RAPD PCR-based differentiation of Xanthomonas campestris pv. Phaseoli and Xanthomonas campestris pv. phaseoli var. fuscans

Abstract

A RAPD PCR-based method was used to differentiate between isolates of Xanthomonas campestris pv. Phaseoli and Xanthomonas campestris pv. phaseoli var. fuscans. Using random primer OP-G11, a single, high intensity band of 820 bp was amplified from DNAs of all X. c. pv. phaseoli var. fuscans isolates, while multiple amplification products of varying sizes were generated from X. c. pv. phaseoli DNAs. Whereas RAPD PCR differentiation gave an unambiguous result in under 4 h, standard differentiation by recording the production of a brown pigment by X. c. pv. phaseoli var. fuscans isolates took up to 7 days and showed variation both between isolates and between media. The unequivocal nature of the RAPD PCR method was demonstrated when isolate 408, originally classified as X. c. pv. phaseoli var. fuscans, failed to produce the 820 bp band typical of X. c. pv. phaseoli var. fuscans isolates,and after also failing to produce a brown pigment, was re-classified as X. c. pv. phaseoli.