Molecular approaches for assessing fungal diversity in marine substrata

A range of molecular techniques is available to assess fungal diversity in natural environments. These include denaturing gradient gel electrophoresis (DGGE), temperature gradient gel electrophoresis (TGGE), single-strand conformation polymorphism ( SSCP), terminal restriction fragment length polymorphism (T-RFLP), amplified rDNA restriction analysis (ARDRA), amplified ribosomal intergenic spacer analysis (ARISA) and cloning. The techniques, polymerase chain reaction (PCR) primers for environmental work and their potential limitations are discussed in this review, together with applications for describing the diversity of filamentous marine fungi associated with their substrata. It is unlikely that a single technique can be used to assess all aspects of diversity, however the limited number of studies with a molecular approach has demonstrated that marine substrata contain a greater fungal diversity than previously recognised. These studies have also shown that it is important to combine mycological and molecular approaches to accurately assess diversity.