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

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.

Detection of Dickeya spp. latent infection in potato seed tubers using PCR or ELISA and correlation with disease incidence in commercial field crops under hot‐climate conditions

“…Pectobacterium atrosepticum and P. carotovorum reference strains (obtained from A. Lees, SCRI, Scotland) were also included in these tests because these pathogens may also be present on potato crops. PCR was conducted according to Nassar et al (1996), using ADE1 ⁄ ADE2 primers (ADE1: 5¢-GATCAGAAA GCCCGCAGCCAGAT-3¢, ADE2: 5¢-CTGTGGCCGA TCAGGATGGTTTTGTCGTGC-3¢). PCR was performed with ReddyMix PCR MasterMix (Thermo Scientific) with initial denaturation at 94°C for 4 min, followed by 40 cycles consisting of incubations for 30 s at 94°C and 1 min at 72°C, with a final extension of 10 min at 72°C.…”

Succession of microbial communities during a biostimulation process as evaluated by DGGE and clone library analyses

“…However, since it is dif®cult to culture most of the bacteria in environmental samples (Jannasch and Jones 1959;Amann et al 1995;Suzuki et al 1997), evaluation of the changes in the bacterial community structure by culturing alone is inadequate. Recently, bacterial community structure analyses that do not depend on culture have been widely carried out (Amann et al 1995;Pace 1997;Head et al 1998), including the use of denaturing gradient gel electrophoresis (DGGE) (Muyzer et al 1993),¯uorescence in situ hybridization (FISH) (Delong et al 1989;Amann et al 1990) and PCR-restriction fragment length polymorphism (RFLP) (Nassar et al 1996;Weidner et al 1996). DGGE is an effective method which enables analysis of many samples simultaneously, and can sequence DNA bands in a distinct pattern.…”

Caractérisation par PCR de deux souches d’Erwinia carotovora isolées de la rhizosphère de la pomme de terre dans la région du grand Casablanca au Maroc

“…Des résultats similaires ont été obtenus par d'autres auteurs (Hélias et al, 1998;Yahiaoui et al, 2003). D'une manière générale cette méthode ainsi que d'autres techniques moléculaires sont actuellement utilisées et associées à des tests microbiologiques et physiologiques pour la détection et la caractérisation des Erwinia pectinolytiques (Nassar et al, 1996;Hyman et al, 2000;Yap et al, 2004).…”