Carbonaceous preservation of Cambrian hexactinellid sponge spicules

Early fossil sponges offer a direct window onto the evolutionary emergence of animals, but insights are limited by the paucity of characters preserved in the conventional fossil record. Here, a new preservational mode for sponge spicules is reported from the lower Cambrian Forteau Formation (Newfoundland, Canada), prompting a re-examination of proposed homologies and sponge inter-relationships. The spicules occur as wholly carbonaceous films, and are interpreted as the remains of robust organic spicule sheaths. Comparable sheaths are restricted among living taxa to calcarean sponges, although the symmetries of the fossil spicules are characteristic of hexactinellid sponges. A similar extinct character combination has been documented in the Burgess Shale fossil Eiffelia. Interpreting the shared characters as homologous implies complex patterns of spicule evolution, but an alternative interpretation as convergent autapomorphies is more parsimonious. In light of the mutually exclusive distributions of these same characters among the crown groups, this result suggests that sponges exhibited an early episode of disparity expansion followed by comparatively constrained evolution, a pattern shared with many other metazoans but obscured by the conventional fossil record of sponges.     

Giving the early fossil record of sponges a squeeze

Twenty candidate fossils with claim to be the oldest representative of the Phylum Porifera have been re‐analysed. Three criteria are used to assess each candidate: (i) the diagnostic criteria needed to categorize sponges in the fossil record; (ii) the presence, or absence, of such diagnostic features in the putative poriferan fossils; and (iii) the age constraints for the candidate fossils. All three criteria are critical to the correct interpretation of any fossil and its placement within an evolutionary context. Our analysis shows that no Precambrian fossil candidate yet satisfies all three of these criteria to be a reliable sponge fossil. The oldest widely accepted candidate, Mongolian silica hexacts from c. 545 million years ago (Ma), are here shown to be cruciform arsenopyrite crystals. The oldest reliable sponge remains are siliceous spicules from the basal Cambrian (Protohertzina anabarica Zone) Soltanieh Formation, Iran, which are described and analysed here in detail for the first time. Extensive archaeocyathan sponge reefs emerge and radiate as late as the middle of the Fortunian Stage of the Cambrian and demonstrate a gradual assembly of their skeletal structure through this time coincident with the evolution of other metazoan groups. Since the Porifera are basal in the Metazoa, their presence within the late Proterozoic has been widely anticipated. Molecular clock calibration for the earliest Porifera and Metazoa should now be based on the Iranian hexactinellid material dated to c. 535 Ma. The earliest convincing fossil sponge remains appeared at around the time of the Precambrian‐Cambrian boundary, associated with the great radiation events of that interval.     

Eine neue Poriferen-Fauna aus dem Septarienton (Oligozän, Rupelium) von Bad Freienwalde (NE-Deutschland) und der erste fossil erhaltene Vertreter der nicht-rigiden Hexactinelliden-GattungAsconema

The hexactinellid sponge fauna, which is described here for the first time, originates from the “Septarienton” (Rupelium, Oligocene) of the “Kirchenziegeleigrube” near Bad Freienwalde (NE Germany). This fauna includes lyssacinosan, as well as hexactinosan species, represented byAsconema oligocaena n. sp. (Hexasterophora, Lyssacinosa), the first occurrence of this genus from the fossil record,Aphrocallistes sp. (Hexasterophora, Hexactinosa), andHyalonema sp. (Amphidiscophora). The three-dimensional, pyritic body-preservation of these non-rigid sponges suggests fossiliyzation by rapid burial. Episodic mudflows are suggested to be responsible for this. The environment of deposition is suggested to have been a moderately shallow shelf, possibly in the distal range of a delta, characterised by relatively low sedimentation rates and no turbulence, except when disturbed by episodic mudflows. Low energy conditions are a precondition for the settlement of the described Hexactinellida.     

