On the taxonomic position of Phaenomenella Fraussen & Hadorn, 2006 (Neogastropoda, Buccinoidea) with description of two new species

Directeur De la publication : Bruno David Président du Muséum national d’Histoire naturelle réDactrice en chef / Editor-in-chiEf : Laure Desutter-Grandcolas assistants De réDaction / AssistAnt Editors : Anne Mabille ([email protected]) Mise en page / PAgE lAyout : Anne Mabille coMité scientifique / sciEntific boArd : James Carpenter (AMNH, New York, États-Unis) Maria Marta Cigliano (Museo de La Plata, La Plata, Argentine) Henrik Enghoff (NHMD, Copenhague, Danemark) Rafael Marquez (CSIC, Madrid, Espagne) Peter Ng (University of Singapore) Norman I. Platnick (AMNH, New York, États-Unis) Jean-Yves Rasplus (INRA, Montferrier-sur-Lez, France) Jean-François Silvain (IRD, Gif-sur-Yvette, France) Wanda M. Weiner (Polish Academy of Sciences, Cracovie, Pologne) John Wenzel (The Ohio State University, Columbus, États-Unis) couverture / covEr : Shells of Phaenomenella nicoi n. sp. 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Diffusion – Publications scientifiques Muséum national d’Histoire naturelle CP 41 – 57 rue Cuvier F-75231 Paris cedex 05 (France) Tél. : 33 (0)1 40 79 48 05 / Fax : 33 (0)1 40 79 38 40 [email protected] / http://sciencepress.mnhn.fr © Publications scientifiques du Muséum national d’Histoire naturelle, Paris, 2020 ISSN (imprimé / print) : 1280-9551/ ISSN (électronique / electronic) : 1638-9387 33 ZOOSYSTEMA • 2020 • 42 (3) © Publications scientifiques du Muséum national d’Histoire naturelle, Paris. www.zoosystema.com urn:lsid:zoobank.org:pub:27C94F0F-BB9B-40A3-B615-4DB19C94F042 Kantor Y., Kosyan A., Sorokin P. & Fedosov A. 2020— On the taxonomic position of Phaenomenella Fraussen & Hadorn, 2006 (Neogastropoda, Buccinoidea) with description of two new species. Zoosystema 42 (3): 33-55. https:// doi.org/10.5252/zoosystema2020v42a3. http://zoosystema.com/42/3 AbstrAct This contribution provides novel information on the anatomy, radula and phylogeny of several species of Phaenomenella Fraussen& Hadorn, 2006, a genus of Buccinoidea Rafinesque, 1815 with unclear affinities. Molecular phylogenetic analysis based on sequences of mitochondrial COI and nuclear 28S rRNA genes of different representatives of Buccinoidea revealed close relationships of Phaenomenella with Siphonalia A.Adams, 1863 both taxa forming a clade with maximal support. The anatomy of two species of the latter genus was examined for the first time for comparative purposes. The subfamily Siphonaliinae Finlay, 1928 was erected for several Recent and fossil genera of Southern Hemisphere Buccinidae Rafinesque, 1815, and is still recognized by current taxonomists (Bouchet etal. 2017). Species of all Recent genera of Siphonaliinae were included in our analysis and the monophyly of the subfamily Siphonaliinae in its original scope is rejected. Molecular and morphological data revealed two still unnamed species of Phaenomenella from the lower bathyal zone of the South China Sea. These species, Phaenomenella nicoi n.sp. and P. samadiae n.sp. are described in the present study. Yuri KANTOR Alisa KOSYAN Pavel SOROKIN Alexander FEDOSOV A.N. Severtsov Institute of Ecology and Evolution of Russian Academy of Sciences, Leninski prospect 33, 119071 Moscow, Russian Federation (Russia) kantor[email protected] [email protected] [email protected] [email protected] Submitted on 28 January 2019 | Accepted on 8 April 2019 | Published on 4 February 2020 On the taxonomic position of Phaenomenella Fraussen & Hadorn, 2006 (Neogastropoda, Buccinoidea) with description of two new species Key words Molecular phylogeny, COI, 28S, classification, Siphonaliinae, new species. 