Primer set details

ORGANISM
Complete name : Armillaria mellea
Database ID : 567
INTERNET RESOURCES
Google Wikispecies, the free species directory Wikipedia, The Free Encyclopeda
NCBI Taxonomy Browser CABI Index Fungorum CBS Anamorph/Teleomorph

PRIMER SET (5’→3′)
Forward Primer Select to Blast
Name : Amel-F165 Sequence :
Reverse Primer
Name : Amel-R401 Sequence :
TARGET DNA (Amplicon size)
Translation Elongation Factor 1 alpha gene (236 bp)
PCR TECHNIQUE
Conventional PCR
REFERENCE
Authors : Mulholland V., MacAskill G. A., Laue B. E., Steele H., Kenyon D., Green S.
Year : 2012
Title : Development and verification of a diagnostic assay based on EF-1 a for the identification of Armillaria species in Northern Europe
Journal : Forest Pathology
Volume : 42
Pages : 229-238
DOI: 10.1111/j.1439-0329.2011.00747.x
Abstract : The overall aim of this study was to develop a new, reliable and rapid diagnostic assay for differentiating six European Armillaria species based on variation in their elongation factor-1 alpha (EF-1 a) gene sequences and to verify a set of species-specific primers on 61 Armillaria isolates from Europe. Partial sequences of the EF-1 a gene obtained in Armillaria borealis, Armillaria cepistipes, Armillaria gallica, Armillaria mellea, Armillaria ostoyae and Armillaria tabescens revealed sufficient interspecific variation to distinguish among species using nested primers. These primers gave unambiguous bands when tested on representative isolates of five of these species. However, the EF-1 a sequences of European A. borealis isolates clustered into two distinct clades, termed here AbX and AbY. Specific primers were subsequently designed and tested successfully on both AbX-type and AbY-type A. borealis isolates. The taxonomy of A. borealis needs to be elucidated to determine whether a new, as yet unnamed Armillaria taxon exists in Europe. Three A. borealis isolates were also found to have heterozygous sites in their EF-1 a sequences, which suggests that the gene could exist in more than one copy or that these isolates contain hybrid sequences. A pyrosequencing method was also developed, targeting a small region of EF-1 a intron 4, which was able to differentiate European Armillaria isolates to the species level and additionally could distinguish AbX-type and AbY-type A. borealis isolates.
SUBMITTER
Dr. Stefano Ghignone (Sistem Administrator)
stefano.ghignone@ipsp.cnr.it
Consiglio Nazionale delle Ricerche (CNR)
Istituto per la Protezione delle Piante, Sez. Torino
V.le P.A. Mattioli 25, I-10125 Turin
Italy
Record creation : 2012-08-31 Last record update : 2012-08-31

* Disclaimer *

No guarantee can be given for the correctness of the data presented.
Although the DNA sequences were reviewed for accuracy,
original pubblications should be consulted to verify their composition.

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