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Appl. Environ. Microbiol., Jul 1996, 2228-2235, Vol 62, No. 7
Copyright © 1996, American Society for Microbiology

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

A Nassar, A Darrasse, M Lemattre, A Kotoujansky, C Dervin, R Vedel and Y Bertheau
Station de Pathologie Vegetale, Institut National de la Recherche Agronomique, Versailles, France.

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.


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