Slowly evolving proteins support the monophyly of Craspedida (Choanoflagellatea) and a marine origin of choanoflagellates

Choanoflagellate species have been taxonomically divided upon the morphological and developmental basis of their extracellular coat (periplast). Species within the order Craspedida possess a purely organic periplast, whereas taxa of the order Acanthoecida have an additional silica based periplast termed the lorica. Whilst small-scale phylogenetic studies have recovered the two orders as monophyletic, recent phylogenomic analyses have rejected the monophyly of the craspedids. These analyses suggest that the freshwater craspedid Codosiga hollandica falls within the earliest branching choanoflagellate lineage; however, it has also been noted that phylogenomic studies result in inconsistent phylogenies, with unstable long-branched species being recovered as basal choanoflagellates. Presented here are phylogenetic analyses, based upon slowly evolving ribosomal proteins, which consistently recover both craspedid monophyly and a derived placement of C. hollandica. The addition of further, long-branched, ribosomal proteins to phylogenetic analyses are shown to generate longer terminal branches and a weakening of the support for Craspedida. The analyses highlight the requirement for an increase in data, in terms of both taxa and gene coverage, to successfully resolve the earliest craspedid branches. In addition, phylogenetically unstable species are identified that are recommended to be omitted from phylogenomic studies as they have the potential to disrupt recovered relationships.