Notes: Lobose amoebae are abundant free-living protists and important pathogenic agents, yet their evolutionary history and position in the universal tree of life are poorly known. Molecular data for lobose amoebae are limited to a few species, and all phylogenetic studies published so far lacked representatives of many of their taxonomic groups. Here we analyse actin and small-subunit ribosomal RNA (SSU rRNA) gene sequences of a broad taxon sampling of naked, lobose amoebae. Our results support the existence of a monophyletic Amoebozoa clade, which comprises all lobose amoebae examined so far, as well as the amitochondriate pelobionts and entamoebids, and the slime molds. Both actin and SSU rRNA phylogenies distinguish two well-defined clades of amoebae, the “Gymnamoebia sensu stricto” and the Archamoebae (pelobionts+entamoebids), and one weakly supported and ill-resolved group comprising some naked, lobose amoebae and the Mycetozoa.

Notes: . Large miliolid foraminifers of the subfamily Soritinae bear symbiotic dinoflagellates morphologically similar to the species of the “Symbiodinium” complex, commonly found in corals and other marine invertebrates. Soritid foraminifers are abundant in coral reefs and it has been proposed that they share their symbionts with other dinoflagellate-bearing reef dwellers. In order to test this hypothesis, we have analysed partial large subunit ribosomal DNA sequences from dinoflagellates symbionts obtained from 28 foraminiferal specimens, and compared them to the corresponding sequences of Symbiodinium-like endosymbionts from various groups of invertebrates. Phylogenetic analysis of our data shows that all soritid symbionts belong to the “Symbiodinium” species complex, within which they form seven different molecular types (Fr1–Fr7). Only one of these types (Fr1) branches within a group of invertebrate symbionts, previously described as type C. The remaining six types form sister groups to coral symbionts previously designed as types B, C, and D. Our data indicate a high genetic diversity and specificity of Symbiodinium-like symbionts in soritids. Except for type C, we have found no evidence for the transmission of symbionts between foraminifers and other symbiont-bearing invertebrates from the same localities. However, exchanges must have occurred frequently between the different species of Soritinae, as suggested by the lack of host specificity and some biogeographical patterns observed in symbiont distribution. Our data suggest that members of the subfamily Soritinae acquired their symbionts at least three times during their history, each acquisition being followed by a rapid diversification and independent radiation of symbionts within the foraminiferal hosts.

Notes: Reticulomyxa filosa is a freshwater protist possessing fine granular, branching and anastomosing pseudopodia and therefore traditionally placed in the class Granuloreticulosea, order Athalamida, as a sister group to the order Foraminiferida. Recent studies have revealed remarkable similarities in pseudopodial motility and ultrastructure between R. filosa and foraminifera (e.g. Allogromia laticollaris), prompting us to conduct a molecular phylogenetic analysis of these seemingly disparate organisms. We sequenced the complete small-subunit of the ribosomal DNA of the cultured strain of R. filosa and compared it to the corresponding sequences of other protists including 12 species of foraminifera. We also sequenced and analyzed the actin coding genes from R. filosa and two species of foraminifera, Allogromia sp. and Ammonia sp. the analysis of both data sets clearly shows that R. filosa branches within the clade of foraminifera, suggesting that R. filosa is in fact a freshwater naked foraminiferan.

Notes: In his grand monography of Radiolaria, Ernst Haeckel originally placed Phaeodarea within the class Radiolaria, together with Acantharea and Polycystinea. Cytological and ultrastructural studies, however, questioned the monophyly of Radiolaria, suggesting the independent evolutionary origin of the three taxa. Some recent molecular analyses based on small subunit ribosomal RNA (SSU rRNA) sequences challenged this classification, supporting the sisterhood of Acantharea and Polycystinea. In order to further test the monophyly of all Haeckel's Radiolaria, and as no data on Phaeodarea were available, we sequenced the complete SSU rRNA gene of three Phaeodarea and three Polycystinea. Our analyses show that the monophyletic Phaeodarea clearly branch among Cercozoa, and confirm that Acantharea and Polycystinea share a common history. This result enhances the morphological variability within Cercozoa, a phylum already containing very heterogeneous groups of protists. Our study suggests that the ability to secrete SrSO4 and the organization of microtubules inside axopodia are better phylogenetic markers than the simple presence of a central capsule and axopodia.

