sign in sign up
<<
Home , Acanthella (sponge), Agelasida, Amorphinopsis, Auletta (sponge), Axinellida, Axinellidae

Org Divers Evol (2012) 12:57–70 DOI 10.1007/s13127-011-0068-9

ORIGINAL ARTICLE

The phylogeny of halichondrid : past and present re-visited with DNA-barcoding data

Dirk Erpenbeck & Kathryn Hall & Belinda Alvarez & Gabriele Büttner & Katharina Sacher & Simone Schätzle & Astrid Schuster & Sergio Vargas & John N. A. Hooper & Gert Wörheide

Received: 16 June 2011 /Accepted: 28 November 2011 /Published online: 1 March 2012 # Gesellschaft für Biologische Systematik 2012

Abstract Halichondrid play a pivotal role in the Keywords Sponges . Porifera . Demospongiae . classification of demosponges as changes in their classification Halichondrida . Phylogeny. Barcoding . CO1 has had direct consequences for the classification of Demo- spongiae. Historically, the systematics of halichondrids has been unstable. During the 1950s, the order was divided into Introduction two subclasses, which were based on empirical and assumed reproductive data. Subsequent morphological and biochemical Sponges, particularly demosponges, which represent approxi- analyses postulated the re-merging of halichondrid families, mately 85% of all species, are among the most difficult but recent molecular data indicate their polyphyly. Here we Metazoa in terms of classification. Indeed, Lévi (1957) dubbed review the classification history of halichondrid taxa, compare the Porifera as the last major group of Metazoa in which the it with the current and predominantly ribosomal molecular orders were still not clearly defined—a situation that still data, and support the new phylogenetic hypotheses with mito- permeates Porifera today. The difficulties are due largely to chondrial data from DNA barcoding. the primitive bauplan of sponges from which few characters can be assessed rigorously; in the absence of robust character : : : : : assessments, robust phylogenetic reconstruction is impossible. D. Erpenbeck: G. Büttner K. Sacher S. Schätzle A. Schuster This situation is evident for all sponges, but is apparent S. Vargas G. Wörheide particularly in halichondrid demosponges. This group of Department of Earth and Environmental Sciences, Ludwig-Maximilians-Universität München, demosponges inhabits all world oceans, in almost all latitudes, Richard-Wagner-Str. 10, from the surface down to great depths. Halichondrid sponges 80333 Munich, Germany are named after the type , Fleming, 1828, which translates from ancient Greek as “cartilage of the sea”, D. Erpenbeck : K. Hall : J. N. A. Hooper Biodiversity Program, Queensland Museum, South Brisbane, in reference to the amorphous shape and cartilaginous consis- Qld 4101, Australia tency of many taxa (Hansson 1994). The “breadcrumb- sponge”, Pallas, 1766, is perhaps the * : D. Erpenbeck ( ) G. Wörheide best studied of all sponges—although also possibly still the GeoBio-CenterLMU Ludwig-Maximilians-Universität München, Richard-Wagner-Str. 10, most ambiguous in terms of decisive morphometric features. 80333 Munich, Germany Halichondrids, as for many other sponge groups, lack e-mail: [email protected] unambiguous synapomorphic characters to indicate their monophyly such as e.g. chelae in s.s., B. Alvarez Museum and Art Gallery of the Northern Territory, verticillate acanthostyles in , or incubated cincto- Darwin, NT 0820, Australia blastula larvae for Homosclerophorida. In the latest compre- hensive classification of sponges (Hooper and Van Soest G. Wörheide 2002), Order Halichondrida Gray, 1867 comprised 45 valid Bayerische Staatssammlung für Paläontologie und Geologie, Richard-Wagner-Str. 10, genera, arranged in five families: Gray 1867, 80333 Munich, Germany Carter 1875; Bubaridae Topsent 1894a; 58 D. Erpenbeck et al.

Dictyonellidae Van Soest, Diaz & Pomponi 1990 and Hetero- org, Wörheide and Erpenbeck 2007 for details). The DNA xyidae Dendy, 1905. Their definitions of the families are extraction was conducted following a plate based extraction generally based upon combinations of absent characters or method, developed for the barcoding project (Vargas et al. character sets that did not include autapomorphies, and con- 2010). Fragments of the mitochondrial cytochrome oxidase sequently the diagnoses of the families overlapped. For in- subunit 1 (standard barcoding fragment) were amplified using stance, according to Hooper and Van Soest (2002), the degenerated version of universal barcoding primers: Axinellidae, Bubaridae and all lack a special- dgLCO1490 (GGT CAA CAA ATC ATA AAG AYA TYG ized ectosomal skeleton. Discord and overlap in the familial G) and dgHCO2198 (TAA ACT TCA GGG TGA CCA AAR diagnoses of halichondrid groups are clear. Bubaridae are AAY CA) (Meyer et al. 2005) with an annealing temperature defined as being entirely encrusting, and bearing a skeleton of 43°C. The PCR products were purified with ExoSAP-IT that is differentiated into a basal layer of interlacing spicules (Affymetrix; http://www.affymetrix.com) or standard Amo- and a perpendicular layer of monactines with bases embedded nium acetate-Ethanol precipitation before cycle sequencing in the basal skeleton (Alvarez and Van Soest 2002). Axinelli- using the BigDye-Terminator Mix v3.1 (Applied Biosystems, dae bear ascending spiculo-fibers connected irregularly by Foster City, CA) following the manufacturer’s protocol. Both loose spicules, or are plumoreticulated with connected ascend- strands of the template were sequenced on an ABI 3730 ing plumose tracts (Alvarez and Hooper 2002). Dictyonellidae automated sequencer. The poriferan origin of the sequences have a dense organic ectosomal layer, giving the sponge a was checked by a BLAST search against the NCBI Genbank fleshy appearance and the choanosomal skeleton is built database (http://www.ncbi.nlm.nih.gov/). Sequences were predominantly of styles, though occasionally oxeas or basecalled, trimmed and assembled in CodonCode Aligner v strongyles are known to occur (Van Soest et al. 2002). 3.7.1.1 and subsequently aligned in Sea-View 4 (Galtier et al. Heteroxyids (Hooper 2002a, better known under the junior 1996) using Muscle 3.6 (Edgar 2004). Sequences are depos- synonym of Desmoxyidae) and halichondriids both bear an ited in GenBank under accession numbers JQ034526– ectosomal skeleton (except in Lendenfeld, 1897 and JQ034587, and in the Sponge Barcoding Database (http:// Laminospongia Pulitzer-Finali, 1987). In the heteroxyids, the www.spongebarcoding.org). For phylogenetic analyses, addi- ectosomal skeleton consists of microxeote microscleres or tional samples representing all major taxa of the modified forms, while in halichondrids the ectosomal skeleton "G4" clade (the major subgroup of all primarily spiculate consists of a densely confused tangentially arranged crust of demosponges (excluding marine Haplosclerida), Borchiellini styles and/or oxeote megascleres. The choanosomal skeleton in et al. 2004) were included in the alignment. both families is a confused arrangement of spicules. In hetero- Bayesian analyses of nucleotide sequences were run with the xyids, a widely spaced reticulation of bundles occasionally parallel version of MrBayes 3.1.2 (Ronquist and Huelsenbeck occurs, whereas in halichondrids, only vague tracts of spicules 2003) on a Linux cluster with one processor assigned to each may lead to the surface (Erpenbeck and Van Soest 2002). Markov chain under the most generalising model (GTR+G+I) In the following, firstly we summarise, review and docu- because overparametrisation has been shown to not negatively ment the pivotal position of halichondrid demosponges within affect Bayesian analyses (Huelsenbeck and Ranala 2004). Each the greater demosponge classification and phylogeny. Numer- Bayesian analysis comprised at least two simultaneous ous sponge classifications have been erected in the past, and runs of four Metropolis-coupled Markov-chains at the only a few selected schemas will be highlighted. Revisions of default temperature (0.2). Analyses were terminated after the halichondrid classifications up to the end of the nineteenth chains converged significantly, as indicated by the average century have been deduced mostly from the historical system- standard deviation of split frequencies<0.01. Maximum like- atic of sponges outlined in Vosmaer (1887), which provides a lihood reconstructions were performed using RAxML 7.2.5 very detailed recapitulation of the history of sponge science up (Stamatakis 2006) under the GTR model of nucleotide substi- to that time. Subsequently, we compare the past and current tution with CAT approximation of rate heterogeneity and fast morphologically based classifications with analyses from the bootstrap (1,000 bootstrap replicates). reconstruction of a mitochondrial gene tree from current DNA barcoding data and compare this tree to previously published nuclear ribosomal gene trees. Results and discussion

Historic (pre-molecular) overview of halichondrids Material and methods within the Demospongiae

