The rapid identification of European Armillaria species from soil samples by nested PCR
Language English Country England, Great Britain Media print
Document type Journal Article
PubMed
15268944
DOI
10.1016/j.femsle.2004.06.019
PII: S0378109704004380
Knihovny.cz E-resources
- MeSH
- Agaricales classification genetics isolation & purification MeSH
- DNA, Fungal analysis isolation & purification metabolism MeSH
- DNA, Ribosomal Spacer analysis isolation & purification metabolism MeSH
- Polymerase Chain Reaction * MeSH
- Polymorphism, Restriction Fragment Length * MeSH
- Soil Microbiology * MeSH
- Deoxyribonucleases, Type II Site-Specific metabolism MeSH
- Sequence Analysis, DNA MeSH
- Sensitivity and Specificity MeSH
- Chromatography, High Pressure Liquid MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- DNA, Fungal MeSH
- GANTC-specific type II deoxyribonucleases MeSH Browser
- DNA, Ribosomal Spacer MeSH
- Deoxyribonucleases, Type II Site-Specific MeSH
New specific primers AR1 and AR2 were successfully used for the amplification of a specific part of internal transcribed spacer (ITS) of rDNA of Armillaria isolated from soil samples. DNA was isolated from 0.5 g of forest soil and ITS region was amplified by nested PCR reaction with external primers ITS1 and ITS4 and internal primers AR1 and AR2. The individual species were distinguished by restriction fragment length polymorphisms (RFLPs) analysis with restriction endonuclease HinfI. The fragments were analysed by ion-exchange HPLC that is more sensible and more rapid than electrophoresis. The amplicons were sequenced to improve the discrimination between the species. The method enables the identification of Armillaria species within one day directly from soil samples without the need for previous isolation and cultivation of mycelium of Armillaria.
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