Sea anemones (order Actiniaria) are among the most diverse and successful members of the anthozoan subclass Hexacorallia, occupying benthic marine habitats across all depths and latitudes. Actiniaria comprises approximately 1,200 species of solitary and skeleton-less polyps and lacks any anatomical synapomorphy. Although monophyly is anticipated based on higher-level molecular phylogenies of Cnidaria, to date, monophyly has not been explicitly tested and at least some hypotheses on the diversification of Hexacorallia have suggested that actiniarians are para- or poly-phyletic. Published phylogenies have demonstrated the inadequacy of existing morphological-based classifications within Actiniaria. Superfamilial groups and most families and genera that have been rigorously studied are not monophyletic, indicating conflict with the current hierarchical classification. We test the monophyly of Actiniaria using two nuclear and three mitochondrial genes with multiple analytical methods. These analyses are the first to include representatives of all three currently-recognized suborders within Actiniaria. We do not recover Actiniaria as a monophyletic clade: the deep-sea anemone Boloceroides daphneae, previously included within the infraorder Boloceroidaria, is resolved outside of Actiniaria in several of the analyses. We erect a new genus and family for B. daphneae, and rank this taxon incerti ordinis. Based on our comprehensive phylogeny, we propose a new formal higher-level classification for Actiniaria composed of only two suborders, Anenthemonae and Enthemonae. Suborder Anenthemonae includes actiniarians with a unique arrangement of mesenteries (members of Edwardsiidae and former suborder Endocoelantheae). Suborder Enthemonae includes actiniarians with the typical arrangement of mesenteries for actiniarians (members of former suborders Protantheae, Ptychodacteae, and Nynantheae and subgroups therein). We also erect subgroups within these two newly-erected suborders. Although some relationships among these newly-defined groups are still ambiguous, morphological and molecular results are consistent enough to proceed with a new higher-level classification and to discuss the putative functional and evolutionary significance of several morphological attributes within Actiniaria.

Snorkelers in mangrove forest waters inhabited by the upside-down jellyfish Cassiopea xamachana report discomfort due to a sensation known as stinging water, the cause of which is unknown. Using a combination of histology, microscopy, microfluidics, videography, molecular biology, and mass spectrometry-based proteomics, we describe C. xamachana stinging-cell structures that we term cassiosomes. These structures are released within C. xamachana mucus and are capable of killing prey. Cassiosomes consist of an outer epithelial layer mainly composed of nematocytes surrounding a core filled by endosymbiotic dinoflagellates hosted within amoebocytes and presumptive mesoglea. Furthermore, we report cassiosome structures in four additional jellyfish species in the same taxonomic group as C. xamachana (Class Scyphozoa; Order Rhizostomeae), categorized as either motile (ciliated) or nonmotile types. This inaugural study provides a qualitative assessment of the stinging contents of C. xamachana mucus and implicates mucus containing cassiosomes and free intact nematocytes as the cause of stinging water.

Cnidae are complex intracellular capsules made by all cnidarians. The most diverse of these capsules are nematocysts, which are made by all members of the phylum; spirocysts and ptychocysts are made only by members of some lineages, and they show less functional and structural diversity. In nematocysts, the apex has been shown to be either a hinged cap (operculum) or three flaps that flex outward during discharge. The operculum is known only from medusozoan nematocysts; flaps are known only from nematocysts of members of the hexacorallian order Actiniaria, although they have been inferred to be characteristic of Anthozoa, the group to which Actiniaria belongs. Using scanning and transmission electron microscopy, we discover a third apical morphology in nematocysts, an apical cap, which we find in all nonactiniarian anthozoans examined. This apical cap is identical structurally to the apical cap of spirocysts, and it resembles the apical structure of ptychocysts, whose apex is documented here for the first time. Additionally, a full survey of nematocysts from all body structures of two actiniarians demonstrates that a particular type of nematocyst, the microbasic p-mastigophore of the mesenterial filaments, does not have apical flaps. The observed variation does not correspond to conventional categorization of capsule morphology and raises questions about the function and structure of capsules across Cnidaria. Despite some ambiguity in optimization of ancestral states across cnidae, we determine that the apical cap is the plesiomorphic structure for anthozoan cnidae and that apical flaps are a synapomorphy of Actiniaria. At present, the operculum is interpreted as a synapomorphy for Medusozoa, but either it or an apical cap is the ancestral state for nematocysts.