A new genus and species of deep-sea glass sponge (Porifera, Hexactinellida, Aulocalycidae) from the Indian Ocean

New hexactinellid sponges were collected from 2589 m depth on the Carlsberg Ridge in the Indian Ocean during deep-sea dredging. All fragments belong to a new genus and species, Indiellagen. n.ridgenensissp. n., a representative of the family Aulocalycidae described here. The peculiar features of this sponge, not described earlier for other Aulocalycidae, are: longitudinal strands present in several layers and epirhyses channelization.     

Bathyal sponges from the late Early Miocene of the Vienna Basin (central Paratethys,Slovakia)

Here we report, for the first time, a very rich and diversified sponge assemblage from late Early Miocene deposits of a central part of the Vienna Basin (Paratethys) in Slovakia. Bodily preserved sponges are described as a new genus and species Paracinachyrella fossilis (Tetiliidae, Demospongiae). Dissociated spicules reveal the presence of the “soft” demosponges that belong to families Tetillidae, Theneidae, Geodiidae, Samidae, Thrombidae, Thoosidae, Agelasidae, Myxillidae, Bubaridae, and Tedaniidae, the lithistid family Pleromidae, and an undetermined rhizoclone-bearing lithistid. Fragments of dictyonal skeleton indicate the presence of hexactinellid sponges that belong to the families Farreidae and Euretidae, and lychniscosan sponges. We estimate that at least 16–19 different species of siliceous sponges inhabited this region of the Central Paratethys during the latest Burdigalian. Most of these sponges are reported for the first time from the Miocene of the Paratethys. This sponge fauna has clear Tethyan affinities and indicates the existence of connection between Paratethys and Tethys during the latest Burdigalian, as well as the presence of open marine, deep-water, bathyal conditions in this part of the Vienna Basin.     

Sponge communities from the Lower Liassic of Adnet (Northern Calcareous Alps, Austria)

The lower slope of the drowned Alpine Adnet Reef was recolonized in Hettangian time by sponge communities of hexactinellid (hexactinosid and lyssacinosid) taxa and a few demosponges. Special taphonomic processes caused an excellent preservation of these sponges. The preservation allows to define several growth forms and to study original spicule configurations of the mainly non-rigid skeletons. Sponge faunas of presumably similar associations are known from adjacent basins, but only by isolated spicules of completely collapsed specimens. In Adnet the sponges are embedded in biodetrital limestones of the Schnöll Formation. Orientation and distribution of the sponges reflect autochthonous faunas that have been mixed with dislocated individuals by local water currents. The predominance of erect sponge types indicates intermediate sedimentation rates and/or occasional high-energy events. Sponge types and community structures are comparable with those ones from Middle Paleozoic mud mounds. Several hiatuses, mostly characterized by ferromanganese crusts have been kept free of sponge settlement. Carbon stable isotopes of the sponge-rich sequence show a small negative δ¹³C_carb excursion that covers the period from Lower Hettangian to Lower Sinemurian.This revised version was published online in May 2005 because several displayed passages had been inadvertently deleted in the original published version.     

‘Cambrian’ demosponges in the Ordovician of Morocco: Insights into the early evolutionary history of sponges

Cambrian spicular sponge faunas are dominated by a distinctive assemblage of demosponges and hexactinellids that are known from Burgess shale-type faunas worldwide. Most of these are previously unknown outside the Lower-Middle Cambrian (and perhaps Tremadoc) and have no obvious close relatives in later sequences. This paper describes examples of Choia sp., Pirania auraeum sp. nov. and Hamptonia christi sp. nov. from the Arenig of Morocco, associated with isolated hexactinellid spicules. A summary of the stratigraphic ranges of the major Cambrian sponge lineages is provided. These indicate an environmental contrast in the Lower Palaeozoic evolution of hexactinellids and non-lithistid demosponges, with demosponges probably undergoing cryptic diversification in nearshore environments during the Upper Ordovician.     