Molecular data were obtained during the barcoding of the Aristotle was probably the first to describe sponges scien- Queensland Museum sponge collection in the course of the tifically [as Spongia and “Aplysina” (0 Sarcotragus cf. Sponge Barcoding Project (see http://www.spongebarcoding. Schmidt 1862)] and among the first to assume that they The phylogeny of halichondrid demosponges 59 are (cf. Johnston 1842). Since Aristotle, the system- disappeared in his system, which consisted of: (1) Hexactinel- atic placement of sponges has undergone somewhat of an lidae, (2) Calcispongiae, (3) Ceraospongiae, (4) Halisarcinae & odyssey through the classification of organisms. Belon Gumminaceae, (5) Tetractinellidae and (6) Monactinellidae. (1553), for example, placed sponges as a new kingdom be- The last order (“monactin”0spicules with only one ray and tween plants and animals. Linnaeus (1759) put them in the one axis) comprised the current halichondrid taxa among Algae (Cryptogamae) and later returned them to Animalia others: Linnaeus (1767). Donati (1750) discovered the presence (1) Renierinae (with reticulate skeleton, with oxeas and of spicules as skeletal elements in sponges and used them in strongyles), including Halichondria (Eumastia), his first classification. Then followed the pioneering work of de Schmidt 1870 and a mixture of Haplosclerida Monet JBPA (1813), who placed the marine sponges into his and Poecilosclerida) “Polypiers polymorphs” (see Vosmaer 1887). Nardo separated (2) Chalinopsidinae (with styles, but no chelae and sigma): sponges into three orders: (1) a group comprising horny , , Acanthella and Dictyonellidae, in sponges, (2) sponges with siliceous spicules, and (3) calcareous addition to Raspailia Nardo 1833 and Clathria sponges. He expanded this system in 1833, independently (cf. Schmidt 1862 (now Poecilosclerida) and others Vosmaer 1887) from Hogg (1842) to include two more orders (3) Desmacidinae (with chelae, sigma and toxa): mostly comprising siliceous-horny and calcareous-horny sponges. Poecilosclerida and Scopalina Grant (1836) published, also independently from Nardo (cf. (4) Chalinae (with anisotropic skeleton, with oxeas and Vosmaer 1882; 1886), a similar three-order system in which strongyles) sponges were called “Porifera” for the first time. (5) Suberitinae (with tylote spicules). Grant’s system was refined by Bowerbank (1864, 1866, 1874), who named the three orders “Silicea” (for the sili- Gray (1867) published a system of “Poriphora calcarea” and ceous sponges), “Calcarea” (for the calcareous sponges) and “Poriphora silicea” independently and obviously ignored the “” (for the "horny" sponges). Halichondrid genera work of Schmidt (cf. Vosmaer 1887). He divided the siliceous were distributed widely throughout Bowerbank’ssystem.Hal- sponges by assumed reproductive features (“ovisacs”), and ichondria, Bowerbank, 1858, and therefore was one of the first to use this type of character in Bowerbank, 1862 have been placed variously in three (out of sponge science. Gray founded the family “Halichondriadae” as seven) separate groups of the order “Silicea”, which they a mixture of Halichondriidae (Halichondria, Ciocalypta), and shared with members of several other currently valid orders. many Haplosclerida, Hadromerida and Poecilosclerida. His Phakellia Bowerbank, 1862 and Halicnemia Bowerbank 1864 families “Chalinidae” (including Axinella and Acanthella) were placed with Ciocalypta. Already halichondrids were off and the newly erected “Phakelliadae” (monotypic for Phakel- to an early, shaky start. lia) were placed closer to keratose sponges than to the Hal- Schmidt (1866) criticized Bowerbank's system sharply for ichondriadae. Gray’s system was criticised strongly by not being genealogical and therefore unnatural. In an earlier Bowerbank (1874), Schmidt (1868, who did not use (1862) monograph on Adriatic sponges, Schmidt had divided the taxon Halichondriadae in his work), Haeckel (1872)and all sponges into six groups, which can be defined roughly as: Vo sm ae r (1882–1886). Subsequently, Gray (1872)published a modified system, which united “Halichondriidae”, “Chali- (1) Calcispongiae0calcareous sponges; nidae”,and“Phakellidae” with six other families in the taxon (2) Ceraospongiae0horny-fibrous sponges “Keratospongia” (Vosmaer 1887). (3) Gumminaeae0globular siliceous sponges with cortex The system of Carter (e.g. 1875) comprised eight orders. (4) Halisarcinae0sponges without skeleton He erected the taxon “” as a family inside his order (5) Corticatae0true siliceous sponges “Echinonemata” (with mostly styles and tylostyles in fibers), (6) Halichondriae0Corticatae "with negative characters" as sister group to "Ectyonida", which comprise actual raspailid Schmidt admitted (a century before Hennigian philosophy) taxa (among others). The Halichondrina were distant in his that his Halichondriae were based on negative characters and Family Renierida of the order Holoraphidota (with various therefore indefensible. He placed his (Adriatic) samples of spicules, minimum spongin) besides other hadromerid fami- Axinella Schmidt 1862, Acanthella Schmidt 1862, Halichon- lies such as . dria (as Reniera Schmidt 1862)andScopalina Schmidt 1862 in The distinction between siliceous and calcareous sponges the Halichondriae together with different species currently in was more important for Vosmaer (1886). He based his system Poecilosclerida, Hadromerida, and Haplosclerida. In Schmidt on a dichotomous separation of sponges in “Porifera calcarea” 1869, he published one of the first phylogenetic trees of and the “Porifera non-calcarea”, which were organised as sponges based on a 12-order system (see also Schmidt 1870), follows: whichwaslaterreducedto6inorder to equalise their ranks (Schmidt 1880, emended by Zittel). The taxon Halichondriae (1) Hyalospongiae (0 Hexactinellida), 60 D. Erpenbeck et al.

(2) Spiculispongiae (no spongin to cement the spicules, Von Lendenfeld (1887) adopted Sollas’ system of oppos- spicules loose). Suborders: ing Calcarea and non-calcarea (“Silicea”), but, in contrast to Vosmaer, he regarded microsclere shape as a more valuable (A) Lithistina character. His classification (congruent with Ridley and Dendy (B) Tetractina (Plakinidae etc.); 1886, 1887) divides the “Subclassis Silicea” into the following (C) Oligosilicina (cf. Chondrosida, Halisarcidae, etc.) orders: (D) Pseudotetraxonina (cf. Hadromerida). (3) Cornacuspongiae (with spongin to cement the spicules) (1) Hexactinellida and the suborders: (2) Chondrospongiae (no spongin to cement the spicules; (A) Halichondrina (basically every Cornacuspongiae asteroid microsleres) except keratose sponges) (3) Cornacuspongiae (spongin cements the spicules; menis- coid microscleres) with the suborder Halichondrina (a) Halichondridae (with Halichondria, Axinella, including: Acanthella, Dictyonella Schmidt 1868,Phakellia, Auletta, Eumastia Schmidt 1870 and several (A) Spongillidae (freshwater sponges) Haplosclerida) (B) Homorrhaphidae: including Family Renieridae (b) Ectyonidae (with Carter, 1883, Schmidt 1870 (with Halichondria,andmostly Carter, 1883, Leucophloeus Carter, haplosclerids) 1883, Ciocalypta, Hymeraphia Bowerbank 1864, (C) Heterorrhaphidae Higgin, 1877) (D) Desmacionidae (cf. Ridley and Dendy 1887) (with (c) Desmacidonidae ) (d) Spongillidae (E) Axinellidae: (with Dendropsis (0 Higginsia) Ridley (B) Ceratina (keratose sponges). and Dendy, 1886, Hymeniacidon, Phakellia, Cioca- lypta, Acanthella, Axinella, Hallmann, With Vosmaer’s “Spiculispongiae” and “Cornacuspon- 1917 and Raspailia (now Poecilosclerida)). giae” began a long tradition of dividing non-hexactinellid siliceous sponges (0 demosponges) among two clades. With Vosmaer, Sollas and Lendenfeld a “certain period of These two clades were based on different definitions by stability” (Lévi 1957) was reached in sponge systematics. Lendenfeld, Sollas, Hentschel, Dendy and Lévi until the Calcarea were distinguished from non-Calcarea, and Hexacti- recent past (see below), but their contents were more-or- nellida obtained a specific position among siliceous sponges. less comparable. Further classificational work was done around that time by Sollas (1885) introduced the name “Demospongiae” into Schulze, Ridley, Dendy, Carter, Thiele, Lundbeck, Kirkpatrick, sponge systematics. He gave calcareous sponges class-level Wilson, Minchin and Hallmann. Their studies lead to multiple status and opposed them to non-Calcarea (“Plethospongiae”). changes of taxon names but were without major impact on the Those “Plethospongiae” were divided into the orders: system. Lévi (1957) remarked that the only “original contribu- tion” at that time was Dendy's (1922)classificationofthe (1) Hexactinellida (glass sponges) siliceous demosponges, by giving microscleres a higher prior- (2) Myxospongiae (all skeleton-lacking sponges) ity in the classification than megascleres. Dendy’s system of (3) Demospongiae siliceous sponges is as follows: (A) Monaxonidae (spicules with one axis) (1) Triaxonida (Hexactinellidae) (B) Tetractinellidae (spicules with four axes) (2) Myxospongida (no skeleton) (C) Cerospongiae (keratose sponges, assumed to be (3) Euceratosa (horny sponges) polyphyletic by Sollas). (4) Tetraxonida, which were divided into the following groups Ridley and Dendy (1886) divided the Monaxonidae in based on Hentschel's (1909, 1911) suggestions: “Clavulina” (with radiate skeleton and/or cortex) and “Hal- (A) Astrotetraxonida (with asterose microscleres, if ichondrina” (with reticulated skeleton and without cortex). not secondarily lost) Halichondrina were divided into: (B) Sigmatotetraxonida (with sigmatose microscleres, if not secondarily lost) including families: (1) Homorrhaphidae (with Halichondria) cylindrical spicules (2) Heterorrhaphidae (a) Lithistidae (3) Desmacidonidae (with Agelas Duchassaing & Michelotti, (b) Tetillidae 1864) (c) Haploscleridae with Halichondria, and most actual (4) Axinellidae (with Hymeniacidon, Axinella, Phakellia). Haplosclerida The phylogeny of halichondrid demosponges 61

