Genus Profundiconus Kuroda, 1956 (Gastropoda, Conoidea): Morphological and molecular studies, with the description of five new species from the Solomon Islands and New Caledonia

Abstract. The genus Profundiconus Kuroda, 1956 is reviewed. The morphological characters of the shell, radular tooth and internal anatomy of species in Profundiconus are discussed. In particular, we studied Profundiconus material collected by dredging in deep water during different scientific campaigns carried out in the Solomon Islands, Madagascar, Papua New Guinea and New Caledonia. We reconstructed a phylogeny of 55 individuals based on partial mitochondrial cox1 gene sequences. The phylogeny shows several clades containing individuals that do not match any of the known species of Profundiconus based on their shell and radular morphologies, and are introduced here as five new species: Profundiconus maribelae sp. nov. from the Solomon Islands; P. virginiae sp. nov. from Chesterfield Plateau (New Caledonia); P. barazeri sp. nov. from Chesterfield Plateau and the Grand Passage area (New Caledonia); P. puillandrei sp. nov. from Norfolk Ridge (New Caledonia), Kermadec Ridge (New Zealand) and possibly Balut Island (Philippines); and P. neocaledonicus sp. nov. from New Caledonia. Furthermore, Profundiconus teramachii forma neotorquatus (da Motta, 1984) is raised to specific status as P. neotorquatus (da Motta, 1984).

Apart from the known extant species, the genus Profundiconus has a long fossil record ranging from the Cretaceous (Profundiconus primitivus (Collignon, 1949)) to the Pliocene (i.e., Profundiconus yanuyanuensis (Ladd, 1945)).None of the extant species of Profundiconus have been reported as fossils (Tucker & Tenorio 2009).The extant species included in the genus Profundiconus occur in the Indo-Pacific region except for P. emersoni (Hanna, 1963) (Fig. 1F), which occurs in the East Pacific region (Tucker & McLean 1993;Tenorio et al. 2012).However, the inclusion of this species in Profundiconus is only provisional (Tenorio et al. 2012).The fossil species are known from the Indo-Pacific region and North America.
As their name indicates, species of Profundiconus normally live in deep to very deep water.P. teramachii has been found at depths of 1134 m (dead) and 977 m (live) (von Martens 1901).However, not all Profundiconus are restricted to deep water: Röckel et al. (1995a) reported 20-300 m for P. ikedai (Ninomiya, 1987) and 75-560 m for P. lani.The value of 20 m corresponding to the minimal depth for P. ikedai seems too shallow in spite of being quoted in the original description for this species.Okutani (2000) corrected these depth ranges to 250-300 m for P. ikedai and 50-560 m for P. lani.In general, the deep-sea habitat makes these species difficult to sample.The most commonly collected one is P. teramachii (Fig. 1B), which surfaces in the nets of deep-water fishing trawlers.Most of the other species are rare in collections.However, species of Profundiconus are not uncommon among the material resulting from dredging carried out by research vessels in deep waters.In this context, the Muséum national d'Histoire naturelle (MNHN) has been carrying out a series of oceanographic expeditions in the deep waters surrounding New Caledonia and beyond since 1980 (Bouchet et al. 2008).Many of those research cruises were surveying the seamounts of the Norfolk Ridge (Castelin et al. 2011), dredging and trawling from 80 m to a depth of 3000 m.Other areas covered have included the Plateau des Chesterfield and the Grand Passage area.More recently, deep-water surveys have been carried out in the Fiji Islands, Philippines, Vanuatu, Madagascar, Papua New Guinea and other locations, with more missions in preparation.These research cruises produced large lots of deep-water cone snails, both in live and dead condition, with exact depth and locality data.Species of the genus Profundiconus are well represented, and their study by an international network of taxonomists has already produced new species (Tenorio 2015a(Tenorio , 2015b;;Rabiller & Richard 2014;Moolenbeek et al. 2008;Röckel et al. 1995b).
Here, we reconstruct the phylogeny of the genus Profundiconus, including a total of 55 individuals for which a fragment of the cox1 gene was sequenced.The phylogeny, in conjunction with comparative analyses of shell and radula characters, provides useful insights into the taxonomy of Profundiconus.The phylogeny shows several clades containing individuals that do not match any of the known species of Profundiconus according to their shell and radular morphologies, and they are introduced here as new species.

