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Article

Aestipascuomyces dupliciliberans gen. nov, sp. nov., the First Cultured Representative of the Uncultured SK4 Clade from Aoudad Sheep and Alpaca

1
Process Engineering in Life Sciences 2: Technical Biology, Karlsruhe Institute of Technology, 76131 Karlsruhe, Germany
2
Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, OK 74074, USA
3
Bavarian State Research Center for Agriculture, Central Department for Quality Assurance and Analytics, Micro- and Molecular Biology, Lange Point 6, 85354 Freising, Germany
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Microorganisms 2020, 8(11), 1734; https://doi.org/10.3390/microorganisms8111734
Received: 13 October 2020 / Revised: 29 October 2020 / Accepted: 30 October 2020 / Published: 5 November 2020
(This article belongs to the Special Issue Unleashing the Hidden Potential of Anaerobic Fungi)
We report on the isolation of the previously-uncultured Neocallimastigomycota SK4 lineage, by two independent research groups, from a wild aoudad sheep rumen sample (Texas, USA) and an alpaca fecal sample (Baden-Württemberg, Germany). Isolates from both locations showed near-identical morphological and microscopic features, forming medium-sized (2–5 mm) white filamentous colonies with a white center of sporangia, on agar roll tubes and a heavy biofilm in liquid media. Microscopic analysis revealed monocentric thalli, and spherical polyflagellated zoospores with 7–20 flagella. Zoospore release occurred through an apical pore as well as by sporangial wall rupturing, a duality that is unique amongst described anaerobic gut fungal strains. Isolates were capable of growing on a wide range of mono-, oligo-, and polysaccharide substrates as the sole carbon source. Phylogenetic assessment based on the D1–D2 28S large rRNA gene subunit (D1–D2 LSU) and internal transcribed spacer-1 (ITS-1) regions demonstrated high sequence identity (minimum identity of 99.07% and 96.96%, respectively) between all isolates; but low sequence identity (92.4% and 86.7%, respectively) to their closest cultured relatives. D1–D2 LSU phylogenetic trees grouped the isolates as a new monophyletic clade within the OrpinomycesNeocallimastixPecoramycesFeramycesGhazallamyces supragenus group. D1–D2 LSU and ITS-1 sequences recovered from the obtained isolates were either identical or displayed extremely high sequence similarity to sequences recovered from the same aoudad sheep sample on which isolation was conducted, as well as several sequences recovered from domestic sheep and few other herbivores. Interestingly, members of the SK4 clade seem to be encountered preferably in animals grazing on summer pasture. We hence propose accommodating these novel isolates in a new genus, Aestipascuomyces (derived from the Latin word for “summer pasture”), and a new species, A. dupliciliberans. The type strain is Aestipascuomycesdupliciliberans strain R4. View Full-Text
Keywords: anaerobic gut fungi; sequence-guided isolation; Neocallimastigomycota SK4 lineage anaerobic gut fungi; sequence-guided isolation; Neocallimastigomycota SK4 lineage
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MDPI and ACS Style

Stabel, M.; Hanafy, R.A.; Schweitzer, T.; Greif, M.; Aliyu, H.; Flad, V.; Young, D.; Lebuhn, M.; Elshahed, M.S.; Ochsenreither, K.; Youssef, N.H. Aestipascuomyces dupliciliberans gen. nov, sp. nov., the First Cultured Representative of the Uncultured SK4 Clade from Aoudad Sheep and Alpaca. Microorganisms 2020, 8, 1734. https://doi.org/10.3390/microorganisms8111734

AMA Style

Stabel M, Hanafy RA, Schweitzer T, Greif M, Aliyu H, Flad V, Young D, Lebuhn M, Elshahed MS, Ochsenreither K, Youssef NH. Aestipascuomyces dupliciliberans gen. nov, sp. nov., the First Cultured Representative of the Uncultured SK4 Clade from Aoudad Sheep and Alpaca. Microorganisms. 2020; 8(11):1734. https://doi.org/10.3390/microorganisms8111734

Chicago/Turabian Style

Stabel, Marcus, Radwa A. Hanafy, Tabea Schweitzer, Meike Greif, Habibu Aliyu, Veronika Flad, Diana Young, Michael Lebuhn, Mostafa S. Elshahed, Katrin Ochsenreither, and Noha H. Youssef 2020. "Aestipascuomyces dupliciliberans gen. nov, sp. nov., the First Cultured Representative of the Uncultured SK4 Clade from Aoudad Sheep and Alpaca" Microorganisms 8, no. 11: 1734. https://doi.org/10.3390/microorganisms8111734

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