34 ZOOSYSTEMA • 2020 • 42 (3) Kantor Y. et al. INtROduCtION The genus Phaenomenella Fraussen& Hadorn, 2006, with the type species Manaria inflata Shikama, 1971, was established for three species of “Buccinidae” Rafinesque, 1815 from taiwan and South China Sea (Fraussen& Hadorn 2006). Several additional species were described since and presently the genus includes nine species, all except one from off South East Asia (Fraussen 2008; Fraussen& Stahlschmidt 2012; Fraussen& Stahlschmidt 2013). The anatomy of the genus has never been examined, but the radula was illustrated for two species, Phaenomenella inflata (Shikama, 1971) and Phaenomenella angusta Fraussen& Hadorn, 2006. The radula is of general buccinid appearance, which gives no clues of the relationships of Phaenomenella to other buccinid genera. The position of the genus within Buccinidae was not addressed in previous publications, probably due to still unesolved intrafamiliar classification of the family. The intrageneric shell variability of Phaenomenella is high that is hampering providing reliable generic diagnosis. Among South-East Asian buccinids several genera bear some conchological resemblance to Phaenomenella, ie., Manaria Smith, 1906, Eosipho Thiele, 1929, Gaillea Kantor, Puillandre, Fraussen, Fedosov & Bouchet, 2013 (all three genera dwelling on biogenic substrates; Kantor etal. 2013) and Siphonalia A.Adams, 1863. Buccinidae from biogenic substrates constitute a clade that is well-supported by molecular data, and are characterized by bicuspid lateral teeth (Bouchet& Warén 1986; Kantor etal. 2013), while the radula of both Phaenomenella and Siphonalia is similar and has tricuspid lateral teeth. Relationships of Siphonalia with other Buccinidae are not clear. Its isolated position was recognized by Finlay (1928), who proposed a new subfamily Siphonaliinae in the newly established family Buccinulidae. Having been proposed without diagnosis or discussion, the subfamily originally included several Recent and fossil genera, which were later synonymized with Penion Fischer, 1884 (=†Austrosipho Cossmann, 1906, Verconella Iredale, 1914, Berylsma Iredale, 1924), Aeneator (=†Ellicea Finlay in Marwick 1928, †Pittella Marwick, 1928), as well as Glaphyrina Finlay, 1926 (presently attributed to Fasciolariidae Gray, 1853; Couto etal. 2016) and †Pomahakia Finlay, 1927. All genera except the type one are confined to southern hemisphere, mostly to Australian-New Zealand region. The validity and scope of the subfamily have not been revised, although it is recognized in the current taxonomy (Bouchet etal. 2017). A recent molecular phylogeny of some southern hemisphere Buccinulinae based on whole mitochondrial genome and nuclear ribosomal sequence data (Vaux etal. 2017) revealed that Recent genera included by Finlay into Siphonaliinae do not constitute a monophyletic group. But Siphonalia itself was not included in the analysis and therefore its relationships remained unresolved. In the course of expeditions organized by the Muséum national d’Histoire naturelle, Paris (MNHN) to the South China Sea several specimens of different species of Phaenomenella and one species of Siphonalia were collected and preserved for dNA sequencing. The examination of this material revealed two still unnamed species. The purpose of the present paper is to provide formal description of the revealed new species, and to clarify relationships of Phaenomenella based both on anatomy and on molecular data of extended dataset of Buccinoidea, including groups that are conchologically similar to Phaenomenella. MAtERIAL ANd MEtHOdS The material was collected mostly in the research cruises in the South China Sea (dONGSHA 2014, ZHONGSHA 2015) and in the Philippines (AuRORA 2007) that were organized respectively by the National taiwan university and the MurÉsUMÉ Position taxonomique de Phaenomenella Fraussen& Hadorn, 2006 (Neogastropoda: Buccinoidea) et description de deux nouvelles espèces. Cette contribution fournit de nouvelles informations sur l’anatomie, la radula et la phylogénie de plusieurs espèces de Phaenomenella Fraussen& Hadorn, 2006, un genre de Buccinoidea Rafinesque, 1815 aux affinités peu claires. une analyse phylogénétique moléculaire basée sur des séquences des gènes mitochondriaux COI et d’ARNr 28S nucléair de différents représentants de Buccinoidea a révélé des relations étroites de Phaenomenella avec Siphonalia A.Adams, 1863 qui forment un clade avec un soutien maximal. L’anatomie de deux espèces de ce dernier genre a été examinée pour la première fois à des fins de comparaison. La sous-famille Siphonaliinae Finlay, 1928 a été érigée pour plusieurs genres récents et fossiles de Buccinidae Rafinesque, 1815 de l’hémisphère sud, et est toujours reconnue par les taxonomistes actuels (Bouchet etal. 2017). Les espèces de tous les genres récents de Siphonaliinae ont été incluses dans notre analyse et la monophylie de la sous-famille Siphonaliinae dans sa définition originelle est rejetée. Les données moléculaires et morphologiques ont révélé deux espèces encore inconnues de Phaenomenella de la zone bathyale inférieure du sud de la mer de Chine. Ces espèces, Phaenomenella nicoi n.sp. et P. samadiae n.sp. sont décrites dans la présente étude. Mots clÉs Phylogénie moléculaire, COI, 28S, classification, Siphonaliinae, espèces nouvelles. 