(d) Desmacioidae with Bubaris Gray, 1867,Axi- the halichondrid taxa and re-redefined Hentschel’s Bubaridae, nella, Phakellia, Auletta, Hymeniacidon, Leuco- including the four valid species: phloeus, Higginsia, Spongosorites, Halicnemia (1) Tetractinellida (four-rayed spicules: orders Lithistida and most actual Poecilosclerida s.s. and Choristida with Homosclerophora) (e) Clavulidae with Didiscus Dendy, 1922 and many (2) Monaxonellida (one-rayed spicules): actual Hadromerida (A) Hadromerina: family Spongosoritidae with Hallmann (1917) erected the taxon “Desmoxyinae” in- Spongosorites cluding its actual taxa Halicnemia, Higginsia (Higginsia) (B) Halichondrina, with the families: and Higginsia (Desmoxya) (Hallmann 1917)(asDesmoxya). Comprehensive classifications were proposed by Hentschel (a) Axinellidae with most modern Halichondriidae: (1923), Topsent (1928) and de Laubenfels (1936). Hentschel Halichondria, Hymeniacidon, Ciocalypta, Axi- (1923) maintained Vosmaer's and Von Lendenfeld's groups, nella, Ptilocaulis, Phakellia, Dragmacidon, but placed Dendroceratida as separate order equivalent to Microxistyla Topsent, 1928, , Higgin- Calcarea, Hexactinellidae, and the demosponge orders. Addi- sia, Ptilocaulis, Dragmacidon, Auletta) tionally, he modified the taxon “Bubaridae”,whichwas (b) Astraxinellidae (Halicnemia,andsome founded by Topsent, (1894b, containing the currently valid Hadromerida) genera Bubaris, Cerbaris Topsent, 1898 and Hymerhabdia (c) Heteroxyidae with Heteroxya Topsent, 1898 Topsent, 1892), by adding (poecilosclerid) taxa. Hentschel’s (d) Bubaridae (Bubaris, Cerbaris, Monocrepidium, system was structured as follows: Hymerhabdia) (C) Poecilosclerina with Didiscus (1) Calcarea (D) Haplosclerina (2) Hexactinellidae (3) Ceratellida (horny sponges, no spicules, orders: Dic- (3) Tetraxonida with the suborders: tyoceratina, Dendroceratina). (A) Homosclerophora De Laubenfels (1936) modified Topsent’s system and (B) Sigmatophora published a monograph that aimed to unite all known (C) Astrophora sponge genera known at that time. He placed Topsent’s (D) Desmophora Astraxinellidae over various taxa and put Halichondria (E) Astromonaxonellina (most hadromerids) and Hymeniacidon in different families. The resulting fam- (4) Cornacuspongida with the suborders: ily Hymeniacidonidae was based on its fleshy appearance. (A) Protorhabdina (microscleres and monactine De Laubenfels' system was structured as follows: megascleres) (1) Keratosa (B) Poikilorhabdina (various megascleres, often (2) Haplosclerina (with Desmacionidae: Liosina) microscleres) including: (3) Poecilosclerina: with Agelasidae and Phorbasinae (a) Coelosphaeridae with Topsentia Berg, 1899, (Didiscus, Myrmekioderma Ehlers, 1870, Heteroxya) (b) Crellidae with Didiscus (4) Halichondrina (ectosomal skeleton, no specially local- (c) Bubaridae with Bubaris, Hymerhabdia, Cerbaris, ized megascleres) with the families Monocrepidium Topsent, 1898, but also actual (A) Axinellidae (with hispid ectosome and axial core), Poecilosclerida. which were separated in: (C) Phthinorhabdina: (simple megascleres, barely microscleres) (a) Axinellinae: (no microscleres) Ptilocaulis, Auletta, Axinella, Dragmacidon, Acanthella, (a) Axinellidae (no ectosomal skeleton), with Auletta, Pararhaphoxya Burton, 1934, Phakellia, Pha- Phakellia, Acanthella kettia de Laubenfels, 1936, Phycopis, Spongo- (b) Ciocalyptidae (with ectosomal skeleton) with sorites, Hallmann, 1914, Bubaris Halichondria, Eumastia, Axinyssa, Hymeniacidon, (b) Higginsiinae: (with spined micoscleres) Des- Ciocalypta, Liosina Thiele, 1899, Spongosorites moxya, Higginsia, Negombo Topsent, 1896 and actual Haplosclerida, (B) Halichondriidae (simple spiculation, smooth spi- (D) Aporhabdina (cf. ) cules, ectosomal skeleton) Ciocalypta, Ciocalapata (5) Dendroceratida (with spongin spicules). de Laubenfels, 1936, Leucophloeus, Halichondria Topsent (1928) divided the demosponges into three sub- (C) Semisuberitidae as a Poecilosclerida/hadromerid classes based on megasclere morphology. He united most of assemblage 62 D. Erpenbeck et al.

(D) Hymeniacidonidae: (simple spiculation, smooth spi- order Axinellida, with nine (assumed oviparous) families, and cules, fleshy appearance) Hymeniacidon, Acanthella, combined Axinellida, Hadromerida, and Epipolasida in the Dictyonella superorder Clavaxinellida - presuming their close relationship (E) Monanthidae (Lévi 1956; 1955; 1957). The Homosclerophorida subse- (5) Hadromerina quently received subclass status, equal to Ceractinomorpha (6) Epipolasida (like Hadromerina, but with oxeas and styles and Tetractinomorpha (Lévi 1973), together with the re- instead: Negombata de Laubenfels, 1936, Parahigginsia discovered (Hartman 1969)Sclerospongiae. Dendy, 1924, Topsentia, Epipolasis de Laubenfels, Lévi’s system of the demosponges (1973) was structured 1936,Axinyssa) as follows: (7) Choristida (8) Carnosa (1) Homosclerophorida (viviparous) (2) Tetractinomorpha (oviparous) ’ Lévi (1951) compared Topsent s findings with his own (A) Astrophorida ontogenetic observations. The orders Halisarca, Poecilosclerida (B) Spirophorida and Haplosclerida were found to be viviparous, and (C) Desmophorida (and other lithistid sponges) Hadromerida, Homosclerophoridae (Lévi 1953a) to be ovipa- (D) Hadromerida “ ” rous. The order Halichondrida was artificiel et hétérogène (E) Axinellida (choanosomal skeleton axial condensed (Lévi 1951) and the viviparous groups (such as Halichondria and indiscriminate presence of styles and oxea) and Hymeniacidon) were recommended to be separated from the oviparous (such as Axinella). (a) Axinellidae: Axinella, Phakellia, Auletta, Ptilo- Lévi (1957) stated that three classes Hexactinellida, Cal- caulis, Acanthella, Dragmacidon carea and Demospongiae in their entities were well defined, (b) Bubaridae: Bubaris, Monocrepidium but their reciprocal relationships were completely unknown. (c) Desmoxyidae: Higginsia, Parahigginsia, He remarked on the congruence of most previous authors in Halicnema dividing the demosponges into two groups, but criticized the (d) Trachycladidae poor internal division, especially of the Cornacuspongiae/ (e) Rhabderemiidae Sigmaxinellidae group (cf. Halichondrida, Poecilosclerida, (f ) Sigmaxinellidae Haplosclerida and keratose sponges). He found sponges with- (g) Raspailiidae out skeletons impossible to classify, the position of the Plakini- (h) Euryponidae dae (now separated as Homosclerophorida) ambiguous, and the (i ) Hemiasterellidae Halichondrina (sensu Topsent), especially the family Axinelli- (3) Ceractinomorpha (viviparous) dae, a pitfall in the sponge systematics. Lévi regarded Axinelli- (A) Dictyoceratida dae and the skeleton-lacking Myxospongida [consisting of (B) Dendroceratida Halisarca Johnston, 1842 (now order Halisarcida) and Oscar- (C) Haplosclerida ella Vosmaer, 1884 (now class Homoscleromorpha)] as inter- (D) Poecilosclerida with families Agelasidae (Agelas) mediate morphological forms connecting the two demosponge and Latrunculidae (Didiscus) — — — groups (Dendroceratida Dictyoceratida Haplosclerida (E) Halichondrida (defined by the ectosomal tangen- — — Poecilosclerida) and (Homosclerophorida Tetractinellida tial skeleton and confused choanosomal skeleton) Hadromerida—Epipolasida). Based on his ontogenetic data, which was subsequently (a) Halichondriidae: (primarily oxeas): Halichondria, extrapolated for the entire class, Lévi divided demosponges Trachyopsis Dendy, 1905 (0 Topsentia), Cioca- into two groups (Lévi 1953b). These groups were defined lypta, Carter, 1887 primarily by reproductive features, based on his own observa- (b) Hymeniacidonidae: (primarily styles): Hymenia- tions. Halichondrida and Myxospongida were split. The first cidon, Leucophloeus group comprised the viviparous “Ceractinomorpha” with their (4) Sclerosponges [raised to class status (Hartman and solid, incubated parenchymella larvae, consisting of the Den- Goreau 1970)] droceratida, Dictyoceratida, Haplosclerida, Poecilosclerida, and Halichondrida s.s. (without Axinellidae), with Halisarca Lévi’s system was extended by Bergquist (1978)and (now Halisarcida) as the most primitive taxon. Opposed to the Hartman (1982), who explored additional supporting (mostly Ceractinomorpha were the oviparous Tetractinomorpha with biochemical, reproductive and ultra-histological) characters. Oscarella (viviparous, now Homosclerophorida) as the most Bergquist, however, expressed doubts about the monophyly of primitive taxon, and Hadromerida, Tetractinellidae, and Axi- Tetractinomorpha (1978). Nevertheless, she placed Bubaridae nellidae separated from Halichondrida. Lévi resurrected the within Axinellidae. The phylogeny of halichondrid demosponges 63

