title: Amplification of soil fungal community DNA using the ITS86F and ITS4 primers.
abstract: Internal transcribed spacer (ITS) 86F and ITS4 and the ITS1-F and ITS86R primer pairs were tested to specifically amplify fungal community DNA extracted from soil. Libraries were constructed from PCR-amplified fragments, sequenced and compared against sequences deposited in GenBank. The results confirmed that the ITS86F and ITS4 primer pair was selectively specific for the Ascomycetes, Basidiomycetes and Zygomycetes fungal clades. Amplified products generated by the ITS1F and ITS86R primer pair also aligned with sequences from a range of species within the Ascomycete and Basidiomycete clades but not from the Zygomycete. Both primer sets demonstrated fungal specificity and appear to be well suited for rapid PCR-based (fingerprinting) analysis of environmental fungal community DNA. This is the first reported use and assessment of the ITS86F and ITS4 and the ITS1-F and ITS86R primer pairs in amplifying fungal community DNA from soil.
authors.affiliation: NSW Department of Primary Industries, Wollongbar, NSW, Australia. tony.vancov@dpi.nsw.gov.au
authors.forename: Tony
authors.initials: T
authors.lastname: Vancov
authors.forename: Brad
authors.initials: B
authors.lastname: Keen
mesh_terms: D001203:Ascomycota / Q000235:genetics; D001487:Basidiomycota / Q000235:genetics; D017931:DNA Primers / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D021903:DNA, Ribosomal Spacer / Q000235:genetics / Q000302:isolation & purification*; D015723:Gene Library; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer