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Evolution of the Radular Apparatus in Conoidea (Gastropoda: Neogastropoda) As Inferred from a Molecular Phylogeny

Evolution of the Radular Apparatus in Conoidea (Gastropoda: Neogastropoda) As Inferred from a Molecular Phylogeny

MALACOLOGIA, 2012, 55(1): 55−90

EVOLUTION OF THE RADULAR APPARATUS IN (: ) AS INFERRED FROM A MOLECULAR PHYLOGENY

Yuri I. Kantor1* & Nicolas Puillandre2

ABSTRACT

The anatomy and evolution of the radular apparatus in predatory marine gastropods of the superfamily Conoidea is reconstructed on the basis of a molecular phylogeny, based on three mitochondrial genes (COI, 12S and 16S) for 102 . A unique feeding mecha- nism involving use of individual marginal radular teeth at the proboscis tip for stabbing and poisoning of prey is here assumed to appear at the earliest stages of evolution of the group. The initial major evolutionary event in Conoidea was the divergence to two main branches. One is characterized by mostly hypodermic marginal teeth and absence of an odontophore, while the other possesses a with primarily duplex marginal teeth, a strong subradular membrane and retains a fully functional odontophore. The radular types that have previously been considered most ancestral, “prototypic” for the group (flat marginal teeth; multicuspid lateral teeth of ; solid recurved teeth of and Duplicaria), were found to be derived conditions. Solid recurved teeth appeared twice, independently, in Conoidea – in and . The Terebridae, the sister group of , are characterized by very high radular variability, and the transformation of the marginal radular teeth within this single clade repeats the evolution of the radular apparatus across the entire Conoidea. Key words: Conoidea, , radula, molecular phylogeny, evolution, feeding mechanisms, morphological convergence, character mapping.

INTRODUCTION the subradular membrane, transferred to the proboscis tip (Figs. 2, 4), held by sphincter(s) Gastropods of the superfamily Conoidea in the buccal tube (Figs. 2, 4 – bts) and used (= Toxoglossa) constitute a hyperdiverse for stabbing and envenomating the prey. Use group of predatory marine snails that includes of marginal teeth at the proboscis tip was in particular the well-studied Conus. observed directly and studied in detail in vari- Conoideans are notable for the possession of ous species of Conus that possess elongate, a large venom gland (Figs. 1–4, vg), together barbed, harpoon-like, hollow marginal teeth with a highly modified radula. (Kohn, 1956; Olivera et al., 1990; Kohn et al., An unusual peculiarity of Conoidea foregut 1999), through which the venom is injected anatomy is that the buccal mass with the radu- into the prey (following Kantor & Taylor, 2000 lar sac is situated at the proboscis base (Fig. 1 we refer to these hollow marginal teeth as – bm, rsod) and the radula cannot be protruded hypodermic). The prey is swallowed whole, through the mouth and used for grabbing and sometimes being similar in size to the preda- rasping the prey. tor itself (e.g., Kantor, 2007). In these cases, The most outstanding character of Conoidea the radular apparatus underwent profound is the unique mechanism of envenomation of transformation and the odontophore completely the prey. Some conoideans were long known to disappeared. Another important character is use individual teeth at the proboscis tip for stab- that the anterior part of the radular diverticulum, bing and injecting into prey (e.g., which is homologous to the sublingual pouch of Kohn, 1956). A marginal is detached from other gastropods, is transformed into a caecum

1A. N. Severtsov Institute of Ecology and Evolution, Russian Academy of Sciences, Leninski Prosp. 33, Moscow 119071, Russia. 2UMR 7138, Muséum National d’Histoire Naturelle, Departement Systematique et Evolution, 55 Rue Buffon, 75231 Paris, France. *Corresponding author: [email protected] 55 56 KANTOR & PUILLANDRE

(“short-arm of the radular sac”), where fully 1). In these conoideans, the marginal teeth formed marginal teeth are stored prior to their can be of very different anatomy, but with few use at the proboscis tip (Taylor et al., 1993). exceptions are not hollow, non-hypodermic. Conversely, in many conoideans the radular In conoideans with non-hypodermic marginal apparatus includes well-developed subradular teeth (and a functional odontophore), a tooth membrane and a fully functional odontophore separated from the rest of the radula was very with muscles, thus suggesting that the radula often (in most preserved specimens examined) still has some (although maybe limited) function found held at the proboscis tip (Figs. 1, 2). Such as a complete organ. As in conoideans with teeth were first recorded in serial histological hypodermic teeth, the radula and odontophore sections of proboscises in several species are situated at the proboscis base and normally of (Cochlespiridae) (Sysoev & Kantor, cannot be protruded through the mouth (Fig. 1987), Drilliidae (Sysoev & Kantor, 1989), and