35 Taxonomic position of Phaenomenella ZOOSYSTEMA • 2020 • 42 (3) séum national d’Histoire naturelle, Paris (MNHN) with the Philippines Bureau of Fisheries and Aquatic Resources (BFAR). Specimens collected were processed with a microwave oven (Galindo etal. 2014): the living molluscs in small volumes of sea water were exposed to microwaves for 10-30 s, depending on specimen size. Bodies were immediately removed from shells and dropped in 96% ethanol. Specimens processed in this way are suitable for further anatomical studies after soaking them in 70% ethanol. Specimens are registered in the MNHN collection and specimens and sequences are deposited in BOLd (Barcode of Life datasystem) and GenBank (table1). due to technical reasons, the sequenced specimens of Siphonalia spadicea (Reeve, 1847) were not available for anatomical examination and the material (not suitable for sequencing) on two other species of Siphonalia was used for anatomical study. Dna extraction and pcr total dNA was extracted from the piece of foot using either the dNeasy96 tissue kit or Investigator Kit (Qiagen), following the manufacturer’s recommendations. The barcode fragment of the Cytochrome Oxidase I (COI) gene (658bp) and a 28S rRNA fragment were amplified using the universal primers LCO1490 and HCO2198 (Folmer etal. 1994) and C1 and d2 (Jovelin& Justine 2001), respectively. PCRs were performed in 20µl final volume containing approximately 3ng template dNA, 1.5mM MgCl2, 0.26mM of each nucleotide, 0.3µl of each primer, 5% dMSO and 0.75µl of either taq Polymerase (Qbiogene) or BioHYtaq dNA polymerase (dialat). The PCR profile for the COI started with 5min at 95°C followed by 40cycles with the denaturation at 95°C (35sec), annealing at 50°C (35s) and elongation at 72°C (1min), with final elongation phase at 72°C (10min). Similar PCR profiles were set for 28S (annealing at 56°C). COI and 28S genes were sequenced in both directions to confirm accuracy of each sequence. The sequencing was performed by Eurofins or in the SIEE RAS molecular facility on an ABI 3500 Genetic analyser. Morphology and radula Radulae were cleaned using diluted bleach (NaOCl), air– dried, coated with gold and examined by scanning electron microscope teScan tS5130MM in the Institute of Ecology and Evolution of Russian Academy of Sciences, Moscow (IEE RAS). Anatomy was examined on manual dissections. phylogenetic analysis COI and 28S sequences were aligned using ClustalW implemented in BioEdit v. 7.0.9.0 (Hall 1999); the accuracy of each alignment was checked by eye and if needed modified. COI and 28S sequences of additional 32 buccinoidean taxa, mainly from the datasets of Kantor etal. (2013) and Vaux etal. (2017) were accessed from GenBank. Three datasets were analyzed, a single-gene COI dataset (57taxa) with three codon positions coded as three independent partitions, a nuclear 28S (45taxa) as single partition, and a concatenated COI–28S dataset (45taxa), where four partitions corresponded to three codon positions of COI and to the 28S fragment respectively. The single gene datasets were mainly used to evaluate primary species hypotheses (PSHs), proposed based on the shell morphology, whereas the analysis of concatenated dataset allowed us to estimate relationships of Siphonalia and remaining genera originally included in Siphonaliinae. The lineage comprising buccinids from the biogenic substrates (Kantor etal. 2013) was used to root the tree based on the topology of Buccinidae tree recovered by Galindo etal. (2016). In the RAxML analyses (Stamatakis 2006) robustness of nodes was assessed using the Thorough Bootstrapping algorithm (Felsenstein 1985) with 1000iterations. The Bayesian inference analyses (BI) were performed using MrBayes (Huelsenbeck& Ronquist 2001), running two parallel analyses, consisting each of six Markov