Strains of Xanthomonas campestris pv. vignicola (Xcv), isolated from cowpea leaves with blight or minute pustules and collected from various geographic areas, were selected on the basis of pathological and physiological features. All strains were analyzed for genotypic markers by two methods: ribotyping with EcoRI endonuclease, and RFLP analysis with a plasmid probe (pthB) containing a gene required for pathogenicity from Xanthomonas campestris pv. manihotis. Ribotyping revealed a unique pattern for all the strains that corresponded to the previously described ribotype rRNA7. Based on polymorphism detected by pthB among Xcv strains, nine haplotypes were defined. The observed genetic variation was independent of the geographic origin of the strains and of pathogenic variation. Some haplotypes were widely distributed, whereas others were localized. In some cases, we could differentiate strains isolated from blight symptoms and pustules according to haplotypic composition. However, in most cases, no significant differences were observed. Our results and the previous pathogenic and biochemical characterizations suggest that the strains isolated from leaves with blight symptoms or minute pustules belong to the same pathovar. We provide information on pathogen diversity that can be used to identify and characterize resistant germplasm. 相似文献
Six isolates of Mycoplasma bovoculi obtained from cattle herds with bovine keratoconjunctivitis were analyzed by gel electrophoresis and immunoblotting techniques. All six strains showed similarity in their protein profiles although no two patterns were identical. Antigenic differences between strains were detected in immunoblots reacted with post-exposure calf serum. A common 94 kDa protein band designated p94 was detected in all six strains reacted with monoclonal antibody MA25.5 developed to one of the strains. The p94 was also recognized in these strains by the calf serum. Trypsin treatment of intact mycoplasma cells resulted in the removal of p94 from immunoblots reacted with MA or hyperimmune rabbit serum. Other trypsin-resistant antigens shared between strains or being strain-specific in nature were identified when trypsin-treated mycoplasma cells were reacted with hyperimmune rabbit serum. The p94 antigen was shown to be of mycoplasmal origin by radio-immunoprecipitation using the MA or hyperimmune rabbit serum. These studies identify the presence of a surface antigen (p94) on M. bovoculi membrane in all strains examined that is trypsin sensitive by the use of monoclonal antibody, calf serum and hyperimmune rabbit serum.
Résumé
Six souches de Mycoplasma bovoculi, d'origine bovine (animaux affectés de keratoconjunctivite) ont été analysées par des techniques d'éléctrophorèse et par ‘immunoblotting’. Les six souches ont montré des similitudes dans leurs profils proteiniques. Cependant, aucun profil n'a été identique. Des differences antigéniques entre les souches ont étés detectées suite au traitement des immunoblots avec un sérum bovin infecté par M. bovoculi. L'anticorps monoclonal MA25.5 dirigé contre une des souches a réagi avec une bande proteinique (94 kDa) commune aux six souches. Cette bande a été designee p94. Cette bande p94 a été également identifiée chez tous les souches en utilisant le sérum bovin. Le traitement des mycoplasmes intactes par la trypsine a éliminé la bande p94 des immunoblots: cela a été démontr é par l'usage d'un anticorps monoclonal, ou par un sérum hyperimmun de lapin. D'autres antigènes résistants à la trypsine, communs ou spécifiques aux souches ont été identifiés quand les mycoplasmes traités par le trypsine ont été mélangés au sérum hyperimmun de lapin. La radioimmunoprécipitation par l'anticorps monoclonal ou le sérum hyperimmun de lapin a demontré que l'antigène p94 est d'origine mycoplasmique. Ces études identifient la présence d'un antigène de surface (le p94) localisé sur la membranc de M. bovoculi chez toutes les souches examinées. Cet antigène est sensible à la trypsine. Cela a été montré avec l'anticorps monoclonal, le sérum bovin et le sérum hyperimmune du lapin. 相似文献