Differentiation of anaerobic polycentric fungi by rDNA PCR-RFLP

Date: Dec 31, 2006

Pub: Folia Microbiologica. 51(4) 273-277

Author(s): Fliegerova, K., Mrázek, J., & Voigt, K.

Abstract:

The suitability of restriction fragment length polymorphism (RFLP) analysis of the ribosomal DNA cluster for discriminating two genera of anaerobic polycentric fungi,Orpinomyces andAnaeromyces, was determined. Three PCR-amplified DNA fragments — nuclear small subunit (SSU; 18S rDNA), the nuclear large subunit (LSU; 28S rDNA) and internal transcribed spacer (ITS) — were restricted with endonucleasesAluI,DraI,HinfI andMboI. Although the SSU DNA fragment could be restricted successfully by all four enzymes, no differences were observed between restriction patterns ofOrpinomyces andAnaeromyces. The most polymorphic restriction pattern betweenOrpinomyces andAnaeromyces resulted from cleavage of LSU rDNA fragments cut byAluI andHinfI and ITS fragment cut byDraI andHinfI. Genus-specific RFLP patterns were determined forOrpinomyces andAnaeromyces genera; the results showed that the PCR-RFLP analysis of rDNA offers an easy and rapid tool for differentiation of two polycentric genera of anaerobic fungi, which could be hardly separated on the basis of morphology.