Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1996 Jul;62(7):2228–2235. doi: 10.1128/aem.62.7.2228-2235.1996

Characterization of Erwinia chrysanthemi by pectinolytic isozyme polymorphism and restriction fragment length polymorphism analysis of PCR-amplified fragments of pel genes.

A Nassar 1, A Darrasse 1, M Lemattre 1, A Kotoujansky 1, C Dervin 1, R Vedel 1, Y Bertheau 1
PMCID: PMC168003  PMID: 8779560

Abstract

Conserved regions about 420 bp long of the pelADE cluster specific to Erwinia chrysanthemi were amplified by PCR and used to differentiate 78 strains of E. chrysanthemi that were obtained from different hosts and geographical areas. No PCR products were obtained from DNA samples extracted from other pectinolytic and nonpectinolytic species and genera. The pel fragments amplified from the E. chrysanthemi strains studied were compared by performing a restriction fragment length polymorphism (RFLP) analysis. On the basis of similarity coefficients derived from the RFLP analysis, the strains were separated into 16 PCR RFLP patterns grouped in six clusters, These clusters appeared to be correlated with other infraspecific levels of E. chrysanthemi classification, such as pathovar and biovar, and occasionally with geographical origin. Moreover, the clusters correlated well with the polymorphism of pectate lyase and pectin methylesterase isoenzymes. While the pectin methylesterase profiles correlated with host monocot-dicot classification, the pectate lyase polymorphism might reflect the cell wall microdomains of the plants belonging to these classes.

Full Text

The Full Text of this article is available as a PDF (393.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Bertheau Y., Madgidi-Hervan E., Kotoujansky A., Nguyen-The C., Andro T., Coleno A. Detection of depolymerase isoenzymes after electrophoresis or electrofocusing, or in titration curves. Anal Biochem. 1984 Jun;139(2):383–389. doi: 10.1016/0003-2697(84)90022-8. [DOI] [PubMed] [Google Scholar]
  3. Darrasse A., Priou S., Kotoujansky A., Bertheau Y. 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. 1994 May;60(5):1437–1443. doi: 10.1128/aem.60.5.1437-1443.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Favey S., Bourson C., Bertheau Y., Kotoujansky A., Boccara M. Purification of the acidic pectate lyase and nucleotide sequence of the corresponding gene (pelA) of Erwinia chrysanthemi strain 3937. J Gen Microbiol. 1992 Mar;138(3):499–508. doi: 10.1099/00221287-138-3-499. [DOI] [PubMed] [Google Scholar]
  6. Hinton J. C., Sidebotham J. M., Gill D. R., Salmond G. P. Extracellular and periplasmic isoenzymes of pectate lyase from Erwinia carotovora subspecies carotovora belong to different gene families. Mol Microbiol. 1989 Dec;3(12):1785–1795. doi: 10.1111/j.1365-2958.1989.tb00164.x. [DOI] [PubMed] [Google Scholar]
  7. Keen N. T., Tamaki S. Structure of two pectate lyase genes from Erwinia chrysanthemi EC16 and their high-level expression in Escherichia coli. J Bacteriol. 1986 Nov;168(2):595–606. doi: 10.1128/jb.168.2.595-606.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kelemu S., Collmer A. Erwinia chrysanthemi EC16 Produces a Second Set of Plant-Inducible Pectate Lyase Isozymes. Appl Environ Microbiol. 1993 Jun;59(6):1756–1761. doi: 10.1128/aem.59.6.1756-1761.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Nassar A., Bertheau Y., Dervin C., Narcy J. P., Lemattre M. Ribotyping of Erwinia chrysanthemi Strains in Relation to Their Pathogenic and Geographic Distribution. Appl Environ Microbiol. 1994 Oct;60(10):3781–3789. doi: 10.1128/aem.60.10.3781-3789.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Reverchon S., Huang Y., Bourson C., Robert-Baudouy J. Nucleotide sequences of the Erwinia chrysanthemi ogl and pelE genes negatively regulated by the kdgR gene product. Gene. 1989 Dec 21;85(1):125–134. doi: 10.1016/0378-1119(89)90472-1. [DOI] [PubMed] [Google Scholar]
  11. Ried J. L., Collmer A. Comparison of pectic enzymes produced by Erwinia chrysanthemi, Erwinia carotovora subsp. carotovora, and Erwinia carotovora subsp. atroseptica. Appl Environ Microbiol. 1986 Aug;52(2):305–310. doi: 10.1128/aem.52.2.305-310.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Roberts K. The plant extracellular matrix: in a new expansive mood. Curr Opin Cell Biol. 1994 Oct;6(5):688–694. doi: 10.1016/0955-0674(94)90095-7. [DOI] [PubMed] [Google Scholar]
  13. Tamaki S. J., Gold S., Robeson M., Manulis S., Keen N. T. Structure and organization of the pel genes from Erwinia chrysanthemi EC16. J Bacteriol. 1988 Aug;170(8):3468–3478. doi: 10.1128/jb.170.8.3468-3478.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. van Gijsegem F. Relationship between the pel genes of the pelADE cluster in Erwinia chrysanthemi strain B374. Mol Microbiol. 1989 Oct;3(10):1415–1424. doi: 10.1111/j.1365-2958.1989.tb00124.x. [DOI] [PubMed] [Google Scholar]
  15. van der Wolf J. M., van Beckhoven J. R., de Vries P. M., Raaijmakers J. M., Bakker P. A., Bertheau Y., van Vuurde J. W. Polymerase chain reaction for verification of fluorescent colonies of Erwinia chrysanthemi and Pseudomonas putida WCS358 in immunofluorescence colony staining. J Appl Bacteriol. 1995 Nov;79(5):569–577. doi: 10.1111/j.1365-2672.1995.tb03178.x. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES