Genomic evidence for the widespread presence of GH45 cellulases among soil invertebrates.

Lignocellulose is a major component of vascular plant biomass. Its decomposition is crucial for the terrestrial carbon cycle. Microorganisms are considered primary decomposers, but evidence increases that some invertebrates may also decompose lignocellulose. We investigated the taxonomic distribution and evolutionary origins of GH45 hydrolases, important enzymes for the decomposition of cellulose and hemicellulose, in a collection of soil invertebrate genomes. We found that these genes are common in springtails and oribatid mites. Phylogenetic analysis revealed that cellulase genes were acquired early in the evolutionary history of these groups. Domain architectures and predicted 3D enzyme structures indicate that these cellulases are functional. Patterns of presence and absence of these genes across different lineages prompt further investigation into their evolutionary and ecological benefits. The ubiquity of cellulase genes suggests that soil invertebrates may play a role in lignocellulose decomposition, independently or in synergy with microorganisms. Understanding the ecological and evolutionary implications might be crucial for understanding soil food webs and the carbon cycle.

The MetaInvert soil invertebrate genome resource provides insights into below-ground biodiversity and evolution.

Soil invertebrates are among the least understood metazoans on Earth. Thus far, the lack of taxonomically broad and dense genomic resources has made it hard to thoroughly investigate their evolution and ecology. With MetaInvert we provide draft genome assemblies for 232 soil invertebrate species, representing 14 common groups and 94 families. We show that this data substantially extends the taxonomic scope of DNA- or RNA-based taxonomic identification. Moreover, we confirm that theories of genome evolution cannot be generalised across evolutionarily distinct invertebrate groups. The soil invertebrate genomes presented here will support the management of soil biodiversity through molecular monitoring of community composition and function, and the discovery of evolutionary adaptations to the challenges of soil conditions.

Ecology, genetics and distribution of Punctoribates zachvatkini, an oribatid mite so far overlooked in Germany.

Punctoribates is one of few genera in Poronota (Acari: Oribatida) containing species with porose areas and species with saccules, the two types of the octotaxic system. These porose organs are the main difference between two morphologically similar species, P. punctum with porose areas and P. zachvatkini with saccules. As the octotaxic system can vary within species, species separation solely based on this trait might be insufficient. To assess the species status of P. zachvatkini, we investigated additional differences from P. punctum by comparing habitat preferences of the two species regarding nature reserves and agricultural landscapes during a field study in the German Eifel region, and by examining Punctoribates material from four large German natural history museums. We also performed scanning electron microscopy (SEM) and a genetic analysis using the D3 marker of the nuclear 28S rDNA gene. In the field study, P. zachvatkini had higher densities in the nature reserves and P. punctum in the agricultural landscapes. Evaluation of the museum material revealed P. punctum occurred more regularly in disturbed sites such as urban, agricultural and post-mining areas compared to P. zachvatkini. Pairwise distances of the 28S D3 genetic marker as well as an additional base pair in P. zachvatkini further support the separation of the two species, and SEM investigations revealed new details regarding the punctulation of P. zachvatkini. The review of the museum material showed that P. zachvatkini already occurred in Germany in 1967 and has a wider distribution than previously known.

Shotgun metagenomics of soil invertebrate communities reflects taxonomy, biomass, and reference genome properties.

Metagenomics - shotgun sequencing of all DNA fragments from a community DNA extract - is routinely used to describe the composition, structure, and function of microorganism communities. Advances in DNA sequencing and the availability of genome databases increasingly allow the use of shotgun metagenomics on eukaryotic communities. Metagenomics offers major advances in the recovery of biomass relationships in a sample, in comparison to taxonomic marker gene-based approaches (metabarcoding). However, little is known about the factors which influence metagenomics data from eukaryotic communities, such as differences among organism groups, the properties of reference genomes, and genome assemblies.We evaluated how shotgun metagenomics records composition and biomass in artificial soil invertebrate communities at different sequencing efforts. We generated mock communities of controlled biomass ratios from 28 species from all major soil mesofauna groups: mites, springtails, nematodes, tardigrades, and potworms. We shotgun sequenced these communities and taxonomically assigned them with a database of over 270 soil invertebrate genomes.We recovered over 95% of the species, and observed relatively high false-positive detection rates. We found strong differences in reads assigned to different taxa, with some groups (e.g., springtails) consistently attracting more hits than others (e.g., enchytraeids). Original biomass could be predicted from read counts after considering these taxon-specific differences. Species with larger genomes, and with more complete assemblies, consistently attracted more reads than species with smaller genomes. The GC content of the genome assemblies had no effect on the biomass-read relationships. Results were similar among different sequencing efforts.The results show considerable differences in taxon recovery and taxon specificity of biomass recovery from metagenomic sequence data. The properties of reference genomes and genome assemblies also influence biomass recovery, and they should be considered in metagenomic studies of eukaryotes. We show that low- and high-sequencing efforts yield similar results, suggesting high cost-efficiency of metagenomics for eukaryotic communities. We provide a brief roadmap for investigating factors which influence metagenomics-based eukaryotic community reconstructions. Understanding these factors is timely as accessibility of DNA sequencing and momentum for reference genomes projects show a future where the taxonomic assignment of DNA from any community sample becomes a reality.

