Phylogenomics and Morphological Reconstruction of Arcellinida Testate Amoebae Highlight Diversity of Microbial Eukaryotes in the Neoproterozoic

Life was microbial for the majority of Earth's history, but as very few microbial lineages leave a fossil record, the Precambrian evolution of life remains shrouded in mystery. Shelled (testate) amoebae stand out as an exception with rich documented diversity in the Neoproterozoic as vase-shaped microfossils (VSMs). While there is general consensus that most of these can be attributed to the Arcellinida lineage in Amoebozoa, it is still unclear whether they can be used as key fossils for interpretation of early eukaryotic evolution. Here, we present a well-resolved phylogenomic reconstruction based on 250 genes, obtained using single-cell transcriptomic techniques from a representative selection of 19 Arcellinid testate amoeba taxa. The robust phylogenetic framework enables deeper interpretations of evolution in this lineage and demanded an updated classification of the group. Additionally, we performed reconstruction of ancestral morphologies, yielding hypothetical ancestors remarkably similar to existing Neoproterozoic VSMs. We demonstrate that major lineages of testate amoebae were already diversified before the Sturtian glaciation (720 mya), supporting the hypothesis that massive eukaryotic diversification took place in the early Neoproterozoic and congruent with the interpretation that VSM are arcellinid testate amoebae

Growth rate modulation enables coexistence in a competitive exclusion scenario between microbial eukaryotes

Coexistence usually are exceeding the explicable rate by competitive exclusion principle. Since the pioneer Gause, many studies have used protist microcosm systems to study competitive exclusion. We explored a two-species system with the testate-amoebae: (Arcella intermedia and Pyxidicula operculata), where competitive exclusion is expected to occur. We determined their growth curves individually and under competitive interaction. We used a state-space model to represent system dynamics and calculated posterior population sizes simulating competition dynamics. Contrarily to our expectation, Arcella and Pyxidicula showed similar growth rates (1.37 and 1.46 days–1 respectively) and only different carrying capacity (1,997 and 25,108 cells cm–2 respectively). The maximum number of cells of both species when growing in competition was much lower if compared to the monospecific cultures (in average, 73% and 80% less for Arcella and Pyxidicula respectively). However, our competition experiments always resulted in coexistence. According to the models, the drop in growth rates and stochasticity mainly explains our coexistence results. We propose that a context of ephemeral resources can explain these results. Additionally, we propose generating factors of stochasticity as intraspecific variation, small population effects, toxicity of waste products and influence of the bacterial community

Deconstructing Difflugia: The tangled evolution of lobose testate amoebae shells (Amoebozoa: Arcellinida) illustrates the importance of convergent evolution in protist phylogeny

Protists, the micro-eukaryotes that are neither plants, animals nor fungi build up the greatest part of eukaryotic diversity on Earth. Yet, their evolutionary histories and patterns are still mostly ignored, and their complexity overlooked. Protists are often assumed to keep stable morphologies for long periods of time (morphological stasis). In this work, we test this paradigm taking Arcellinida testate amoebae as a model. We build a taxon-rich phylogeny based on two mitochondrial (COI and NADH) and one nuclear (SSU) gene, and reconstruct morphological evolution among clades. In addition, we prove the existence of mitochondrial mRNA editing for the COI gene. The trees show a lack of conservatism of shell outlines within the main clades, as well as a widespread occurrence of morphological convergences between far-related taxa. Our results refute, therefore, a widespread morphological stasis, which may be an artefact resulting from low taxon coverage. As a corollary, we also revise the groups systematics, notably by emending the large and highly polyphyletic genus Difflugia. These results lead, amongst others, to the erection of a new infraorder Cylindrothecina, as well as two new genera Cylindrifflugia and Golemanskia