FIGS. 1−4. Diagrammatic sections through the anterior foregut of Conoidea. FIG. 1: Anterior foregut of the Conoidea with non-hypodermic marginal radular teeth and odontophore (generalized representa- tive of the clade B). A duplex marginal tooth detached from the subradular membrane is used at the proboscis tip for stabbing and envenomating the prey; FIG. 2: Section of the tip of the proboscis with the duplex marginal tooth held by sphincters of the buccal tube (actual specimen of Aforia kupriyanovi Sysoev & Kantor, 1988 – Cochlespiridae); FIG. 3: Anterior foregut of the Conoidea with hypodermic marginal radular teeth and lacking odontophore (generalized representative of clade A). A hypodermic marginal tooth detached from the subradular membrane is used at the proboscis tip; FIG. 4: Section of the tip of the proboscis with the hypodermic marginal tooth held by a sphincter of the buccal tube (actual specimen of wareni Sysoev & Kantor, 1995 – ). Abbreviations: bm – buccal mass; bt – vuccal tube; bts – buccal tube sphincter; dmt – duplex marginal tooth at the proboscis tip; hmt – hypodermic marginal tooth at the proboscis tip; mb – muscular bulb of the venom gland; oe – oesophagus; pr – proboscis; rhs – rhynchostomal sphincter; rs – radular sac without odon- tophore; rsod – radular sac with odontophore; sg – salivary gland; vg – venom gland. EVOLUTION OF RADULA IN CONOIDEA 57 in three additional families, here referred to as Since about 1990, anatomical investigations Turridae, and Pseudomelatomi- of conoideans have revealed great variability dae (Kantor & Taylor, 1991). The base of the in foregut anatomy, and characters defined in tooth was held by special sphincter(s) and/or these studies have been used to unravel phylo- an epithelial pad of the buccal tube. Thus, the genetic relationships (Taylor, 1990; Taylor et al., presence of marginal teeth detached from the 1993; Kantor et al., 1997). Various hypotheses radular membrane and of different morpholo- have been proposed concerning the evolution- gies, from solid duplex to specialized hypoder- ary transformations in radular morphology of mic, used one by one at the proboscis tip for Conoidea (Shimek & Kohn, 1981; Kantor & stabbing the prey can be inferred from anatomi- Taylor, 2000; Kantor, 2006). However, one of cal characters (presence of the sphincters in the reasons for the lack of a clear understand- the buccal tube). ing of major transformations of the radula is Therefore, the feeding mechanism of Conoidea that radula evolution was inferred from phyloge- involves use of single marginal tooth detached netic hypotheses themselves based partially on from the rest of the radula apparatus held at radular morphology (e.g., Taylor et al., 1993). the proboscis tip for stubbing and poisoning As a consequence, many parallel evolutionary the prey either through the central cavity of the transformations cannot be traced. hollow tooth (in hypodermic ones) or through the The rapid development of molecular phyloge- laceration made by the non-hypodermic tooth. netics provided new insight and revolutionary Peculiarities of the feeding have been dis- changes in our understanding of conoidean cussed for different groups of Conoidea (e.g., evolution. DNA sequences were first used to Kantor & Sysoev, 1989; Taylor et. al., 1993; infer phylogenetic relationships within genera Kantor et al., 1997; Kantor & Taylor, 2002), or subfamilies (e.g., Duda & Palumbi, 1999; and several subtypes of feeding mechanisms Espiritu et al., 2001; Duda & Kohn, 2005; Her- have been suggested. Based on the foregut alde et al., 2007; Holford et al., 2009) and then anatomy, the use of the teeth at the proboscis among most of the families and subfamilies (as tip was found improbable in only two groups of erected by Powell, 1942, 1966; McLean, 1971; radular bearing Conoidea that possess the pro- Taylor et al., 1993) of the Conoidea (Puillandre boscis – Strictispiridae, and the clade formed by et al., 2008). An updated molecular phylogeny the genera Pseudomelatoma, Hormospira and based on three mitochondrial genes (COI, 12S Tiariturris, previously recognized as a separate and 16S) and including representatives of 102 (sub)family Pseudomelatomidae (Kantor, 1988; genera was recently proposed (Puillandre et Kantor & Taylor, 1991, 1994). In all others, the al., 2011). The single recognized taxon missing marginal teeth are used at the proboscis tip. from the analysis was the (sub)family Stric- Despite the fact that the Conoidea are one of tispiridae McLean, 1971. Most of the clades the most well-known groups of Neogastropoda inferred have robust support that allowed the from the point of view of anatomy and lately status of the different families and subfamilies molecular phylogeny, data on their feeding and previously proposed to be clarified and led to a diet are still very limited. With the exception of new classification of the group into 15 families Conus, information on diet is available for fewer (Bouchet et al., 2011). than 50 species and involved much less direct The molecular framework provides an op- observation (e.g., Shimek, 1983a–c, Heralde portunity to reconstruct the transformation of et al., 2010). Most of the conoideans (other the morphological characters and to test pre- than Conus) feed on sedentary and errant viously proposed hypotheses. This approach polychaetes, although feeding on other worms has demonstrated the independent loss of (sipunculans and nemerteans) and even mol- the venom gland in two independent lineages luscs has been recorded (Miller, 1989, 1990). of Terebridae (Holford et al., 2009), but is still This information is derived mainly from gut not widely used in Conoidea. Here we attempt content analysis. for the first time a reconstruction of the major Radular anatomy of the Conoidea is highly morphological transformations of the radular variable both in terms of the number of teeth in apparatus in Conoidea based on the molecular a transverse row and in the shape of the teeth. phylogeny. Understanding the transformations For a long time, radula morphology together of the radular apparatus is important not only with shell characters constituted the basis of the for understanding the evolution of the group in higher classification of the group (e.g., Thiele, general, but also because it may provide new 1929; Powell, 1942, 1966; McLean, 1971). insight into the factors leading to hyperdiversifi- 58 KANTOR & PUILLANDRE cation of the group that led to the appearance of The larger radulae were cleaned in a watch probably the most species-rich marine mollusk glass, while most tiny ones – in a drop of water taxon. Furthermore, clarifying the evolution placed on a cover-slip. For latter ones, the of the group, and in particular the evolution bleach was added either with a syringe or a of characters linked to the venom apparatus, minute plastic pipette. After dissolving the soft should be of great value in the discovery of tissues, the radula or separate radular teeth new venom compounds with pharmacologi- were transferred with a needle or single hair into cal applications (Olivera, 2006; Puillandre & a drop of clean water on the same cover-slip. Holford, 2010). This minimizes the chance of losing the small radulae. Two changes of water were usually enough to rinse the radula. After rinsing, the MATERIAL AND METHODS radula was partially pulled out of the drop so that the extruded part adheres to the glass by surface Radula Preparation tension. This permits the radula membrane to be more easily unfolded with a single hair, and Of the 102 ingroup species in the molecular allows individual teeth to be placed in the desired analysis (Puillandre et al., 2011), the radula of position prior to drying. The radula was then 51 species was examined (in most cases using completely pulled out of the water drop and al- the same specimens as used for the molecular lowed to dry. The cover-slip was then mounted analysis), ten species were radula-less, and for on the stub. Although simple, this method pro- 13 species published data were used (Table 1). vides excellent results, allowing manipulating In most cases, unavailability of the radula was objects smaller then 100 μm in length. explained by destruction of the body during DNA extraction (usually for very small speci- Tree Mapping mens). For six species, radular characters were examined using congeners. The generic posi- Within the Neogastropoda, the sister-group tion of most of these was confirmed by other of Conoidea is the rachiglossate group of molecular data. These species are marked by superfamilies (Oliverio & Modica, 2010). Con- an asterisk on the molecular trees (e.g., Figs. sequently, we included in our analyses several 5, 6). The complete range of variability of the outgroups from four families of Rachiglossa: radula in Conoidea is not, however, covered by ( costatum), Harpidae the species in our tree (only a single species of (Harpa kajiyamai), Buccinidae (Belomitra each genus was used in the analysis). Some ) and Fasciolariidae (Turrilatirus species with important or unique radular mor- turritus). Two other distant outgroups were phology are not included in the current analysis, also included: Xenophora solarioides (Xeno- but we inferred their phylogenetic position from phoridae, ) and Laevistrombus separate molecular analyses and therefore guidoi (, Littorinimorpha). mention some of them in the discussion. For the The details of the phylogenetic analysis is outgroups, radula was examined in conspecific provided in Puillandre et al. (2011). specimens or congeners. Photographs of the Nine radular morphological characters coded radulae are arranged in plates on the taxonomi- as 31 character states (Table 2) were used to cal basis to make the comparison easier. reconstruct the radular transformation. Charac- The radulae were cleaned with diluted bleach ters were mapped on the tree of Puillandre et (1 part of commercially available bleach to 3–4 al. (2011) using Mesquite Version 2.74 (Mad- parts of distilled water). Cleaning radulae in dison & Maddison, 2007–2010), with the option bleach does not damage radular teeth or the “tracing character history” and the parsimony subradular membrane if used in the correct ancestral reconstruction method. Most of the concentration and if the radulae are not ex- characters were treated as unordered. For the posed to bleach for a long time. Furthermore, characters describing central and lateral teeth soft tissues are diluted in bleach rapidly (usu- (characters 1 and 2 in Table 2), the stepmatrix ally within a few minutes), allowing continuous model was tried in addition to the unordered; it observation under the microscope that reveals allows interdicting some of the transformation many important features that otherwise can sequences, in our case from absent to present, easily be overlooked, for example, folding of that is interdicting re-appearance of central and the radular membrane, attachment of radular lateral teeth after they had been lost. teeth to the membrane, and presence of a The familiar classification accepted here is ligament. that of Bouchet et al. (2011). For convenience, EVOLUTION OF RADULA IN CONOIDEA 59 (continues) - - - - + + + + + + + + + Radula-less species source of information Radula preparation or 15°140’S, 12°290’E, 50 m 10°510’S, 14°230’E, 5–10 m 20°060’S, 160°230’E, 490–550 m 8°780’S, 13°230’E, 40–60 m 9°33.5’N, 123°49.50’E, 97–120 m 15°31’S, 167°09’E, intertidal 9°41.70’N, 123°50.90’E, 4–12 m 9°43.30’N, 123°48.80’E, 123–135 m 22°47’S, 167°22’E, 395 m 7°45.10’S, 156°56.30’E, 768–890 m 21°060’S, 158°350’E, 637–650 m 9°29.30’N, 123°51.50’E, 95–128 m 8°24.40’S, 159°22.50’E, 897–1,057 m 9°38.20’N, 123°43.50’E, 584–596 m Institutional registration number station and depth Expedition/locality, MNHN IM200911056 Musorstom 4, DW226, MNHN IM200717833AF10, Angola, MNHN IM200717849 EBISCO, DW2619, MNHN IM200717829AF1, Angola, MNHN IM200717904T9, Panglao 2004, MNHN IM200742607 Santo 2006, VM53, MNHN IM200717869 Panglao 2004, S21, MNHN IM200717875T26, Panglao 2004, MNHN IM200717857 EBISCO, CP2551, MNHN IM200742293MNHN IM200717887AF7, Pta. Das Lagostas Angola, T36, Panglao 2004, + Strebel, 1914 MNHN IM200717830AF3, Angola, sp. MNHN IM200717931 Salomon 2, CP2269, sp. MNHN IM200717932 Salomon 2, CP2197 tenera optima (W. Wood, 1828) Wood, (W. cf. cf. (Thiele, 1925) neocaledonica (Hedley, 1922) (Hedley, (Hedley, 1899) (Hedley, (Martens, 1902) (Montrouzier, 1872) (Montrouzier, (C. B. Adams, 1850) (C. B. Puillandre et al., 2010 Nolf & Verstraeten, 2006 Nolf & Verstraeten, Sysoev & Bouchet, 2001 (Sysoev & Bouchet, 2001) mitriformis (cf.) compacta subspirata Gemmuloborsonia colorata xanteni Etrema ravella Zemacies excelsa nigrotincta Tomopleura reevii Typhlomangelia gen. 1 sp. MNHN IM200717911 Panglao 2005, CP2333, Family Genus Species orsonidae Clavatuldae TABLE 1. List of specimens (+ data available, - unavailable). TABLE B Clathurellidae 60 KANTOR & PUILLANDRE (continues) - - + + + + + + McLean, 1971 McLean, 1971 McLean, 1971 source of information Radula preparation or Tucker & Tenorio, 2009 Tenorio, & Tucker 2009 Tenorio, & Tucker 2009 Tenorio, & Tucker 2009 Tenorio, & Tucker 2009 Tenorio, & Tucker 22°50’S, 167°16’E, 496 m South China Sea 8°41.30’N, 123°17.80’E, 150–163 m 8°36.41’N, 79°09.70’W, 73 m 8°36.41’N, 79°09.70’W, 21 m 8°31.83’N, 79°05.09’W, 25–32 m 8°19.50’N, 78°47.71’W, 24–25 m 8°31.37’N, 79°05.79’W, m 9°29.70’N, 123°50.20’E, 110–112 66–68 m 8°26.24’N, 79°09.14’W, 14°59.70’S, 166°55.00’E, 630–705 m 15°32.10’S, 167°16.10’E, 235–271 m 9°29.40’N, 123°44.40’E, 271–318 m 9°29.40’N, 123°56.00’E, 6–8 m Off southern Hong Kong, Off Gulf of Panama, JTD-00-51, Gulf of Panama, JTD-00-47, Gulf of Panama, JTD-00-18, Gulf of Panama, JTD-00-46, Gulf of Panama, JTD-00-34, 2 6 8 3 Institutional registration number station and depth Expedition/locality, MNHN IM200742331 Norfolk 2, DW2147, MNHN IM200717914 Panglao 2005, CP2380, MNHN IM200717920 Panglao 2005, CP2340, MNHN IM200717939 AT87, Santo 2006, MNHN IM200717858 Panglao 2004, S12, HMUK MOEA 20100549 HMUK MOEA 20100541 HMUK MOEA HMUK MOEA 2010055 HMUK MOEA 2010054 HMUK MOEA 2010054 HMUK MOEA NHMUK MOEA 2010054 NHMUK MOEA N N N N N

spurius

(Hinds, 1844) California Monterey, sp. sp. MNHN IM200717701 BOA1, CP2432, (Kuroda, 1956) Philippines Hinds, 1844 MNHN IM200717889T41, Panglao 2004, Azuma, 1972 Philippines (Hinds, 1843) (Kiener, 1845) (Kiener, (Reeve, 1843) (Hedley, 1922) (Hedley, (Roeding, 1798) (M. Smith, 1946) (Schepman, 1913) G. B. Sowerby I, 1833 (Pilsbry & Lowe, 1932) Sysoev & Bouchet, 2001 pudica Benthofascis lozoueti TurriculaCochlespira nelliae pulchella ConusProfundiconusTaranteconus pagoda teramachii chiangi consors ClavusCruziturricula canalicularis arcuata Calliclava CerodrilliaClathrodrillia cybele walteri californicus Sibogasyrinx striata Family Genus Species Drilliidae (continued) Cochlespiridae Conorbidae EVOLUTION OF RADULA IN CONOIDEA 61 (continues) - + + + + + + + + + + + + McLean, 1971 source of information Shimek & Kohn, 1981 Radula preparation or m 9°29.30’N, 123°51.50’E, 95–128 m 20°060’S, 160°220’E, 427–505 m 15°37.7’S, 167°11.3’E, 10 m 15°37.7’S, 167°11.3’E, 21°030’S, 160°440’E, 372–404 m 15°32’S, 167°17’E, 100–105 m 9°29.40’N, 123°56.00’E, 17 m 8°19.50’N, 78°47.71’W, 25–32 m 8°19.50’N, 78°47.71’W, 24–25 m 8°31.37’N, 79°05.79’W, 7°58’S, 157°34’E, 650–836 m 9°36’N, 123°42’E, 609–673 m 8°47’S, 159°38’E, 762–1060 m 9°37.10’N, 123°46.10’E, 3 m 16°37.50’S, 167°57.40’E, 618–641 9°38.80’N, 123°45.90’E, 396–418 m 9°57.53’N, 79°07.71’W, 268–270 m 9°57.53’N, 79°07.71’W, Gulf of Panama, JTD-00-18, Gulf of Panama, JTD-00-46, 0 Institutional registration number station and depth Expedition/locality, MNHN IM200717888T36, Panglao 2004, MNHN IM200742556 Santo 2006, DB25, MNHN IM200717840 EBISCO, DW2631, MNHN IM200742614 Santo 2006, DS99, MNHN IM200717868 Panglao 2004, B19, MNHN IM200717835 BOA1, CP2462, MNHN IM200717900 Panglao 2004, B8, INVEMAR MOL-1929 Colombia, E-73, HMUK MOEA 2010055 HMUK MOEA NHMUK MOEA 20100527 NHMUK MOEA N

