytic Erwinia species. Can. J. Microbiol., 48: 387-398. Go to original source... Go to PubMed...
- HAUBEN L., MOORE E.R.B., VAUTERIN L., STEENACKERS M., VERDONCK L., S WINGS J. (1998): Phylogenetic position of phytopathogens within the Enterobacteriacae. System. Appl. Microbiol., 21: 384-397. Go to original source... Go to PubMed...
- NASSAR A., DARRASSE A., LEMATTRE M., KOTOUJANSKY A., DERVIN C., VEDEL R., BERTHEAU Y. (1996): Characterisation of Erwinia chrysanthemi by pectynolitic isozyme polymorphism and Restriction Fragment Length Polymorphism analysis of PCR-amplified fragments of pel genes. Appl. Environ. Microbiol., 62: 2228-2235. Go to original source... Go to PubMed...
- S PRÖER C., M ENDROCK U., S WIDERSKI J., L ANG E., STACKEBRANDT E. (1999): The phylogenetic position of Serratia, Buttiauxella and some other genera of the family Enterobacteriaceae. Int. J. Syst. Bacteriol., 49: 1433-1438. Go to original source... Go to PubMed...
- SUTRA L., CHRISTEN R., BOLLET C., SIMONEAU P., GARDAN L. (2001): Samsonia erythrinae gen. nov., sp. Nov., isolated from bark necrotic lesions of Erythrina sp., and discrimination of plant pathogenic Enterobacteriaceae by phenotypic features. Int. J. Syst. Evol. Microbiol., 51: 1291-1304. Go to original source... Go to PubMed...
- TOTH I.K., AVOROVA A.O., HYMAN L.J. (2001): Rapid identification and differentiation of the soft rot erwinias by 16S-23S Intergenic Transcribed Space-PCR and restriction fragment length polymorphism analyses. Appl. Environ. Microbiol., 67: 4070-4076. Go to original source... Go to PubMed...
- VANTOMME R., SWINGS J., GOOR M., KERSTERS K., DE LEY J. (1982): Phytopathological, serological, biochemical and protein electrophoretic characterization of Erwinia amylovora strains isolated in Belgium. Phytopathol. Z., 103: 349-360. Go to original source...
- VERDONCK L., MERGAER J. RIJCKAERT C., SWINGS J., KERSTERS K., DE LEY J. (1987): The genus Erwinia: a numerical analysis of phenotypic features. Int. J. Syst. Bacteriol., 37: 4-18. Go to original source...
- WALERON M., WALERON K., PODHAJSKA A.J., £OJKOWSKA E. (2002): Genotyping of bacteria belonging to the former Erwinia genus by PCR-RFLP analysis of a recA gene fragment. Microbiology, 148: 583-595. Go to original source... Go to PubMed...
- ZHANG Y., GEIDER K. (1997): Differentiation of Erwinia amylovora strains by pulsed-field gel electrophoresis. Appl. Environ. Microbiol., 63: 4421-4426. Go to original source... Go to PubMed...
- ZHANG Y., MERIGHI M., BAZZI C., GEIDER K. (1998): Genomic analysis by pulsed-field gel electrophoresis of Erwinia amylovora strains from the Mediterranean region including Italy. J. Appl. Plant. Pathol., 80: 225-232.
This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.