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The Phanerozoic diversification of silica-cycling testate amoebae and its possible links to changes in terrestrial ecosystems

, Lahr, Daniel J.G, Bosak, Tanja, Lara, Enrique, Mitchell, Edward

The terrestrial cycling of Si is thought to have a large influence on the terrestrial and marine primary production, as well as the coupled biogeochemical cycles of Si and C. Biomineralization of silica is widespread among terrestrial eukaryotes such as plants, soil diatoms, freshwater sponges, silicifying flagellates and testate amoebae. Two major groups of testate (shelled) amoebae, arcellinids and euglyphids, produce their own silica particles to construct shells. The two are unrelated phylogenetically and acquired biomineralizing capabilities independently. Hyalosphenids, a group within arcellinids, are predators of euglyphids.We demonstrate that hyalosphenids can construct shells using silica scales mineralized by the euglyphids. Parsimony analyses of the current hyalosphenid phylogeny indicate that the ability to “steal” euglyphid scales is most likely ancestral in hyalosphenids, implying that euglyphids should be older than hyalosphenids. However, exactly when euglyphids arose is uncertain. Current fossil record contains unambiguous euglyphid fossils that are as old as 50 million years, but older fossils are scarce and difficult to interpret. Poor taxon sampling of euglyphids has also prevented the development of molecular clocks. Here, we present a novel molecular clock reconstruction for arcellinids and consider the uncertainties due to various previously used calibration points. The new molecular clock puts the origin of hyalosphenids in the early Carboniferous (~370 mya). Notably, this estimate coincides with the widespread colonization of land by Si-accumulating plants, suggesting possible links between the evolution of Arcellinid testate amoebae and the expansion of terrestrial habitats rich in organic matter and bioavailable Si.

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NAD9/NAD7 (mitochondrial nicotinamide adenine dinucleotide dehydrogenase gene): A new “Holy Grail” phylogenetic and DNA-barcoding marker for Arcellinida (Amoebozoa)?

, Blandenier, Quentin, Lara, Enrique, Mitchell, Edward, Alcantara, Daniel M.C, Siemensma, Ferry J, Todorov, Milcho, Lahr, Daniel J.G

Molecular phylogeny is an indispensable tool for assessing evolutionary relationships among protists. The most commonly used marker is the small subunit ribosomal RNA gene, a conserved gene present in many copies in the nuclear genomes. However, this marker is not variable enough at a fine-level taxonomic scale, and intra-genomic polymorphism has already been reported. Finding a marker that could be useful at both deep and fine taxonomic resolution levels seemed like a utopic dream. We designed Amoebozoa-specific primers to amplify a region including partial sequences of two subunits of the mitochondrial nicotinamide adenine dinucleotide dehydrogenase gene (NAD9/NAD7). We applied them to arcellinids belonging to distantly related genera (Arcella, Difflugia, Netzelia and Hyalosphenia) and to Arcellinid-rich environmental samples to obtain additional Amoebozoa sequences. Tree topology was congruent with previous phylogenies, all nodes being highly supported, suggesting that this marker is well-suited for deep phylogenies in Arcellinida and perhaps Amoebozoa. Furthermore, it enabled discrimination of close-related taxa. This short genetic marker (ca. 250 bp) can therefore be used at different taxonomic levels, due to a fast-varying intergenic region presenting either a small intergenic sequence or an overlap, depending on the species.

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Soil protistology rebooted: 30 fundamental questions to start with

, Geisen, Stefan, Mitchell, Edward, Wilkinson, David M, Adl, Sina, Bonkowski, Michael, Brown, Matthew W, Fiore-Donno, Anna Maria, Heger, Thierry, Jassey, Vincent E.J, Krashevska, Valentyna, Lahr, Daniel J.G, Marcisz, Katarzyna, Mulot, Matthieu, Payne, Richard, Singer, David, Anderson, Roger O, Charman, Dan J, Ekelund, Flemming, Griffiths, Bryan S, Rønn, Regin, Smirnov, Alexey, Bass, David, Belbahri, Lassaâd, Berney, Cédric, Blandenier, Quentin, Blandenier, Quentin, Chatzinotas, Antonis, Clarholm, Marianne, Dunthorn, Micah, Feest, Alan, Fernández, Leonardo D, Foissner, Wilhelm, Fournier, Bertrand, Gentekaki, Eleni, Hájek, Michal, Helder, Johannes, Jousset, Alexandre, Koller, Robert, Kumar, Santosh, La Terza, Antonietta, Lamentowicz, Mariusz, Mazei, Yuri, Santos, Susana S, Seppey, Christophe V.W, Spiegel, Frederick W, Walochnik, Julia, Winding, Anne, Lara, Enrique

Protists are the most diverse eukaryotes. These microbes are keystone organisms of soil ecosystems and regulate essential processes of soil fertility such as nutrient cycling and plant growth. Despite this, protists have received little scientific attention, especially compared to bacteria, fungi and nematodes in soil studies. Recent methodological advances, particularly in molecular biology techniques, have made the study of soil protists more accessible, and have created a resurgence of interest in soil protistology. This ongoing revolution now enables comprehensive investigations of the structure and functioning of soil protist communities, paving the way to a new era in soil biology. Instead of providing an exhaustive review, we provide a synthesis of research gaps that should be prioritized in future studies of soil protistology to guide this rapidly developing research area. Based on a synthesis of expert opinion we propose 30 key questions covering a broad range of topics including evolution, phylogenetics, functional ecology, macroecology, paleoecology, and methodologies. These questions highlight a diversity of topics that will establish soil protistology as a hub discipline connecting different fundamental and applied fields such as ecology, biogeography, evolution, plant-microbe interactions, agronomy, and conservation biology. We are convinced that soil protistology has the potential to be one of the most exciting frontiers in biology.