Hartman (1982) elevated Agelasida in his demosponge formed a putative sister-group relationship to Heteroxyidae classification inside the Tetractinomorpha to ordinal level: (as Desmoxyidae) based on the possession of a fleshy ecto- some (Van Soest et al. 1990) (see also Carballo et al. 1996 for 1) Homoscleromorpha: Order Homosclerophorida subsequently suggested emendations for Dictyonellidae). The 2) Tetractinomorpha: Orders Astrophorida (Choristida), combination of non-localized interchangeable monaxons Spirophorida, Lithistida, Hadromerida, Axinellida, and (styles and oxeas), sinuous strongyles, plumoreticulate archi- Agelasida tecture, and collagenous mesohyl united the three families 3) Ceractinomorpha: Orders Dendroceratida, Dictyoceratida, with Axinellidae into the order Halichondrida. Previously, a Verongida, Haplosclerida, Petrosida, Poecilosclerida, series of halichondrid genera with intermediate forms between Halichondrida an axinellid plumo-reticulated skeleton and a halichondrid Several morphological (Vacelet 1985; Van Soest 1984; confused skeleton has been shown (Diaz 1997). The segrega- Reitner 1992), molecular (Chombard et al. 1997) and che- tion of families based on the predominance of either styles or motaxonomic data-sets (see review in Wörheide 1998) have oxeas (e.g. Halichondriidae against Hymeniacidonidae) was demonstrated unequivocally the polyphyly of Sclerospongia no longer upheld. Additionally, the placement of the hetero- and that its sole family, Astroscleridae, is in fact closely xyid genera Myrmekioderma and Didiscus back in the hal- related to Agelasidae. ichondrids was proposed. This classification comprised the Van Soest (1984) and Hooper (1984) pointed out incon- following halichondrid genera: sistencies in the current classifications in their independent Halichondriidae: (high density of spicules in confusion analyses on Poecilosclerida and Axinellida, respectively. They and in ill-defined tracts) Halichondria, Hymeniacidon, found the Levi-Bergquist-Hartman classification with its dis- Amorphinopsis, Myrmekioderma, Didiscus, Epipolasis, tinction between Ceractinomorpha and Tetractinomorpha Spongosorites, Axinyssa Lendenfeld, 1897; unparsimonious and subsequently suggested the re-merging Axinellidae: (choanosomal skeleton axially condensed, of Axinellida with Halichondrida. Van Soest, after introducing extra-axially plumoreticulated): Axinella, Phakellia, cladistic character analyses in sponge systematics (1985), Auletta, Bubaris, Monocrepidium, Hymerhabdia, Cerbaris; found the taxon Tetractinomorpha to be based only on the Dictyonellidae: (spongin reinforced tracts, fleshy- absence of primitive reproductive features (1987). Further- conulose surface) Dictyonella, Scopalina, Acanthella, more, the placement of Axinellida in Tetractinomorpha was Dactylella Thiele, 1898, Tethyspira Topsent, 1890, based solely on the possession of hemiasterellid-type asters Liosina; and plesiomorphic oviparity, and therefore problematic. Such a Desmoxyidae: (0Heteroxyidae; reticulate-fasciculate classification, if natural, would mean that the plumoreticulate choanosomal skeleton of spongin-enveloped tracts with architecture must have evolved twice. The placement of hal- a fleshy surface): Higginsia, Halicnemia, Ptilocaulis. ichondrids in Ceractinomorpha was also problematic, as the larval morphology of these sponges differs from the remaining Van Soest (1991) supported his classification with a cla- Ceractinomorpha, and a diversity of types of larvae had been distic analysis. The resulting consensus combined Poecilo- observed in halichondrids (Wapstra and Van Soest 1987). As sclerida and Haplosclerida, Halichondrida (with Axinellida) an additional complication to reproduction-based classifica- and the keratose sponges in a clade separated from Tetracti- tion, several ceractinomorph taxa were found to be oviparous nellida Hadromerida, Homoscleromorpha and Calcarea. The (verongids, agelasids, petrosiids, desmacellids, (e.g. Bergquist axial compression of the skeleton, a synapomorphy for the 1978;Bergquist1980b; Hoppe and Reichert 1987). Conse- Order Axinellida sensu Lévi, was suggested to be homoplas- quently, it seemed unlikely that viviparity evolved only ious in demosponges. once in sponges as reported for several other invertebrate Although this new classification gained broader acceptance groups such as molluscs and ascidians. Independently, Hooper in the ensuing years (e.g. Lévi 1997), morphological and observed closer relationships of the tetractinomorph Family biochemical data indicated that Axinellidae and Heteroxyidae Raspailiidae to the ceractinomorph Family , might not be homogeneous groups and the genera might not than to other axinellid groups, supporting the concept of de be easily differentiated (Bergquist and Hartman 1969;Sole- Laubenfels (1936) (Hooper 1990, 1991). Consequently, Cava et al. 1991;HooperandBergquist1992;Hooperetal. (Clav-) Axinellida sensu Lévi was abandoned and its families 1992). In subsequent revisions, the genera nominated for were partially (Heteroxyidae (as Desmoxyidae) and Axinelli- Axinellidae were reduced to 11 (Alvarez et al. 1998;Alvarez dae) redistributed within Halichondrida (Van Soest et al. de Glasby 1996; Hooper and Lévi 1993) and morphological as 1990). In this schema, Family Dictyonellidae was erected to well as molecular phylogenies of this family were inferred cope with genera of the polyphyletic order Axinellida. Dic- (Alvarez and Crisp 1994; Alvarez et al. 2000). Myrmekio- tyonellidae was predicted to be close to Halichondriidae, derma and Didiscus were returned to Desmoxyidae (subse- based on the loss of the reticulate skeleton. Both families quently Heteroxyidae, see Van Soest and Hooper 2005), 64 D. Erpenbeck et al. together with the genera Higginsia, Halicnemia, Heteroxya, Fig. 1 Bayesian inference phylogram on CO1 barcoding samples.b Values at branches are support values. Below: Maximum-likelihood Myrmekioderma, Didiscus,andJulavis de Laubenfels 1936 bootstrap values (>70), above: Bayesian posterior probabilities (> 0.9). (Van Soest and Lehnert 1997). Finally, in Systema Porifera Support values for some branches of lesser importance of this analysis (Hooper and Van Soest 2002), Halichondrida were clas- (inside outgroups, Spongillina, Poecilosclerida s.s.) may not be pro- sified as an order containing five families: Halichondriidae vided to maintain legibility. Grey boxes indicate halichondrid taxa sensu Hooper and Van Soest (2002). The taxon names are followed (Erpenbeck and Van Soest 2002), Axinellidae (Alvarez and by Genbank accession numbers. In case the sequences have not been Hooper 2002), Dictyonellidae (Van Soest et al. 2002), Heter- published earlier, collection numbers of the Queensland Museum Bris- oxyidae (Hooper 2002a), and the Bubaridae (Alvarez and Van bane (Gxxxxxx) are provided Soest 2002), which re-attained family status (Van Soest and Hooper 2002). However, with the advent of molecular tech- paratangential forms may be present in Hymeniacidon and niques in sponge systematics (Kelly Borges et al. 1991), early other Halichondriidae, and species of the suberitid genus indications were that the strictly morphological classification Caulospongia Kent, 1871 possess a tangential ectosomal of the halichondrids was in many aspects likely to be incon- skeleton. Caulospongia and other taxa with overlapping mor- gruent with molecular hypotheses (Lafay et al. 1992). The phological traits (e.g., Pseudosuberites) represent pivotal taxa molecular hypotheses will be in the highlighted in the follow- to investigate to fully understand the evolution of this clade. ing section, which is based on phylogenetic reconstructions In our reconstruction, the non-monophyly of Halichon- incorporating our new halichondrid barcoding data. drida and halichondrid families is demonstrated in other aspects. Stylissa carteri (Dendy, 1889) and S. massa (Carter, The molecular view on halichondrid sponges 1887) are distant to other Dictyonellidae and join the sister group to Order Agelasida with several axinellid taxa (termed Halichondrida (and several of its constituent families) has as Hymerhabdiidae, Morrow et al. 2011). This corroborates never been recovered as monophyletic; there is no analysis, one of the first molecular phylogenies on sponges (Lafay et al. be it based on morphological, biochemical (e.g., Erpenbeck 1992) and biochemical analyses (Van Soest and Braekman and Van Soest 2005) or molecular datasets. This is in accord 1999) and subsequent independent data sets (see Erpenbeck et with the phylogenetic trees we reconstructed from the CO1 al. 2005a, 2006; Nichols 2005;Morrowetal.2011). mtDNA barcoding fragment. Our tree comprises 584 charac- Dictyonellidae were classically defined based mostly on ters from a total of 163 halichondrid and selected other demo- negative characters, and were therefore likely candidates to sponges to obtain the most representative possible data set. Of form an assemblage of unrelated sponges (e.g. Van Soest et these, 62 sequences were newly generated (Identifier: al. 2002). Dictyonellidae possess predominantly styles and Gxxxxxx in Fig. 1). CO1 cannot support the deeper splits; display a fleshy appearance due to a comparatively low spicule however, it is evident that halichondrid CO1 sequences fall in content. Sponges from several demosponge lineages can re- distant clades (Fig. 1, grey boxes) and no halichondrid family semble dictyonellids after a (presumable) reduction of their is recovered as monophyletic (with the exception of Bubar- skeletal elements. Consequently, we find dictyonellid taxa idae, which are not sampled here). scattered all over the demosponge phylogeny (Fig. 1). For Most Halichondriidae, the nominal family of the hali- example, CO1 recovers Liosina paradoxa Thiele, 1899, clas- chondrids, form a clade with suberitid taxa. A close hal- sified "tentative[ly] in the absence of any definite synapomor- ichondrid+suberitid relationship was previously inferred phies with either Halichondrida and Poecilosclerida" (Van using ribosomal data (e.g. Chombard and Boury-Esnault Soest et al. 2002), in a clade with tethyid and other hemi- 1999; McCormack and Kelly 2002; Erpenbeck et al. 2004; asterellid sequences. This result agrees with earlier findings on 2005b), and also using CO1 data Erpenbeck et al. (2007a), Liosina based on nuclear ribosomal DNA sequence data as well as recent combined analyses (Morrow et al. 2011). (Kober and Nichols 2007), and supports the placement of Morphologically, and Halichondriidae share the Liosina in a redefined family Tethyidae (Morrow et al. 2011). presence of a (mostly) confused choanosomal skeleton, Likewise, analyses of molecular data (CO1 and ribosomal) although in suberitids it can tend to form radial arrange- infer the positions of Scopalina lophyropoda Schmidt 1862 ments. Consequently the (approximate) distinction into taxa (and Svenzea Alvarez, VanSoest & Rützler, 2002) as unrelated with oxeas (halichondriids) and tylostyles (suberitids) is to most other halichondrids (Nichols 2005; Erpenbeck et al. invalid. This is particularly obvious in the genus Vosmaeria 2007a) and with uncertain relationship to other taxa; this result Fristedt, 1885, which combines both spicule types (see triggered the proposal of a separate family (see Morrow et al. Gerasimova et al. 2008 for a molecular analysis). 2011). We also observe long interspecific branches in Scopa- Halichondriids and suberitids both possess an ectosomal lina CO1 preventing the formation of single Scopalina clade, skeleton with overlapping traits, but a distinction based on its unlike in nuclear ribosomal data (see also Morrow et al. 2011). (mostly) tangential shape in Halichondriidae and its brush- or Several species allocated to former axinellid/dictyonellid palisade-type in Suberitidae is not warranted. For example, genera display "fuzzy boundaries" due to overlapping The phylogeny of halichondrid demosponges 65

