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DNA‐based species level detection of Glomeromycota : one PCR primer set for all arbuscular mycorrhizal fungi
Author(s) -
Krüger Manuela,
Stockinger Herbert,
Krüger Claudia,
Schüßler Arthur
Publication year - 2009
Publication title -
new phytologist
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.742
H-Index - 244
eISSN - 1469-8137
pISSN - 0028-646X
DOI - 10.1111/j.1469-8137.2009.02835.x
Subject(s) - biology , primer (cosmetics) , glomeromycota , internal transcribed spacer , ribosomal dna , phylogenetic tree , dna barcoding , ribosomal rna , field pea , genetics , botany , evolutionary biology , gene , arbuscular mycorrhizal , symbiosis , bacteria , chemistry , organic chemistry , pisum
Summary• At present, molecular ecological studies of arbuscular mycorrhizal fungi (AMF) are only possible above species level when targeting entire communities. To improve molecular species characterization and to allow species level community analyses in the field, a set of newly designed AMF specific PCR primers was successfully tested. • Nuclear rDNA fragments from diverse phylogenetic AMF lineages were sequenced and analysed to design four primer mixtures, each targeting one binding site in the small subunit (SSU) or large subunit (LSU) rDNA. To allow species resolution, they span a fragment covering the partial SSU, whole internal transcribed spacer (ITS) rDNA region and partial LSU. • The new primers are suitable for specifically amplifying AMF rDNA from material that may be contaminated by other organisms (e.g., samples from pot cultures or the field), characterizing the diversity of AMF species from field samples, and amplifying a SSU‐ITS‐LSU fragment that allows phylogenetic analyses with species level resolution. • The PCR primers can be used to monitor entire AMF field communities, based on a single rDNA marker region. Their application will improve the base for deep sequencing approaches; moreover, they can be efficiently used as DNA barcoding primers.