chains of 20 000 000generations with default parameters. Parameters of the substitution model were estimated during the analysis (sixsubstitution categories, a gamma-distributed rate variation across sites approximated in four discrete categories and a proportion of invariable sites). The trees from the first 5 000 000generations (25% from total number of generations) were discarded as burn-in prior to the calculation of consensus trees. Convergence of each analysis was evaluated using tracer 1.4.1 (Rambaut etal. 2014) to check that all ESS values exceeded 200 (with default burning). All analyses were performed on the Cipres Science Gateway (http://www. phylo.org/portal2), using MrBayes on XSEdE (3.2.6) and RAxML-HPC2 on XSEdE (8.2.10) (Miller etal. 2010). The matrix of K2P pairwise genetic distances for COI, was calculated for the Phaenomenella and Siphonalia taxa in MEGA 6 (tamura etal. 2013) (table2). abbreviations and conventions adg opening of anterior duct of digestive gland; AL aperture length; ao anterior aorta; aoe anterior oesophagus; ba buccal artery; bm buccal mass; bn buccal nerves; cep.t cephalic tentacles; cg capsule gland; cm columellar muscle; ct ctenidium; dd dead shell; dg digestive gland; eye eye; ft foot; gl gland of Leiblein; gon gonad; hd head; hg hypobranchial gland; int intestine; kd kidney; lfl longitudinal fold on inner stomach wall; lv live collected specimen; mrr medial retractor of radula; n nerves; nr nerve ring; od odontophore; odn odontophore nerves; odr odontophore retractors; oeo oesophageal opening; 36 ZOOSYSTEMA • 2020 • 42 (3) Kantor Y. et al. Table 1 . — List of sequenced material used in phylogenetic reconstructions. Exact collection localities of specimens from MNHN (specimen code starts with MNHNIM-) can be found at MNHN site at https://science.mnhn.fr/institution/mnhn/collection/im/item/****-***** (asterisks correspond to the digital number of specimen). Accession numbers Specimen code Genus species Collection data BOLD Genbank (COI) Genbank (28S) Source IM-2007-32673 Siphonalia spadicea TAIWAN 2004, st. CP264 NEOGA544-10 MK567642 – Present study IM-2007-32674 spadicea TAIWAN 2004, st. CP264 NEOGA545-10 MK567658 – Present study IM-2007-32762 spadicea TAIWAN 2001 st, CP79 NEOGA589-10 MK567643 – Present study IM-2007-32856 spadicea TAIWAN 2004, st. CP244 NEOGA601-10 MK567646 – Present study IM-2007-32995 spadicea TAIWAN 2001 st, CP109 NEOGA666-10 MK567652 – Present study IM-2007-32997 spadicea TAIWAN 2001 st, CP109 NEOGA668-10 MK567664 – Present study IM-2007-32996 spadicea TAIWAN 2001 st. CP109 NEOGA667-10 MK567656 MK567635 Present study IM-2009-11271 spadicea TAIWAN 2004 st. CH257 BUCC001-19 MK567649 MK567632 Present study IM-2013-41068 Phaenomenella cf thachi TAIWAN 2013 st. CP4090 BUCC007-19 MK567653 – Present study IM-2013-41072 inflata TAIWAN 2013 st. CP4090 BUCC008-19 MK567650 – Present study IM-2013-41073 inflata TAIWAN 2013 st. CP4090 BUCC009-19 MK567660 – Present study IM-2013-50012 cf thachi DONGSHA 2014 st. CP4120 BUCC006-19 MK567654 MK567634 Present study IM-2013-50203 insulapratasensis DONGSHA 2014 st. CP4129 BUCC002-19 MK567648 MK567631 Present study IM-2013-50204 insulapratasensis DONGSHA 2014 st. CP4129 BUCC003-19 MK567661 – Present study IM-2013-50205 insulapratasensis DONGSHA 2014 st. CP4129 BUCC004-19 MK567655 – Present study IM-2013-50260 cf callophorella DONGSHA 2014 st. CP4130 BUCC005-19 MK567665 MK567639 Present study IM-2013-61674 samadiae n. sp. ZHONGSHA 2015 st. CP4134 BUCC013-19 MK567662 MK567638 Present study IM-2013-61617 samadiae n. sp. ZHONGSHA 2015 st. CP4133 BUCC015-19 MK567644 MK567628 Present study IM-2007-34644 nicoi n. sp. AURORA 2007 st. CP2685 NEOGA783-10 MK567663 – Present study IM-2013-59398 nicoi n. sp. ZHONGSHA 2015 st. CP4157 BUCC016-19 MK567645 MK567629 Present study IM-2013-61585 nicoi n. sp. ZHONGSHA 2015 st. CP4132 BUCC014-19 MK567641 MK567627 Present study IM-2013-61637 nicoi n. sp. ZHONGSHA 2015 st. CP4133 BUCC010-19 MK567647 MK567630 Present study IM-2013-61639 nicoi n. sp. ZHONGSHA 2015 st. CP4133 BUCC011-19 MK567651 MK567633 Present study IM-2013-61673 nicoi n. sp. ZHONGSHA 2015 st. CP4134 BUCC012-19 MK567659 MK567637 Present study IM-2007-34639 nicoi n. sp. AURORA 2007 st. CP2685 NEOGA781-10 MK567657 MK567636 Present study