Material and methods
Most of the material studied here was previously deposited in institutional repositories.Descriptions and measurements are based on shells oriented in the traditional way; spire up with the aperture facing the viewer.The taxonomy used in the present work follows Tucker & Tenorio (2009) with the updates and modifications included in Tucker & Tenorio (2013).Specimens were collected by dredging in deep water during different campaigns carried out by the MNHN in New Caledonia and the Solomon Islands, most of them aboard the R/V Alis between 1985 and 2008, at depth ranges of 270 to 1100 m.Some specimens were taken off Curtis Island, Kermadec Ridge and west of northern New Zealand from a depth of 900-1100 m by R/V Tangaroa during a dredging campaign by the New Zealand Oceanographic Institute in 1979.Distribution maps were generated using GeoMapApp (http://www.geomapapp.org) with the general bathymetric map of the oceans as the default basemap.
We describe shell morphology using the terminology established in Röckel et al. (1995a).We also used their procedure for counting the number of protoconch whorls.For morphometric comparisons, adult shells randomly selected among available specimens in the collections of the MNHN and other sources (private collections) were measured with a digital caliper and the measurements rounded to 0.1 millimeter.All the measurements are in a spreadsheet, deposited as electronic supporting information (Appendix).For comparisons of shell morphometry, we performed analyses of the covariance (ANCOVA) for different shell parameters, namely maximum diameter (MD), height of the maximum diameter (HMD) and spire height (SH), using species hypotheses as factor and shell length (S L ) as covariate.Additionally, we statistically compared the mean values of S L using t-and U-tests.Statistical tests were carried out using STATGRAPHICS 5.1 or PAST3 (Hammer et al. 2001) once all the measurement sets passed the normality tests.We used the terminology for radular morphology of Tucker & Tenorio (2009) and the abbreviations in Kohn et al. (1999).The number of individuals for which the entire radula was examined is indicated in the description of each new taxon.Specimens of shells containing the dried animal inside were digested in concentrated aqueous KOH for 24 h.The contents were flushed out of the shell by injecting distilled water through the aperture of the shell by means of a syringe with an incurved needle.The resulting mixture was then placed in a Petri dish and examined with the binocular microscope.The entire radula was removed with fine tweezers and rinsed with distilled water, then mounted on a slide using Aquatex (Merck) Mounting Medium and examined under the optical microscope.Photos were obtained with a CCD camera attached to the microscope.Samples of individual radular teeth for scanning electron microscopy (SEM) were allowed to dry in the air upon rinsing with distilled water and then mounted on stubs covered with double-sided carbon tape.SEM studies were carried out at the MNCN-CSIC on a FEI Inspect scanning electron microscope, equipped with a secondary and retro-dispersed electron detector, and an Oxford Instruments analytical-INCA integrated analysis system.
Forty-one partial DNA sequences of the mitochondrial cox1 gene (Folmer et al. 1994) were extracted from GenBank (from the study of Puillandre et al. 2014; GenBank accession numbers in Table 1).This corresponded to a selection of the DNA sequences of the targeted species of Profundiconus as defined by Tucker & Tenorio (2013), which includes members of the "teramachii/smirna/aff.profundorum/n.sp.g" complex mentioned in Puillandre et al. (2014).An additional set of 14 cox1 sequences corresponding to Profundiconus species were kindly supplied by Dr. Nicolas Puillandre (submitted to GenBank and BOLD) and included in the present study, increasing the number to 55 specimens.Within Conoidea, the sister group of the cone snails is Borsoniidae (Tucker & Tenorio 2009: fig. 16;Puillandre et al. 2011Puillandre et al. , 2014)).Consequently, we included in our analyses one member of the family Borsoniidae: Bathytoma Table 1 (Moolenbeek, Röckel & Richard, 1995), were selected to form a closer outgroup in order to provide a broader phylogenetic context for the species complex that we were interested in.
The cox1 gene sequences were translated into amino-acids using MEGA v. 4.0 (Tamura et al. 2007) and the invertebrate mitochondrial genetic code to check for stop codons.DNA sequences were aligned using the MUSCLE Server (Edgar 2004).The accuracy of DNA sequence alignment was confirmed by eye.Identical DNA sequences were identified using DnaSP v. 5 (Librado & Rozas 2009) and excluded from posterior analyses.The best-fit nucleotide substitution model (HKY + I + G) for the cox1 dataset was determined using JModeltest 2 (Guindon & Gascuel 2003;Darriba et al. 2012) and the Bayesian Information Criterion (BIC; Posada & Buckley 2004).A maximum likelihood (ML) tree was built using RAxML HPC2 (Stamatakis 2006) on Teragrid v. 7.2.7,implemented in the Cyber Infrastructure for Phylogenetic Research (CIPRES) portal version 3.1 (http://www.phylo.org/portal2).The best-scoring ML tree was estimated from 100 independent searches, each starting from distinct random trees.Robustness of nodes was assessed using the rapid bootstrapping algorithm (1000 replicates) (Felsenstein 1985;Stamatakis et al. 2008).Bayesian analyses (BA) were performed running two parallel analyses in MrBayes (Huelsenbeck & Ronquist 2001), each consisting of four Markov chains of 5 000 000 generations, each with a sampling frequency of 1 tree each 100 generations.The number of swaps was set to 5, and the chain temperature at 0.05.Convergence and mixing of the chains of each analysis was evaluated using Tracer v. 1.4.1 (Rambaud & Drummond 2007) to check that effective sample size (ESS) values were all greater than 200.A consensus tree was then calculated after omitting the first 25% of trees as burn-in.We considered a clade to be 'moderately supported' if it had a bootstrap support value (BP) between 75 and 89% and posterior probability (PP) between 0.95 and 0.97, and 'highly supported' when BP ≥ 90% and PP ≥ 0.98 (Fig. 2).
The following abbreviations are used for museums and institutions:

Description
Shell characterS (Fig. 1).Conical to narrowly conical shell, usually thin, very small to very large in size; shoulders become rounded in outer whorls, although a ridge is present in some cases; a few cords present on early whorls and become numerous and smaller in outer whorls; nodules obsolete early; anal notch shallow; larval shell either paucispiral or multispiral; operculum large and serrate; periostracum smooth.radular tooth (Fig. 2).Blade and barb present (may be poorly differentiated); blade pointed, moderate in length, up to half length of anterior section of tooth; serrations absent; adapical opening large; a structure that we will refer to as external cusp (non-homologue of a posterior blade) starting at base of adapical opening and extending towards waist; external cusp often laterally expanded and serrated, with several small denticles (Fig. 2C, F); external cusp may appear partially covered by rolled sheet, which conforms to anterior portion of tooth; barb, blade and external cusp arranged in three different planes, which form angle of c. 120º between them; waist evident; characteristic fringe composed of closely spaced projections pointing towards apex present immediately below waist (Fig. 2C, F); anterior section of tooth shorter than posterior section; shaft fold present; slanted base with large basal spur.4-5).The internal anatomy of Profundiconus tuberculosus (Tomlin, 1937) (Fig. 1G) has been studied in detail by Taki (1937), and can be considered representative for other members of the genus.Taki's work was reviewed by Röckel (1994).Some details of the external anatomy and radular apparatus of several other species of Profundiconus are presented in Rolán & Raybaudi-Massilia (1994).We hereby reproduce some of the figures from Taki (1937) (Figs 4-5) to illustrate the details of the internal anatomy in Profundiconus.Taki (1937) remarked that the anatomical features of P. tuberculosus indicate in many aspects the ancestral nature of this species.The proboscis sheath (Fig. 3A: RS) has longitudinal folds on the inner side; the inner wall of the respiratory siphon (Fig. 3A-B: SI) is smooth, lacking the furrows or invaginations that have been reported in other species of cone snails; the osphradium (Fig. 3B: OS) is simple and not divided, with the small lobules arranged like feather banners on both sides of the longitudinal axis; there is only one salivary gland (Fig. 3B: SD), consisting of a multitude of small lobules.