Parthenogenetic vs. sexual reproduction in oribatid mite communities.

The dominance of sex in Metazoa is enigmatic. Sexual species allocate resources to the production of males, while potentially facing negative effects such as the loss of well-adapted genotypes due to recombination, and exposure to diseases and predators during mating. Two major hypotheses have been put forward to explain the advantages of parthenogenetic versus sexual reproduction in animals, that is, the Red Queen hypothesis and the Tangled Bank/Structured Resource Theory of Sex. The Red Queen hypothesis assumes that antagonistic predator-prey/ parasite-host interactions favor sex. The Structured Resource Theory of Sex predicts sexual reproduction to be favored if resources are in short supply and aggregated in space. In soil, a remarkable number of invertebrates reproduce by parthenogenesis, and this pattern is most pronounced in oribatid mites (Oribatida, Acari). Oribatid mites are abundant in virtually any soil across very different habitats, and include many sexual and parthenogenetic (thelytokous) species. Thereby, they represent an ideal model group to investigate the role of sexual versus parthenogenetic reproduction across different ecosystems and habitats. Here, we compiled data on oribatid mite communities from different ecosystems and habitats across biomes, including tropical rainforests, temperate forests, grasslands, arable fields, salt marshes, bogs, caves, and deadwood. Based on the compiled dataset, we analyzed if the percentage of parthenogenetic species and the percentage of individuals of parthenogenetic species are related to total oribatid mite density, species number, and other potential driving factors of the reproductive mode including altitude and latitude. We then interpret the results in support of either the Red Queen hypothesis or the Structured Resource Theory of Sex. Overall, the data showed that low density of oribatid mites due to harsh environmental conditions is associated with high frequency of parthenogenesis supporting predictions of the Structured Resource Theory of Sex rather than the Red Queen hypothesis.

Slow-moving soil organisms on a water highway: aquatic dispersal and survival potential of Oribatida and Collembola in running water.

BACKGROUND: Oribatida and Collembola are an important part of the soil food web and increase soil fertility by contributing to the recycling of nutrients out of dead organic matter. Active locomotion enables only limited dispersal in these tiny, wingless arthropods, while passive dispersal plays an important role for long-distance dispersal. Previous investigations have focused on passive transport by wind, other animals, or sea currents, whereas studies on transport via running water are missing. However, previous observation of the long survival of submerged terrestrial microarthropods makes passive dispersal with running water very likely. METHODS: By combining field and lab experiments, we studied the potential for passive dispersal of oribatid mites with running water. We investigated terrestrial Oribatida and Collembola: (1) along a stream taking soil and moss samples, (2) in a stream using sticky covers and aquarium nets, and (3) studied their ability to colonise new soil after aquatic transport with the help of floating islands. Furthermore, we investigated the survival of submerged Oribatida species and their floating capabilities in lab experiments to predict dispersal distances with running water. We tested for differences between species using Kruskal-Wallis test for equal medians and Mann-Whitney pairwise-comparison and χ2-test for the influence of body size on aquatic dispersal. RESULTS: Soil and moss samples revealed a pool of 52 oribatid mite species at the stream bank. Within the stream, we caught 180 individuals from 36 oribatid mite species. Only 14 of those species were also found in the soil and moss samples, whereas the remaining 22 were of unknown origin. Based on material caught on sticky covers, an average of 63.9 (± 54.6) oribatid mite individuals fell on one m2 stream water per week. Four species of Collembola (27 individuals) and 21 species of oribatid mites (47 individuals) were collected with aquarium nets. Eight microarthropod species (Oribatida + Collembola) successfully colonised new soil in the floating islands after aquatic dispersal. Lab experiments showed that Oribatida can float for at least 14 hours at the surface of running water and may survive for more than 365 days when submerged. The floating abilities and survival rates were largely species-specific. CONCLUSION: This is the first study to demonstrate successful passive dispersal with running water for two groups of terrestrial soil microarthropods, including subsequent colonisation of new soil. We show that submersion survival, as well as floating abilities, and therefore dispersal capability, are not only high in oribatid mites, but also species-specific. Running waters obviously serve as long-distance dispersal highways for many of these less mobile soil-living animals.

