IDEAS home Printed from https://ideas.repec.org/a/caa/jnljfs/v68y2022i5id161-2021-jfs.html
   My bibliography  Save this article

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

Author

Listed:
  • Tomáš Tonka
  • 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)

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.

Suggested Citation

  • Tomáš Tonka & Dagmar Stehlíková & Lucie Walterová & Vladislav Čurn, 2022. "Development of loop mediated isothermal amplification for rapid species detection of Armillaria ostoyae using assimilating probe," Journal of Forest Science, Czech Academy of Agricultural Sciences, vol. 68(5), pages 163-169.
    • Handle: RePEc:caa:jnljfs:v:68:y:2022:i:5:id:161-2021-jfs
    DOI: 10.17221/161/2021-JFS
    as

    Download full text from publisher

    File URL: http://jfs.agriculturejournals.cz/doi/10.17221/161/2021-JFS.html
    Download Restriction: free of charge
    File URL: http://jfs.agriculturejournals.cz/doi/10.17221/161/2021-JFS.pdf
    Download Restriction: free of charge
    File URL: https://libkey.io/10.17221/161/2021-JFS?utm_source=ideas
    LibKey link: if access is restricted and if your library uses this service, LibKey will redirect you to where you can use your library subscription to access this item
    ---><---

    As the access to this document is restricted, you may want to search for a different version of it.

    Corrections

    All material on this site has been provided by the respective publishers and authors. You can help correct errors and omissions. When requesting a correction, please mention this item's handle: RePEc:caa:jnljfs:v:68:y:2022:i:5:id:161-2021-jfs. See general information about how to correct material in RePEc.

    If you have authored this item and are not yet registered with RePEc, we encourage you to do it here. This allows to link your profile to this item. It also allows you to accept potential citations to this item that we are uncertain about.

    We have no bibliographic references for this item. You can help adding them by using this form .

    If you know of missing items citing this one, you can help us creating those links by adding the relevant references in the same way as above, for each refering item. If you are a registered author of this item, you may also want to check the "citations" tab in your RePEc Author Service profile, as there may be some citations waiting for confirmation.

    For technical questions regarding this item, or to correct its authors, title, abstract, bibliographic or download information, contact: Ivo Andrle (email available below). General contact details of provider: https://www.cazv.cz/en/home/ .

    Please note that corrections may take a couple of weeks to filter through the various RePEc services.

    IDEAS is a RePEc service. RePEc uses bibliographic data supplied by the respective publishers.