Specificity, sensitivity and discrimination of primers for PCR-RFLP of larger basidiomycetes and their applicability to identification of ectomycorrhizal fungi in Eucalyptus forests and plantations

M. Glen; I.C. Tommerup; N.L. Bougher; P.A. O'Brien

doi:10.1017/S0953756200003361

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Specificity, sensitivity and discrimination of primers for PCR-RFLP of larger basidiomycetes and their applicability to identification of ectomycorrhizal fungi in Eucalyptus forests and plantations

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Specificity, sensitivity and discrimination of primers for PCR-RFLP of larger basidiomycetes and their applicability to identification of ectomycorrhizal fungi in Eucalyptus forests and plantations

M. Glen, I.C. Tommerup, N.L. Bougher and P.A. O'Brien

Mycological Research, Vol.105(2), pp.138-149

2001

DOI: https://doi.org/10.1017/S0953756200003361

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Abstract

Techniques to rapidly identify the basidiomycete fungal partner of ectomycorrhizal associations would be a major advantage for ecological, fungal population dynamics and life history studies of epigeous and hypogeous forms in plantations, forests, wild lands and other native or natural vegetation. PCR - RFLP (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism) identification of DNA regions is an available technique; however, primers which have a high probability of amplifying only the basidiomycete DNA are needed. Here we have assessed the specificity, sensitivity and discrimination of six different primer pairs, three targeting nuclear and three mitochondrial regions, for use in identification of Australian basidiomycete fungi from Eucalyptus forests by matching PCR - RFLP patterns to morphologically defined species. Two sets of primers, one newly designed and targeting the nuclear ribosomal DNA internal transcribed spacers (ITS) and the other amplifying a fragment of mitochondrial large subunit ribosomal DNA met the requirements of high specificity and sensitivity, amplifying DNA from a broad range of larger basidiomycetes, with no amplification of plant, bacterial or ascomycete DNA. The specificity of the ITS primer pair was compared with that of ITS1-F/ITS4-B. PCR - RFLP of the two regions discriminated fungi to species level for 91 fungal species from 28 families. Hence these two DNA regions and the specific primers are a potential practical PCR - RFLP tool for identifying basidiomycetes associated with plants from field samples.

Details

Title: Specificity, sensitivity and discrimination of primers for PCR-RFLP of larger basidiomycetes and their applicability to identification of ectomycorrhizal fungi in Eucalyptus forests and plantations
Authors/Creators: M. Glen (Author/Creator) - Murdoch University
I.C. Tommerup (Author/Creator) - Commonwealth Scientific and Industrial Research Organisation
N.L. Bougher (Author/Creator) - Commonwealth Scientific and Industrial Research Organisation
P.A. O'Brien (Author/Creator)
Publication Details: Mycological Research, Vol.105(2), pp.138-149
Publisher: Cambridge University Press
Identifiers: 991005541283207891
Copyright: The British Mycological Society 2001
Murdoch Affiliation: School of Biological Sciences and Biotechnology
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration
Citation topics: 3 Agriculture, Environment & Ecology; 3.97 Plant Pathology; 3.97.488 Mycorrhizal Symbiosis
Web Of Science research areas: Mycology
ESI research areas: Plant & Animal Science