sp. MNHN IM200717847 EBISCO, DW2617, sp. MNHN IM200717908 Panglao 2005, CP2332, sp. MNHN IM200742513 Salomon 2, CP2182, coronata trophonoidea cf. sp. (juvenile) MNHN IM200742475 Panglao 2005, CP2396, sp. (juvenile) MNHN IM200742501 Salomon 2, CP2219, (Smith, 1884) (Hinds, 1843) (Garrett, 1873) (Bartsch, 1934) cf. (A. Adams, 1867) (H. Adams, 1872) (Sowerby I, 1834) (Schepman, 1913) (G. B. Sowerby III, 1870) (Mellvill & Standen, 1896) IredaleaSplendrillia pupoidea clavata Anacithara lita Horaiclavidae gen. 1 Anguloclavus Ceritoturris pupiformis enae Horaiclavus splendidus Anticlinura minutissima Imaclava unimaculata Family Genus Species (continued) Horaiclavidae 62 KANTOR & PUILLANDRE (continues) - - + + + + + + + + + + + (Dautzenberg, 1887) used Radula of source of information Radula preparation or powisiana 15°35.7’S, 167°07.4’E, 11 m 15°35.7’S, 167°07.4’E, 11 9°37.10’N, 123°46.10’E, 3 m 9°30.90’N, 123°41.20’E, 90–110 m 9°30.90’N, 123°41.20’E, 90–110 9°41.50’N, 123°51.00’E, 21 m 9°43.30’N, 123°48.80’E, 123–135 m 6°37.20’S, 156°12.70’E, 508–522 m 20°050’S, 160°190’E, 627–741 m 8°19.50’N, 78°47.71’W, 25–32 m 8°19.50’N, 78°47.71’W, South China Sea, Sta. 70 South China Sea, Sta. 70 m 117–148 24°33.47’N, 81°07.72’W, 25–32 m 8°19.50’N, 78°47.71’W, 9°26.90’N, 123°34.50’E, 762–786 m Gulf of Panama, JTD-00-18, southern Hong Kong, Off southern Hong Kong, Off Lower Florida Keys, JTD-01-15, Gulf of Panama, JTD-00-18, 9°30.90’N, 123°41.20’E, 90–110 m 9°30.90’N, 123°41.20’E, 90–110 Institutional registration number station and depth Expedition/locality, MNHN IM200742552 Santo 2006, NR8, MNHN IM200717898 Panglao 2004, B8, MNHN IM200717925 Salomon 2, CP2227, MNHN IM200717755 Panglao 2004, L46, HMUK MOEA 20100533 HMUK MOEA NHMUK MOEA 20100530 NHMUK MOEA 20100556 NHMUK MOEA 20100533 NHMUK MOEA 20100525 NHMUK MOEA N

sp. MNHN IM200717884 Panglao 2004, L46, sp. MNHN IM200742325 Hornsund, Svalbard, 1184–2001 - sp. MNHN IM200717918 Panglao 2005, CP2388, sp. MNHN IM200717851 EBISCO, DW2625, (Dall, 1889) (Hinds, 1843) (Hinds, 1839) (Smith, 1877) (Reeve, 1849) (E. A. Smith, 1882) (E. Pilsbry & Lowe, 1932 Sysoev & Kantor, 1990 Sysoev & Kantor, (Broderip & Sowerby, 1829) (Broderip & Sowerby, metula Heterocithara Mangeliidae gen. 2Lovellona sp.Carinodrillia MNHN IM200717872 Panglao 2004, S26, atramentosa dichroa Mangeliidae gen. 1 sp. pagoda Toxicochlespira MNHN IM200717874T26, Panglao 2004, CheungbeiaComitas robusta HindsiclavaInquisitor alesidota tuberculifera quadrilirata incerta Family Genus Species (continued) Pseudomelato- midae EVOLUTION OF RADULA IN CONOIDEA 63

sp. (continues) - - - - + + + + + + irretita Raphitoma used (Kuroda & Habe mitrellaformis in Habe, 1962) used (Hedley, 1899) used (Hedley, Radula of (Nomura, 1940) used Radula of source of information Radula preparation or Radula of Radula of yoshidai 9°29.40’N, 123°56.00’E, 6–8 m 20°050’S, 160°190’E, 627–741 m 9°36.50’N, 123°45.30’E, 90 m 9°29.40’N, 123°56.10’E, 16 m 9°30.10’N, 123°50.40’E, 100–138 m 21°320’S, 162°300’E, 750–458 m 9°38.50’N, 123°49.20’E, 2–4 m 19°380’S, 158°460’E, 603–630 m Vietnam Vietnam 25–32 m 8°19.50’N, 78°47.71’W, 66–68 m 8°26.24’N, 79°09.14’W, 25–32 m 8°19.50’N, 78°47.71’W, 8°03.50’S, 156°54.50’E, 399–427 m 9°29.60’N, 123°52.50’E, 196–216 m Luong SoN, 15 km N. Nha Trang, Trang, Luong SoN, 15 km N. Nha Gulf of Panama, JTD-00-18, Gulf of Panama, JTD-00-34, Gulf of Panama, JTD-00-18, Institutional USNM 857830 Belize, intertidal Carrie Bow Cay, + registration number station and depth Expedition/locality, MNHN IM200717903 Panglao 2004, S12, MNHN IM200717890 Panglao 2004, L49, MNHN IM200717878 Panglao 2004, B25, MNHN IM200717861 Panglao 2004, B14, NHMUK MOEA 20100539 NHMUK MOEA 20100536 NHMUK MOEA NHMUK MOEA 20100560 NHMUK MOEA 20100540 NHMUK MOEA

Berry, 1958 Berry, sp. MNHN IM200717848 EBISCO, DW2625, sp. MNHN IM200717892T39, Panglao 2004, sp. MNHN IM200717841 EBISCO, CP2648, sp. MNHN IM200717846 EBISCO, CP2600, sp. MNHN IM200717905 Panglao 2005, CP2348, sp. sp. MNHN IM200717927 Salomon 2, CP2260, (Carpenter, 1865) (Carpenter, California 1988 Kantor, (Dall, 1919) (Gray, 1838) (Gray, (C. B. Adams, 1850) (C. B. (Hervier, 1899) (Hervier, (Hervier, 1897) (Hervier, (E. A. Smith, 1885) (E. (Melvill & Standen, 1895) jayana (cf.) Raphitoma rubroapicata cymatodes Gymnobela Ptychobela suturalis Pseudomelatoma moesta Kermia aureotincta Kermia aureotincta Glyphostomoides Hemilienardia calcicincta Otitoma PyrgospiraTiariturrisZonulispira spectabilis aenone Daphnella Family Genus Species (continued) Raphitomidae 64 KANTOR & PUILLANDRE (continues) Veprecula - - - + used (Hedley, 1903) (Hedley, Radula-less species Radula-less species Radula-less species Radula-less species Radula-less species Radula-less species Radula-less species Radula-less species Radula-less species Radula of Kantor & Taylor, 2002 Taylor, Kantor & source of information Radula preparation or vepratica 9°37.4’N, 123°54.5’E, 4–5 m 15°31’S, 167°09’E, intertidal 15°35.40’S, 166°58.70’E, 3–8 m 15°28.7’S, 167°15.2’E, 19 m 9°35.7’N, 123°44.4’E, 0–2 m 9°36.2’N, 123°43.8’E, 382–434 m 9°30.10’N, 123°50.40’E, 100–138 m m 9°30.90’N, 123°41.20’E, 90–110 15°35.2’S, 167°59.4’E, intertidal 8°16.90’S, 159°59.70’E, 464–523 m 15°36.9’S, 167°10.5’E, 6–33 m 9°39.2’N, 123°47.5’E, 255–268 m 15°57’N, 121°50’E, 743 m 21°290’S, 162°360’E, 883–957 m 16°37.50’S, 167°57.40’E, 618–641 m Institutional registration number station and depth Expedition/locality, MNHN IM200730449 Panglao 2004, S2, MNHN IM200730558 Santo 2006, ED5, MNHN IM200717938 Santo 2006, LD28, MNHN IM200730371 Santo 2006, NR5, MNHN IM200716735 Panglao 2005, CP2395, MNHN IM200730459 Panglao 2004, S18, MNHN IM200730401 Panglao 2005, CP2331, MNHN IM200730420 Santo 2006, VM24, MNHN IM200717924 Salomon 2, CP2184, MNHN IM200730480 Santo 2006, FR10, MNHN IM200717845 EBISCO, CP2651,

