Use of PCR-RFLP for assessment of genetic diversity among Xanthomonas spp.

The genus Xanthomonas is formed by a complex groupof bacterial species with diverse physiological andphytopathological traits. This group of phytopathogensinfects many economically important plants such ascotton, beans, rice, crucifers and therefore, thesebacteria represent a threat to agriculture around theworld. Some of the most important diseases caused byXanthomonas species include citrus canker and blackrot, additionally Xanthomonas is also important inbiotechnology due to the production of a polysaccharideknown as a xanthan gum, which is used as a stabilizerand emulsifier in the food and cosmetic industry, amongother applications. In this study genetic diversity among15 strains of Xanthomonas spp. were clarified by PCRRestriction Fragment Length Polymorphism (RFLP)analysis of 16S-23S rDNA internal transcribed spacer(ITS) region using four restriction enzymes including AluI,MboI, DdeI, BsnI. Total bacterial DNA was prepared byusing phenol- chloroform method. PCR was performedby using ITS primer, PCR products were digested withrestriction enzymes. Three enzymes including AluI, DdeI,BsnI showed two different profiles among all strains butone enzyme, MboI, showed three different profiles. PCRRFLPanalysis of ITS is a simple and rapid technique toevaluate biodiversity among Xanthomonas spp. and canuse for partially differentiation among Xanthomonasspecies

Keywords:

PCR- RFLP , Xanthomonas spp. , biodiversity