The nuclear 28S gene fragment D3 as species marker in oribatid mites (Acari, Oribatida) from German peatlands.

To make oribatid mites an applicable tool in monitoring programs it is necessary to find a molecular species marker that allows distinct, rapid and easy species identification. In previous studies, the common barcoding sequence COI showed to be too variable to serve as species marker in oribatid mites. The aim of the present study is to evaluate the potential use of the D3 region of the nuclear 28S rDNA gene for species identification. Therefore, we generated a reference DNA library of 28S D3 to identify specimens of the Oribatida from Germany, with focus on species occurring in peatlands being one of the most endangered habitats in Europe. New DNA sequences were obtained from 325 individuals and 64 species (58 genera, 34 families). By adding 28S D3-sequences from GenBank we altogether analysed 385 sequences from 89 German species, 32 of them restricted to peatlands and further 42 occurring in peatlands occasionally, representing 46 and 33% of the oribatids in German peatlands, respectively. P-distances were measured between species within families as well as for intraspecific divergence. 28S D3 showed low intraspecific genetic p-distances between 0 and 0.5%, interspecific distances within families varied between 0 and 9.7%. Most species pairs within families were further separated by one to four indels in addition to substitutions. Altogether, 93% of all analysed species are clearly delineated by 28S D3. Our study emphasises that 28S D3 rDNA is a useful barcode for the identification of oribatid mite specimens and represents an important step in building-up a comprehensive barcode library to allow metabarcoding analyses of environmental peatland samples for Oribatida in Germany as well as in Central Europe.

Convergent evolution of aquatic life by sexual and parthenogenetic oribatid mites.

Convergent evolution is one of the main drivers of traits and phenotypes in animals and plants. Here, we investigated the minimum number of independent colonisations of marine and freshwater habitats in derived oribatid mites (Brachypylina), a mainly terrestrial taxon. Furthermore, we investigated whether the reproductive mode (sexual vs. thelytokous) is associated with the habitat type (marine, freshwater) where the animals live. We hypothesized that continuous resource availability in freshwater systems fosters asexual reproduction. We used 18S rDNA sequences to construct a molecular phylogeny of oribatid mites from terrestrial, marine and freshwater habitats. The results indicate that aquatic life in oribatid mites evolved at least 3×: once in Limnozetoidea (including only freshwater taxa) and at least twice in Ameronothroidea. In Ameronothroidea the taxon Ameronothridae n. gen. (nr. Aquanothrus) colonized fresh water independently from Selenoribatidae and Fortuyniidae (mainly marine Ameronothroidea). Reproductive mode was associated neither with marine nor with freshwater life; rather, in both habitats sexual and parthenogenetic taxa occur. However, the reproductive mode was related to the stability of the habitat. Species that live underwater permanently tend to be parthenogenetic whereas taxa whose life cycle is often interrupted by flooding, such as marine oribatid mites, or by desiccation, e.g., freshwater-living Ameronothridae n. gen. (nr. Aquanothrus) (Ameronothroidea) species, are mainly sexual, indicating that continuous access to resources indeed favours parthenogenetic reproduction. Findings of our study therefore suggest that parthenogenetic reproduction is not selected for by disturbances but by unlimited access to resources.

Checklist of earthworms (Oligochaeta: Lumbricidae) from Germany.

A checklist of the German earthworm fauna (Oligochaeta: Lumbricidae) is presented, including published data, data from reports, diploma- and PhD- theses as well as unpublished data from museum collections, research institutions and private persons. Overall, 16,000 datasets were analyzed to produce the first German checklist of Lumbricidae. The checklist comprises 46 earthworm species from 15 genera and provides ecological information, zoogeographical distribution type and information on the species distribution in Germany. Only one species, Lumbricus badensis Michaelsen, 1907, is endemic to Germany, whereas 41% are peregrine. As there are 14 species occurring exclusively in the southern or eastern part of Germany, the species numbers in German regions increase from north to south.