Callaghan, T. | Search bioRxiv

bioRxiv2019

Seven new Neocallimastigomycota genera from fecal samples of wild, zoo-housed, and domesticated herbivores: Description of Ghazallomyces constrictus gen. nov., sp. nov., Aklioshbomyces papillarum gen. nov., sp. nov., Agriosomyces longus gen. nov., sp. nov., Capellomyces foraminis gen. nov., sp. nov. and Capellomyces elongatus sp. nov., Joblinomyces apicalis gen. nov., sp. nov., Khoyollomyces ramosus gen. nov., sp. nov., and Tahromyces munnarensis gen. nov., sp. nov.

We isolated and characterized sixty-five anaerobic gut fungi (AGF, Neocallimastigomycota) strains from fecal samples of five wild (W), one zoo-housed (Z), and three domesticated (D) herbivores in the US states of Texas (TX) and Oklahoma (OK), Wales (WA), and the Indian states of Kerala (KE) and Haryana (HA). Phylogenetic assessment based on D1-D2 region of the large rRNA subunit (LSU) identified seven distinct lineages, with strains recovered from Axis Deer (W-TX) clustering within the Orpinomyces-Neocallimastix-Pecoramyces-Feramyces clade; Boer Goat-domesticated Goat strains (W-TX, D-KE) clustering within the Oontomyces-Anaeromyces-Liebetanzomyces clade; and domesticated Goat and Sheep strains (D-HA) as well as Nilgiri Tahr strains (W-KE) forming two distinct clades associated with genus Buwchfawromyces. The remaining three lineages, represented by strains recovered from Mouflon-Boer Goat (W-TX), White Tailed Deer (W-OK), and Zebra-Horse (Z-OK, and D-WA), displayed no specific suprageneric affiliation. All strains displayed monocentric thalli and produced mono/uniflagellate zoospores with the exception of Axis Deer strains, which produced polyflagellate zoospores. Isolates displayed multiple interesting microscopic features including sporangia with tightly constricted necks and fine septa at the base (Axis Deer), papillated and pseudo-intercalary sporangia (White-Tailed Deer), swollen sporangiophores and zoospores with long flagella (Mouflon-Boer Goat), zoospore release through an apical pore followed by either sporangial wall collapse (Axis Deer and Boer Goat-domesticated Goat) or sporangial wall remaining intact after discharge (Zebra-Horse), multi-sporangiated thalli with branched sporangiophores (Zebra-Horse), and short sporangiophores with subsporangial swellings (Nilgiri Tahr). Internal transcribed spacer-1 region (ITS-1) sequence analysis indicated that Zebra-Horse strains are representatives of the AL1 lineage, frequently encountered in culture-independent surveys of the alimentary tract and fecal samples from hindgut fermenters. The other six lineages, five of which were isolated from wild herbivores, have not been previously encountered in such surveys. Our results significantly expand the genus level diversity within the Neocallimastigomycota, and strongly suggest that wild herbivores represent a yet-untapped reservoir of AGF diversity. We propose the creation of seven novel genera and eight novel Neocallimastigomycota species to accommodate these strains, for which we propose the names Agriosomyces longus (Mouflon and wild Boer Goat), Aklioshbomyces papillarum (White tailed Deer), Capellomyces foraminis (wild Boar Goat) and C. elongatus (domesticated Goat), Ghazallomyces constrictus (Axis Deer), Joblinomyces apicalis (domesticated Goat and Sheep), Khoyollomyces ramosus (Zebra-Horse), and Tahromyces munnarensis (Nilgiri Tahr). The type species are strains Axs-31, WT-2, MS-4, BGB-11, GFKJa1916, GFH683, ZS-33, and TDFKJa193, respectively.

microbiology↗

bioRxiv2018

Horizontal gene transfer as an indispensible driver for Neocallimastigomycota evolution into a distinct gut-dwelling fungal lineage

Survival and growth of the anaerobic gut fungi (AGF, Neocallimastigomycota) in the herbivorous gut necessitate the possession of multiple abilities absent in other fungal lineages. We hypothesized that horizontal gene transfer (HGT) was instrumental in forging the evolution of AGF into a phylogenetically distinct gut-dwelling fungal lineage. Patterns of HGT were evaluated in the transcriptomes of 27 AGF strains, 22 of which were isolated and sequenced in this study, and 4 AGF genomes broadly covering the breadth of AGF diversity. We identified 283 distinct incidents of HGT in AGF transcriptomes, with subsequent gene duplication resulting in an HGT frequency of 2.1-3.6% in AGF genomes. The majority of HGT events were AGF specific (91.5%) and wide (70.7%), indicating their occurrence at early stages of AGF evolution. The acquired genes allowed AGF to expand their substrate utilization range, provided new venues for electron disposal, augmented their biosynthetic capabilities, and facilitated their adaptation to anaerobiosis. The majority of donors were anaerobic fermentative bacteria prevalent in the herbivorous gut. This work strongly indicates that HGT indispensably forged the evolution of AGF as a distinct fungal phylum and provides a unique example of the role of HGT in shaping the evolution of a high rank taxonomic eukaryotic lineage. ImportanceThe anaerobic gut fungi (AGF) represent a distinct basal phylum lineage (Neocallimastigomycota) commonly encountered in the rumen and alimentary tracts of herbivores. Survival and growth of anaerobic gut fungi in these anaerobic, eutrophic, and prokaryotes dominated habitats necessitates the acquisition of several traits absent in other fungal lineages. This manuscript assesses the role of horizontal gene transfer as a relatively fast mechanism for trait acquisition by the Neocallimastigomycota post sequestration in the herbivorous gut. Analysis of twenty-seven transcriptomes that represent the broad Neocallimastigomycota diversity identified 283 distinct HGT events, with subsequent gene duplication resulting in an HGT frequency of 2.1-3.6% in AGF genomes. These HGT events have allowed AGF to survive in the herbivorous gut by expanding their substrate utilization range, augmenting their biosynthetic pathway, providing new routes for electron disposal by expanding fermentative capacities, and facilitating their adaptation to anaerobiosis. HGT in the AGF is also shown to be mainly a cross-kingdom affair, with the majority of donors belonging to the bacteria. This work represents a unique example of the role of HGT in shaping the evolution of a high rank taxonomic eukaryotic lineage.

microbiology↗