Discovery of a “Living Dinosaur”: Globally unique modern hexactinellid sponge reefs off British Columbia, Canada

Summary Globally unique hexactinellid sponge reefs occur on the continental shelf off British Columbia, Canada. They cover about 425 km² of seafloor on the continental shelf off British Columbia (Canada) in water depths between 165 and 240 metres and occur on a low-angle deep shelf, iceberg scoured seafloor, characterized by very low sedimentation rates and very stable environmental conditions. The sponge bioherms are up to 19 metres high with steep flanks, whereas the biostromes are 2–10 metres thick and many kilometres wide. They all consist of dense populations of only seven hexactinellid species. Three of them, all hexactinosan species (Aphrocallistes vastus, Heterochone calyx, Farrea occa) are the main frambuilders, composing a true rigid framework of sponge skeletons encased in a organic rich matrix of modern clay baffled by the sponges. Growth rates of hexactinosan sponges range in the order of 0–7 centimetres per year. The base of the oldest sponge reefs date from approximately 9000 years b.p. Different invertebrate and fish faunas occupy the reefs than occur on adjacent seafloor areas and some species appear to use the sponge reef complex structures as refugia where they can hide. Sidescan sonar data and direct observation by manned submersible clearly show that large areas of sponge reefs have been heavily damaged by seafloor trawling in the past decade. These unique extant siliceous sponge reefs can be used as a modern analogue for a better understanding and interpretation of fossil siliceous sponge reefs, known from many ages and many locations world wide.     

Early sponge evolution: A review and phylogenetic framework

Sponges are one of the critical groups in understanding the early evolution of animals. Traditional views of these relationships are currently being challenged by molecular data, but the debate has so far made little use of recent palaeontological advances that provide an independent perspective on deep sponge evolution. This review summarises the available information, particularly where the fossil record reveals extinct character combinations that directly impinge on our understanding of high-level relationships and evolutionary origins. An evolutionary outline is proposed that includes the major early fossil groups, combining the fossil record with molecular phylogenetics. The key points are as follows. (1) Crown-group sponge classes are difficult to recognise in the fossil record, with the exception of demosponges, the origins of which are now becoming clear. (2) Hexactine spicules were present in the stem lineages of Hexactinellida, Demospongiae, Silicea and probably also Calcarea and Porifera; this spicule type is not diagnostic of hexactinellids in the fossil record. (3) Reticulosans form the stem lineage of Silicea, and probably also Porifera. (4) At least some early-branching groups possessed biminerallic spicules of silica (with axial filament) combined with an outer layer of calcite secreted within an organic sheath. (5) Spicules are homologous within Silicea, but also between Silicea and Calcarea, and perhaps with Homoscleromorpha. (6) The last common ancestor of extant sponges was probably a thin-walled, hexactine-bearing sponge with biminerallic spicules. (7) The stem group of sponges included tetraradially-symmetric taxa that grade morphologically into Cambrian fossils described as ctenophores. (8) The protomonaxonid sponges are an early-branching group, probably derived from the poriferan stem lineage, and include the problematic chancelloriids as derived members of the piraniid lineage. (9) There are no definite records of Precambrian sponges: isolated hexactine-like spicules may instead be derived from radiolarians. Early sponges had mineralised skeletons and thus should have a good preservation potential: the lack of sponge fossils in Precambrian strata may be due to genuine absence of sponges. (10) In contrast to molecular clock and biomarker evidence, the fossil record indicates a basal Cambrian diversification of the main sponge lineages, and a clear relationship to ctenophore-like ancestors. Overall, the early sponge fossil record reveals a diverse suite of extinct and surprising character combinations that illustrate the origins of the major lineages; however, there are still unanswered questions that require further detailed studies of the morphology, mineralogy and structure of early sponges.     