Internal anatomy (Figs
The anterior lobe of the midgut gland (Fig. 4A: L 1 , L 2 ) has a bifurcated excretory duct.The male genitalia (Fig. 4B) have been described in detail.The organs that Bergh (1896) described as testes are considered to be the prostate in the opinion of Taki (1937).The testis consists of two lobes (Fig. 4B: HA, HP), which are spirally coiled and reach the tip of the spire.The prostate (Fig. 4B: PG) is separated into three parts (PGA, PGD, PGS), with irregular shallow grooves on its surface.Both cerebral ganglia (Fig. 4C: C) largely merge with each other and can be distinguished only by a slight constriction in the middle; the right parietal and right visceral nerves (Fig. 4C: PA, V) go together as a single nerve from the subintestinal (IN) ganglion.They separate only after a prolonged course.

Geologic range
Cretaceous to Recent.

Geographic distribution
The Holocene species included in the genus (Table 2) occur in the Indo-Pacific region, except for Profundiconus emersoni (Fig. 1F), which occurs in the East Pacific region.Extinct species are known from the Indo-Pacific region and North America (Tucker & Tenorio 2009).For a listing of fossil species placed in the genus Profundiconus, see Tucker & Tenorio (2009).

Remarks
The shells of species in Profundiconus are morphologically related to species included in the fossil genus Conilithes Swainson, 1840.Both taxa contain shells with square nodules that are interconnected by carinae on the body whorl, which constitutes a plesiomorphic trait.However, the anal notch is deep in Conilithes and shallow in Profundiconus.Furthermore, Rolán & Raybaudi-Massilia (1994) have suggested that Conilithes antidiluvianus (Bruguière, 1792) (Fig. 1E) and P. teramachii (Fig. 1B) are close relatives.Based upon similarities in shell morphology Tucker & Tenorio (2009) placed both genera, Conilithes and Profundiconus, within the family Conilithidae and separate from Conidae.However, shell traits can be ambiguous.The serrated operculum and the morphology of the radular tooth are more robust proxies for placing a given specimen in Profundiconus.The presence on the radular tooth of a laterally widened, often serrated external cusp, along with a characteristic fringe located immediately below the waist composed of closely spaced projections pointing towards the apex in addition to other morphological features (i.e., barb, pointed blade, shaft fold, etc.; see Fig. 2), allow the immediate identification of an individual as a member of Profundiconus.The function of the coronated fringe on the tooth in species of Profundiconus is unknown, but it resembles a similar structure (collar-shaped band of tubercles) observed on the radular teeth of several members of the genus Lienardia Jousseaume, 1884, family Clathurellidae, such as Lienardia tagaroae Fedosov, 2011, L. jousseaumei (Hervier, 1896) or L. cf.rosella Hedley, 1922 (Fedosov 2011;Bouchet et al. 2011).
Nothing is known about the diet of Profundiconus cone snails, nor about the families of conotoxins which might be present in the species of the genus.Whereas most of the toxinological studies on cone snails carried out during the last three decades have focused on species that belong to only a few lineages (Puillandre et al. 2012), several lineages remain largely understudied or even not studied at all, as is the case for Profundiconus (Puillandre et al. 2014).Radular morphology suggests a most likely vermivorous diet.However, Marshall (1981) reported finding the beaks of a small cephalopod in the stomach of an adult specimen of P. smirnoides (identified as smirna) from Wanganella Bank, New Zealand.This suggests that this species might produce a conotoxin of sufficient potency to rapidly immobilize a fastmoving prey.

Phylogenetic analyses
Ingroup sequences included 657 bp containing 192 variable sites, of which 127 were phylogenetically informative.Excluding redundant sequences, 27 sequences were unique in the ingroup.The ML and BA tree topologies were congruent (Fig. 5) and supported Profundiconus as a monophyletic group (PP = 1; BP = 87%).Within Profundiconus, several clades corresponding to different species were recovered, although their phylogenetic relationships were poorly resolved (Fig. 5).The individuals belonging to P. teramachii were split into two separate monophyletic groups corresponding to different geographic regions, one with specimens from the Indian Ocean (Madagascar) and another with specimens from the Pacific Ocean (China, Papua New Guinea and Solomon Islands).This splitting has previously been reported and discussed by Puillandre et al. (2014).The specimens of P. teramachii from the Indian Ocean belong to the forma neotorquatus da Motta, 1985 (Tucker & Tenorio 2013).In the Indian Ocean this species is widely distributed, from Somalia to South Africa (Natal), including Madagascar.
According to the phylogeny and the genetic distances, the specimens from Madagascar deserve specific status, i.e., Profundiconus neotorquatus (da Motta, 1984), rather than consideration as a mere form of P. teramachii.In spite of the molecular divergence that exists between P. neotorquatus and P. teramachii, the morphological differences in their shells (Fig. 1B-C) and radulae (Fig. 6) are slight.
The genetic differentiation of P. smirnoides and P. teramachii was not supported.Moreover, the monophyly of the group was not supported by the ML and BA analyses.Reciprocal monophyly between this group and P. neotorquatus was therefore not demonstrated.This is usually the case for recently diverged species, due to the lack of time needed to coalesce (Knowles & Carstens 2007).It is interesting to note that one of the cox1 sequences of P. smirnoides in GenBank (KJ550448) is essentially identical to that of P. teramachii, which represents the haplotype for 18 specimens (Table 1).As the voucher specimen (MNHN IM-2009-18244) seems to be properly identified, this could be due to contamination.However, there also exists the possibility that the cox1 can simply not separate these two species, which on the other hand can be easily separated based on shell and radular morphologies.There are many examples in the literature of morphologically distinct species having identical or almost identical cox1 sequences (e.g., Mengual et al. 2006;McGuire et al. 2007;April et al. 2011;Chee 2014).In most cases this has been attributed to hybridization-mediated mitochondrial introgression, as well as incomplete lineage sorting.
Specimens attributed a priori to the species Profundiconus loyaltiensis (Röckel & Moolenbeek, 1995) based on shell morphology were segregated into two distinct lineages.One of these lineages formed a monophyletic group (PP = 1; BP = 83%) along with specimens assigned to P. vaubani (Röckel & Moolenbeek, 1995) and P. kanakinus (Richard, 1983).Furthermore, some specimens of P. vaubani and P. loyaltiensis share the same haplotype.It is not clear at this stage whether this indicates one single polymorphic species, contamination, or simply the failure to separate the three closely related species based upon the cox1 gene fragment only.All of these specimens come from the same area (Isle of Pines, South New Caledonia).The group of specimens from the Solomon Islands labelled as "Profundiconus n. sp.cf.loyaltiensis" was monophyletic and highly supported by BA and ML analyses (PP = 0.97; BP = 99).If we assume that the specimens from New Caledonia are the genuine representatives of a population of the taxon P. loyaltiensis, we have to postulate that the specimens from the Solomon Islands belong to a separate group deserving recognition at the species level.The new species is introduced here under the name P. maribelae sp.nov.This taxon had previously been labelled as "Profundiconus n. sp.c cf. loyaltiensis" (Puillandre et al. 2014).
There are two groups containing specimens from Plateau des Chesterfield, and one with specimens from Norfolk Ridge and Loyalty Ridge, which do not match any known species of Profundiconus described to date.They are respectively introduced here as P. virginiae sp.nov., P. barazeri sp.nov.and P. puillandrei sp.nov.(previously considered in Puillandre et al. 2014 as "Profundiconus n. sp.h", "n.sp.b" and "n.sp.g", respectively).Additionally, we found that in spite of the lack of support at the corresponding nodes in the tree, the specimens (3 individuals sharing identical haplotype) corresponding to P. cf. profundorum from Norfolk Ridge exhibit significant conchological differences with the nominal taxon P. profundorum known from Japan and China.Based upon these constant differences and the independent branch in the tree, we describe it here as P. neocaledonicus sp.nov.

