Species-specific PCR primers for the detection of poorly distinguishable Xanthomonas euvesicatoria

New species-specific PCR primers for detection of bacterium Xanthomonas euvesicatoria, the causal agent of bacterial spot of tomato and pepper, were designed in this work. Developed PCR primer pair ZnDoF/ZnDoR partially targets the coding sequence of Zn-dependent oxidoreductase gene. The primer pair was tested on various bacterial strains including bacteria from the bacterial spot-causing xanthomonads (BSX) group. Designed primers showed to be species-specific for the samples containing DNA of X. euvesicatoria. Specificity of the designed primers was also compared with the primer pairs already published. Subsequently, sensitivity of ZnDoF/ZnDoR primers was determined by ten-fold dilution of standardized sample. Detection capability of developed primers was also investigated on artificially infected pepper plants and seeds collected from bacterial spot-like peppers. Primers proved to be species-specific for in planta and in seed detection as well as to distinguish X. euvesicatoria from other bacterial strains used in this assay.