Phylogenetic analyses of marine sponges within the order Verongida: a comparison of morphological and molecular data

Abstract. Because the taxonomy of marine sponges is based primarily on morphological characters that can display a high degree of phenotypic plasticity, current classifications may not always reflect evolutionary relationships. To assess phylogenetic relationships among sponges in the order Verongida, we examined 11 verongid species, representing six genera and four families. We compared the utility of morphological and molecular data in verongid sponge systematics by comparing a phylogeny constructed from a morphological character matrix with a phylogeny based on nuclear ribosomal DNA sequences. The morphological phylogeny was not well resolved below the ordinal level, likely hindered by the paucity of characters available for analysis, and the potential plasticity of these characters. The molecular phylogeny was well resolved and robust from the ordinal to the species level. We also examined the morphology of spongin fibers to assess their reliability in verongid sponge taxonomy. Fiber diameter and pith content were highly variable within and among species. Despite this variability, spongin fiber comparisons were useful at lower taxonomic levels (i.e., among congeneric species); however, these characters are potentially homoplasic at higher taxonomic levels (i.e., between families). Our molecular data provide good support for the current classification of verongid sponges, but suggest a re-examination and potential reclassification of the genera Aiolochroia and Pseudoceratina . The placements of these genera highlight two current issues in morphology-based sponge taxonomy: intermediate character states and undetermined character polarity.     

Molecular and functional analysis of the (6-4) photolyase from the hexactinellid Aphrocallistes vastus

The hexactinellid sponges (phylum Porifera) represent the phylogenetically oldest metazoans that evolved 570-750 million years ago. At this period exposure to ultraviolet (UV) light exceeded that of today and it may be assumed that this old taxon has developed a specific protection system against UV-caused DNA damage. A cDNA was isolated from the hexactinellid Aphrocallistes vastus which comprises high sequence similarity to genes encoding the protostomian and deuterostomian (6-4) photolyases. Subsequently functional studies were performed. It could be shown that the sponge gene, after transfection into mutated Escherichia coli, causes resistance of the bacteria against UV light. Recombinant sponge photolyase was prepared to demonstrate that this protein binds to DNA treated with UV light (causing the formation of thymine dimers). Finally, it is shown that the photolyase gene is strongly expressed in the upper part of the animals and not in their middle part or their base. It is concluded that sponges not only have an excision DNA repair system, as has been described earlier by us, but also a photolyase-based photo-reactivating system.     

Sponge paraphyly and the origin of Metazoa

In order to allow critical evaluation of the interrelationships between the three sponge classes, and to resolve the question of mono‐ or paraphyly of sponges (Porifera), we used the polymerase chain reaction (PCR) to amplify almost the entire nucleic acid sequence of the 18S rDNA from several hexactinellid, demosponge and calcareous sponge species. The amplification products were cloned, sequenced and then aligned with previously reported sequences from other sponges and nonsponge metazoans and variously distant outgroups, and trees were constructed using both neighbour‐joining and maximum parsimony methods. Our results suggest that sponges are paraphyletic, the Calcarea being more related to monophyletic Eumetazoa than to the siliceous sponges (Demospongiae, Hexactinellida). These results have important implications for our understanding of metazoan origins, because they suggest that the common ancestor of Metazoa was a sponge. They also have consequences for basal metazoan classification, implying that the phylum Porifera should be abandoned. Our results support the upgrading of the calcareous sponge class to the phylum level.     

Kieselschwämme aus dem unterjurassischen Misonekalk der Trento-Plattform (Südalpen): Taxonomie und phylogenetische Relevanz

The Lower Jurassic Misone Limestone of the Trento Platform (Southern Alps, Italy) contains a siliceous sponge fauna which is here described. Besides the well-known Moroccan sponge carbonates, these Lower Jurassic spongioliths from the Trento Platform are presently the second mass occurrence of siliceous sponges, which is known from the southern margin of the Tethys. They differ from each other in regard of the composition of the sponge fauna and the absence of microbial crusts in the spongioliths of the Trento Platform. There, hexactinosans and lithistid demosponges occur in equal proportions. Sphinctozoans are another very characteristic element. Because of the richness in both sphinctozoans and siliceous sponges, the Trento occurrences may be considered as a transitional fauna between the late Palaeozoic-Triassic sponge fauna dominated by sphinctozoans and the post-Liassic sponge fauna dominated by more modern groups of siliceous sponges. Two new siliceous sponge genera with their species are established:Misonia baldensis n. gen. n. sp. (Hexactinosa) andBenacia princeps n. gen. n. sp. (lithistid Demospongiae). The rarity of siliceous sponge dominated spongioliths in the Early Jurassic is due to the restricted occurrence of low energy, deeper shelf areas.     