sp. MNHN IM200717883 Panglao 2004, L46, sp. MNHN IM200742296 Aurora 07, CP2749, sp. MNHN IM200717836 BOA1, CP2462, hyalina cf. (Link, 1807) Hinds, 1844 (Born, 1778) (Deshayes, 1859) MNZ New-Zealand Rudman, 1969 (Hinds, 1844) (Burch, 1965) (Angas, 1877) (Lamarck, 1822) (Linnaeus, 1758) (Deshayes, 1857) (Quoy & Gaimard, 1833) Sysoev & Bouchet, 2001 (cf.) sp. MNHN IM200717891T39, Panglao 2004, TerebraTerenolla textilis pygmaea Strioterebrum plumbum Cinguloterebra cingulifera Oxymeris areolata Clathroterebra fortunei Tritonoturris Veprecula HastulopsisMyurella pertusa kilburni EuterebraHastula tristis strigilata Teretiopsis mirabilis Family Genus Species Terebridae (continued) EVOLUTION OF RADULA IN CONOIDEA 65 sp. Latirus - - + + + + + + + used unpublished Bandel, 1984 Bandel, 1984 Bandel, 1984 A. E. Fedosov, A. E. Fedosov, Radula of source of information Radula preparation or Hughes & Emerson, 1987 15°27.6’S, 167°14.3’E, 6–35 m 15°35.8’S, 167°06.1’E 15°42.7’S, 167°15.1’E, 7 m 15°36.2’S, 167°02.5’E, 80–82 m 15°37.3’S, 167°05.8’E, 78–91 m 15°38.5’S, 167°15.1’E, 13 m 24834.24’N; 81816.64’W, 30–34 m 24834.24’N; 81816.64’W, 22°180’S, 159°280’E, 360–370 m 15°33.6’S, 167°16.6’E, 8–9 m 9°33.30’N, 123°43.90’E, 2 m 9°06’N, 123°25’E, 437–439 m 8°17’S, 160°00’E, 464–523 m 19°330’S, 158°400’E, 400–413 m 9°37.10’N, 123°52.60’E, 8–22 m 38°14’N, 70°29.28’W, 3,188–5,300 m 38°14’N, 70°29.28’W, S. of Bahia Honda Key, Florida Keys, S. of Bahia Honda Key, 20110066 Institutional USNM 832922Atlantic, North NHMUK MOEA NHMUK MOEA registration number station and depth Expedition/locality, MNHN IM200911059 Santo 2006, FB52, MNHN IM200911061 AT55, Santo 2006, MNHN IM200911060 Santo 2006, LR3, MNHN IM200911057 Salomon 2, CP2184, MNHN IM200740569 Santo 2006, EP22, MNHN IM200742448 Panglao 2005, CP2363, MNHN IM200717838 EBISCO, DW2533, MNHN IM200717685 Santo 2006, FS84, MNHN IM200717860 Panglao 2004, R44, MNHN IM200717684 Santo 2006, DR87, MNHN IM200717850 EBISCO, DW2607, MNHN IM200717754 Panglao 2004, R42,

casearia armillata (Gmelin, 1791) MNHN IM200911058 Santo 2006, DR64, (Man in’t Veld & (Man in’t Veld cf. cf. (Perry, 1811) (Perry, 1811) (Perry, (Haas, 1949) Iredale, 1929 (Reeve, 1845) (Reeve, 1845) (Gmelin, 1791) Linnaeus, 1758 De Turck, 1998) De Turck, (Lamarck, 1822) (Schepman, 1913) Habe & Kosuge, 1970 Kuroda & Oyama, 1971 (Hedley & Petterd, 1906) Xenophora solarioides LaevistrombusTurrilatirus guidoi turritus Vexillum costatum Harpa kajiyamai Belomitra brachytoma Ptychosyrinx carynae IotyrrisLophiotomaLucerapex Polystira cingulifera acuta albida Gemmula rarimaculata TurrisXenuroturris babylonia legitima Family Genus Species Outgroups Turridae (continued) 66 KANTOR & PUILLANDRE

TABLE 2. List of characters states used in the analysis.

Character states

No. Charater description 01 2 34

1 morphology of central absent multicuspid of multicuspid of unicuspid unicuspid tooth caenogastro- neogastropod narrow broad pod type 2 morphology of the absent cuspidate of cuspidate of unicuspid plate-like lateral teeth caenogastro- neogastropod of neogastro- pod type type pod type 3 morphology of the absent non-duplex duplex or solid, recurved hypodermic marginal teeth (flat) semi-enrolled 4 duplex marginal teeth marginal teeth with un- teeth with sub- teeth of semi-enrolled teeth absent equal limbs equal limbs type 5 details of hypodermic absent present hypodermic marginal teeth - spur teeth absent 6 details of hypodermic absent one barb two barbs three or more hypodermic marginal teeth – present present barbs present teeth absent barbs 7 details of hypoder- absent present hypodermic mic marginal teeth teeth absent - blade 8 details of hypodermic absent present hypodermic marginal teeth – liga- teeth absent ment 9 use of marginal teeth separate tooth separate tooth marginal teeth at proboscis tip used at the not used at the absent proboscis tip proboscis tip in addition to the families recognized by RESULTS Bouchet et al. (2011), we refer to major clades A and B (without attributing a taxonomic status Although the radulae of Conoidea have been to them) that are different in many aspects of described in many publications, the thorough anatomy and radular morphology. use of scanning electron microscopy revealed many previously overlooked characters and al- Acronyms for Depositories of Voucher Speci- lowed new interpretations of structures already mens described. Therefore, we provide here a much updated overview of the radular characters. INVEMAR Instituto de Investigaciones Marinas y The evolution of each of the nine characters Costeros, Santa Marta, Colombia analysed is described. MNHN Muséum National d’Histoire Natu- relle, Paris, France Character 1: Central Tooth of the Radula (Figs. MNZ Museum of New Zealand Te Papa 5, 6) Tongareva, Wellington, New Zea- land A central tooth is present in all outgroups. It is NHMUK The Natural History Museum, Lon- absent in clade A and present in some groups don, U.K. of clade B (Figs. 7–41). The morphology of the USNM National Museum of Natural History, central tooth is rather variable in gastropods. Smithsonian Institution, Washington Two major types are found in the outgroups, D.C., U.S.A. both multicuspid. Describing in detail the EVOLUTION OF RADULA IN CONOIDEA 67

FIG. 5. Evolution of the central radular tooth morphology (character 1) mapped on the conoidean molecular phylogeny (Puillandre et al., 2011). Character states are treated as unordered. The different shading and tiling of the branches corresponds to the most parsimonious ancestral state for the corresponding clade. The mixed shading and/or tiling of the branch indicates that analysis was not able to resolve the single most parsimonious state. 68 KANTOR & PUILLANDRE

FIG. 6. Evolution of the central radular tooth morphology mapped on the conoidean molecular phylogeny (character 1). A stepmatrix parsimony model was used, interdicting reversion of the character states. Since central teeth are absent in the entire clade A, only clade B is illustrated. morphology of the teeth of groups other than tively three teeth – vestigial central and vestigial Conoidea (e.g., Bandel, 1984) is outside the laterals (see below) partially or completely fused scope of the current paper. In Conoidea with a (Figs. 17, 18). Vestigial structures were found central tooth, its form varies widely. Two major in some Pseudomelatomidae, Clavatulidae and types of well-defined central teeth can be identi- Turridae (Iotyrris Medinskaya & Sysoev, 2001). fied: narrow unicuspid (shield-like with a small Narrow unicuspid central teeth are found cusp and sometimes with additional serrations) in most Drilliidae and some Turridae (in our (Figs. 7–9, 12, 13); and broad unicuspid, with dataset exemplified by species of Xenuroturris a large curved cusp and well-defined lateral Iredale, 1929, and Turridrupa Hedley, 1922 – flaps (Fig. 14). Figs. 33, 34, as well as Gemmula and ). A Conversely, in a number of Conoidea from broad, well-defined central tooth was recorded clade B, vestigial rather indistinct structure(s) in three clades – in some Pseudomelatomidae occupy the middle portion of the subradular (Pseudomelatoma, Hormospira Berry, 1958, membrane (Figs. 28, 30 – marked with arrows). and Tiariturris Berry, 1958 – Fig. 14), Co- In some cases, they can be hardly seen without chlespiridae (Fig. 39), and Gemmuloborsonia staining the radular membrane or observing Shuto, 1989 (not currently attributed to any under SEM. These structures may be either a family – Fig. 32). In these genera, the posterior much reduced broad central tooth with lateral edge of the tooth, bearing the cusp, is well flaps, with or without a central cusp, or alterna- elevated over the membrane. EVOLUTION OF RADULA IN CONOIDEA 69

FIGS. 7−13. Radulae of Drilliidae. If not otherwise mentioned, data for the specimens are given in Table 2. FIGS. 7, 8: sp., MNHN IM200717847; FIG. 9: exasperatus (Reeve, 1843), MNHN, New-Caledonia, LIFOU 2000, st. 1420, 20°47.7’S, 167°09.35’E, 4−5m; FIG. 10: Imaclava pilsbryi (Bartsch, 1950), after Kantor & Taylor, 2000; FIG. 11: Cruziturricula arcuata (Reeve, 1843), NHMUK MOEA 20100541. Semi-enrolled marginal teeth; FIGS. 12, 13: Clavus sp. 3, MNHN uncataloged, BA- THUS 2, DW714; FIG. 12: Bending plane of the radula, arrows indicate strong wear of the teeth; FIG. 13: Central part of the same radula, showing intact central and lateral teeth. Abbreviations: ct – central tooth; lt – lateral tooth, mt – marginal tooth. 70 KANTOR & PUILLANDRE EVOLUTION OF RADULA IN CONOIDEA 71