Notes: Percolomonas cosmopolitus is a common free-living flagellate of uncertain phylogenetic position that was placed within the Heterolobosea on the basis of ultrastructure studies. To test the relationship between Percolomonas and Heterolobosea, we analysed the primary structure of the actin and small-subunit ribosomal RNA (SSU rRNA) genes of P. cosmopolitus as well as the predicted secondary structure of the SSU rRNA. Percolomonas shares common secondary structure patterns of the SSU rRNA with heterolobosean taxa, which, together with the results of actin gene analysis, confirms that it is closely related to Heterolobosea. Phylogenetic reconstructions based on the sequences of the SSU rRNA gene suggest Percolomonas belongs to the family Vahlkampliidae. The first Bayesian analysis of a large taxon sampling of heterolobosean SSU rRNA genes clarifies the phylogenetic relationships within this group.

Notes: . Xenophyophorea are giant deep-sea rhizopodial protists of enigmatic origins. Although species were described as Foraminifera or sponges in the early literature, the xenophyophoreans are currently classified either as a class of Rhizopoda or an independent phylum. To establish the phylogenetic position of Xenophyophorea, we analysed the small subunit (SSU) rRNA gene sequence of Syringammina corbicula Richardson, a newly described xenophyophorean species from the Cape Verde Plateau. The SSUrDNA analyses showed that S. corbicula is closely related to Rhizammina algaeformis, a tubular deep-sea foraminiferan. Both species branch within a group of monothalamous (single-chambered) Foraminifera, which include also such agglutinated genera as Toxisarcon, Rhabdammina, and Saccammina, and the organic-walled genera Gloiogullmia and Cylindrogullmia. Our results are congruent with observations of similar cytoplasmic organisation in Rhizammina and Syringammina. Thus, the Xenophyophorea appear to be a highly specialised group of deep-sea Foraminifera.

Notes: Over the past few years, the use of cultivation-independent techniques to detect eukaryotic diversity has proven to be a powerful approach. Based on small-subunit ribosomal RNA (SSU rRNA) gene analyses, these studies have revealed the existence of an unexpected variety of new phylotypes. Some of them do not seem to be related to any molecularly described lineage, and have been proposed to represent novel eukaryotic kingdoms. In order to critically review the evolutionary importance of this novel high-level eukaryotic diversity and to test the potential technical and analytical pitfalls and limitations of eukaryotic environmental DNA surveys (EES), we analysed 484 environmental SSU rRNA gene sequences, including 81 new sequences from sediments of the river Seymaz (Geneva, Switzerland). Based on a detailed screening of an exhaustive alignment of SSU rRNA gene sequences and the phylogenetic re-analysis of previously published sequences using Bayesian methods, our results suggest that the number of novel higher-level taxa revealed by previous EES was over-estimated. Three main sources of errors are responsible for this situation, namely (1) the presence of undetected chimeric sequences; (2) the misplacement of several fast evolving sequences; and (3) the incomplete sampling of described, but yet unsequenced eukaryotes. EES undoubtedly contribute to unravel many novel eukaryotic lineages, but there is no clear evidence for a spectacular increase of the diversity at a megaevolutionary level. After our re-analysis, we found only five candidate lineages of possible novel high-level eukaryotic taxa. To ascertain their taxonomic status, however, the organisms themselves have to be identified now.

Notes: Molecular sampling of the taxonomic diversity of the living world is nowadays a task of paramount importance. Heliozoa represents one of the major eukaryotic taxa, which remain significantly underrepresented in molecular databases. The term Heliozoa was coined to embrace organisms with a rounded body and stiff pseudopodia. Despite evidences from ultrastructural studies, which conclusively show the polyphyly of selected heliozoan groups, contemporary morphological systems retain Heliozoa as a monophyletic taxon. From the perspective of reconstructing the true phylogeny of Eukaryota, molecular approaches to analyse relationships within this large protist group are evidently necessary. Phylogenetic analysis of our data shows that the four heliozoan taxa branch either independently or within different eukaryotic phyla. The actinophryids (Actinosphaerium, Actinophrys) appear as a lineage of stramenopiles, while the desmothoracids (Clathrulina, Hedriocystis) branch within “core Cercozoa”. The position of both groups is strongly supported in all analyses and is congruent with ultrastructure-based taxonomic revisions. The centrohelids (Chlamydaster, Heterophrys, Pterocystis, and Raphidiophrys) do not seem to be related to any particular eukaryotic phylum, in agreement with a recent molecular study. The taxopodid Sticholonche was found to branch between Polycystinea and Acantharea, two classes of radiolarians. Results obtained in this study suggest that the heliozoan body form cannot be used as a diagnostic argument to unite Heliozoa. Instead, we discriminate between the three heliomorphic taxa of independent origin, Actinophryida, Desmothoracida and Sticholonche, and propose the novel higher rank taxon Centrohelida. The term Heliozoa should thus be used solely in historical context.