0.98 Artemisina melana EF519575 89 Clathria prolifera AJ704978 (as Microciona) Clathria oxeota EF519605 1 Negombata magnifica AM076981 99 Sceptrella biannulata JF440339 Diacarnus spinipoculum AY561975 Iotrochota baculifera G306386 0.99 Iotrochota sp. G300236 100 Iotrochota birotulata AY561963 1 Lissodendoryx isodictyalis EF519638 0.99 100 Mycale fibrexilis AJ843890 1 76 Tedania ignis DQ133896 98 Mycale laxissima EF519649 Strongylacidon bermuda AJ843889 Eurypon pilosella JF440337 0.98 Monanchora arbuscula EF519645 71 1 Scopalina blanensis AM498642 1 100 Scopalina lophyropoda HQ379411 1 97 1 Scopalina canariensis AM498644 99 Scopalina ceutensis AM498645 83 Scopalina ruetzleri EF519669 1 domuncula AM690374 1 HQ379429 0.96 99 99Suberites ficus AJ843891 0.94 Ciocalypta sp. G301042 0.96 Protosuberites sp. AY561979 .98 Hymeniacidon heliophila EF519629 1 Hymeniacidon sinapium EF217329 0.97 99 100 Hymeniacidon flavia EF217333 1 Amorphinopsis fenestrata G303287 1 88 Amorphinopsis sp. G313108 1 99 Ciocalypta sp. EF519604 100 1 Halichondria magniconulosa EF519615 82 Halichondria melanadocia EF519617 1 1 91Halichondria okadai EF217340 98 96 .94 Amorphinopsis maculosa G300854 sp. AY561983 1 Suberites aurantiacus EF519676 93 sp. G303294 Ephydatia fluviatilis DQ176777 0.99 Swartschewskia papyracea EU000571 93 Baikalospongia bacillifera EU000570 Lubomirskia baicalensis DQ167169 Echinospongilla brichardi EU000573 1 Trochospongilla pennsylvanica DQ087503 95 Eunapius fragilis AJ843882 Corvomeyenia sp. DQ176781 0.94 Tethya actinia NC006991 0.99 71 Tethya citrina HQ379427 0.91 79 Tethya aurantium EF584565 1 Tethya coccinea G303083 99 1 Adreus fascicularis HQ379428 93 Liosina paradoxa G307744 0.99 Cliona celata HM999031 Pione vastifica EF519665 0.94 Clionaopsis platei HM999043 1 77 Polymastia sp. AY561971 100 Placospongia sp. AY094604 Pseudotrachya sp. AY561965 0.97 Polymastia janeirensis EU076813 1 Sphaerotylus sp. HQ379425 1 82 Tentorium semisuberites HQ379426 100 Polymastia sp. AY561982 Scopalina lophyropoda AM498640 Svenzea zeai EF519682 1 Auletta sp. G300636 100 Axinella sp. G300253 Acanthella sp. G306117 Acanthella sp. G306118 1 Acanthella sp. G300772 100 Acanthella sp. G300619 Acanthella sp. G303176 Acanthella sp. G305548 1 Acanthella sp. G305639 Phycopsis epakros G300016 0.92 97 Acanthella acuta HQ379408 Phakellia ventilabrum HQ379409 1 concentrica G306736 1 Cymbastela concentrica G307850 98 98Cymbastela vespertina G300002 1 1 1 Phakettia euctimena G300580 100 100 100 Phakettia euctimena G300758 Dictyonella sp. AM498649 Acanthella carduus G301212 Stylissa carteri G306142 73 Stylissa carteri G306987 1 Stylissa massa G306438 Stylissa massa G307272 100 Axinella corrugata NC006894 1 Prosuberites laughlini AY561960 97 EF519538 1 Astrosclera willeyana AY561969 100 0.99 EF519542 97 Agelas dispar EF519546 96 Neofibularia hartmani G306606 Neofibularia hartmani G306628 1 Neofibularia nolitangere EF519653 1 95 Neofibularia irata G307266 100 Biemna variantia HQ379424 G307149 0.98 Cymbastela coralliophila G307525 1 100 1 Cymbastela coralliophila G307723 99 Ciocalypta sp. G301298 100 Topsentia ophiraphidites EU237482 Raspailia elegans G300224 Trikentrion flabelliforme G306125 Thrinacophora cervicornis G300850 1 0.93 Thrinacophora cervicornis G304943 100 Axechina raspailioides G300288 1 Axechina raspailioides G300597 97 Axechina raspailioides G303088 0.99 cf. Rhaphoxya systremma G307173 86 Ectyoplasia ferox EF519612 0.91 0.99 Endectyon delaubenfelsi HQ379412 91 Endectyon fruticosum aruense G300254 1 Didiscus aceratus G312916 100 Didiscus sp. AY561972 0.99 Epipolasis sp. G307709 100 Epipolasis sp. G307785 1 Ptilocaulis marquezi EF519668 100 Ptilocaulis walpersi EU237488 Ptilocaulis sp. G306291 73 Echinodictyum cancellatum G300863 1 Echinodictyum mesenterinum (Gr.Bar.Reef) G307750 1 100 1 Echinodictyum clathrioides G300559 97 100 Echinodictyum mesenterinum (W-Australia) G306004 1 .95 Raspailia hispida HQ379416 1 96 90 Raspailia ramosa HQ379417 Raspailia sp. G300385 1 97 Pandaros acanthifolium EF519662 98 Eurypon clavigerum HQ379413 Raspaciona aculeata HQ379415 Tethyspira spinosa HQ379418 1 rigida HQ379420 0.91 Stelligera stuposa HQ379421 93 96Paratimea constellata HQ379419 Halicnemia patera HQ379422 87 Higginsia robusta G300227 Higginsia sp. G300611 Hymeraphia verticillata HQ379414 Axinella aruensis G300581 Axinella aruensis G300595 1 Axinella aruensis G300635 100 Axinella aruensis G301092 0.96 Axinella aruensis G312935 1 83 Axinella infundibuliformis HQ379410 98 Dragmacidon sp. G300605 1 Dragmacidon reticulatum AJ843894 substitutions / site 94 Myrmekioderma granulatum G306335 1 Myrmekioderma granulatum G313288 100 Myrmekioderma granulatum G314109 0.0 0.2 Myrmekioderma gyroderma EF519652 Tetractinellida (10 species) 66 D. Erpenbeck et al. characters and phenotypic plasticity (Alvarez and Hooper Our CO1 reconstructions, including the holotype of Cym- 2009). In the clade containing Acanthella acuta (type spe- bastela vespertina Hooper and Bergquist, 1992, corrobo- cies of Acanthella) and Dictyonella we recover representa- rates independent nuclear ribosomal data that Cymbastela tives from the erstwhile axinellids Auletta, Phakellia, and shares a close relationship to Acanthella and Dictyonella (e. Phycopsis. Phakellia is represented by its type species P. g. Alvarez et al. 2000; Erpenbeck et al. 2005b; Gazave et al. ventilabrum and its affinity to a Dictyonellidae clade has 2010; Morrow et al. 2011) although skeleton and spiculation been shown earlier (Morrow et al. 2011). Several species of of both groups is relatively different (Alvarez and Hooper Acanthella have been mistaken consistently for Phakellia. 2010). Our CO1 data also supports, for the first time with a For example, Acanthella carduus has been a suggested new mitochondrial marker, that Cymbastela coralliophila Hooper combination for Phakellia carduus (Alvarez and Hooper and Bergquist, 1992 is in fact unrelated to other Cymbastela 2010), nevertheless, its molecular affinities are equivocal species (see the ribosomal data of Alvarez et al. 2000). Cym- in this data set. Acanthella, Phakellia plus species from bastela coralliophila differs from C. vespertina by the pos- other genera might form a species complex, the resolution session of a paratangential ectosomal skeleton (absent in C. of which requires additional data. vespertina)andtwosizecategoriesofoxeasintheprimary Phakettia euctimena (Hentschel, 1912) was initially skeleton (only one in C. vespertina). assigned to Acanthella before being reclassified on the Cymbastela coralliophila forms a clade with an as yet grounds of the general skeletal architecture and the spicule unidentified oxea-bearing species of Ciocalypta (resulting composition (Alvarez and Hooper 2010). Our molecular tree paraphyly of Ciocalypta), and a GenBank specimen submit- lacks the comparison to the type species P. cactoides (Burton, ted as Topsentia ophiraphidites (Halichondriidae). Cymbas- 1928) but corroborates a genealogical distance to Acanthella. tela coralliophila and Topsentia share the possession of The skeletal organisation of Auletta is comparable to oxeas in several different size classes, and the possession Phakellia (see Alvarez and Hooper 2002) and discussed as of a paratangential skeleton, but differ in choanosomal skel- homologous (Morrow et al. 2011). Auletta, Phycopsis and etal architecture (confused vs plumo-reticulate). A closer Phakellia all share the flexuous strongyles with Acanthella examination of this Topsentia specimen is required. It dis- and our analyses raise further evidence that the possession plays the same position distant to Halichondriidae in 18S of strongyles is of greater taxonomic relevance than pres- analyses (see http://www.spongegenetrees.org, Erpenbeck et ently recognised (see also Morrow et al. 2011). For exam- al. 2008), but is different to another specimen of T. ophir- ple, close relationships between the genera Ptilocaulis and aphidites, which is related to Halichondriidae in 28S and Reniochalina are well established (e.g. Hooper and Lévi CO1 analyses (Erpenbeck et al. 2005b; Raleigh et al. 2007). 1993, 1994; Alvarez et al. 2000), but molecular and mor- The non-monophyly of Axinella—the nominal genus for phological data rejected a monophyletic Ptilocaulis. There- the Axinellidae—has been shown earlier (Alvarez et al. fore, the strongyle-bearing species Ptilocaulis fusiformis 2000; Gazave et al. 2010). Our reconstruction recovers a Lévi, 1967 and P. epakros Hooper and Lévi, 1993 were clade of Dragmacidon, A. aruensis (Hentschel, 1912) and A. assigned to Phycopsis opposed to the strongyle-lacking type infundibuliformis (Linnaeus, 1759). D. reticulatum (Ridley species, Ptilocaulis walpersi (Duchassaing & Michelotti, and Dendy, 1886)andA. aruensis have nearly identical 1864) and Ptilocaulis marquezi (Duchassaing & Michelotti, choanosomal skeletons, spicule compositions and sizes. 1864) (Alvarez et al. 1998; Alvarez and Hooper 2002). Our Nevertheless, Axinella and Dragmacidon are distinguished data support this placement within Phycopsis, as the type largely by the relative density of the spiculation and the specimen of P. epakros falls in a clade with Acanthella and degree of axial compression in the skeleton, with the skel- Phakellia, distant to P. walpersi and P. marquezi. eton of Axinella being less dense and more compressed. Nevertheless, the affinities of other strongyle bearing-taxa Dragmacidon is further distinguished from Axinella by its Rhaphoxya systremma Hooper and Lévi 1993 (represented growth form and surface characteristics. These character- here only by a putative specimen) or certain raspilid taxa, e.g. istics tend to grade into one another and can be difficult to Ceratopsion rugosum (Schmidt 1870, not represented here) separate. await confirmation. The possession of strongyles is not an Our CO1 data corroborates the closer relationship of obligatory feature for this clade, because Axinyssa, previously Ptilocaulis (s.s.) to Raspailiidae (previously classified to classified as the Halichondriidae (Erpenbeck and Van Soest the order Poecilosclerida, Hooper 2002b), as has been indi- 2002)islikelytofallintothisclade,aswasindicatedbyrDNA cated previously by several ribosomal and mitochondrial analyses (Erpenbeck et al. 2005b;Morrowetal.2011). Axi- analyses (Erpenbeck et al. 2007a; Erpenbeck et al. 2007b; nyssa also lacks the halichondriid ectosomal skeleton, which Erpenbeck, et al. 2007c; Holmes and Blanch 2007). Species makes its different phylogenetic position plausible. of Raspailiidae (mostly) possess a typical ectosomal skeleton, As with Axinyssa, species of the axinellid genus Cym- consisting of small thin spicules forming bouquets around bastela Hooper and Bergquist, 1992 only possess oxeas. long styles, or oxeas that penetrate the surface, in combination The phylogeny of halichondrid demosponges 67 with echinating acanthostyles in the choanosomal skeleton features (e.g. Lévi 1951). These different viewpoints were (Hooper 2002b). However, these features are found neither likely to be triggered by unsatisfying earlier classifications in Ptilocaulis nor in Epipolasis, for which specimens are that originated from the paucity of phylogenetically relevant recovered among raspailiids. Epipolasis was assigned to the sponge characters, and reflect the struggles of early workers to Halichondriidae based on its confused choanosomal skeleton find these characters to establish a robust sponge classifica- and an ectosomal crust of intercrossing spicules (Erpenbeck tion. Nevertheless, it is difficult to extract particular historic and Van Soest 2002). The molecular results lack a satisfactory schemes of the authors, which would fit the current molecular morphological explanation, yet they support earlier hypothe- hypotheses in more than just a few higher taxa. ses on the homoplasious nature of extra-axially plumo- The phylogeny inferred by Morrow et al. (2011) unites most reticulate and axially compressed skeletons, as found in axi- of the currently proposed molecular hypotheses as far as it nellids, raspailiids and other (e.g. hadromerid) taxa (Hooper concerns the halichondrids and summarizes several hypothe- 1991). ses, which have never before been discussed. Their study Our reconstruction corroborates the polyphyly of hetero- agrees in large part with the findings presented here. Together, xyid sponges (Erpenbeck et al. 2005a, b;Morrowetal.2011). these molecular studies provide strong evidence that molecular Didiscus is related more closely to Raspailiidae than to other methods, when they are associated with good and correct heteroxyid taxa Erpenbeck et al. (2007b). This tree also indi- taxonomic identifications (strengthened by the process of typ- cates, for the first time, the phylogenetic position of Higginsia ification and morphological vouchering), can provide alterna- (close to Halicnemia and former hemiasterellid taxa, recently tive ways of interpreting morphological data. Importantly, combined as Stelligeridae, Morrow et al. 2011). these studies also (as frequently underestimated) demonstrate In conclusion, CO1 underlines the polyphyletic nature of that thorough and careful morphological classifications can be the halichondrids, one of the most pivotal and controversial “right” and natural. groups in demosponge classification. This result supports previous, predominantly ribosomal DNA-based, analyses. Acknowledgements The Sponge Barcoding Project thanks The Marine Despite the fact that the lower resolution power of CO1 Barcode of Life initiative (MarBol), funded by the Alfred P. Sloan Foun- LMU for deeper splits has been described in detail (Lavrov et al. dation and the GeoBio-Center , for financing the subsampling and extraction of 17,000 sponge specimens of the Queensland Museum. We 2008), CO1 gene trees clearly contain sufficient signal to thank the DNA Bank at the Zoologische Staatssammlung München (http:// analyse the extend of halichondrid polyphyly. Indeed, this is www.zsm.mwn.de/dnabank) for collaboration in DNA subsampling and particularly emphasised when our COI gene tree is congruent storage. G.W. acknowledges funding by the German Science Foundation “ ” to trees based on independent genes such as nuclear ribosomal (DFG) through the Deep Metazoan Phylogeny Priority Program (Project Wo896/6). We furthermore thank the editors and two anonymous sequences (18S, 28S, ITS). Independent loci likely reflect reviewers for their constructive comments. different evolutionary histories and substitution patterns (see e.g. Moore 1995), therefore congruent topologies provide strong evidence for “correctness” and should be regarded as References more important than high branch support, which solely meas- ures the power of the underlying characters (e.g. Felsenstein Alvarez, B., & Crisp, M. D. (1994). A preliminary analysis of the 1985). phylogenetic relationships of some axinellid sponges. In R. W. M. The results from our study highlight the need for stringent Van Soest, T. M. Gv. Kempen, & J. C. Braekman (Eds.), Sponges typification in phylogenetic studies. Molecular phylogenetic in Time and Space (pp. 117–122). Rotterdam: Balkema. studies that aim to contribute to the classification of sponges Alvarez, B., Crisp, M. D., Driver, F., Hooper, J. N. A., & VanSoest, R. W. M. (2000). Phylogenetic relationships of the family Axinellidae require clear definition of the type species. This should not (Porifera: Demospongiae) using morphological and molecular data. necessarily be based solely on the type material, which is often Zoologica Scripta, 29(2), 169–198. unavailable because of historical factors (specimens lost, Alvarez, B., & Hooper, J. N. A. (2002). Family Axinellidae Carter. In destroyed, inadequately or inappropriately fixed) or cultural J. N. A. Hooper & R. W. M. Van Soest (Eds.), Systema Porifera, A guide to the classification of Sponges (Vol. 1, pp. 724–747). reasons (museum regulations), but also on genetic and mor- New York: Kluwer/Plenum. phometric variation of the type species in their type localities. It Alvarez, B., & Hooper, J.N.A. (2009). Taxonomic revision of the order is certainly insufficient to raise new taxa based on single speci- Halichondrida (Porifera: Demospongiae) from northern Australia. mens, without proper comparisons of the intra-population Family Axinellidae. The Beagle, Records of the Museums and Art Galleries of the Northern Territory, 25,17–42. variability and the likelihoods of misidentifications. Alvarez, B., & Hooper, J. N. A. (2010). Taxonomic revision of the order Importantly, through this study, we see that the evolution of Halichondrida (Porifera: Demospongiae) of northern Australia. demosponge, and particularly halichondrid taxa, underwent Family Dictyonellidae. Beagle, 26,13–36. scrutiny from several different viewpoints. Early views fo- Alvarez, B., & Van Soest, R. W. M. (2002). Family Bubaridae Topsent. In J. N. A. Hooper & R. W. M. Van Soest (Eds.), Systema cused on megascleres, with more importance on microscleres Porifera, A guide to the classification of Sponges (Vol. 1, pp. (e.g. von Lendenfeld 1887), with later views stressing other 748–754). New York: Kluwer/Plenum. 68 D. Erpenbeck et al.