Four analyses were performed. In the first, the Character 2: Lateral Teeth of the Radula (Fig. inconspicuous central teeth, when recognized, 42) were considered as unicuspid narrow (lateral flaps were considered as vestigial plate-like Lateral teeth are present in all outgroups. lateral teeth), and character states were unor- They are absent in Clade A and present in some dered (Fig. 5). The tree was 13 steps long and groups of Clade B. suggested that the plesiomorphic condition of There are two major types of lateral teeth the character in clade B is the absence of the among the ingroup species in our tree. In central tooth and the central teeth originated Drilliidae, they are well formed and multicuspid, independently in eight clades. Since central completely separate from the central tooth teeth are present in the outgroups, this can be (Figs. 7–10). In all others (some Pseudome- considered a reversion. However, the presence latomidae, Turridae and Clavatulidae), they of numerous reversions within clade B seems are very weak, plate-like, non-cuspidate and rather unlikely. usually completely or partially fused with the Therefore, the second analysis was per- central tooth (when it is present), forming the formed with reversions interdicted (stepmatrix “central formation” (Kantor, 2006) (see discus- parsimony model) (Fig. 6). The tree was 18 sion below). In some groups, the laterals are so steps long (five steps longer), and the analy- weak that their presence can be revealed only sis did not allow reconstruction of the single by staining of the subradular membrane. This most parsimonious state in Clade B, since a is particularly characteristic in Clavatulidae, multicuspid tooth (characteristic for Neogas- in which they were first revealed by Kilburn tropoda), a narrow unicupsid tooth and a broad (1985). unicuspid tooth are equally parsimonious. This A first analysis with character states unor- analysis suggested independent losses of dered suggested the absence of lateral teeth central teeth in several clades: most species is ancestral for the Conoidea and independent of Pseudomelatomidae, Leucosyrinx Dall 1889, appearance of the lateral teeth occurred inde- Horaclavidae and Terebridae, as well as in pendently in five clades (all in clade B). Since some species of Turridae and Clavatulidae. lateral teeth are present in outgroups, these The third and fourth analyses were with events would be considered as a reversions. alternative coding of the characters and with The second analysis was performed with unordered and stepmatrix parsimony models reversions interdicted (Fig. 42) and resulted in correspondingly. Species with vestigial central a longer tree (17 steps vs. nine in the previous structures were coded as having the broad analysis). The analysis did not allow recon- unicuspid teeth. The reconstruction produced struction of the single most parsimonious state longer trees (15 and 20 steps, respectively), for the entire Conoidea nor for Clade B, since which were therefore rejected. multicuspid neogastropod type teeth, unicuspid

→ FIGS. 14−26. Radulae of Pseudomelatomidae. If not otherwise mentioned, data for the specimens are given in Table 2. FIG. 14: Tiariturris spectabilis Berry, 1958, NHMUK MOEA 20100540. Radula with solid recurved marginal teeth and broad central teeth; FIG. 15: Comitas onokeana vivens Dell, 1956, MNHN, New-Caledonia, MONTROUZIER, st. 1269, after Kantor & Taylor, 2000. Radula with paired plate-like lateral teeth; FIGS. 16, 17: Comitas sp., MNHN IM200717918; FIG. 17: Enlarged central segment of the radula; FIG. 18: Knefastia tuberculifera (Broderip & Sowerby, 1829), NHMUK MOEA 20100533. Enlarged central segment of the radula; FIGS. 19, 20: (Sowerby, 1834). Costa Rica, Off Nacascola, west side of Bahia Culebra, after Kantor et al., 1997. Radula with paired plate-like lateral teeth; FIG. 21: sp., NHMUK MOEA 20100536. Radula with semi- enrolled marginal teeth; FIG. 22: Ptychobela suturalis (Gray, 1838), NHMUK MOEA 20100560. Radula with semi-enrolled marginal teeth; FIGS. 23−26: Diagrammatic transverse sections of different duplex marginal teeth. Black horizontal line represents the subradular membrane; FIG. 23: Comitas-type; FIG. 24: Typical duplex tooth; FIG. 25: Semienrolled tooth of Zonulispira; FIG. 26: Semienrolled tooth of Ptychobela. Abbreviations: al – accessory limb; ct – central tooth; lt – lateral tooth; ml – major limb; vlt – vestigial lateral tooth. 72 KANTOR & PUILLANDRE

FIGS. 27−32. Radulae of Clavatulidae. If not otherwise mentioned, data for the specimens are given in Table 2. FIGS. 27, 28: Pusionella compacta Strebel, 1914, MNHN IM200717830; FIG. 28: Enlarged central segment of the radula with vestigial lateral teeth; FIGS. 29, 30: Clavatula xanteni Nolf & Verstra- eten, 2006, MNHN IM200717829; FIG. 30: Enlarged central segment of the radula. Arrow indicates the narrow central tooth; FIG. 31: Toxiclionella tumida (Sowerby, 1870), South Africa, after Kantor &Taylor (2000). Semi-enrolled marginal teeth. Left upper corner – diagrammatic section of the tooth. Subradular membrane was strongly damaged during the preparation of the radula; FIG. 32: Gemmuloborsonia colorata (Sysoev & Bouchet, 2001), MNHN IM200717849. Abbreviations: vlt – vestigial lateral teeth; ct – central tooth. EVOLUTION OF RADULA IN CONOIDEA 73

FIGS. 33, 34: Radulae of Turridae; FIGS. 35−38: Radulae of Pseudomelatomidae; FIGS. 39−41: Radulae of Cochlespiridae. All with different duplex marginal teeth. If not otherwise mentioned, data for the speci- mens are given in Table 2; FIG. 33: Xenuroturris legitima Iredale, 1929, MNHN IM200717684; FIG. 34: Turridrupa acutigemmata (E. A. Smith, 1877), MNHN uncataloged, . Radula with narrow central and plate-like lateral teeth; FIG. 35: Carinodrillia dichroa Pilsbry & Lowe, 1932, NHMUK MOEA 20100530; FIG. 36: Funa incerta (Smith, 1877), NHMUK MOEA 20100554; FIG. 37: Cheungbeia robusta (Hinds, 1839), NHMUK MOEA 20100557; FIG. 38: sp., MNHN IM200717851; FIGS. 39, 40: Cochlespira radiata (Dall, 1889), MNHN, SE Brazil, after Kantor & Taylor, 2000; FIG. 41: Sibogasyrinx sp., MNHN IM200717701. Abbreviations: al – accessory limb of the marginal duplex tooth; ct – central tooth; lt –lateral tooth; ml – major limb of the marginal duplex tooth. 74 KANTOR & PUILLANDRE

FIG. 42. Evolution of the lateral radular tooth morphology (character 2) mapped on the conoidean molecular phylogeny (Puillandre et al., 2011). A stepmatrix parsimony model was used, interdicting reversion of the character states. EVOLUTION OF RADULA IN CONOIDEA 75

FIG. 43. Evolution of marginal tooth morphology (character 3) mapped on the conoidean molecular phylogeny (Puillandre et al., 2011). Character states are treated as unordered. 76 KANTOR & PUILLANDRE neogastropod type teeth and plate-like lateral nearly equally developed and the teeth teeth were equally parsimonious. attain a trough-shape, becoming “semi- For most of clade B conoideans (except Co- enrolled” (see below, Figs. 25, 26). In the chlespiridae) the most parsimonious state was analysis, this condition was also coded plate-like teeth, while multicuspid teeth seem to as “duplex teeth”. Very similar teeth were appear in the branch that combines Drilliidae recorded in one species of Terebridae, and Pseudomelatomidae. Lateral teeth are Pelifronia jungi (Fig. 44), not present in independently lost in several lineages – in most our analysis (see below). species of Pseudomelatomidae, Horaiclavidae, (b) flat simple plate-like teeth (some Drilliidae) Terebridae and others. (Fig. 7). In all the species in our tree, the presence of (c) solid, recurved teeth, attached to the mem- the lateral teeth was combined with the pres- brane along part of the length, sometimes ence of the central tooth, which is not the case with a slightly broadened base that is for all conoideans (see discussion). actually attached to the membrane (some Pseudomelatomidae – Pseudomelatoma, Character 3: Morphology of the Marginal Teeth Hormospira Berry, 1958, and Tiariturris (Fig. 43) Berry, 1958 – Fig. 14; some Terebridae – Euterebra and Duplicaria – Fig. 45, the Despite great variability of the marginal teeth in latter not represented in our tree). Conoidea, four major types can be recognized: (d) hypodermic teeth. These are hollow en- (a) duplex teeth, consisting of a major element rolled teeth attached to the subradular (limb) attached to the subradular mem- membrane only by a narrow base or brane along most of its length (Figs. 23, through a flexible stalk, the ligament (some 24 – ml) and the accessory limb, that is the Borsoniidae, Conidae and others). thickened edge of the major element, usu- The analysis was not able to resolve the ally more or less elevated above the mem- single most parsimonious state for the entire brane (Figs. 23–25 – al). These teeth vary Conoidea, but suggested that a duplex tooth is greatly in shape (Kantor & Taylor, 2000) the most parsimonious state for clade B. Flat (Figs. 27, 29, 32–41) and have often been teeth are characteristic only for some Drilliidae referred to as “wishbone” (e.g., Powell, and according to the tree they are an autapo- 1966). The term was coined based on the morphy of several species, thus suggesting misconception that the limbs are separate their derivation from duplex teeth. Similarly, and the tooth is actually bifurcating, as it solid recurved teeth originated from duplex appears under the light microscope (most teeth twice independently in the evolutionary of Clade B). In some cases, the limbs are history of Conoidea – in some Pseudomelato-

FIGS. 44, 45. Radula of Terebridae with non-hypodermic teeth. If not otherwise mentioned, data for the specimens are given in Table 1. FIG. 44: Pellifronia jungi (Lai, 2001), MNHN 30591, Salomon 2, CP2195; FIG. 45: Duplicaria bernardi (Deshayes, 1857), Venus Bank, off NE end of Moreton Island, Moreton Bay, , Australia, 27°02’069”S, 153°19’00”E, 3.5−4.8 m, leg. Glover, Taylor, 2008. EVOLUTION OF RADULA IN CONOIDEA 77 midae and Terebridae. Hypodermic teeth are not allow more detailed analysis, although the a synapomorphy of clade A but also appeared general patterns can be traced. independently in Terebridae. The marginal We recognize four subtypes of duplex teeth, teeth have been also lost several times inde- although much more variation can be found in pendently (at least three times in clade B and other Conoidea not included in our study. twice in Terebridae). The first subtype is characterized by equal or nearly subequal development of major and Character 4: Morphology of Duplex Marginal accessory limbs. This type of tooth is found in Teeth (Fig. 46) Cochlespiridae (Figs. 39, 40) (represented only by two genera in our tree) in which the teeth Duplex teeth are very variable in morphology. are characterized by relatively large size of the The difference in appearance is mainly deter- accessory limb (Fig. 40 – al) that is of nearly the mined by relative size and shape of the acces- same size as the major limb (Fig. 40 – ml). This sory limb, as well as the degree of its elevation produces the appearance of the tooth folded above the surface of the subradular membrane. lengthwise. The analysis suggested that it is The representation of the taxa in our tree does an apomorphy of the clade.

