Development of loop mediated isothermal amplification for rapid species detection of Armillaria ostoyae using assimilating probe

Tomáš Tonka, Dagmar Stehlíková, Lucie Walterová and Vladislav Čurn
Additional contact information
Dagmar Stehlíková: Department of Plant Production, Faculty of Agriculture and Technology, University of South Bohemia in České Budějovice, České Budějovice, Czech Republic
Lucie Walterová: Department of Plant Production, Faculty of Agriculture and Technology, University of South Bohemia in České Budějovice, České Budějovice, Czech Republic
Vladislav Čurn: Department of Plant Production, Faculty of Agriculture and Technology, University of South Bohemia in České Budějovice, České Budějovice, Czech Republic

Journal of Forest Science, 2022, vol. 68, issue 5, 163-169

Abstract: We introduced here the first loop mediated isothermal amplification (LAMP) assay for the identification of honey fungus, Armillaria ostoyae, a basidiomycote playing an important role in spruce declines in the Palaearctic region. In total, 101 isolates, representing three Armillaria species, were used to develop a new LAMP assay to determine species specific identification. We have here described LAMP primers enhanced with fluorescent dye that are able to amplify A. ostoyae DNA and detect fungi in a fast single step reaction. The detection limit of LAMP was 1 pg of genomic DNA per reaction. We optimized a new LAMP assay for the rapid detection of A. ostoyae using the translation elongation factor 1-α (tef1) marker and fluorescence labelled oligonucleotide assimilating probe. The LAMP assay does not require any specialized equipment, hence it can be used in the field for the rapid detection of A. ostoyae even using the portable and mobile device. The specificity of the assay was confirmed by the use of A. ostoyae strains and Armillaria cepistipes and Armillaria gallica strains, respectively. In conclusion, the assay could be a rapid, specific, sensitive and low-cost tool for identification of A. ostoyae as well as the first step for expansion of this method in practical applications.

Keywords: detection of fungi; molecular diagnosis; forest pathogen; root rot fungi; spruce decline (search for similar items in EconPapers)
Date: 2022
References: View complete reference list from CitEc
Citations:

Downloads: (external link)
http://jfs.agriculturejournals.cz/doi/10.17221/161/2021-JFS.html (text/html)
http://jfs.agriculturejournals.cz/doi/10.17221/161/2021-JFS.pdf (application/pdf)
free of charge

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:caa:jnljfs:v:68:y:2022:i:5:id:161-2021-jfs

DOI: 10.17221/161/2021-JFS

Access Statistics for this article

Journal of Forest Science is currently edited by Mgr. Ilona Procházková

More articles in Journal of Forest Science from Czech Academy of Agricultural Sciences
Bibliographic data for series maintained by Ivo Andrle ().