IM-2007-34635 Calagrassor aldermenensis AURORA 2007 st. CP2673 KC756037 KC755997 Kantor et al. 2013 IM-2007-32864 Manaria clandestina PANGLAO 2005 st. CP2389 KC756058 KC756004 Kantor et al. 2013 IM-2007-32952 brevicauda SALOMON 2 st. CP2219 KC756055 KC756003 Kantor et al. 2013 IM-2009-7079 Enigmaticolus monnieri MIRIKY st. CP3279 KC756034 KC755996 Kantor et al. 2013 20140783 Buccinum undatum Reykjanesskagi, Iceland MK558051 MK543285 Vaux et al. 2017 KK12 Kelletia kelletii Santa Barbara, California, USA MH198161 MH277543 Vaux et al. 2017 KL2 lischkei Kansai, Mie Prefecture, Japan MH198160 MH277544 Vaux et al. 2017 SFKH-TMP005 Pararetifusus carinatus Chatham Rise, NZ MK583342 MK543289 Vaux et al. 2017 M.190082/2 Penion chathamensis Chatham Rise, NZ MH140429 MH277545 Vaux et al. 2017 M.190085 chathamensis Chatham Rise, NZ MH140428 MH277546 Vaux et al. 2017 M.183792/1 cuvierianus Red Mercury Island, NZ MH140431 MH277548 Vaux et al. 2017 M.183927 cuvierianus Coromandel, NZ MH140432 MH277549 Vaux et al. 2017 C.456980 mandarinus Gabo Island, Victoria, Australia MG211145 MH277553 Vaux et al. 2017 C.487648 maximus Terrigal, NSW, Australia MG211144 MH277554 Vaux et al. 2017 Phoenix1 fairfieldae Otago Peninsula, NZ MH198165 MH277547 Vaux et al. 2017 Phoenix9 sulcatus Auckland, NZ MG098232 MG194426 Vaux et al. 2017 M.274111 Aeneator benthicolus Cape Palliser, NZ MK577960 MK543272 Vaux et al. 2017 SFKH-TMP015 elegans Chatham Rise, NZ MH198157 MH277534 Vaux et al. 2017 M.279437 otagoensis Tasman Bay, NZ MK577961 MK543274 Vaux et al. 2017 M.190119 recens Cape Turnagain, NZ MH198159 MH277535 Vaux et al. 2017 SFKH-TMP013 valedictus TAN 616/83, NZ MK577962 MK543276 Vaux et al. 2017 M.183832 Antarctoneptunea benthicola Chatham Rise, NZ MH198156 MH277537 Vaux et al. 2017 SFKH-TMP014 Austrofusus glans Island Bay, Wellington, NZ MK558053 MK543277 Vaux et al. 2017 M.302907/2 Buccinulum fuscozonatum Ariel Bank, Gisborne, NZ MH198158 MH277540 Vaux et al. 2017 M.258277/6 pallidum Stewart Island, NZ MK577963 MK543280 Vaux et al. 2017 M.302870/2 pertinax finlayi Point Gibson, NZ MH198162 MH277541 Vaux et al. 2017 M.314755/1 robustum Oneroa Bay, Bay of Islands, NZ MK577965 MK543282 Vaux et al. 2017 SFKH-TMP012 vittatum vittatum Mahia Peninsula, NZ MK577964 MK543284 Vaux et al. 2017 SFKH-TMP009 Cominella adpsersa Urupukapuka Bay, NZ MH198163 MH277542 Vaux et al. 2017 SFKH-TMP010 virgata Spirits Bay, NZ MK558054 MK543287 Vaux et al. 2017 SFKH-TMP004 Glaphyrina caudata Farwell Spit, Golden Bay, NZ MK558055 MK543288 Vaux et al. 2017 37 Taxonomic position of Phaenomenella ZOOSYSTEMA • 2020 • 42 (3) op operculum; os osphradium; p penis; pdg opening of posterior duct of digestive gland; pma posterior mixing area; poe posterior oesophagus; pr proboscis; prp propodium; prpg propodial groove; prr proboscis retractors; pw proboscis wall; r radula; rd rhynchodaeum; re rectum; s siphon; sd salivary duct; sg salivary gland; SL shell length; sp seminal papilla; st stomach; st. station; SW shell width; va vagina; vd vas deferens; vl valve of Leiblein. Institution MNHN Muséum national d’Histoire naturelle, Paris. RESuLtS phylogenetic analysis Phylogenetic analysis of both the the COI and 28S datasets recovered a well-supported clade comprising sequences of Siphonalia and Phaenomenella ( Figs1; 2 ). The K2P pairwise distances distribution for Phaenomenella and Siphonalia revealed two modes: below 1.1% and above 7.2% that we interpret as corresponding to the intraand interspecific comparisons respectively ( table2 ). The seven revealed clusters also corresponded to the well-supported monophyletic or single specimen lineages on the COI-based tree but relationships among them are mostly poorly resolved. Specimens of only six cluster recognized based on the COI were present in the 28S data set, and they formed five reciprocally monophyletic lineages ( Fig.2 ). Thus the 28S-based clusters mostly corresponded to those revealed with COI, except for the unresolved relationship of the specimen MNHN-IM-2013-50260. Based on the analysis of single-gene datasets, we conclude that the analyzed specimens of Phaenomenella and Siphonalia comprise seven MOtus. two of the MOtu can be identified as Phaenomenella inflata Shikama, 1971 (Fig.1 (1)) and Phaenomenella insulapratasensis (Okutani& Lan, 