FIG. 46. Evolutionary transformations of duplex marginal tooth morphology (character 4) mapped on the conoidean molecular phylogeny (Puillandre et al., 2011). Character states are treated as unordered. Since duplex marginal teeth are absent in the entire clade A, only clade B is illustrated. 78 KANTOR & PUILLANDRE

A similar subtype, although having a differ- Bartsch, 1950, and McLean, 1971 ent appearance, is the so-called semi-enrolled – Fig. 21, and independently in Ptychobela tooth (Taylor et al., 1993; Kantor & Taylor, Thiele, 1925 (Fig. 22), Cruziturricula Marks, 2000). In this type, the accessory limb is also 1951 (Fig. 11), Imaclava Bartsch, 1944, and subequal in size to the major limb (Figs. 25, 26), Iotyrris (Turridae). but the lengthwise folding is much less tight and The most parsimonious plesiomorphic state the teeth attain a trough-like shape. According for most of clade B (except Cochlespiridae) is to the analysis, this type of tooth appeared the duplex marginal tooth with unequal sizes several times independently in clade B – in of the major, larger limb and smaller accessory some genera of Pseudomelatomidae (in the limb (“unequal limbs” in Fig. 46). Depending on clade Pilsbryspira McLean, 1971, Zonulispira the degree of difference, the tooth may look

FIGS. 47, 48: Radula of Terebridae; FIGS: 49−52: Various hypodermic teeth from species of clade. If not otherwise mentioned, data for the specimens are given in Table 2; FIG. 47: Cinguloterebra cingulifera (Lamarck, 1822), MNHN 30565, Panglao 05, st. CP2340; FIG. 48: Impages hectica (Linnaeus, 1758), MNHN, uncataloged, Philippines, Panglao Island, Alona Beach, intertidal, 2004; FIG. 49: Mangelia powi- siana (Dautzenberg, 1887). Plymouth, England, after Taylor et al. (1993); FIG. 50: Bathytoma neocale- donica Puillandre et al., 2010, MNHN IM200717857; FIG. 51: Genota mitriformis (Wood, 1828), MNHN IM200742293; FIG. 52: Benthomangelia trophonoidea (Schepman, 1913), MNHN IM200717835. EVOLUTION OF RADULA IN CONOIDEA 79 very different, in its most extreme state being limb thickened at the tip and along one side, nearly flat with a narrow and very slightly raised while the accessory limb is represented by the accessory limb (e.g., Funa Kilburn, 1988 – Fig. narrow thickened margin of the tooth that does 36). In most groups, the accessory limb is not reach the tip of the tooth but is inserted in a comparatively large and the tooth edge adjoin- shallow and narrow socket, slightly overlaying ing the limb is significantly raised above the the thickened part of the major limb. membrane, so that the accessory limb occupies the dorsal position on the major limb (e.g., 20, Several Characters (5–8) that Apply to Hypo- 38). Different teeth of this subtype have been dermic Teeth Only (Figs. 47–52) thoroughly illustrated by Kantor et al. (1997) and Taylor et al. (1993). Hypodermic teeth are hollow enrolled margin- A characteristic type of duplex tooth is found als, usually with overlapping edges (exemp- in the genera Comitas Finlay, 1926, and Kne- tions are some representatives of Mangeliidae fastia Dall, 1919 (Figs. 16, 23). The teeth are – Fig. 49, not present in our dataset) and open nearly flat, broadly elongate, with the major at both the tooth base and near the tip. Teeth

FIG. 53. Presence or absence of a spur on marginal teeth (character 5) mapped on the conoidean molecular phylogeny (Puillandre et al., 2011). Character states are treated as unordered. The spur is absent in most of clade B, except some Terebridae. Therefore for clarity only Terebridae from clade B are shown. 80 KANTOR & PUILLANDRE of this morphology are found mostly among tip during feeding and thus to prevent pre- representatives of clade A and in most radulate mature loss of the tooth from the proboscis Terebridae. Nevertheless, in at least one genus (Kohn et al., 1999). Our analysis suggested of Clavatulidae (Toxiclionella Powell, 1966) and several independent origins of this character in Cruziturricula, the marginal teeth are very – in Conidae, Borsoniidae, Mangeliidae and similar in anatomy (Figs. 11, 31). The major Terebridae (Fig. 53). difference between hypodermic teeth in clade A and Terebridae on one hand and in Toxiclio- Character 6: Presence of a Barb(s) (Fig. 54) nella and Cruziturricula on the other is the form of attachment to the radular membrane. In the A barb is a projection from the shaft of the former, the teeth are attached only by the base, tooth that has a cutting edge and joins the while in the latter along most of their length. shaft at an acute angle (Fig. 51). There can be This suggests different evolutionary origins of from 0 to 5 barbs (Conus californicus) (Kohn such teeth (Kantor & Taylor, 2000). et al., 1999). The analysis suggests that the High congruence was found between feeding barbs appeared independently in every family type and hypodermic tooth anatomy in Conus of clade A except Mitromorphidae, in which (e.g., Duda et al., 2001; Espino et al., 2008). they are absent. At the same time, there is a limited number of characters that are widespread across multiple Character 7: Presence of a Blade (Fig. 55) families possessing hypodermic teeth. The blade is a projection from the shaft of the Character 5: Presence of a Spur (Fig. 53) tooth that has a cutting edge and joins the shaft at an obtuse angle (Kohn et al., 1999) (Fig. 52). The basal spur is an anterior projection on In some cases, the distinction between a barb the base of the tooth (Fig. 52). Its function is and a blade is subtle. The analysis suggested probably to tighten the grasp of the proboscis that a blade originated independently twice – in

FIG. 54. Presence or absence of barbs on marginal teeth (character 6) mapped on the conoidean molecular phylogeny (Puillandre et al., 2011). Character states are treated as unordered. The barb(s) are present only in clade A and therefore clade B is omitted. EVOLUTION OF RADULA IN CONOIDEA 81 clade A and in Terebridae. Presence of a blade A ligament has so far only been recorded in was the most parsimonious ancestral state for clade A, and its presence is the most parsimo- clade A (Fig. 55). nious ancestral state for the clade. It is pres- ent in Conidae, Borsoniidae, Mitromorphinae Character 8: Presence of a Ligament (Fig. and at least in one species of Raphitomidae 56) ( – see Taylor et al., 1993: fig. 23 c). The ligament is an elongate, flexible stalk, attached to the base of the tooth and to the Character 9: Use of Marginal Teeth at the Pro- membrane, when the latter is present (Fig. 50). boscis Tip for Stabbing Prey (Fig. 57) In fresh radulae, the ligament can be subcir- cular in cross-section, but when air-dried the The analysis suggested that the use of ligament is usually flat and membrane-like. The marginal teeth at the proboscis tip is the most presence of a ligament is often not recorded parsimonious plesiomorphic state for the entire during radula description. Conoidea.

FIG. 55. Presence or absence of blade on marginal teeth (character 7) mapped on the conoidean molecular phylogeny (Puillandre et al., 2011). Character states are treated as unordered. The blade is absent in most of clade B, except some Terebridae. Therefore for clarity only Terebridae from clade B are shown. 82 KANTOR & PUILLANDRE

FIG. 56. Presence or absence of ligament of marginal teeth (character 8) mapped on the conoidean molecular phylogeny (Puillandre et al., 2011). Character states are treated as unordered. The ligament is present only in clade A and therefore clade B is omitted.

DISCUSSION to extrapolate to the remaining members of the clade. The analyses clearly suggested that Origin of the Conoidean Feeding Mechanism contrary to the hypothesis of Taylor et al. (1993) and General Evolutionary Trends the origin of the peculiar feeding mechanism was not a gradual process but is an apomorphy Use of separate marginal teeth one at a time of Conoidea in general, and it appeared before after their detachment from the rest of the the divergence of the two major clades (A and radular apparatus at the proboscis tip is one B) (Fig. 57). of the most intriguing characters of conoidean The initial divergence of Conoidea into clades evolution. Taylor et al. (1993) suggested that A with primarily hypodermic, and B, with pri- conoidean feeding mechanisms gradually marily duplex marginal teeth is an unexpected evolved within the group, but appeared already inference from the conoidean molecular phy- at early stages of conoidean evolution. We logeny. In previous cladistic analyses based on traced on a molecular tree the possibility of use anatomical characters mainly of the anterior of individual marginal teeth at the proboscis foregut, the representatives of clade A (referred tip using available published and unpublished to as family Conidae by Taylor et al., 1993) anatomical data. As was mentioned in the appeared as a terminal clade, suggesting the introduction, the base of the tooth is held by gradual transformation of radular morphology. special sphincter(s) and/or an epithelial pad The molecular-based results contradict this of the buccal tube. Thus, use of teeth at the hypothesis. proboscis tip can be inferred from anatomical The marginal teeth used one at a time after characters (presence of the sphincters of the their detachment from the rest of the radular buccal tube). Although we do not have ana- apparatus at the proboscis tip both in clades A tomical data for every species included in our and B and therefore there is no fundamental analysis, they are available for species of most difference in feeding mechanism between of the genera and for every family, allowing us these clades. Nevertheless, there are impor- EVOLUTION OF RADULA IN CONOIDEA 83

