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Open Access Bringing Laboulbeniales into the 21st century: enhanced techniques for extraction and PCR amplification of DNA from minute ectoparasitic fungi

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Laboulbeniales is one of the most peculiar orders of Ascomycota. These fungi are characterized by an ectoparasitic life-style on arthropods, determinate growth, lack of an asexual stage, high species richness, and intractability to culture. The order Laboulbeniales, sister to Pyxidiophorales, has only recently been assigned a separate class, the Laboulbeniomycetes, based on very few ribosomal DNA sequences. So far, DNA isolations and PCR amplifications have proven difficult. Here, we provide details of isolation techniques and the application of commercially available kits that enable efficient and reliable genetic analyses of these fungi. We provide 43 newly generated Laboulbeniales ribosomal DNA sequences, among which are the first published sequences for species in the genera Gloeandromyces, Herpomyces, Laboulbenia, Monoicomyces, and Polyandromyces. DNA extractions were possible using from 1 to 30 thalli from hosts preserved in ethanol (70–100 %). In two cases, we successfully isolated DNA from thalli on dried insect collections. Laboulbeniales molecular systematics could be substantially enhanced through these improved methods by allowing more complete sampling of both taxa and gene regions.
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Keywords: ASCOMYCOTA; DNA ISOLATION; INSECT COLLECTIONS; LABOULBENIALES-SPECIFIC PRIMERS; RIBOSOMAL DNA; UNCULTURABLE FUNGI

Document Type: Research Article

Publication date: 01 December 2015

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  • Scope: All aspects of pure and applied mycological research and news.
    Aims: The flagship journal of the International Mycological Association. IMA Fungus is an international, peer-reviewed, open-access, full colour, fast-track journal. Papers on any aspect of mycology are considered, and published on-line with final pagination after proofs have been corrected; they are then effectively published under the International Code of Nomenclature for algae, fungi, and plants. The journal strongly supports good practice policies, and requires voucher specimens or cultures to be deposited in a public collection with an online database, DNA sequences in GenBank, alignments in TreeBASE, and validating information on new scientific names, including typifications, to be lodged in MycoBank. News, meeting reports, personalia, research news, correspondence, book news, and information on forthcoming international meetings are included in each issue.

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