1994) (Fig.1(5)). One MOtu was represented by a single specimen only in the COI data set (Fig.1(3), MNHN-IM-2013-50260). It has strong resemblance to Phaenomenella callophorella (Fraussen, 2004), described off taiwan based on a single specimen from similar depths (500-900m vs 795-822m in our specimen) (Fraussen 2004), although it differs in having higher spire and less inflated whorls. Since we are not able to identify the variability of this species we attribute our specimen to P.callophorella with some reservation. two specimens (Fig.1(2), MNHNIM-2013-50012 and MNHN-IM-2013-41068) are subadults (with shell length less than 15mm) and have some resemblance to P.thachi Fraussen& Stahlschmidt, 2012. Although they may represent a different species, the limited material available to us prevents us presently from further taxonomic consideration of the species. two MOtus were represented by several well-preserved adult specimens and cannot be attributed to any existing species of Phaenomenella. They are described herein as new species, Phaenomenella samadiae n.sp. and P. nicoi n.sp. Finally one MOtu can be unambiguously identified as Siphonalia spadicea (Reeve, 1847). The analysis of the COI–28S data set (Fig.3) recovered a well-supported Phaenomenella-Siphonalia cluster consistent with the one in the COI tree, except for P.inflata, which was not represented in the concatenated data set. Whereas Phaenomenella cf. thachi, P. cf. calloporella and P. nicoi n.sp. formed a well-supported subclade (BI posterior probability = 0.97), P. samadiae n.sp. showed weakly supported affinity to Siphonalia spadicea (BI posterior probability = 0.75). This result questions monophyly of Phaenomenella in relation to Siphonalia. In the absence of the data on Siphonalia cassidariaeformis, the type species of the genus, and bearing in mind the conchological differences between the genera, we accept the conservative approach and do not synonymize Phaenomanella with Siphonalia. The two major clades of the Southern hemisphere buccinids included in the present study formed a weakly supported clade comprising a well-supported Buccinulum – Aeneator grouping (BI posterior probability = 1.00), and a weakly supported Penion-Kelletia-Antarctoneptunea clade (BI posterior probability = 0.73). This clade showed no supported relationship to the Phaenomenella-Siphonalia cluster. As mentioned above, Penion and Aeneator were originally included by Finlay (1928) in Siphonaliinae. SYStEMAtIC ACCOuNt Order NEOGAStROPOdA Wenz, 1938 Family buccinidae Rafinesque, 1815 Genus Phaenomenella Fraussen& Hadorn, 2006 type species.— Manaria (?) inflata Shikama, 1971 (Od). reMarks The intrageneric shell variability of Phaenomenella is high and a few general characters can be mentioned – “broader than high protoconch with flattened tip and (…) a rather sharp angulation just above the suture. The upper teleoconch whorls are shouldered, a shape which is accentuated by the axial knobs or ribs, or have the appearance of being by the presence of obviously convex axial ribs” (Fraussen& Stahlschmidt 2013:82). Radula with a tricuspid central tooth with rectangular base and laterals with 3 or 4cusps. Anterior foregut with well-defined valve of Leiblein and large gland of Leiblein. 38 ZOOSYSTEMA • 2020 • 42 (3) Kantor Y. et al. Table 2. — The matrix of K2P pairwise genetic distances in % (e.g. 133 = 0.133) for COI for the Phaenomenella Fraussen & Hadorn, 2006 and Siphonalia A. Adams, 1863 species. Within species: min = 0.000, mean = 0.018, max = 0.011. Between species: min = 0.072, mean = 0.157, max = 0.198. IM-2013-41072 Ph. inflata IM-2013-41073 Ph. inflata IM-2013-50203 Ph. insulapratasensis IM-2013-50204 Ph. insulapratasensis IM-2013-50205 Ph. insulapratasensis IM-2013-41068 Ph. cf thachi IM-2013-50012 Ph. cf thachi IM-2013-50260 Ph. cf callophorella IM-2007-34639 Ph. nicoi IM-2007-34644 Ph. nicoi IM-2013-59398 Ph. nicoi IM-2013-61585 Ph. nicoi IM-2013-61637 Ph. nicoi IM-2013-61639 Ph. nicoi IM-2013-61673 Ph. nicoi IM-2007-32673 S. spadicea IM-2007-32674 S. spadicea IM-2007-32762 S. spadicea IM-2007-32856 S. spadicea IM-2007-32995 S. spadicea IM-2007-32996 S. spadicea IM-2009-11271 S. spadicea IM-2007-32997 S. spadicea IM-2013-61674 Ph. samadiae IM-2013-61617 Ph. samadiae IM-2013-41072 Ph. inflata