Alvarez, B., Van Soest, R. W. M., & Rützler, K. (1998). A revision of elongation factor-1 alpha fragment in demosponges (Metazoa, Axinellidae (Porifera: Demospongiae) of the Central West Atlanic Porifera, Demospongiae). Zoologica Scripta, 34(4), 437–445. Region. Smithonian Contributions to Zoology, 598,1–47. Erpenbeck, D., Breeuwer, J. A. J., & Van Soest, R. W. M. (2005b). Alvarez de Glasby, B. (1996). The Phylogenetic Relationships of the Implications from a 28S rRNA gene fragment for the phylogenetic Family Axinellidae (Porifera: Demospongiae). Canberra: Australian relationships of halichondrid sponges (Porifera: Demospongiae). National University. Journal of Zoological Systematics and Evolutionary Research, 43 Belon, P. (1553). De aquatilibus (Vol. Libri II). Parisiis. (2), 93–99. Bergquist, P. R. (1978). Sponges. London: Hutchinson University Erpenbeck, D., Duran, S., Rützler, K., Paul, V., Hooper, J. N. A., & Library. Wörheide, G. (2007a). Towards a DNA of Caribbean Bergquist, P. R. (1980). A revision of the supraspecific classification of demosponges: a gene tree reconstructed from partial mitochon- the orders Dictyoceratida, Dendroceratida, and Verongida (class drial CO1 gene sequences supports previous rDNA phylogenies Demospongiae). New Zealand Journal of Zoology, 7,443–503. and provides a new perspective on the systematics of Demospon- Bergquist, P. R., & Hartman, W. D. (1969). Free amino acid patterns giae. Journal of the Marine Biological Society of the United and the classification of the Demospongiae. Marine Biology, 3, Kingdom, 87, 1563–1570. 247–268. Erpenbeck, D., Hooper, J. N. A., List-Armitage, S. E., Degnan, B. M., Borchiellini, C., Chombard, C., Manuel, M., Alivon, E., Vacelet, J., & Wörheide, G., & Van Soest, R. W. M. (2007b). Affinities of the Boury-Esnault, N. (2004). Molecular phylogeny of Demospongiae: family Sollasellidae (Porifera, Demospongiae). II. Molecular evi- implications for classification and scenarios of character evolution. dence. Contributions to Zoology, 76(2), 95–102. Molecular Phylogenetics and Evolution, 32(3), 823–837. Erpenbeck, D., List-Armitage, S. E., Alvarez, B., Degnan, B. M., Bowerbank, J.S. (1864, 1866, 1874). Monograph of the British Spon- Hooper, J. N. A., & Wörheide, G. (2007c). The systematics of giadae (Vol. I–III). London: Ray Society. Raspailiidae (Demospongiae, Poecilosclerida, Microcionina) Bowerbank, J.S. (1874). A monograph of the British Spongiadae (Vol. reanalysed with a ribosomal marker. Journal of the Marine Bio- 3). London: Ray Society. logical Society of the United Kingdom, 87, 1571–1576. Carballo, J. L., Uriz, M. J., & GarciaGomez, J. C. (1996). Halichon- Erpenbeck, D., McCormack, G. P., Breeuwer, J. A. J., & Van Soest, R. drids or axinellids? Some problematic genera of sponges with W. M. (2004). Order level differences in the structure of partial descriptions of new species from the Strait of Gibraltar (southern LSU across demosponges (Porifera): new insights into an old Iberian Peninsula). Journal of Zoology, 238, 725–741. taxon. Molecular Phylogenetics and Evolution, 32(1), 388–395. Carter, H.J. (1875). Notes Introductory to the Study and Classification Erpenbeck, D., & Van Soest, R. W. M. (2002). Family Halichondriidae of the Spongida. Part II. Proposed Classification of the Spongida. Gray, 1867. In J. N. A. Hooper & R. W. M. Van Soest (Eds.), Annals and Magazine of Natural History, (4) 1(6(92)), 126–145, Systema Porifera, A guide to the classification of Sponges (Vol. 1, 177–200. pp. 787–815). New York: KluwerAcademic/Plenum Publishers. Chombard, C., & Boury-Esnault, N. (1999). Good congruence be- Erpenbeck, D., & Van Soest, R. W. M. (2005). A survey for biochemical tween morphology and molecular phylogeny of Hadromerida, or synapomorphies to reveal phylogenetic relationships of halichon- how to bother sponge taxonomists. Memoirs of the Queensland drid demosponges (Metazoa: Porifera). Biochemical Systematics Museum, 44, 100. and Ecology, 33,585–616. Chombard, C., Boury-Esnault, N., Tillier, A., & Vacelet, J. (1997). Erpenbeck, D., Voigt, O., Gültas, M., & Wörheide, G. (2008). The Polyphyly of "sclerosponges" (Porifera, Demospongiae) sup- sponge genetree server-providing a phylogenetic backbone for ported by 28S ribosomal sequences. The Biological Bulletin, poriferan evolutionary studies. Zootaxa, 1939,58–60. 193(3), 359–367. Felsenstein, J. (1985). Confidence limits on phylogenies: an approach De Laubenfels, M.W. (1936). A discussion of the sponge fauna of the using the bootstrap. Evolution, 39(4), 783–791. Dry Tortugas in particular and the West Indies in general, with Galtier, N., Gouy, M., & Gautier, C. (1996). SEAVIEWand PHYLO_WIN: material for a revision of the families and orders of the Porifera. Two graphic tools for sequence alignment and molecular phylogeny. Publications Carnegie Institute Washington 467 (Papers Tortugas Computer Applications in the Biosciences, 12(6), 543–548. Laboratory), 30,1–225. Gazave, E., Carteron, S., Chenuil, A., Richelle-Maurer, E., Boury- Dendy, A. (1922). Report on the Sigmatotetraxonida collected by H.M. Esnault, N., & Borchiellini, C. (2010). Polyphyly of the genus S. ‘Sealark’ in the Indian Ocean. In Reports of the Percy Sladen Axinella and of the family Axinellidae (Porifera: Demospongiae Trust Expedition to the Indian Ocean in 1905, Volume 7. Trans- (p)). Molecular Phylogenetics and Evolution, 57(1), 35–47. actions of the Linnean Society of London (2) (Vol. 18(1). pp. Pp. doi:10.1016/j.ympev.2010.05.028. 1–164, pls 161–118.). Gerasimova, E., Erpenbeck, D., & Plotkin, A. (2008). Vosmaeria Diaz, M.C. (1997). Molecular detection and characterization of specif- Fristedt, 1885 (Porifera, Demospongiae, Halichondriidae): revi- ic bacterial groups associated with tropical sponges. Proceedings sion of species, phylogenetic reconstruction and evidence for of the eighth international coral reef symposium, Panama, June split. Zootaxa(1694), 1–37. 24–29, 1996. Smithsonian Tropical Research Institute, Balboa, Grant, R. E. (1836). Kingdom. In R. B. Todd (Ed.), The Panama, 2, 13399–11402. Cyclopaedia of Anatomy and Physiology 1–813 (Vol. 1, pp. Donati, V. (1750). Della storia naturale marina dell’ Adriatico. Venezia: 107–118). London: Sherwood, Gilbert, and Piper. Saggio. Gray, J. E. (1867). Notes on the arrangement of sponges, with the Edgar, R. C. (2004). MUSCLE: multiple sequence alignment with high descriptions of some new genera. Proceedings Zoological Society accuracy and high throughput. Nucleic Acids Research, 32(5), London, Lit. IV (1868), 492–558. 1792–1797. Gray, J. E. (1872). Notes on the Classification of the Sponges. Annals Erpenbeck, D., Breeuwer, J. A. J., Parra-Velandia, F. J., & Van Soest, R. and Magazine of Natural History, 4(9 (54)), 442–461. W. M. (2006). Speculation with spiculation?—Three independent Haeckel, E.H.P.A. (1872). Die Kalkschwaemme (Vol. 1): Atlas. gene fragments and biochemical characters versus morphology in Hallmann, E.F. (1917 (1916)). A revision of the genera with micro- demosponge higher classification. Molecular Phylogenetics and scleres included, or provisionally included, in the family Axinel- Evolution, 38,293–305. lidae; with descriptions of some Australian species. Part III. Erpenbeck, D., Breeuwer, J. A. J., & Van Soest, R. W. M. (2005a). Proceedings of the Linnean Society of New South Wales, 41 Identification, characterization and phylogenetic signal of an (164), 634–675. The phylogeny of halichondrid demosponges 69