FIG. 57. Presence or absence of use of the separate marginal teeth on the proboscis tip (character 9) mapped on the conoidean molecular phylogeny (Puillandre et al., 2011). Character states are treated as unordered. 84 KANTOR & PUILLANDRE tant differences in the anatomy of the radular The solid recurved teeth were previously apparatus. In clade B, the radular apparatus considered as the prototype marginal teeth consists of a more or less well-developed in Conoidea (e.g., Kantor & Sysoev, 1990) odontophore with supporting musculature (it is and with the exception of three genera, which absent only in a few species that lack a radula, were united in the Pseudomelatomidae (sensu e.g., Horaiclavus phaeocercus Sysoev, 2008, Taylor et al., 1993), were also found in some Horaiclavidae – Fedosov & Kantor, 2008), mod- Terebridae (genera Duplicaria and Euterebra). erately strong continuous radular membrane Plotting the character on a molecular tree and (often) presence of central and/or lateral clearly indicated that this tooth type appeared teeth. An important character of the radula independently twice in Conoidea. Analysis sug- is that the marginal teeth are attached to the gested that this type of marginal tooth devel- membrane along a significant or even most of oped secondarily from duplex teeth. Despite the their length. Exceptions are some Terebridae general similarity of shape of the solid recurved (discussed below). teeth in Pseudomelatomidae and Terebridae, In clade A an odontophore with muscles is some important differences can also be men- absent, and the subradular membrane is very tioned. In Pseudomelatomidae sensu Taylor et thin to vestigial, although keeps the integrity of al., 1993, the radula is long (about 100 rows the radula in the radular diverticulum. The teeth of teeth in Tiariturris) and possesses large and are attached to the membrane only by the very broad, unicuspid central teeth; in Terebridae narrow base of the tooth, sometimes through a with these solid recurved teeth the radula is flexible stalk – the ligament. The attachment of short (about 20 rows of teeth) and lacks the the marginal tooth (of hypodermic type) to the central teeth. membrane only by the base facilitates rolling of The functioning of this type of radula remains the tooth, which may be formed by a few com- largely unknown. The shape of the rather pletely overlapping rolls in Conidae (for more strongly recurved teeth precludes their use one details: Kantor & Taylor, 2000), although usually at a time at the proboscis tip. In Duplicaria and the edges of the tooth plate are only slightly Euterebra, the venom gland as well as probos- overlapping (Fig. 50 – cross sections through cis is absent, while the odontophore is present the tooth). This attachment to the membrane (Rudman, 1969; Taylor, 1990). In Pseudome- only by the base of the tooth probably also latoma and Hormospira, the venom gland is facilitates the important process of complete present and well developed, the proboscis is separation of the tooth during feeding. long, but the buccal tube lacks the sphincter Thus, the molecular analysis suggests that ap- that can hold a single tooth (Kantor, 1988). pearance of the conoidean feeding mechanism The three genera comprising Pseudome- was the key apomorphy of the superfamily. We latomidae sensu Taylor et al., 1993 (and suggest that it may have greatly improved prey encompassing only six Recent species), have capture and allowed rapid diversification and very limited distribution – all are found in the species radiation that resulted in the modern Panamic province. It is possible that they hyperdiverse group that includes about 4,600 comprise a local radiation connected to a shift Recent described species and a larger number to some peculiar type of prey, although this of still unnamed taxa (Bouchet et al., 2009). needs further confirmation. The presence of a The splitting of Conoidea into two groups with large odontophore suggests that radulae with different radular types and foregut anatomies solid recurved marginal teeth are primarily was the first major evolutionary event, taking used as an entire organ (probably for tearing place at the earliest stage of evolution of the and rasping the prey). In contrast, Terebridae group after the initial appearance of the unique with this type of radula (Taylor, 1990) have a feeding mechanism. broad distribution in the Indo-Pacific – species are found from South Africa to Japan, includ- Transformation of the Non-Hypodermic Marginal ing the and Oman, and some Teeth (clade B) species have a broad Indo-Pacific distribution. They lack a proboscis and venom gland. There Non-hypodermic (non-hollow) marginal teeth are data for diet of only one species with this are found in clade B and are very variable, type of radula, Terebra nassoides, feeding on although they can be reduced to three major capitellid polychaetes (Taylor, 1990). Poly- morphological types – duplex (including semi-en- chaetes of this family are widely distributed in rolled), solid recurved and flat simple plate-like. the World Ocean. EVOLUTION OF RADULA IN CONOIDEA 85

