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Bringing the Laboulbeniales to the 21st century: Enhanced techniques for extraction and PCR amplification of DNA from minute ectoparasitic fungi

Haelewaters, Danny and Gorczak, Michał and Pfliegler, Walter P. and Tartally, András and Tischer, Marta and Wrzosek, Marta and Pfister, Donald H. (2015) Bringing the Laboulbeniales to the 21st century: Enhanced techniques for extraction and PCR amplification of DNA from minute ectoparasitic fungi. IMA Fungus, 6 (2). pp. 363-372. ISSN 2210-6359

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Abstract

One of the most peculiar groups of Ascomycota are the Laboulbeniales. 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 Laboulbeniales, sister to the Pyxidiophorales, have only recently been assigned their own 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 peculiar fungi. We provide 43 newly generated Laboulbeniales rDNA 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 (80-100%). 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.

Item Type: Article
Subjects: Q Science / természettudomány > QL Zoology / állattan
Q Science / természettudomány > QR Microbiology / mikrobiológia
Depositing User: dr András Tartally
Date Deposited: 26 Sep 2015 14:57
Last Modified: 06 Nov 2015 10:48
URI: http://real.mtak.hu/id/eprint/28382

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