Hansson, H. G. (1994). Sydskandinaviska marina flercelliga ever- Kelly Borges, M., Bergquist, P. R., & Bergquist, P. L. (1991). Phylo- tebrater. Göteborg: Länstyrelsen i Göteborgs och Bohus län. genetic relationships within the order Hadromerida (Porifera, Hartman, W. D. (1969). New genera and species of coralline sponges Demospongiae, Tetractinomorpha) as indicated by ribosomal (Porifera) from Jamaica. Postilla, 137,1–39. RNA sequence comparisons. Biochemical Systematics and Ecology, Hartman, W. D. (1982). Porifera. In S. P. Parker (Ed.), Synopsis 19(2), 117–125. and Classification of Living Organisms (Vol. 1). New York: Kober, K., & Nichols, S. (2007). On the phylogenetic relationships of McGraw-Hill. hadromerid and poecilosclerid sponges. Journal of the Marine Hartman, W. D., & Goreau, T. F. (1970). Jamaican coralline sponges: Biological Association of the United Kingdom, 87(06), 14. Their morphology, ecology and fossil relatives. Symposia of the doi:10.1017/S0025315407058237. Zoological Society of London, 25, 205–243. Lafay, B., Boury Esnault, N., Vacelet, J., & Christen, R. (1992). An Hentschel, E. (1923). Erste Unterabteilung der Metazoa: Parazoa, analysis of partial 28S ribosomal RNA sequences suggests early Porifera0Schwämme. In W. Kükenthal, & T. Krumbach (Eds.), radiations of sponges. Biosystems, 28(1–3), 139–151. Handbuch der Zoologie. Eine Naturgeschichte der Stämme des de Monet JBPA, Lamarck. (1813). Sur les polypiers empates. Ann du Tierreiches. Protozoa, Porifera, Coelenterata, Mesozoa (Vol. 1, Mus., XX, 370, 432. pp. 307–418). Berlin and Leipzig: Walter de Gruyter und Co. Lavrov, D., Wang, X., & Kelly, M. (2008). Reconstructing ordinal Hogg (1842). Remarks on the Horny Sponges with proposed divisions relationships in the Demospongiae using mitochondrial ge- of the Order Spongiae. Ann. And Mag., VIII,3. nomic data. Molecular Phylogenetics and Evolution, 49(1), Holmes, B., & Blanch, H. (2007). Genus-specific associations of 111–124. marine sponges with group I crenarchaeotes. Marine Biology, Lévi, C. (1951). L’oviparité chez les spongiaires. Comptes rendus des 150(5), 759–772. séances de l’Académie des Sciences, 233, 272–274. Hooper, J. N. A. (1984). Sigmaxinella soelae and Desmacella ithystela, Lévi, C. (1953a). Description de Plakortis nigra nov. sp. et remarques two new desmacellid sponges (Porifera, Axinellida, Desmacelli- sur les Plakinidae (Demosponges). Bulletin de Museum, 2e serie,, dae) from the Northwest shelf of Western Australia, with a revi- XXV(3). sion of the family Desmacellidae. Monograph series of the Lévi, C. (1953b). Sur une nouvelle classification des Démosponges. Northern Territory Museum of Arts and Sciences, 2,1–58. Comptes rendus des séances de l’Académie des Sciences, 236, Hooper, J. N. A. (1990). Character stability, systematics and affinities 853–855. between Microcionidae (Poecilosclerida) and Axinellida. In K. Lévi, C. (1955). Les Clavaxinellides, Démosponges Tétractinomor- Rützler (Ed.), New perspectives in Sponge Biology (pp. 284– phes. Archives de Zoologie expérimentale et générale. Notes et 294). Washington DC: Smithsonian Institution Press. Revue., 92(2), 78–87. Hooper, J. N. A. (1991). Revision of the family Raspailiidae (Porifera: Lévi, C. (1956). Étude des Halisarca de Roscoff. Embryologie et systém- Demospongiae) with description of Australian species. Inverte- atique des Démosponges. Archives de Zoologie Expérimentale et brate Taxonomy, 5(6), 1179–1418. Générale, 93,1–181. Hooper, J. N. A. (2002a). Family Desmoxyidae Hallmann, 1917. In J. Lévi, C. (1957). Ontogeny and systematics in sponges. Systematic N. A. Hooper & R. W. M. Van Soest (Eds.), Systema Porifera, A Zoology, 6,1–4. guide to the classification of Sponges (Vol. 1, pp. 755–772). New Lévi, C. (1973). Systématique de la classe des Demospongiaria York: Kluwer/Plenum. (Démosponges). In P. Brien, C. Lévi, M. Sarà, O. Tuzet, & J. Hooper, J. N. A. (2002b). Family Raspailiidae Hentschel, 1923. In J. Vacelet (Eds.), Traité de Zoologie. Anatomie, Systématique, Biol- N. A. Hooper & R. W. M. Van Soest (Eds.), Systema Porifera. ogie. III. Spongiaires (pp. 577–631). Paris: Masson et Cie. Guide to the classification of sponges (Vol. 1, pp. 469–510). New Lévi, C. (1997). La classification des Porifera Grant, 1836, en 1996. York, Boston, Dordrecht, London, Moscow: Kluwer/Plenum. Bulletin de la Societe Zoologique de France, 122(3), 255–259. Hooper, J. N. A., & Bergquist, P. R. (1992). Cymbastela, a new genus Linnaeus, C. (1759). Systema Naturae. Vol.II. Vegetabilia: Holmiae. of lamellate coral reef sponges. Memoirs of the Queensland Linnaeus, C. (1767). Systema Naturae (12th edn revised. ed., Vol. 1): Museum, 32(1), 99–137. Holmiae, Laur. Salvii. Hooper, J. N. A., Capon, R. J., Keenan, C. P., Parry, D. L., & Smit, N. McCormack, G. P., & Kelly, M. (2002). New indications of the phy- (1992). Chemotaxonomy of marine sponges: Families Microcioni- logenetic affinity of Spongosorites suberitoides Diaz et al., 1993 dae, Raspailidae and Axinellidae and their relationships with (Porifera, Demospongiae) as revealed by 28S ribosomal DNA. other families of the orders Poecilosclerida and Axinellida Journal of Natural History, 36(9), 1009–1021. (Porifera: Demospongiae). Invertebrate Taxonomy, 6,261–301. Meyer, C. P., Geller, J. B., & Paulay, G. (2005). Fine scale endemism Hooper, J. N. A., & Lévi, C. (1993). Axinellida (Porifera: Demospon- on coral reefs: Archipelagic differentiation in turbinid gastropods. giae) from the New Caledonia Lagoon. Invertebrate Taxonomy, 6, Evolution, 59(1), 113–125. 1395–1472. Moore, W. S. (1995). Inferring phylogenies from mtDNA variation— Hooper, J. N. A., & Lévi, C. (1994). Biogeography of Indo-west mitochondrial-gene trees versus nuclear-gene trees. Evolution, 49 Pacific sponges: Microcionidae, Raspailidae, Axinellidae. In R. (4), 718–726. W. M. Van Soest, T. M. Gv. Kempen, & J. C. Braekman (Eds.), Morrow, C. C., Picton, B. E., Erpenbeck, D., Boury-Esnault, N., Maggs, Sponges in Time and Space (pp. 191–212). Rotterdam: Balkema. C. A., & Allcock, A. L. (2011). Congruence between nuclear and Hooper, J. N. A., & Van Soest, R. W. M. (2002). Systema Porifera, A mitochondrial genes in Demospongiae: A new hypothesis for rela- guide to the classification of sponges. New York: Kluwer/Plenum. tionships within the G4 clade (Porifera: Demospongiae). Molecular Hoppe, W. F., & Reichert, M. J. M. (1987). Predictable annual mass Phylogenetics and Evolution. doi:10.1016/j.ympev.2011.09.016. release of gametes by the coral reef sponge (Porifera: Demospon- Available online 7 October 2011. giae). Marine Biology, 94(2), 277–285. Nardo, G.D. (1833). Auszug aus einem neuen System der Spongiarien, Huelsenbeck, J. P., & Ranala, B. (2004). Frequentist properties of wonach bereits die Aufstellung in der Universitäts-Sammlung zu Bayesian posterior probabilities of phylogenetic trees under sim- Padua gemacht ist., Isis, oder Encyclopädische Zeitung Coll. ple and complex substitution models. Systematic Biology, 53(6), (Oken: Jena). 519–523. 904–913. Nichols, S. (2005). An evaluation of support for order-level monophy- Johnston, G. (1842). A History of British Sponges and Lithophytes. ly and interrelationships within the class Demospongiae using Edinburgh: W.H. Lizars. partial data from the large subunit rDNA and cytochrome oxidase 70 D. Erpenbeck et al.