Notes: . Large miliolid foraminifers bear various types of algal endosymbionts including chlorophytes, dinoflagellates, rhodophytes, and diatoms. Symbiosis plays a key role in the adaptation of large foraminifera to survival and growth in oligotrophic seas. The identity and diversity of foraminiferal symbionts, however, remain largely unknown. In the present work we use ribosomal DNA (rDNA) sequences to identify chlorophyte endosymbionts in large miliolid foraminifera of the superfamily Soritacea. Partial 18S and complete Internal Transcribed Spacer (ITS) rDNA sequences were obtained from symbionts of eight species representing all genera of extant chlorophyte-bearing Soritacea. Phylogenetic analysis of the sequences confirms the previous fine structure-based identification of these endosymbionts as belonging to the genus Chlamydomonas. All foraminiferal symbionts form a monophyletic group closely related to Chlamydomonas noctigama. The group is composed of seven types identified in this study, including one previously morphologically described species, Chlamydomonas hedleyi. Each of these types can be considered as a separate species, based on the comparison of genetic differences observed between other established Chlamydomonas species. Several foraminiferal species share the same symbiont type, but only one species, Archaias angulatus, was found to bear more than one type.

Notes: The holdfast apparatus fills the posterior half of the larva; it is comprised of cytoplasmic granules called heterosomes and of about twenty collecting canals which converge at a septate chamber, the rosette. The latter opens to the exterior by a secretory appendage called the podite. My hypothesis is that, when the larva attaches to a substrate, the heterosomal contents empty into the collecting canals and are mixed in the rosette. With the aid of the podite, this secretion is applied to the gill substrate where it forms the so-called style of the future adult. The holdfast apparatus of the adult Spirochona comprises two regions of different nature: the pseudostyle, which is the posterior part of the cell, and the style which is an extracellular secretion. The style looks like a truncated cone and adheres firmly to the substrate. The upper face of the style shows 16–18 stylonemes which are inserted in the crypts of the pseudostylar rosette. The latter corresponds to the larval rosette, and the crypts communicate with a chamber and numerous suprastylar acini, which represent the remnants of the larval collecting canals. Except for a few details, the holdfast apparatus appears to be similar among all the chonotrichs studied to date. This apparatus differs from that of the Suctoria and of the Peritricha. It has some analogies with the “mouth-rosette” complex of the Apostomatida. Finally, the presence of a podite in a chonotrich larva is new evidence supporting a closer systematic relationship between the Chonotrichida and the Dysteriidae.〈section xml:id="abs1-2"〉〈title type="main"〉RESUMEL'appareil de fixation occupe la moitié postérieure de la larve; il est composé de granules cytoplasmiques, les hétérosomes, et d'une vingtaine de canaux collecteurs convergeant vers une chambre septée appelée rosette. Celle-la communique avec l'extérieur par un appendice sécréteur, le podite. II est supposé que, lors de la phase d'attachement, le matériel contenu dans les hétérosomes se vide dans les canaux collecteurs et se mélange dans la rosette, puis, grâce au podite, est appliqué sur la branchie où il forme le pied ou style du futur adulte. L'appareil de fixation de l'adulte comprend donc deux parties de nature différentes: le pseudostyle, c'est-à-dire la région postérieure du corps cellulaire, et le style, une sécrétion extracellulaire. Celui-ci forme une masse tronconique intimement fixée au substrat. Sur sa face supérieure, le style porte 16–18 stylonèmes enserrés dans les cryptes de la rosette pseudostylaire. Celle-ci correspond à la rosette larvaire, et les cryptes communiquent avec une chambre et des acini suprastylaires, eux-mêmes vestiges des canaux collecteurs larvaires. A quelques détails près, il semble bien que l'appareil de fixation des Chonotriches actuellement étudiés soit construit sur le même type. Cet appareil de fixation est différent de celui des Suctoria et des Peritricha; il présente peut-être quelques similitudes avec le complexe “bouche-rosette” des Apostomatida. Enfin, la présence d'un podite chez une larve de Chonotriche est un argument nouveau en faveur du rapprochement Chonotrichida–Dysteriidae.