IM-2013-41073 Ph. inflata 000 IM-2013-50203 Ph. insulapratasensis 133 133 IM-2013-50204 Ph. insulapratasensis 133 133 000 IM-2013-50205 Ph. insulapratasensis 139 139 006 006 IM-2013-41068 Ph. cf thachi 130 130 127 127 124 IM-2013-50012 Ph. cf thachi 127 127 127 127 127 002 IM-2013-50260 Ph. cf callophorella 130 130 120 120 123 077 080 IM-2007-34639 Ph. nicoi n. sp. 140 140 130 130 136 072 072 079 IM-2007-34644 Ph. nicoi n. sp. 140 140 130 130 136 072 072 079 000 IM-2013-59398 Ph. nicoi n. sp. 140 140 131 131 136 081 082 079 011 011 IM-2013-61585 Ph. nicoi n. sp. 140 140 131 131 136 081 082 079 011 011 000 IM-2013-61637 Ph. nicoi n. sp. 140 140 130 130 136 076 077 079 004 004 006 006 IM-2013-61639 Ph. nicoi n. sp. 143 143 133 133 139 079 079 077 009 009 002 002 004 IM-2013-61673 Ph. nicoi n. sp. 143 143 133 133 139 079 079 077 009 009 002 002 004 000 IM-2007-32673 S. spadicea 195 195 194 194 191 171 174 165 172 172 170 170 169 173 173 IM-2007-32674 S. spadicea 195 195 194 194 191 171 174 165 172 172 170 170 169 173 173 000 IM-2007-32762 S. spadicea 195 195 194 194 191 171 174 165 172 172 170 170 169 173 173 000 000 IM-2007-32856 S. spadicea 185 185 185 185 182 168 171 162 169 169 166 166 166 169 169 006 006 006 IM-2007-32995 S. spadicea 198 198 198 198 194 174 178 168 176 176 173 173 173 176 176 002 002 002 009 IM-2007-32996 S. spadicea 192 192 191 191 188 168 171 162 169 169 167 167 166 170 170 002 002 002 004 004 IM-2009-11271 S. spadicea 195 195 194 194 191 171 174 165 172 172 170 170 169 173 173 000 000 000 006 002 002 IM-2007-32997 S. spadicea 192 192 191 191 188 168 171 162 169 169 167 167 166 170 170 002 002 002 004 004 000 002 IM-2013-61674 Ph. samadiae n. sp. 157 157 157 157 163 172 171 156 176 176 173 173 179 176 176 186 186 186 176 182 183 186 183 IM-2013-61617 Ph. samadiae n. sp. 160 160 159 159 166 169 168 152 167 167 164 164 170 167 167 179 179 179 170 183 176 179 176 006 39 Taxonomic position of Phaenomenella ZOOSYSTEMA • 2020 • 42 (3) 1.00 Phaenomenella cf thachi IM-2013-50012 (2) Aeneator valedictus S FKH-TMP013 Glaphyrina caudata S FKH-TMP004 Kelletia kelletii KK12 Aeneator elegans SFKH-TMP015 Volutopsius norwegicus 20140783 Pararetifusus carinatus SFKH-TMP005 Phaenomenella insulapratasensis IM-2013-50205 Penion sulcatus Phoenix1 IM-2013-61639 Austrofusus glans SFKH-TMP014 IM-2013-61673 Manaria brevicauda Siphonalia spadicea IM-2007-32997 Siphonalia spadicea IM-2007-32996 IM-2007-34639 Aeneator recens M.190119 Siphonalia spadicea IM-2007-32856 (6) IM-2013-61674 Penion cuvierianus M.183792 Siphonalia spadicea IM-2007-32762 IM-2007-34644 Calagrassor aldermenensis IM-2007-34635 Penion mandarinus C.456980 IM-2013-61585 (4) Penion chathamensis M.190082/2 Enigmaticolus monnieri IM-2009-7079 Penion cuvierianus M.183927 Phaenomenella insulapratasensis IM-2013-50204 Buccinum undatum 20070640 Aeneator otagoensis M.279437 Cominella adspersa S FKH-TMP009 Penion chathamensis M.190085/3 Buccinulum pertinax finlayi M.302870/2 Buccinulum vittatum vittatum SFKH-TMP012 Antarctoneptunea benthicola M.183832 Phaenomenella cf callophorella IM-2013-50260 (3 ) Kelletia lischkei KL2 Penion sulcatus Phoenix9 Phaenomenella insulapratasensis IM-2013-50203 (5) Phaenomenella cf thachi IM-2013-41068 Siphonalia spadicea IM-2007-32673 Cominella virgata brookesi SFKH-TMP010 Phaenomenella inflata IM-2013-41072 (1) IM-2013-61637 Siphonalia spadicea IM-2007-32674 Siphonalia spadicea IM-2009-11271 Buccinulum fuscozonatum M.302907/2 Phaenomenella inflata IM-2013-41073 Buccinulum robustum M.314755/1 IM-2013-59398 Manaria clandestina IM-2007-32864 Buccinulum pallidum M.258277_6 Siphonalia spadicea IM-2007-32995 Aeneator benthicolus M.274111 Penion maximus C.487648 IM-2013-61617 (7) 1.00 1.00 0.99 0.99 0.99 0.96 0.94 0.96 0.75 0.94 0.84 100 1.00 1.00 0.89 0.87 1.00 0.98 0.78 1.00 0.97 Phaenomenella nicoi n.sp. 1.00 1.00 1.00 Phaenomenella samadiae n.sp. 0.82 0.92 0.96 0.4 M a n a ri a b r e vi cauda g M a n a ri a c l a n de s tin a IM-2 00 73 2 86 4 0.82 0 . 4 (1) (2) (3) (4) (5) (6) (7) Fig. 1. — Phylogenetic tree of a dataset of Buccinoidea Rafinesque, 1815 obtained with Bayesian analysis of COI gene sequences. Support values shown for the supported nodes with posterior probability values between 0.70 and 1.00 only. The numbers in brackets following the species names correspond to illustrated specimens at the bottom of the figure. 