Analysis of the rDNA internal transcribed spacer region of the species by polymerase chain reaction-restriction fragment length polymorphism.

The species are a widely spread phytopathogen identified in an extensive variety of hosts. The genus is one of the most heterogeneous fungi and is difficult to classify. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis is a useful method in detection of DNA polymorphism in objective sequences. The aim of the present study was to identify the phylogenetic associations and usefulness of the internal transcribed spacer (ITS) region as a genetic marker within the most clinically important strain of the species. A total of 50 strains of spp. were used in the study, including environmental, clinical and reference isolates. The primers ITS1 and ITS4 were used in the study. Two restriction enzymes, III and I, were assessed for the digestion of PCR products. A PCR product of ~550-base pairs was generated for each species. The digested products with III and I demonstrated that the bands generated for the medically significant species, including , , , and , have different restriction enzyme patterns. In conclusion, it appears that the PCR-RFLP method used in the present study produces a sufficient restriction profile for differentiation of the most medically significant species.