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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

Background: The origin of animals from their unicellular ancestor was one of the most important events in evolutionary history, but the nature and the order of events leading up to the emergence of multicellular animals are still highly uncertain. The diversity and biology of unicellular relatives of animals have strongly informed our understanding of the transition from single-celled organisms to the multicellular Metazoa. Here, we analyze the cellular structures and complex life cycles of the novel unicellular holozoans Pigoraptor and Syssomonas (Opisthokonta), and their implications for the origin of animals. Results: Syssomonas and Pigoraptor are characterized by complex life cycles with a variety of cell types including flagellates, amoeboflagellates, amoeboid non-flagellar cells, and spherical cysts. The life cycles also include the formation of multicellular aggregations and syncytium-like structures, and an unusual diet for single-celled opisthokonts (partial cell fusion and joint sucking of a large eukaryotic prey), all of which provide new insights into the origin of multicellularity in Metazoa. Several existing models explaining the origin of multicellular animals have been put forward, but these data are interestingly consistent with one, the "synzoospore hypothesis." Conclusions: The feeding modes of the ancestral metazoan may have been more complex than previously thought, including not only bacterial prey, but also larger eukaryotic cells and organic structures. The ability to feed on large eukaryotic prey could have been a powerful trigger in the formation and development of both aggregative (e.g., joint feeding, which also implies signaling) and clonal (e.g., hypertrophic growth followed by palintomy) multicellular stages that played important roles in the emergence of multicellular animals.

a b s t r a c tFunctional traits (FT) offer a new framework to understand the ecology of organisms and overcome taxonomic difficulties that currently limit the study of minute soil taxa. FT are likely to be selected by environmental filters and hence they may provide more direct information on ecosystem characteristics than the species composition of a community.We tested the potential of testate amoeba (TA) functional traits as bioindicators of selected ecosystem processes in the context of a restored floodplain in north-western Switzerland. The floodplain was divided into six functional process zones (FPZs) associated to distinct post-restoration successional stages. We selected TA FT and computed three functional indices: functional richness (FRic), divergence (FDiv), evenness (FEve), and dispersion (FDis). We then compared the patterns of functional indices and classical diversity indices such as species richness, diversity and evenness. We assessed whether traits converged or were over-dispersed in the different FPZs using a randomization procedure. Finally, we related environmental variables and functional traits using the "Fourth Corner" statistic. This procedure enabled us to highlight relations that can potentially be used for bioindication. Promising candidates include the relationships between shell biovolume and vegetation structure and between shell compression and plant litter input variables.

16Among the non-pollen micro-fossils commonly encountered in Quaternary sediment samples prepared 17 for pollen analysis are many shells of testate amoebae. Testate amoebae are eukaryotic micro-18 organisms which are increasingly used in ecological and palaeoecological studies, particularly as 19 indicators of hydrological change in Sphagnum-dominated peatlands. In this study we address the 20 extent to which testate amoebae are used in palynological research, the key challenges to more 21 widespread use, and the extent to which ecological information is retained in the testate amoeba 22 assemblages of standard palynological slides. To achieve this we review the literature on the use of 23 testate amoebae in palynology, compare testate amoeba records produced by palynological and water-24 based preparation methods and carry out simulations using previously-derived datasets. Our results 25show that testate amoebae are widely encountered in Quaternary palynological studies, primarily in 26 peatlands, but the information which they can provide is undermined by limited taxonomic knowledge. 27Many taxa are destroyed in pollen preparations, but for taxa that are retained patterns of abundance 28 parallel those determined using water-based preparation methods. Although the loss of sensitive taxa 29 limits the ecological information contained in testate amoeba assemblages the information preserved is 30 likely to be useful in a multiproxy approach to palaeoenvironmental reconstruction. To help improve 31 taxonomic awareness and encourage the use of testate amoebae in palynology we present a basic 32 introduction to testate amoeba taxonomy and a guide to the taxonomic literature. 33

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Aim  The biogeography and global distribution of protists has long been disputed, with two primary, opposing views. To test these two sets of views in greater detail, we have compiled the available data for marine benthic ciliates and assessed the general patterns of their diversity and distribution compared with Metazoa. Location  World‐wide. Methods  A comprehensive database (1342 species, over 350 sources) was used to analyse the diversity, distribution, species occurrences and range size distribution of free‐living ciliates that inhabit marine sediments in 17 geographical regions. Results  Twenty‐five per cent of the species have been found in a single region only, whereas 18% are widespread (they occur in more than half the regions covering both hemispheres). Only 5–7% of regional faunas are endemic, which is much lower than for macroorganisms. Regional diversity depends neither on total area nor on coastline length and does not show any obvious latitudinal trends, but correlates highly with the investigation effort expended in a region and (negatively) with the average salinity. A comparison of species composition reveals distinctions between the Arctic Area (the White, Barents and Kara seas), Laurasian Area (north Atlantic, north Pacific and European seas), Gondwanian Area (Southern Ocean) and the Antarctic. No clear geographical correlations are found for faunistic composition at the genus or family levels. There is the tendency to narrow the latitudinal ranges for species found at high latitudes (reversal of Rapoport's rule). Main conclusions  Undersampling and data insufficiency are the key factors affecting the observed diversity and distribution of microorganisms. Nevertheless, marine benthic ciliates demonstrate certain patterns that generally agree with the ‘moderate endemicity’ model (Foissner, 2004, 2008), but consistently contradict the regularities commonly observed for multicellular taxa. Thus, ciliates do have a biogeography, but their macroecological patterns may be different in some respects from that of macroorganisms.