subunit I. Molecular Phylogenetics and Evolution, 34(1), 81–96. Van Soest, R. W. M. (1984). Marine sponges from Curacao and other doi:10.1016/j.ympev.2004.08.019. Caribbean Localities. Part III. Poecilosclerida. In P. Wagenaar- Raleigh, J., Redmond, N. E., Delahan, E., Torpey, S., Van Soest, R. W. Hummelinck, & L. J. van der Steen (Eds.), Studies on the Fauna M., Kelly, M., et al. (2007). Mitochondrial Cytochrome oxidase 1 of Curaçao and Other Caribbean Islands (Vol. 66). Utrecht: phylogeny supports alternative taxonomic scheme for the marine Foundation for Scientific Research in Suriname and the Nether- Haplosclerida. Journal of the Marine Biological Association of lands Antilles No. 112. the UK, 87(6), 1577–1584. doi:10.1017/S0025315407058341. Van Soest, R. W. M. (1987). Phylogenetic exercises with monophyletic Reitner, J. (1992). ‘Coralline Spongien’ Der Versuch einer phylogenetisch- groups of sponges. In J. Vacelet & N. Boury-Esnault (Eds.), taxonomischen Analyse. Berliner geowissenschaftliche Abhandlun- NATO ASI Series (Vol. G13, pp. 227–241). Berlin Heidelberg: gen Reihe E (Paläobiologie), 1,1–352. Springer. Taxonomy of Porifera. Ridley, S.O., & Dendy, A. (1886). Preliminary Report on the Monaxonida Van Soest, R. W. M. (1991). Demosponge higher taxa classification re- collected by the H.M.S. ‘Challenger’. Annals and Magazine of examined. In J. Reitner & H. Keupp (Eds.), Fossil and Recent Natural History, 5(18), 325–351, 470–493. Sponges (pp. 54–71). Berlin: Springer. Ridley, S.O., & Dendy, A. (1887). Report on the Monaxonida collected Van Soest, R. W. M., & Braekman, J. C. (1999). Chemosystematics of by H.M.S. ‘Challenger’ during the years 1873–1876. Report on the Porifera: A review. Memoirs of the Queensland Museum, 44, scientific Results Voyage HMS ‘Challenger’,1873–76. Zoology, 20, 569–589. 1–275. VanSoest, R. W. M., Diaz, M. C., & Pomponi, S. A. (1990). Phylogenetic Ronquist, F., & Huelsenbeck, J. P. (2003). MrBayes 3: Bayesian classification of the halichondrids (Porifera, Demospongiae). phylogenetic inference under mixed models. Bioinformatics, 19 Beaufortia, 40(2), 15–62. (12), 1572–1574. Van Soest, R. W. M., Erpenbeck, D., & Alvarez, B. (2002). Family Schmidt, E. O. (1862). Die Spongien des adriatischen Meeres. Leipzig: Dictyonellidae Van Soest, Diaz & Pomponi 1990. In J. N. A. Wilhelm Engelmann. Hooper & R. W. M. Van Soest (Eds.), Systema Porifera, A guide Schmidt, E. O. (1866). Zweites Supplement der Spongien des adriat. to the classification of Sponges (Vol. 1, pp. 773–786). New York: Leipzig: Meeres. Kluwer/Plenum. Schmidt, E. O. (1868). Drittes Supplement der Spongien des adriat. Van Soest, R. W. M., & Hooper, J. N. A. (2002). Order Halichondrida Meeres. Leipzig: Wilhelm Engelmann. Gray, 1867. In J. N. A. Hooper & R. W. M. Van Soest (Eds.), Schmidt, E.O. (1869). Das natürliche System der Spongien. Mitth. Systema Porifera, A guide to the classification of Sponges (Vol. 1, Naturw. Verein Steiermark, II, 261. pp. 721–723). New York: Kluwer/Plenum. Schmidt, E. O. (1870). Grundzüge einer Spongien-Fauna des Atlantischen Van Soest, R. W. M., & Hooper, J. N. A. (2005). Resurrection of Gebietes.Leipzig. Desmoxya (Porifera: Halichondrida), with the description of a Schmidt, E.O. (1880). Die Spongien des Meerbusen von Mexico (Und new species from Rockall Bank bathyal coral reefs, North Atlantic. des caraibischen Meeres). Abtheilung II. Hexactinelliden. In Journal of the Marine Biological Association of the United Reports on the dredging under the supervision of Alexander Kingdom, 85(6), 1367–1371. Agassiz, in the Gulf of Mexico, by the USCSS ‘Blake’. (Vol. 2, Van Soest, R. W. M., & Lehnert, H. (1997). The genus Julavis de pp. 33–90 pls V–X). Jena: Gustav Fischer:. Laubenfels (Porifera: Halichondrida). Proceedings of the Biolog- Sole-Cava, M. A., Thompson, J. D., & Manconi, R. (1991). A new ical Society of Washington, 110(4), 502–510. mediterranean species of Axinella detected by biochemical genet- Vargas, S., Erpenbeck, D., Schuster, A., Sacher, K., Büttner, G., ic methods. In J. Reitner & H. Keupp (Eds.), Fossil and recent Schätzle, S., et al. (2010). A high-throughput, low-cost Porifera sponges (pp. 313–321). Berlin: Springer. DNA barcoding pipeline. In VIII. World Sponge Conference (pp. Sollas, D. (1885). A classification of the sponges. Annals and Magazine 242). Girona, Spain. of Natural History, 5(16 (95)), 395. von Lendenfeld, R. (1887). On the systematic position and classifica- Stamatakis, A. (2006). RAxML-VI-HPC: Maximum likelihood-based tion of sponges. Proc. Soc. London, 558–662. phylogenetic analyses with thousands of taxa and mixed models. Vosmaer, G. C. J. (1882-1886). Porifera. In Bronn’sKlassenund Bioinformatics, 22(21), 2688–2690. Ordnungen des Tierreichs II. Topsent, E. (1894a). Application de la taxonomie actuelle à une col- Vosmaer, G.C.J. (1886 (1887)). Porifera. In H. G. Bronn (Ed.), Die lection de spongiaires du Banc de Campêche et de la Guadeloupe Klassen und Ordnungen des Thierreichs (Vol. 2, pp. i-xii, 1–496). décrite précédemment. Mémoirs de Societé Zoologique de Wapstra, M., & Van Soest, R. W. M. (1987). Sexual reproduction, France, 7,27–36. larval morphology and behaviour in demosponges from the south- Topsent, E. (1894b). Une réforme dans la classificaiton des Halichon- west of the Netherlands. In J. Vacelet & N. Boury-Esnault (Eds.), drina. Mémoires de la Société Zoologique de France, 7,5–26. Taxonomy of Porifera NATO ASI Series (Vol. G13, pp. 281–307). Topsent, E. (1928). Spongiaires de l'Atlantique et de la Méditerranée Berlin: Springer. provenant des croisières du Prince Albert Ier de Monaco. Résul- Wörheide, G. (1998). The reef cave dwelling ultraconservative coral- tats des Campagnes Scientifiques Accomplies sur son Yacht par line demosponge Astrosclera willeyana Lister 1900 from the Albert Ier Prince Souverain de Monaco, 74,1–376. Indo-Pacific - Micromorphology, ultrastructure, biocalcification, Vacelet, J. (1985). Coralline sponges and the evolution of Porifera. In isotope record, taxonomy, biogeography, phylogeny. Facies, 38, S. C. Morris, J. D. George, R. Gibson, & H. M. Platt (Eds.), The 1–88. Origins and Relationships of Lower Invertebrates. 28 (Vol. i–ix, Wörheide, G., & Erpenbeck, D. (2007). DNA taxonomy of sponges— 1–397, pp. 1–13). The Systematics Association, Oxford: progress and perspectives. Journal of the Marine Biological As- Clarendon sociation of the United Kingdom, 87, 1629–1633.