Journal Article

Improved PCR primers for the detection and identification of arbuscular mycorrhizal fungi

Young, J. Peter W., Lee, Jaikoo, Lee, Sangsun

2008

Overview

A set of PCR primers that should amplify all subgroups of arbuscular mycorrhizal fungi (AMF, Glomeromycota), but exclude sequences from other organisms, was designed to facilitate rapid detection and identification directly from field-grown plant roots. The small subunit rRNA gene was targeted for the new primers (AML1 and AML2) because phylogenetic relationships among the Glomeromycota are well understood for this gene. Sequence comparisons indicate that the new primers should amplify all published AMF sequences except those from Archaeospora trappei. The specificity of the new primers was tested using 23 different AMF spore morphotypes from trap cultures and Miscanthus sinensis, Glycine max and Panax ginseng roots sampled from the field. Non-AMF DNA of 14 plants, 14 Basidiomycota and 18 Ascomycota was also tested as negative controls. Sequences amplified from roots using the new primers were compared with those obtained using the established NS31 and AM1 primer combination. The new primers have much better specificity and coverage of all known AMF groups.

• Publisher: Oxford, UK : Blackwell Publishing Ltd,Blackwell Publishing Ltd,Oxford University Press
• Subject: AML1 protein / Arbuscular mycorrhizas / Archaeospora / Ascomycota / Ascomycota - classification / Ascomycota - genetics / Basidiomycota / Basidiomycota - classification / Basidiomycota - genetics / classification / Coverage / Deoxyribonucleic acid / DNA / DNA primers / DNA Primers - genetics / Ecology / Fungi / Fungi - classification / Fungi - genetics / Fungi - isolation & purification / genes / Genes, rRNA / genetics / Ginseng / Glomeromycota / Glycine max / isolation & purification / methods / Microbiology / Miscanthus sinensis / Molecular Sequence Data / Mycorrhizae / Mycorrhizae - classification / Mycorrhizae - genetics / Mycorrhizae - isolation & purification / mycorrhizal fungi / Panax ginseng / PCR primer / Phylogeny / Plant roots / Plants / Plants - classification / Plants - microbiology / Polymerase chain reaction / Polymerase Chain Reaction - methods / Primers / rapid methods / ribosomal RNA / Roots / rRNA / Sensitivity and Specificity / sequence analysis / Sequence Analysis, DNA / spores / SSU rRNA gene / Subgroups