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

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

Comorbidity of eczema, rhinitis, and asthma in IgE-sensitised and non-IgE-sensitised children in MeDALL: a population-based cohort study

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Genetic and taxonomic assessment of the widespread Afrotropical ambrosia beetle Xyleborus principalis (Coleoptera, Scolytinae)

The taxonomy of the widespread Afrotropical ambrosia beetle Xyleborus principalis Eichhoff, 1878 is reviewed based on morphological and molecular data. The species is both morphologically and genetically variable but without any structure corresponding to geography, or between genes and morphology. Examination of type material and other specimens collected across Africa and Madagascar documented the existence of a single species. New synonymies are proposed for Xyleborus principalis (=X. alluaudi Schaufuss, 1897, =X. camerunus Hagedorn, 1910, =X. consobrinus Eggers, 1932, =X. discrepans Schedl, 1950, =X. annectens Schedl, 1957, =X. peramploides Schedl, 1957)

First record of Hesperomyces virescens (Laboulbeniales, Ascomycota) on Harmonia axyridis (Coccinellidae, Coleoptera) in Poland

Hesperomyces virescens Thaxt. is a fungal parasite of coccinellid beetles. One of its hosts is the invasive harlequin ladybird Harmonia axyridis (Pallas). We present the first records of this combination from Poland

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

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