Upper Triassic (Carnian) reef biota from the Sambosan Accretionary Complex, Kyushu, Japan

Calcified sponges, algae, and reef problematica are abundant yet poorly known from the Triassic of Japan. They are abundant in shallow-water carbonate, redeposited blocks of the Sambosan Accretionary Complex, Konosé Group, and southern Kyushu. Based on study of thin-sections from reef limestone exposed along the Kuma River, some important organisms and reef microfacies are described, which seem typical of Upper Triassic reef complexes. The most abundant reef organisms are hypercalcified sponges, including sphinctozoans, inozoans and chaetetids, followed by cyanophycean algae (including “Tubiphytes”-like organisms), and solenoporacean red algae. Loose sponge spicules in one thin-section also indicate the occurrence of rare hexactinellid sponges. Chambered demosponges described from the Konosé carbonate rocks include Solenolmia manon manon (Münster), Colospongia sp., Jablonskyia andrusovi (Jablonsky), several unidentified chambered sponges as well as the inozoid Permocorynella sp. 1 and Permocorynella sp. 2. Also present are chaetetid sponges and solenoporacean red algae belonging to Parachaetetes cassianus (Flügel) and Parachaetetes? sp. or Solenopora? sp. Especially abundant in thin-sections are cyanophyceans and “Tubiphytes”-like organisms. Among the organisms is Cladogirvanella Ott and Hedstroemia sp. The composition of the biota and presence of typical problematic organisms increases our knowledge of shallow-water Upper Triassic carbonate rocks in a remote setting in western Panthalassa. The composition of the biota indicates a mostly Carnian age. Most comparable organisms are known from both the northeastern and southern Tethys.     

Recruitment,Growth and Mortality of an Antarctic Hexactinellid Sponge,Anoxycalyx joubini

Polar ecosystems are sensitive to climate forcing, and we often lack baselines to evaluate changes. Here we report a nearly 50-year study in which a sudden shift in the population dynamics of an ecologically important, structure-forming hexactinellid sponge, Anoxycalyx joubini was observed. This is the largest Antarctic sponge, with individuals growing over two meters tall. In order to investigate life history characteristics of Antarctic marine invertebrates, artificial substrata were deployed at a number of sites in the southern portion of the Ross Sea between 1967 and 1975. Over a 22-year period, no growth or settlement was recorded for A. joubini on these substrata; however, in 2004 and 2010, A. joubini was observed to have settled and grown to large sizes on some but not all artificial substrata. This single settlement and growth event correlates with a region-wide shift in phytoplankton productivity driven by the calving of a massive iceberg. We also report almost complete mortality of large sponges followed over 40 years. Given our warming global climate, similar system-wide changes are expected in the future.     

Phylogenetic Position of the Hexactinellida Within the Phylum Porifera Based on the Amino Acid Sequence of the Protein Kinase C from Rhabdocalyptus dawsoni

