CORDIS OpenAire Sygma Bielefeld Academic Search Engine (BASE) UnpayWall SLU publication database Digitala Vetenskapliga Arkivet - Academic Archive On-line Publikationer från Uppsala Universitet Crossref Europe PubMed Central PubMed Central European Union Open Data Portal https://pub.epsilon.slu.se/27699/1/eshghi-sahraei-s-et-al-220505.pdf https://onlinelibrary.wiley.com/doi/pdf/10.1002/ece3.8676 https://doi.org/10.1002/ece3.8676 Ecology and Evolution Open Access 2022-03-08 Abstract<p>Long amplicon metabarcoding has opened the door for phylogenetic analysis of the largely unknown communities of microeukaryotes in soil. Here, we amplified and sequenced the ITS and LSU regions of the rDNA operon (around 1500ￂﾠbp) from grassland soils using PacBio SMRT sequencing. We tested how three different methods for generation of operational taxonomic units (OTUs) effected estimated richness and identified taxa, and how well large￢ﾀﾐscale ecological patterns associated with shifting environmental conditions were recovered in data from the three methods. The field site at Kungsￃﾤngen Nature Reserve has drawn frequent visitors since Linnaeus's time, and its species rich vegetation includes the largest population of Fritillaria meleagris in Sweden. To test the effect of different OTU generation methods, we sampled soils across an abrupt moisture transition that divides the meadow community into a Carex acuta dominated plant community with low species richness in the wetter part, which is visually distinct from the mesic￢ﾀﾐdry part that has a species rich grass￢ﾀﾐdominated plant community including a high frequency of F.ￂﾠmeleagris. We used the moisture and plant community transition as a framework to investigate how detected belowground microeukaryotic community composition was influenced by OTU generation methods. Soil communities in both moisture regimes were dominated by protists, a large fraction of which were taxonomically assigned to Ciliophora (Alveolata) while 30%￢ﾀﾓ40% of all reads were assigned to kingdom Fungi. Ecological patterns were consistently recovered irrespective of OTU generation method used. However, different methods strongly affect richness estimates and the taxonomic and phylogenetic resolution of the characterized community with implications for how well members of the microeukaryotic communities can be recognized in the data.</p> 580 species hypothesis Ekologi 0301 basic medicine 0303 health sciences Ecology rDNA Biological Systematics 15. Life on land 03 medical and health sciences 14. Life underwater ITS Research Articles clustering Eshghi Sahraei, Shadi, Furneaux, Brendan, Kluting, Kerri, Zakieh, Mustafa, Rydin, Håkan, Hytteborn, Håkan, Rosling, Anna, 2022-03-01 journalpaper Abstract<p>Long amplicon metabarcoding has opened the door for phylogenetic analysis of the largely unknown communities of microeukaryotes in soil. Here, we amplified and sequenced the ITS and LSU regions of the rDNA operon (around 1500ￂﾠbp) from grassland soils using PacBio SMRT sequencing. We tested how three different methods for generation of operational taxonomic units (OTUs) effected estimated richness and identified taxa, and how well large￢ﾀﾐscale ecological patterns associated with shifting environmental conditions were recovered in data from the three methods. The field site at Kungsￃﾤngen Nature Reserve has drawn frequent visitors since Linnaeus's time, and its species rich vegetation includes the largest population of Fritillaria meleagris in Sweden. To test the effect of different OTU generation methods, we sampled soils across an abrupt moisture transition that divides the meadow community into a Carex acuta dominated plant community with low species richness in the wetter part, which is visually distinct from the mesic￢ﾀﾐdry part that has a species rich grass￢ﾀﾐdominated plant community including a high frequency of F.ￂﾠmeleagris. We used the moisture and plant community transition as a framework to investigate how detected belowground microeukaryotic community composition was influenced by OTU generation methods. Soil communities in both moisture regimes were dominated by protists, a large fraction of which were taxonomically assigned to Ciliophora (Alveolata) while 30%￢ﾀﾓ40% of all reads were assigned to kingdom Fungi. Ecological patterns were consistently recovered irrespective of OTU generation method used. However, different methods strongly affect richness estimates and the taxonomic and phylogenetic resolution of the characterized community with implications for how well members of the microeukaryotic communities can be recognized in the data.</p> Effects of operational taxonomic unit inference methods on soil microeukaryote community analysis using long-read metabarcoding 10.1002/ece3.8676