Etymology
This species is dedicated to Maribel Albarrán Quintanilla, wife of the first author and a shell-lover, in recognition for her support and constant encouragement to the first author at all times.

Description
Morphometric parameters: S L = 27-32 mm; RD = 0.55-0.62;RSH = 0.21-0.26;PMD = 0.89-0.93.Shell moderately small.Maximum length 31.5 mm.Shell profile conical, with straight to very slightly convex sides and spire moderate to high.Spire profile straight, stepped.Paucispiral protoconch with 1.5-1.75whorls, brownish, glossy and translucent (Fig. 7D).Teleoconch whorls stepped, ridged with small but strong nodules which persist in shoulder in most cases.Sutural ramp concave, with subsutural ridge and 3 to 4 strong spiral cords crossed by thin radial threads.Shoulder carinated, most often covered with small nodules along shoulder angle.Early teleoconch whorls pure white.Late teleoconch whorls in vicinity of shoulder area may exhibit some small, irregular brown blotches.Last whorl with grooves forming flat spiral ribbons, which may extend from shoulder to base.In some specimens sculpture of grooves and flat spiral ribbons in last whorl reduced to subshoulder area and basal half.Ground colour white overlaid with sparse brown, axially arranged flammules or blotches.White ground colour predominates in all specimens studied.Columella and aperture white.Anal notch shallow.Periostracum yellow-brown, thin and translucent.Operculum present, serrated on left border.Radular teeth examined in paratype 1 (Fig. 7G).38 teeth in radular sac.Radular tooth medium-sized, its total length relative to shell length S L /T L = 45.Anterior portion much shorter than posterior section of tooth (T L /AP L = 3.43).With one barb and pointed, well-defined blade which covers 50% of anterior portion of tooth.With external cusp located at approximately lower quarter of anterior portion of tooth, extending between 75% and 90% of length of anterior portion of tooth.External cusp laterally expanded and serrated, with 5 small denticles.Immediately below waist with characteristic fringe composed of closely spaced projections pointing towards apex.Shaft fold present.Large and prominent basal spur present on top of slanted base of tooth.

Distribution and habitat
Known from the Solomon Islands, including the New Georgia Group (Vella Lavella Island), Santa Isabel and Guadalcanal, at depths between 336 and 690 m (Fig. 8).

Remarks
The specimens of P. maribelae sp.nov.from the Solomon Islands form a monophyletic group supported by BI and ML analyses.P. maribelae sp.nov.was initially referred to as P. cf. loyaltiensis in the phylogenetic analysis, given the resemblance of its shell to that of P. loyaltiensis, a species known only from New Caledonia (Fig. 1H).However, the shell of P. maribelae sp.nov.attains a larger size (27-32 mm for maribelae vs. 21.5-26mm for loyaltiensis).The spire outline in P. maribelae sp.nov. is straight rather than deeply concave as occurs in P. loyaltiensis.The sculpture of flat spiral ribbons in the last whorl is much more developed in P. maribelae sp.nov.than in P. loyaltiensis.The most evident difference between the two species is that the shell of P. loyaltiensis is white and patternless, whereas the shell of P. maribelae sp.nov.exhibits a pattern of sparse, axially arranged brown flammules or blotches.The morphology of the radular teeth of P. maribelae sp.nov.and P. loyaltiensis is very similar, but they differ in their relative sizes, being larger in P. loyaltiensis (S L /T L ) = 30-37; versus 45 for P. maribelae sp.nov.).The shell of P. teramachii (Fig. 1B) is easily separated from that of P. maribelae sp.nov.by its much larger size, its pale, straw-yellow, patternless shell, smooth sculpture of the last whorl, usually much less developed nodules, absence of strong cords on the sutural ramp, and by its multispiral instead of paucispiral protoconch.P. maribelae sp.nov. is distantly related to both P. loyaltiensis and P. teramachii in the phylogeny presented here.Bouchet, 2008).

