Ultrastructure of Olkasia polycarbonata (Euglenozoa, Euglenida) demonstrates cytoskeletal innovations associated with the feeding and flagellar apparatuses.

Euglenids are flagellates with diverse modes of nutrition, including the photosynthetic Euglenophyceae, which acquired plastids via secondary endosymbiosis with green algae, and a diverse assemblage of predators of bacteria and other microeukaryotes. Most heterotrophic euglenids have never been cultivated, so their morphology remains poorly characterized and limited to only a few studies. "Ploeotids" are a paraphyletic group representing much of the diversity of heterotrophic euglenids and are characterized by their feeding apparatus and a rigid pellicle of 10-12 longitudinally arranged strips. Ploeotid-like euglenids gave rise to the Spirocuta, a large clade of heterotrophic and photosynthetic euglenids defined by a flexible pellicle of helically arranged strips. Using single-cell approaches, we report the first ultrastructural characterization of Olkasia polycarbonata, a ploeotid that is consistently positioned as the sister lineage to the Spirocuta in multigene phylogenetic analyses. O. polycarbonata shares several morphological characteristics with members of Spirocuta, such as prominent swellings on the paraxonemal rods and a robust feeding apparatus consisting of rods and vanes. These morphological traits are consistent with the phylogenetic position of O. polycarbonata and demonstrate an increase in cytoskeletal complexity that occurred prior to the key strip duplication event in the most recent common ancestor of Spirocuta.