Recent analyses of genes encoding proteins typical for multicellularity, especially adhesion molecules and receptors, favor the conclusion that all metazoan phyla, including the phylum Porifera (sponges), are of monophyletic origin. However, none of these data includes cDNA encoding a protein from the sponge class Hexactinellida. We have now isolated and characterized the cDNA encoding a protein kinase C, belonging to the C subfamily (cPKC), from the hexactinellid sponge Rhabdocalyptus dawsoni. The two conserved regions, the regulatory part with the pseudosubstrate site, the two zinc fingers, and the C2 domain, as well as the catalytic domain were used for phylogenetic analyses. Sequence alignment and construction of a phylogenetic tree from the catalytic domains revealed that the yeast Saccharomyces cerevisiae and the protozoan Trypanosoma brucei are at the base of the tree, while the hexactinellid R. dawsoni branches off first among the metazoan sequences; the other two classes of the Porifera, the Calcarea (the sequence from Sycon raphanus was used) and the Demospongiae (sequences from Geodia cydonium and Suberites domuncula were used), branch off later. The statistically robust tree also shows that the two cPKC sequences from the higher invertebrates Drosophila melanogaster and Lytechinus pictus are most closely related to the calcareous sponge. This finding was also confirmed by comparing the regulatory part of the kinase gene. We suggest, that (i) within the phylum Porifera, the class Hexactinellida diverged first from a common ancestor to the Calcarea and the Demospongiae, which both appeared later, and (ii) the higher invertebrates are more closely related to the calcareous sponges. Received: 6 August 1997 / Accepted: 24 October 1997     

Mechanisms of propagule release in the carnivorous sponge Asbestopluma occidentalis

Carnivorous sponges characteristically inhabit the deep sea, so extensive observations of the biology of living specimens are rare. We report on newly discovered shallow‐water (<30 m depth) populations of the carnivorous sponge Asbestopluma occidentalis and on observations of living adults and larvae from this unique group of sponges. In the Salish Sea, British Columbia, Canada, populations of A. occidentalis exist at depths as shallow as 18 m, where they co‐occur with hexactinellid sponges. Adults with and without embryos (n =127) were collected and easily maintained in the laboratory for several months, allowing continuous examination of live specimens. Parent sponges naturally disassociated their tissue, facilitating larval release and dispersal. Dispersed larvae had actively beating cilia, but no swimming was observed. Larvae settled and attached from several hours to several days post‐release. After larval release, parent sponges reaggregated their disassociated bodies into spherical balls of apparently undifferentiated tissue, which could also disperse and settle. Sexually mature adults were sampled in the field from August to November, with a high proportion of adults containing mature embryos in late November. High‐resolution photography and electron microscopy verified that adults were covered with anisochelae spicules, and used these to capture nauplii of Artemia sp. under experimental conditions; however, time‐lapse photography showed that some captured prey could free themselves with vigorous swimming. The occurrence of abundant shallow‐water populations of A. occidentalis in the Salish Sea provides a rare opportunity to study the evolution and ecology of carnivory in the Porifera.     

Sponge assemblages of the deep Weddell Sea: ecological and zoogeographic results of ANDEEP I–III and SYSTCO I expeditions

The aim of this study is to analyze the community structure, ecology and distribution of deep Weddell Sea sponge faunas. Analysis was performed on the basis of sponges sampled during ANDEEP I–III and SYSTCO I expeditions (2002–2008) by RV Polarstern. The material obtained comprises about 800 sponge specimens, representing 129 species, within these are 95 species of demosponges (including 15 new to science), 25 hexactinellid species (7 new) and 9 calcarean species (5 new). Sponges were sampled at 51 stations in depths between 500 and 5,500 m. At most stations, sponge densities were very low, and many species are represented by one or two specimens only. Community structure by Bray–Curtis similarity was analyzed as well as depth range and spatial distribution of the most common species. Zoogeographic affinities of sampled faunas are analyzed. Three associations of sponges are found in the deep Weddell Sea: (1) The Polymastia/Tentorium community, (including Rossella associations) distributed on the lower shelf and continental slope. (2) The Bathydorus community, distributed on the continental slope and upper abyssal. (3) The Caulophacus community, associated with Cladorhizidae, is characteristic for the abyssal plains. The associations follow each other successively both bathymetrically and geographically, from shallow to deep, from shelf and ridge structures into the open abyssal. A distinct faunistic boundary between shelf and deep sea is not present. In general, the sponge fauna of the deep Weddell Sea is regionally restricted and shows stronger affinities only to the sponge fauna of the subantarctic islands.     