Etymology
This species is dedicated to Virginie Héros, assistant curator of molluscs at the Muséum national d'Histoire naturelle of Paris (MNHN) and an experienced member of the numerous collecting expeditions carried out by this institution.Her contribution to our knowledge of the New Caledonian deep-water cones is recognised by naming this remarkable new species of Profundiconus after her.Shell moderately small to medium sized.Maximum length 42.5 mm.Shell profile ventricosely conical, with spire moderate to high.Spire profile sigmoid.Multispiral protoconch with 3-3.5 whorls, white, glossy and translucent (Fig. 9C).Early 4-5 teleoconch whorls stepped, ridged with small nodules which tend to disappear after fifth whorl.Sutural ramp flat to slightly concave, with 3 to 6 fine spiral cords becoming obsolete in late spire whorls.Shoulder subangulate, forming characteristic ridge, covered with axial costae on last whorl.Early teleoconch whorls creamy white with brown spiral band on periphery, extending over row of nodules.On later whorls, this brown band interrupted by white areas.Spire creamy white with sparse, small brown blotches in areas near suture.Last whorl smooth or with very fine striae and with spiral ribs on basal third.Ground colour creamy white overlaid with orange-brown to purplish brown, irregular blotches or axially arranged flammules, interrupted by ground-colour band at midbody.Columella white.Aperture creamy white.Anal notch shallow.Periostracum and operculum not observed.

Holotype
Radular teeth examined in holotype (Fig. 9G) and in paratype 1.48 to 53 teeth in radular sac.Radular tooth medium to large-sized, its total length relative to shell length S L /T L = 37-45, rather elongated.Waist poorly defined.Anterior portion shorter than posterior section of tooth (T L /AP L = 2.61-2.70).With one barb and pointed, well-defined blade which covers 40-43% of anterior portion of tooth.With external cusp located at approximately lower third of anterior portion of tooth, extending between 60% and 81% of length of anterior portion of tooth.External cusp laterally expanded and serrated, with 5-6 small denticles.With characteristic fringe of closely spaced projections pointing towards apex located immediately below waist.Shaft fold present.Large and prominent basal spur on top of slanted base of tooth.

Distribution and habitat
Only known from the Coral Sea, Plateau des Chesterfield area, New Caledonia (Fig. 10).

Remarks
Profundiconus virginiae sp.nov.was initially misidentified as P. smirnoides (Fig. 1D).The latter has a fusiform shell, larger in size (S L = 52-98 mm; versus 33-43 mm), more slender (RD = 0.46-0.56;versus 0.59-0.63)and with a higher spire (RSH = 0.25-0.36;versus 0.21-0.23)than P. virginiae sp.nov.The shell of P. smirnoides has a pattern consisting of a brown spiral band on each side of centre, interrupted by creamy white axial streaks, but lacks the axial costae on the ridge.The radular teeth of P. virginiae sp.nov.and P. smirnoides also differ.The latter has a very elongated radular tooth (Fig. 9H), with the anterior and posterior sections difficult to separate (Tenorio 2015b).Still, the anterior section of the tooth in adult P. smirnoides is longer than the posterior section (T L /AP L = 1.7-1.8).The strongly pointed blade covers less than 25% of the anterior section.The external cusp is pointed, not expanded laterally and is not serrated, in contrast to P. virginiae sp.nov.Interestingly, in P. smirnoides the external cusp occupies a very high position in the anterior section, extending from 20 to 30% of its length (i.e., almost the same position as the blade, but with a different orientation).
P. virginiae sp.nov.resembles no other species of Profundiconus.Although only two live-taken specimens and the spire of a broken shell of this species have been examined, their shell and radula features, as well as the phylogenetic analysis, warrant its description as a new species.Apart from P. smirnoides, P. virginiae sp.nov.can be compared to P. vaubani from New Caledonia (Fig. 1I), and to P. cakobaui from the Fiji Islands (Fig. 1K) and P. cf. cakobaui (Fig. 9E) from the Philippines.P. vaubani also has axial costae and brown elements on the shell pattern.However, its paucispiral protoconch readily separates this species from P. virginiae sp.nov., which has a multispiral protoconch and appears rather distant from P. vaubani on the tree shown in Fig. 5. P. cakobaui and P. cf. cakobaui exhibit a shell pattern similar to that of P. virginiae sp.nov., including the brown spiral band on the periphery of the early teleoconch whorls extending over the row of nodules.However, both have a paucispiral protoconch instead of multispiral, and both lack the characteristic axial costae present in P. virginiae sp.nov. at the shoulder ridge.The elusive species P. frausseni (Tenorio & Poppe, 2004) (Fig. 9F), known only from a few specimens collected in the Philippines, has a protoconch and early teleoconch resembling that of P. virginiae sp.nov.However, the multispiral protoconch is white in P. virginiae sp.nov., but cream-coloured in P. frausseni (Tenorio & Poppe 2004).The latter is lower spired (RSH = 0.19-0.21;versus 0.21-0.23)and more conical (PMD = 0.87-0.89;versus 0.82-0.83)than P. virginiae sp.nov.Cords on the teleoconch whorls of P. frausseni are more developed, whereas they become obsolete on the late whorls in P. virginiae sp.nov.The shoulder in P. frausseni is rounded (subangulate in juvenile specimens), whereas in P. virginiae sp.nov. it is subangulate and ridged, covered with axial costae which are absent in P. frausseni.The scarce number of specimens of P. virginiae sp.nov.available prevented any statistical comparison of shell morphometry among different taxa.The two individuals of P. virginiae sp.nov.form a monophyletic group in the phylogeny that is the sister group of P. zardoyai, P. vaubani, P. loyaltiensis and P. kanakinus.

Distribution and habitat
Specimens from two separate populations in New Caledonia are known: from NW Bellona Reef, Plateau des Chesterfield (type locality), and from the Grand Passage area; at depths from 277 to 350 m (Fig. 10).

