Characterization of Cladosporium Species by Internal Transcribed Spacer-PCR and Microsatellites-PCR.

Background: This investigation compared genetic similarities and diversities within and among Cladosporium species populations using the two PCR-based markers; Internal Transcribed Spacer (ITS)-PCR and microsatellite-PCR.

Methodology: Nuclear ribosomal DNA internal transcribed spacers have been used to analyze intraspecific and interspecific relationships in various fungi. In the present study, the internal transcribed spacer (ITS)-PCR and microsatellite-PCR were used to identify the genetic diversities in Cladosporium species.

Results: The Internal Transcribed Spacer (ITS) was amplified using polymerase chain reaction combining primers ITS4 and ITS5. The PCR products were digested with three restriction enzymes and separated by agarose gel electrophoresis. Restriction patterns generated by CfoI and Msp I and RsaI were unique for most species assayed. The ITS-PCR fingerprinting methods led to a clear differentiation of the isolates at the species level. Fingerprinting profiles generated readily discriminated between each of the 6 species. Cluster analysis further supported this observation and clusters corresponding to each species were readily identified in the dendrograms. Seven microsatellite primers out of eight primers were unable to generate visible DNA fingerprints.

Conclusion: Amplification experiments demonstrated that microsatellite primer, T3B and (GTG) 5 are technically simple tools for assaying genetic variability in Cladosporium spp.