Flat marginal teeth have also been consid- having two barbs at the tip and a subterminally ered as a prototype for the duplex teeth (Taylor opening tooth canal. Nevertheless, contrary to et al., 1993) and were found in some Drilliidae hypodermic teeth of the species of the clade A, (among the genera used in our analysis, in the marginal teeth in Toxiclionella are attached Agladrillia, Splendrillia and ). The to the membrane along most of their length, analysis demonstrates that this condition is similarly to other species of clade B. autapomorphic and this type of tooth originated by weakening and reduction of accessory limb Transformations of the Central Radular Segment of the duplex teeth, the presence of which is (Clade B) the plesiomorphic state for the entire clade B and Drilliidae in particular. In all Conoidea, the buccal mass with the Within the most common duplex type of mar- radular diverticulum (and odontophore with ginal teeth, many different morphologies can muscles in clade B) is situated at the pro- be recognized (Figs. 16–21, 33–41), although boscis base and often behind proboscis in its they are very similar in mode of formation. contracted state. The odontophore cannot be Kantor & Taylor (2000) studied maturing teeth protruded through the mouth and therefore the along the radular membrane and showed that radula can function in most conoideans as an they develop from a flat plate by thickening of integrated whole organ only in the buccal cav- the tooth edges and elevation of the posterior ity, but not for grasping and tearing the prey. A edge (additional limb) from the membrane. few exceptions occur sporadically in different This thickening of the margins of duplex teeth, clades. In these groups, the buccal mass is folding along the length and partial enrolling secondarily shifted anteriorly or is able to evert (in the semi-enrolled teeth) can be explained through the mouth along with the walls of the as features that provide mechanical strength. buccal tube (e.g., in Funa latisinuata – Taylor In mechanical terms, a simple flat plate is less et al., 1993: fig. 14). stiff and more likely to buckle when subject to The limited functioning of the radula as an a compressive force, than one with thickened integrated organ in adults is indirectly confirmed edges or a hollow (Wainwright et al., by absence of worn teeth (marginal, and central 1976). Teeth used at the proboscis tip need to or laterals when present) in radulae examined be rigid to pierce the prey’s integument. by us (except Drilliidae, see below). Odonto- Although marginal tooth shape appeared to phore in general is rather small (sometimes its be rather homoplastic, some of the well-defined presence can be confirmed only on serial sec- clades have characteristic tooth shapes. For tions), with poorly developed muscles. Thus, example, in Cochlespiridae the accessory the central and lateral teeth are hardly function- limb is large, nearly equal in size to the ma- al in Conoidea (while the marginals detached jor limb. Therefore, the tooth appears to be from the membrane are used individually at the folded lengthwise with a solid tip. Clavatulidae proboscis tip) and therefore the adaptive value also possess rather distinctive duplex teeth of morphological transformations of central and with a sharp-edged major limb and a deep lateral teeth may be reduced. This explains the socket where an accessory limb is inserted, high variability of the morphology of the central often with angulation distal to the socket. The segment of the radula that includes the central well-supported clade including Pilsbryspira, and lateral teeth. Zonulispira, and Pyrgospira is characterized The evolutionary transformation of the by semi-enrolled teeth with a similar shape central (and lateral teeth) in clade B is com- and was previously considered a separate plicated. Analysis of the morphology of the subfamily Zonulispirinae. In some duplex teeth central tooth with unordered character states (e.g., in Funa and Cheungbeia), the secondary produced a shorter tree, but suggested initial limb is minute, nearly obsolete. Kantor & Taylor reduction of the tooth in the entire Conoidea (2000) suggested, based on a morphological and subsequent multiple (eight in our dataset) tree, that this is a derived state. The current independent reappearances in different clades, analysis confirms this hypothesis. that is multiple reversions (Fig. 5). It is espe- In the single genus Toxiclionella Powell, cially difficult to explain these reversions from 1966 (not present in our analysis), referred to a functional point of view due to the limited Clavatulidae and that still possesses an odon- functions of the central and lateral teeth. On tophore, the teeth are hardly distinguishable the contrary, reduced functionality of the central from hypodermic teeth of the clade A (Fig. 31), segment seems to be congruent with reduction 86 KANTOR & PUILLANDRE and complete loss of the central segment in teeth) to a seemingly solid central tooth occur, different lineages, as was suggested by the sometimes within a single genus, for example, stepmatrix analysis (Fig. 6). Gemmula (Kantor, 2006). Similarly, we found the numerous losses of In three clades – in Cochlespiridae, in clade the lateral teeth (stepmatrix analysis – Fig. 42) combining Pseudomelatoma and Tiariturris more probable (although less parsimonious) (Pseudomelatomidae sensu Taylor et al., than initial reduction of the teeth in the entire 1993), and in Gemmuloborsonia, there is no Conoidea and independent reappearance in indication that the central tooth has a composite five clades (analysis with unordered character origin. The posterior margin bearing the cusp is states). Only in Drilliidae do the lateral teeth equally developed along its width and elevated seem to be functional (see below), while in over the radular membrane. Without further other clades they are very weak plate-like information, we conclude that these groups structures, sometimes appearing only as possess the broad unicuspid central tooth, inconspicuous thickenings of the subradular while the lateral teeth are absent. membrane (e.g., in Pusionella compacta – Fig. Thus, the great morphological variability of 28, vlt). the central segment of the radula in all clades The central tooth is thought to be highly (except the three mentioned above) is likely variable in Conoidea, ranging from very nar- to correspond to the different stages of the row unicuspid to broad with a large cusp and process of fusion of lateral and central teeth well-developed lateral flaps. Kantor & Sysoev with simultaneous reduction of either central, (1991) and Taylor et al. (1993) suggested that or lateral or both teeth. the broad central tooth in some groups may One of the unexpected results of our analysis be the result of fusion of paired lateral teeth is the possible secondary origin of the multi- with the narrow unicuspid central tooth. At cuspidate separate and well-formed lateral that time, conclusion was rather hypothetical teeth in Drilliidae. Previously, this type of the and besides general shape of the tooth was tooth was considered prototypic for based on the pattern of radular membrane (Powell, 1966; Kantor & Sysoev, 1991), and folding. In some species, attributed here to this hypothesis was the rationale for placing the Pseudomelatomidae (Crassiclava turricula – Drilliidae as a separate family from the other Figs. 19, 20, sanctiioannis (Smith, Turridae sensu Taylor et al. 1993. According 1875) – Kantor & Sysoev, 1991: figs. 27, 28; to the analysis, the multicuspid lateral teeth Comitas onokeana vivens Dell, 1956 – Fig. may be the ancestral state for clade B; they 15), clearly separate, plate-shaped lateral teeth then disappeared and reappeared again in without cusps were found. In these species, the the Drilliidae plus Pseudomelatomidae clade, central tooth (cusp) is absent. Later, in Comitas though multicuspid teeth are present only in Finlay, 1926, and the related genus Knefastia the former family. However, different runs of the Dall, 1919, intermediate stages were found. analysis with different coding of radular charac- In K. tuberculifera, in addition to a very weak ter states in the outgroups suggested different and reduced central tooth (Fig. 18, ct) vestigial most parsimonious plesiomorphic conditions in lateral plates (teeth) can be observed (vlt); in clade B, but in none of the runs were multicus- Comitas pachycercus Sysoev & Bouchet, 2001, pid teeth the single plesiomorphic state. One Comitas murrawolga (Garrard, 1961), and Co- of the reasons for these inconclusive results mitas sp. (Figs. 16, 17), the central structure may be the fact that the homology of different looks like a well-defined central tooth with a teeth in Neogastropoda (including outgroups) narrow cusp and broad lateral flaps (no lateral is not yet finally established and the relation- teeth present). In our opinion, this transitional ships within the entire Neogastropoda are far row suggests that instead of a morphologically from resolved. Therefore, the sister group of extremely variable central and lateral teeth in Conoidea is not yet identified, and we do not closely related groups, we are observing the know what is the plesiomorphic morphology of different stages of fusion of lateral cuspless the conoidean forerunner. plates with narrow central tooth. Similar gradual From a general point of view, it seems more stages of fusion of lateral and central narrow probable that well-pronounced multicuspid tooth was demonstrated in Turridae, in which lateral teeth in Drilliidae is the plesiomorphic transitional conditions ranging from a clearly condition in Conoidea, retained due to peculiari- tripartite structure with a gap between the cusp ties of their feeding mechanism. Unfortunately, (= central tooth) and the lateral flaps (= lateral there is practically no information on the diet EVOLUTION OF RADULA IN CONOIDEA 87 and radula functioning of drilliids. The only 1990) and therefore in prey capture mecha- published record is that of Maes (1983) of the nism. We may imagine that similar changes gut content of Drillia cydia (Bartsch, 1943). can occur in clade B and that the prey capture Finding intact prey (sipunculid) in the posterior and feeding mechanisms may differ between oesophagus suggested that the radula is used young individuals and adults and that at some for gripping and/or piercing, not rasping or tear- ontogenetic stage the odontophore may be ing. Drilliidae studied by us possess large and fully operational. This supposition needs careful powerful odontophores, which may suggest research to detect any ontogenetic radular and active use of the radula as an integrated whole foregut anatomy changes. organ. To confirm this, we carefully examined the bending plane of the radulae. In many spe- Transformation of the Hypodermic Marginal Teeth cies the laterals and even the central teeth were badly damaged (Fig. 12 – broken parts of the Hypodermic marginal teeth are found in clade teeth are marked by small arrows); damage on A and some Terebridae. The morphology of the the marginal teeth in the same specimens was hypodermic teeth is extremely variable and was not observed. These observations suggest that traditionally used for . Some hypoder- lateral (and even small central) teeth are func- mic teeth are very simple, semi-enrolled. Such tional, but without additional data on feeding teeth are found among Mangeliidae (e.g., Man- in this group, it is impossible to draw any final gelia – Fig. 49). Unfortunately, none of these conclusions about the mode of functioning. species is present in our molecular tree. One important question remains: why in The hypodermic teeth of Conidae s.s. have clade B conoideans, which possess in general been described in great detail and correlation the same feeding mechanism as clade A spe- between tooth morphology and diet has been cies, are the odontophore and its musculature demonstrated (e.g., Nybbaken, 1990). The retained? The odontophore varies in size from representation of genera in our data matrix is large in Drilliidae and some Pseudomelatomi- relatively sparse and therefore we could only dae (Pseudomelatoma) (Taylor et al., 1993) trace a limited number of characters of the hy- to very small or nearly obsolete (e.g., in some podermic teeth. It appeared that the spurs and , Pseudomelatomidae – Kantor et barbs of the teeth are homoplastic and evolved al., 1997). There are no data that explain this independently several times. No clear trends phenomenon. From what has been said above, were obvious, possibly due to the incomplete- it is clear that the functioning of the radula as ness of our dataset. an integrated organ may be limited, possibly to transferring the swallowed prey from the buccal Terebridae Radiation cavity, situated at the proboscis base, further to oesophagus. Only in very few conoideans One of the most remarkable findings of the is the buccal cavity plus radula either shifted molecular analysis (Puillandre et al., 2008, to the proboscis tip or able to protrude through 2011) is that the Terebridae do not represent the mouth due to the evertion of the buccal a totally separate lineage, but are included in tube (for details: Taylor et al., 1993). From the clade B and are sister to the Turridae. point of view of speciation, clade A conoideans Common among all terebrids is complete loss are more diverse, including 202 genera (not of the central and lateral teeth. In our analysis, counting 82 genera recognized in the Conidae, the first taxon to diverge among the Terebridae the phylogeny and taxonomy of which is not is Euterebra tristis (Deshayes, 1859), charac- yet finally revised) versus 180 genera in clade terized by the solid recurved teeth, superficially B. Thus, absence of the odontophore does similar to that in Pseudomelatoma and related not seem to limit prey capture and feeding. genera (Pseudomelatomidae). Our analysis Moreover, clade A conoideans seem to have was not able to resolve the most parsimonious a broader prey range, including other gastro- ancestral state for Terebridae. Before the mo- pods (numerous Conus species) and bivalves lecular analysis of all Conoidea was performed (Phymorhynchus, Raphitomidae – Fujikura et (Puillandre et al., 2011), more detailed analysis al., 2009) and even fish (several species of of Terebridae has been conducted (Holford et Conus). al., 2009). Among other things, it revealed that In some species of Conus, ontogenetic Pellifronia jungi (Lai, 2001) might constitute changes of radular teeth are probably related sister lineage to all the other Terebridae. The to changes in diet (summarized by Nybakken, radula of P. jungi appeared to have a new 88 KANTOR & PUILLANDRE type of marginal teeth for the family, similar Work was conducted in Paris by YK during in general arrangement to the duplex teeth of visits under the MNHN visiting curatorship other families in clade B (Fig. 44). The species program. He expresses his thanks to Virginie possess a venom gland with a muscular bulb, Héros, Barbara Buge, Philippe Maestrati and proboscis, and small odontophore. Pierre Lozouet for assistance during his time Thus, within the Terebridae, the entire tran- in Paris. This work was supported by a grant sition can be found from species with duplex from the Russian Foundation of Basic Re- teeth with a strong subradular membrane (and search (RFBR) 11-04-01284-а “Evolution of an odontophore) through solid recurved teeth to digestive system of carnivorous gastropods: species with typical hypodermic teeth, attached testing of morphologically-based hypotheses only by their bases to a vestigial membrane by molecular data” (PI Y. Kantor). (similar to the arrangement of the marginal The senior author conducted much of the teeth in clade A and similarly lacking an odon- study of the radulae of the Conoidea in co- tophore). There are some species (not included authorship with John Taylor and some of the in our analysis, but the position of which was images have been reproduced here and used inferred in the molecular phylogeny of Holford in discussion. We thank Dr. Alexander Fedosov et al. 2009), for example, Impages hectica for his comments on the manuscript. Dr. Robert (Linnaeus, 1758), that possess hypodermic Cowie assisted in editing of the manuscript. teeth (penetrated by numerous holes – Fig. 48) We want to thank referees, anonymous and that are attached to a rather strong membrane Alan Kohn for very valuable commentaries along their length, a condition similar to that in and corrections. Toxiclionella (Clavatulidae). Within this single clade the radula transforma- tion independently parallels the evolution of the LITERATURE CITED radular apparatus within the entire Conoidea. BANDEL, K., 1984, The radulae of Carribean and This is a remarkable example of the radular other Mesogastropoda and Neogastropoda. evolvability in Conoidea. Zoologische Verhandelingen, 214: 1–188, 22 pls. BOUCHET, P., Y. KANTOR, A. SYSOEV & N. ACKNOWLEDGEMENTS PUILLANDRE, 2011, A new operational classifi- cation of the Conoidea (, Gastropoda). Journal of Molluscan Studies, 77: 273–308. Material for this study was collected during a BOUCHET, P., P. LOZOUET & A. SYSOEV, 2009, number of field trips and expeditions. The PAN- An inordinate fondness for turrids. Deep-sea re- GLAO 2004 Marine Biodiversity Project was search. Part II-Topical Studies in Oceanography, funded by the Total Foundation and the French 56(19–20): 1724–1731. DUDA, T. F. & A. J. KOHN, 2005, Species-level Ministry of Foreign Affairs, the MNHN-IRD-PNI phylogeography and evolutionary history of the Santo 2006 expedition was made possible by hyperdiverse marine gastropod genus Conus. grants, among others, from the Total Founda- Molecular Phylogenetics and Evolution, 34: tion and the Stavros Niarchos Foundation, and 257–272. the AURORA 2007 cruise was made possible DUDA, T. F., A. J. KOHN & S. R. PALUMBI, 2001, through a grant from the Lounsbery Founda- Origins of diverse feeding ecologies within Co- nus, a genus of venomous marine gastropods. tion. The Coral Sea and Solomon Islands Biological Journal of the Linnean Society, 73: cruises took place on board R/V Alis deployed 391–409. from Nouméa by the Institut de Recherche pour DUDA, T. F., JR. & S. R. PALUMBI, 1999, Mo- le Développement (IRD), and Bertrand Richer lecular genetics of ecological diversification: de Forges was cruise leader for the Solomons, duplication and rapid evolution of toxin genes Coral Sea and Vanuatu expeditions. Ellen of the venomous gastropod Conus. Proceed- ings of the National Academy of Sciences, 96: Strong, Marie-Catherine Boisselier and Sarah 6820–6823. Samadi are thanked for their role in molecular ESPINO, S. S., A. J. KOHN, J. A. VILLANUEVA, sampling during these expeditions. Cruises F. M. HERALDE III, P. S. CORNELI, G. P. CON- in the Gulf of Panama (2000) and the Lower CEPCION & B. M. OLIVERA, 2008, Feeding Florida Keys (2001) were on R/V Urraca and behavior, phylogeny, and toxinology of Conus furvus Reeve, 1843 (Gastropoda: Neogastropo- R/V Bellows, respectively, and we thank the da: Conidae). The Nautilus, 122: 143–150. captain and crew of each and the respective ESPIRITU, D. J. D., M. WATKINS, V. DIA-MONJE, expedition leaders H. Lessios (STRI, Panama) G. E. CARTIER, L. E. CRUZ & B. M. OLIVERA, and T. Collins (FIU, Miami). 2001, Venomous cone snails: molecular phy- EVOLUTION OF RADULA IN CONOIDEA 89

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Revised ms. accepted 7 October 2011

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