Remarks
P. barazeri sp.nov.resembles in general aspect a small specimen of Boucheticonus alisi (Fig. 11H).The latter has a larger shell, variably patterned, with a multispiral protoconch, which exhibits a characteristic brown blotch.The protoconch is paucispiral in P. barazeri sp.nov.These two species are phylogenetically distant (Fig. 5) and have very different radular morphologies.In contrast to the tooth of P. barazeri sp.nov., the radular tooth of B. alisi (Fig. 11L) is very large and elongated, with an extremely long anterior section that is more than four times longer than the posterior section of the tooth.It has a small and indistinct barb opposite a blade, which is enlarged and widened laterally.P. barazeri sp.nov.shows some similarities to P. zardoyai (Fig. 11I) and to Continuconus estivali (Moolenbeek & Richard, 1995) (Fig. 11J).The scarce specimens available of the latter species come from the Chesterfield Reef area.C. estivali is also characterized by its small size and conical shape.However, the pattern of C. estivali is quite constant and consists of 6 to 8 fine brown spiral lines on a white background.The shoulder in C. estivali is sharply angulated to carinate instead of angulated, and the teleoconch whorls on its stepped spire are concave.P. barazeri sp.nov.lacks the large, globose protoconch of about 2 whorls, which constitutes one of the most relevant features of C. estivali.The species P. zardoyai, recently described from Grand Passage, North New Caledonia, has a similar size and ground colour, with a variable pattern.However, its shell usually has a higher spire (RSH = 0.14-0.23 versus 0.11-0.19for barazeri) of sigmoid profile rather than straight.Although their radular teeth look superficially similar, the tooth of P. barazeri sp.nov.(Fig. 11K) has a larger relative size (S L /T L = 26-30 versus 33-40 for zardoyai) and bears more denticles in the laterally widened external cusp (7-9 in barazeri sp.nov.versus 5-6 in zardoyai).P. zardoyai and P. barazeri sp.nov.are phylogenetically distant (Fig. 5).NEW ZEALAND: 38.9 × 17.5 mm, R/V Tangaroa, st.K861, Kermadec Ridge, 30°36.5'S, 178°22.5'W, 1030 m (NIWA 99587; paratype 8; Fig. 12J); 44.9 × 19.5 mm, R/V Tangaroa, st.K831, Kermadec Ridge, 29°51.5'S, 178°10.5'W, 965 m (NIWA 99588; paratype 9).
Additionally, we examined 31 more specimens from 15 uncataloged MNHN lots collected at several stations in Norfolk Ridge and Loyalty Ridge, New Caledonia, by the R/V "Alis" in the course of several campaigns.Several specimens of shells in private collections collected in Balut Is., Philippines at 100-150 m, showing the conchological features of Profundiconus puillandrei sp.nov., were also examined.

Description
Morphometric parameters: S L = 29-57 mm; RD = 0.53-0.62;RSH = 0.22-0.29;PMD = 0.81-0.90.Shell moderately small to medium sized (maximum length 57.0 mm).Shell profile ventricosely conical, with high spire.Spire profile sigmoid to slightly concave.Protoconch multispiral, with 3-3.5 whorls, white to yellow-brown.Last whorl of larval shell shows minute axial ridges.Early teleoconch whorls with nodules which are often indistinct after whorls 5 to 6, but may persist, forming nodulose ridge reaching shoulder on last whorl.Sutural ramp flat to slightly concave, with very fine striae and arcuate threads becoming obsolete in late whorls.Shoulder with distinct ridge, usually smooth, although nodulose or even strongly nodulose in some specimens.Last whorl with convex sides adapically, then almost straight and slightly concave abapically.Last whorl smooth or with very fine striae becoming more evident towards base.Spire and last whorl patternless, white to pale straw-yellow in colour.Columella white.Aperture pale yellow or white.Periostracum yellow, thin and translucent.Operculum with serrations.
Radular tooth examined in holotype (Fig. 13A), in paratypes 3 (Fig. 13B-D), 6 and 9, and in an additional non-type specimen.48 to 62 teeth in radular sac.Radular tooth small for size of shell: its total length relative to shell length S L /T L = 75-105.Anterior portion shorter than posterior section of tooth (T L /AP L = 3.1-3.6).With one barb and pointed blade which covers 50-62% of anterior portion of tooth.External cusp present, extending between 64 and 90% of length of anterior portion of tooth.External cusp laterally widened and serrated, with 4-5 small denticles.Large adapical opening occupying most of anterior portion of tooth (100AO L /AP L = 64-75).Fringe of closely spaced projections pointing towards apex immediately below waist.Shaft fold present.Large and prominent basal spur on top of slanted base of tooth.

Distribution and habitat
New Caledonia (Norfolk Ridge and Loyalty Ridge) and New Zealand (Kermadec Ridge), at depths from 380 to 1100 m (Fig. 14).Several empty shells matching P. puillandrei sp.nov.from Balut Is., Mindanao, Philippines, have been examined.The identity of these specimens from the Philippines (allocated in several private collections) could not be confirmed by radular or molecular studies, but the conchological features seem consistent with the identification of these specimens (often labelled as Conus cf.ikedai, or Conus darkini "albinistic") as P. puillandrei sp.nov.This is a feasible possibility given the multispiral protoconch of this species (suggesting a planktonic larval development), and might represent a significant range extension to the Philippines.

Remarks
Profundiconus puillandrei sp.nov.has been dredged alive from 1030-1180 m off Curtis Island, Kermadec Ridge, New Zealand (identified as Conus smirna; see Marshall 1981).This observation makes this species one of the deepest-living ones among the known cone snails.P. puillandrei sp.nov.was initially identified as "giant" P. vaubani (Fig. 1I).Apart from being smaller in size, P. vaubani has a paucispiral protoconch of 1.75 whorls and a ridge at the shoulder with axial costae, which are absent in the case of P. puillandrei sp.nov.The shell pattern of P. vaubani consists of light brown axial streaks from base to spire, whereas the shell of P. puillandrei sp.nov. is patternless.The radular teeth of P. vaubani (Fig. 2A-C) and P. puillandrei sp.nov.(Fig. 13) are superficially similar, but the tooth of P. vaubani has a much larger relative size, with S L /T L = 27-31 compared to 75-105 for P. puillandrei sp.nov.The new species can also be compared with P. profundorum (Fig. 1A, L) and P. smirnoides (Fig. 1D).Like P. puillandrei sp.nov, these two species have multispiral protoconchs.There are no significant differences in shell shape among these species: ANCOVA for MD, HMD and SH, using species hypothesis as a factor and S L as covariate, did not yield statistically significant results.However, they differ significantly in average shell length: puillandrei S L 39.82 mm, profundorum S L 96.93 mm (t = 10.28,p = 2.13 × 10 -12 ; U = 0, p = 5.15 × 10 -7 ), smirnoides S L 76.09 mm (t = -10.20,p = 5.25 × 10 -10 ; U = 0, p = 2.5 × 10 -5 ).The shell of P. puillandrei sp.nov.has a distinct shoulder ridge, usually smooth but some times nodulose, which is absent in P. profundorum and less developed and always smooth in P. smirnoides.The shell of P. puillandrei sp.nov. is patternless, whereas both P. profundorum and P. smirnoides exhibit a characteristic pattern of broad, pale brown spiral bands on each side of centre, often interrupted by creamy white axial streaks in the case of P. smirnoides.The morphology of the radular tooth of P. puillandrei sp.nov.(Fig. 13) and of P. smirnoides (Fig. 9H) is very different.The radular tooth of P. profundorum is unknown, preventing its comparison with the tooth of P. puillandrei sp.nov.
Most of the shells of P. puillandrei sp.nov.examined were not nodulose at the shoulder ridge (Fig. 12).About 10% of the specimens studied had a nodulose spire and shoulder ridge (i.e., paratype 6, Fig. 12G), coming mainly from Loyalty Ridge.These include two of the sequenced individuals, which, however, were genetically similar to the ones with a smooth ridge.Moreover, nodulose and non-nodulose specimens exhibit analogous radular and protoconch morphology.The presence of nodules at the spire and shoulder causes an apparent difference in shape, which is possibly the main source of intraspecific phenotypic variability within this species.Nodulose specimens may resemble a small P. teramachii (e.g., paratype 6, Fig. 12G), a distinct but related species as inferred from the tree in Fig. 5.The shell of P. teramachii is also patternless and has a ground colour and protoconch morphology similar to that of P. puillandrei sp.nov.The radular tooth of P. teramachii (Fig. 2D-F) is also similar to that of P. puillandrei sp.nov.(Fig. 13).However, P. teramachii attains a larger size (S L = 55-111 mm), has a lower spire

