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Abstract
We have designed two taxon-selective primers for the internal transcribed spacer (ITS)
region in the nuclear ribosomal repeat unit. These primers, ITS1-F and ITS4-B, were
intended to be specific to fungi and basidiomycetes, respectively. We have tested
the specificity of these primers against 13 species of ascomycetes, 14 of basidiomycetes,
and 15 of plants. Our results showed that ITS4-B, when paired with either a 'universal'
primer ITS1 or the fungal-specific primer ITS1-F, efficiently amplified DNA from all
basidiomycetes and discriminated against ascomycete DNAs. The results with plants
were not as clearcut. The ITS1-F/ITS4-B primer pair produced a small amount of PCR
product for certain plant species, but the quantity was in most cases less than that
produced by the 'universal' ITS primers. However, under conditions where both plant
and fungal DNAs were present, the fungal DNA was amplified to the apparent exclusion
of plant DNA. ITS1-F/ITS4-B preferential amplification was shown to be particularly
useful for detection and analysis of the basidiomycete component in ectomycorrhizae
and in rust-infected tissues. These primers can be used to study the structure of
ectomycorrhizal communities or the distribution of rusts on alternate hosts.