Sponge cell culture: a comparative evaluation of adhesion to a native tissue extract and other culture substrates

Hexactinellids are deep water sponges that possess syncytial rather than cellular tissues. In order to investigate the syncytial character of the tissue of these unusual sponges, primary cultures were developed using a substrate of acellular tissue extract (ATE) that promotes the adhesion and spreading of sponge tissues. Primary cultures of the hexactinellid sponge Rhabdocalyptus dawsoni, grown on this substrate, form thinly spread, multinucleate, confluent tissue masses which exhibit active cytoplasmic streaming. Sponge tissue adhered equally well to commercial substrates of concanavalin A and poly-l-lysine, but did not adhere to chicken collagen. Acellular tissue extracts prepared from demosponges, which are known to be cellular, also promoted adhesion and spreading of cells from those sponges. Scanning electron microscopy showed adherent Rhabdocalyptus tissue to have an uninterrupted, smooth membrane covering the entire culture, unlike primary cultures of the cellular demosponge, Haliclona sp., which consisted of numerous individual cells. Tissue from freshly collected sponges adhered preferentially to ATE from a conspecific. However, after continued wounding, tissue adhered indescriminately to any substrate. The tissue extract congealed if added to sea water or 10 mM CaCl(2), forming a white, cloudy solid, which could be fixed and sectioned for transmission electron microscopy. Thin sections of the congealed extract showed it to contain membranes but no visible collagen fibrils.     

No speed dating please! Patterns of social preference in male and female house mice

Glass sponges (Class Hexactinellida) are important components of deep-sea ecosystems and are of interest from geological and materials science perspectives. The reconstruction of their phylogeny with molecular data has only recently begun and shows a better agreement with morphology-based systematics than is typical for other sponge groups, likely because of a greater number of informative morphological characters. However, inconsistencies remain that have far-reaching implications for hypotheses about the evolution of their major skeletal construction types (body plans). Furthermore, less than half of all described extant genera have been sampled for molecular systematics, and several taxa important for understanding skeletal evolution are still missing. Increased taxon sampling for molecular phylogenetics of this group is therefore urgently needed. However, due to their remote habitat and often poorly preserved museum material, sequencing all 126 currently recognized extant genera will be difficult to achieve. Utilizing morphological data to incorporate unsequenced taxa into an integrative systematics framework therefore holds great promise, but it is unclear which methodological approach best suits this task. Here, we increase the taxon sampling of four previously established molecular markers (18S, 28S, and 16S ribosomal DNA, as well as cytochrome oxidase subunit I) by 12 genera, for the first time including representatives of the order Aulocalycoida and the type genus of Dactylocalycidae, taxa that are key to understanding hexactinellid body plan evolution. Phylogenetic analyses suggest that Aulocalycoida is diphyletic and provide further support for the paraphyly of order Hexactinosida; hence these orders are abolished from the Linnean classification. We further assembled morphological character matrices to integrate so far unsequenced genera into phylogenetic analyses in maximum parsimony (MP), maximum likelihood (ML), Bayesian, and morphology-based binning frameworks. We find that of these four approaches, total-evidence analysis using MP gave the most plausible results concerning congruence with existing phylogenetic and taxonomic hypotheses, whereas the other methods, especially ML and binning, performed more poorly. We use our total-evidence phylogeny of all extant glass sponge genera for ancestral state reconstruction of morphological characters in MP and ML frameworks, gaining new insights into the evolution of major hexactinellid body plans and other characters such as different spicule types. Our study demonstrates how a comprehensive, albeit in some parts provisional, phylogeny of a larger taxon can be achieved with an integrative approach utilizing molecular and morphological data, and how this can be used as a basis for understanding phenotypic evolution. The datasets and associated trees presented here are intended as a resource and starting point for future work on glass sponge evolution.