Etymology
The epithet of this species makes reference to its distribution in deep water around New Caledonia.KJ550426).This specimen was databased (http://coldb.mnhn.fr/catalognumber/mnhn/im/2007-34866)and sequenced, but the shell was destroyed in the process and is no longer available.

Holotype
Additionally, we examined 32 more specimens from 18 uncataloged MNHN lots collected at several stations in Norfolk Ridge and Loyalty Ridge, New Caledonia, in the course of several campaigns.
Medium-sized to moderately large (maximum length 92.0 mm).Shell profile ventricosely conical to conical, with rounded shoulder and spire low to moderately high.Spire profile sigmoid, occasionally slightly concave.Protoconch multispiral of 3 or more whorls, white to pale violet-brown (Fig. 15F).First 4-7 postnuclear whorls nodulose.Teleoconch sutural ramp flat, slightly concave or sigmoid in later whorls, smooth, with cords absent.Last whorl smooth, with fine spiral ribs at base.Ground colour creamy-white to cream.Last whorl with two broad violet-brown, light brown or tan spiral bands above and below midbody, which exhibits broad ground-coloured spiral band.Colour is darker towards base, usually purplish.Narrow ground-colour spiral band often present at height of shoulder.Spire patternless, of ground colour, occasionally showing diffuse pale violet-brown or light brown on top of teleoconch whorls.Aperture light to pinkish brown.Periostracum olive, thin, translucent, smooth.Operculum with lateral serrations.
Radular tooth examined in paratypes 1 (Fig. 16A), 2 and 7 (Fig. 16B, C). 34-45 teeth in radular sac.Radular tooth of rather small relative size: its total length relative to shell length S L /T L = 61-87.Anterior portion shorter than posterior section of tooth (T L /AP L = 3.1-3.4).With one barb and pointed, prominent blade which covers 43-54% of apical portion of tooth.External cusp present, extending between 65 and 88% of length of anterior portion of tooth.External cusp not much widened laterally and serrations can be indistinct, with only 0-3 small blunt denticles.Large adapical opening occupying most of anterior portion of tooth (100AO L /AP L = 62-75).With characteristic fringe of closely spaced projections pointing towards the apex located immediately below waist (Fig. 16C).Shaft fold present.Large and prominent basal spur on top of slanted base of tooth.

Distribution and habitat
Norfolk Ridge and Loyalty Ridge, New Caledonia, at depths from 290 to 1100 m (Fig. 17).A couple of empty shells resembling P. neocaledonicus sp.nov.from Aliguay Island, Philippines, have been examined.The identity of these specimens from the Philippines (coming from the John K. Tucker collection, now with the INHS, Illinois, USA) could not be confirmed by radular or molecular studies.This observation might suggest an extension of the distribution range to the Philippines, but would require additional substantiation.

Remarks
Profundiconus neocaledonicus sp.nov.has long been considered a local form of P. profundorum from New Caledonia (Röckel et al. 1995a).P. profundorum was originally described based upon material from Japan (type locality: "Off Tosa, Japan; 100+ fathoms") (Fig. 1A).In recent times, specimens of P. profundorum have become available from China (Fig. 1L).These specimens are characterised by their significantly larger mean shell length compared to the Japanese specimens (mean S L profundorum from Japan: 74.3 mm; mean S L profundorum from China: 111.05 mm; t = 13.12,p = 0; U = 0, p = 0.000028).Apart from differences in shell length, ANCOVA indicated no statistically significant differences between the morphometric parameters of P. profundorum from Japan and those from China (ANCOVA on MD : F = 1.94, p = 0.18; on HMD : F = 3.44, p = 0.08; on SH : F = 0.05, p = 0.82).Hence, we consider the Chinese and Japanese populations conspecific.Compared to P. profundorum, the shells of P. neocaledonicus sp.nov.from New Caledonia have a different shape, are consistently smaller in average shell length, and are also paler in colour.Table 3 shows the results of the ANCOVA with the morphometric shell parameters MD, HMD and SH as variables, and using species hypothesis as factor and S L as covariate.Least-squares means are listed, along with average shell lengths for each of the species.
These results confirm that the shell of P. neocaledonicus sp.nov. is significantly broader, more conical (higher HMD) and lower-spired than the shell of P. profundorum, which also has a larger mean S L .A discriminant function analysis (DFA) using MD, HMD, SH and S L as variables and species hypothesis as factor correctly classified 94.4% of the specimens (all correct except two specimens of profundorum A DFA excluding S L from the set of variables correctly classified 92.6% of the specimens (three profundorum were misclassified as neocaledonicus and one specimen of neocaledonicus misclassified as profundorum).These results indicate that P. neocaledonicus sp.nov.can be separated with a high degree of certainty from P. profundorum based on significant differences in size and shell shape.It was not possible to compare the radula of P. neocaledonicus sp.nov.with that of P. profundorum.The radular tooth morphology of P. profundorum is unknown, since all the published information (Rolán & Raybaudi-Massilia 1994) actually corresponds to specimens from New Caledonia, here introduced as P. neocaledonicus sp.nov.No preserved specimens of P. profundorum from Chinese or Japanese localities were available for radular or molecular analyses.
Although the genetic differentiation of P. neocaledonicus sp.nov.from P. smirnoides and P. teramachii was not supported at the nodes on the tree in Fig. 5, P. neocaledonicus sp.nov. is easily separated from the sympatric species P. smirnoides based on shell and radular morphology.The shell of P. smirnoides (Fig. 1D) is significantly narrower, less conical and much more highly spired than that of P. neocaledonicus sp.nov.(ANCOVA with S L as covariate: on MD : F = 103.91,p = 0; on HMD : F = 62.54, p = 0; on SH : F = 28.37,p = 0).It also has a significantly larger mean shell length (76.09 mm for smirnoides versus 57.03 mm for neocaledonicus, t = -4.299,p = 0.00011; U = 285.5,p = 0.00055).P. smirnoides has a pattern of brown spiral bands on each side of centre interrupted by creamy white axial streaks, whereas in P. neocaledonicus sp.nov. is much simpler, consisting of pale purplish-brown broad bands on each side of centre on a creamy-white ground colour.Comparison of the large and elongated radular tooth of P. smirnoides (Fig. 9H) with that of P. neocaledonicus sp.nov.(Fig. 16) also allows the straightforward separation of the two species.P. teramachii (Fig. 1B) has a very different shell, larger in size, and is patternless, with a characteristic stepped spire and with a broadly carinated shoulder, sometimes with densely set rounded tubercles, particularly in smaller adults.
The species P. puillandrei sp.nov., which lives sympatrically with P. neocaledonicus sp.nov., has a significantly smaller shell length (t = 4.296, p = 0.0001; U = 53, p = 0.000066), with differences in morphometric parameters that suggest that P. puillandrei sp.nov.has a narrower shell with a higher spire (ANCOVA with S L as covariate on MD : F = 6.23, p = 0.0168; on HMD : F = 40.19,p = 0; on SH : F = 35.46,p = 0).This species shows a characteristic ridge at the shoulder, sometimes nodulose, whereas in P. neocaledonicus sp.nov. the shoulder is always rounded.The shell of P. puillandrei sp.nov. is patternless and does not exhibit the banding pattern visible in the shell of P. neocaledonicus sp.nov.Röckel et al. (1995a: pl. 73, figs 17-18) illustrated one specimen identified as "Conus species no.32".This specimen measures 50 × 23 mm and comes from Nazca Ridge.Other specimens like the one illustrated in Röckel et al. (1995a) have been collected in deep-water on several seamounts across Sala y Gómez and Nazca Ridges in the course of several campaigns carried out by Russian research vessels in the 70's and 80's.The biota of the Nazca and Sala y Gómez submarine ridges was reviewed in Parin et al. (1997).The fauna of benthic and benthopelagic invertebrates of this area is much more closely related to the Indo-West Pacific than to the Eastern Pacific fauna and is characterized by a very high degree of endemism at the species level (51% among identified bottom invertebrates).We have been unable to examine the specimens from the Nazca Ridge area, but the available photos (Röckel et al. 1995a) show a striking resemblance to the shells of P. neocaledonicus sp.nov.collected on Norfolk Ridge, New Caledonia, at a distance of c. 10 500 km to the east.Further research might eventually prove their conspecificity, which would imply a considerable range extension for this species, or alternatively disclose a new species of Profundiconus most likely endemic to the Sala y Gómez and Nazca Ridge areas.
Appendix.Measurements of shell parameters (see Material and methods).

Taxon
Abbreviations for shell morphometry: AH = aperture height HMD = height of the maximum diameter MD = maximum diameter PMD = relative position of the maximum diameter (= HMD/AH) RD = relative diameter (= MD/AH) RSH = relative spire height (= SH/S L ) SH = spire height S L = maximum shell length

Fig. 5 .
Fig. 5. Likelihood phylogenetic tree of ProfundiconusKuroda, 1956 based on a subsample of the mitochondrial cox1 dataset produced byPuillandre et al. (2014).Posterior probabilities and bootstrap values are indicated for each node (when PP ≥ 0.95% and BP ≥ 75%, respectively).For clarity purposes, external branches of phylogenetically highly supported species are highlighted in red in the consensus tree.Cox1 sequences are labelled using the MNHN voucher identification number, the Genbank accession number (when available), the species name and the number of specimens (or identical haplotypes) when greater than 1.

Fig. 8 .
Fig. 8. Distribution map for Profundiconus maribelae sp.nov.Red circles indicate the points where the species has been collected.

Fig. 10 .
Fig. 10.Distribution map for Profundiconus virginiae sp.nov.(red circles) and P. barazeri sp.nov.(yellow circles).Symbols indicate the points where each of the species have been collected.

Fig. 14 .
Fig. 14.Distribution map for Profundiconus puillandrei sp.nov.Red circles indicate the points where the species has been collected.

Fig. 17 .
Fig. 17.Distribution map for Profundiconus neocaledonicus sp.nov.Red circles indicate the points where the species has been collected.

.
Species vouchers with GenBank and BOLD accession numbers for the individuals and haplotypes included in the present study.[page 1 of 4]

Table 1 .
Species vouchers with GenBank and BOLD accession numbers for the individuals and haplotypes included in the present study.[page 4 of 4] Sysoev, Olivera, Couloux & Bouchet, 2010 was selected as the most external outgroup of the targeted species complex.In addition, sequences from GenBank of other species of cone snails from two different clades, namely Conus marmoreus Linnaeus, 1758, Cylinder textile (Linnaeus, 1758), Bathyconus orbigny (Audoin, 1831), Conasprella pagoda (Kiener, 1847) and Boucheticonus alisi
Appendix.[continued]Measurements of shell parameters (see Material and methods).