46 ZOOSYSTEMA • 2020 • 42 (3) Kantor Y. et al. AB CE D F H G K L JI Fig. 7. — Shells of Phaenomenella nicoi n. sp.: A-C, holotype, MNHN-IM-2013-61585, SL 41.1 mm; D, ZHONGSHA 2015, st. CP4157, sequenced spm, MNHNIM-2013-59398, SL 37.5 mm; E, ZHONGSHA 2015, st. CP4133, sequenced spm, MNHN-IM-2013-61639, SL 43.2 mm; F, AURORA 2007, st. CP2685, sequenced spm, MNHN-IM-2007-34639, SL 34.7 mm; G, AURORA 2007, st. CP2685, not-sequenced spm, MNHN-IM-2007-34644, SL 38.7 mm; H, ZHONGSHA 2015, st. CP4133, sequenced spm, MNHN-IM-2013-61637, SL 30.6 mm (anatomy studied); I, ZHONGSHA 2015, st. CP4134, sequenced spm, MNHN-IM-2013-61673, SL 42 mm; J, ZHONGSHA 2015, st. CP4133, not-sequenced spm, MNHN-IM-2013-61636, SL 40.2 mm (anatomy studied); K, L, ZHONGSHA 2015, st. CP4132, not-sequenced spm, MNHN-IM-2013-61592, 39.1 mm; L, enlarged protoconch. Scale bars: A-K, 20 cm; L, 1 mm. 47 Taxonomic position of Phaenomenella ZOOSYSTEMA • 2020 • 42 (3) A B C D G E F p prpg ft pma eye scm st gon pr rd vl aoe prr sd gl poe ao nr prr sd ba vl aoe poe prr mrr bm aoe odr rd r bn pw gl sg sg sg nr poe gl sd pma st poe dg cep.t hd os sp p ct ht poe kd int pr dg Fig. 8. — Anatomy of Phaenomenella nicoi n. sp.: A-D, MNHN-IM-2013-61636, male; E-G, MNHN-IM-2013-61637, female: A, soft body removed from the shell; B, penis; C, proboscis, opened dorsally; D, foregut, right lateral view; E, foregut, right lateral view, right salivary gland displaced; F, foregut, right lateral view, right salivary gland in original position; G, stomach, outer view. Abbreviations: see material and methods. Scale bars: 5 mm. 48 ZOOSYSTEMA • 2020 • 42 (3) Kantor Y. et al. penultimate whorl, on last whorl 55 cords, of which about 20 on canal. Cords differing slightly in width, with most narrow on subsutural ramp, interspaces between cords about 1/3-½ of cord’s width. Strong axial ribs present on entire shell, suture to suture on uppermost teleoconch whorls, gradually becom - ing obsolete on subsutural ramp and absent on ramp of last and penultimate whorls. On last whorl ribs distinct only on shoulder, producing its angulated appearance. Number of ribs stable, i.e., 15-16per whorl. Aperture ovate, white inside, angulated posteriorly, outer lip thin, evenly convex, concave at transition to canal. Parietal wall and columella with narrow and thin glossy callus. Shell covered with very light olive adhering periostracum, forming densely spaced low axial lamellae visible in interspaces between cords. Operculum partially abraded, when intact (Fig.7 d) spanning most of aperture with distinctly turned leftwards terminal nucleus and rounded upper part. Radula (Fig.5 C-F) Examined in five specimens, including holotype. Rather similar in all specimens; central tooth with rectangular basal part with weakly arcuate anterior margin and three medium long triangular broad cusps, central one shorter and narrower than lateral ones. Lateral teeth normally tricuspate with weakly curved basal side, attached to membrane. Outermost cusp recurved, medium long, inner cusp weakly recurved, about 2/3 of outer cusp length. Intermediate cusp shortest, situated slightly closer to inner cusp. In one sequenced specimen (MNHN-IM-2013-61673, Fig.5F) intermediate cusps of the left lateral teeth of the radula paired, nearly equal in size, while lateral teeth on right side have broader intermediate cusp subdivided on top. Measurements (holotype) Shell length 41.3mm, last whorl length 26.9mm, aperture length (without canal) 16.0mm, diameter 16.5mm. In the largest specimen studied, shell length reaching 43 mm. Anatomy two specimens examined: MNHN-IM-2013-61636, male, and MNHN-IM-2013-61637, female, sequenced paratype (similar in both studied specimens). Soft body partly extracted from the shell. Head medium large, with two long tentacles and large black eyes on lobes at bases of tentacles. Mantle similar to that of Phaenomenella samadiae n. sp. Penis flattened, with seminal papilla situated on its top and surrounded by circle fold of skin (Fig.8B). digestive system. Proboscis almost completely inverted into rhynchodaeum (Fig.8d, pr). Several bands of proboscis retractors muscles attached to middle part on both sides of rhynchodaeum (Fig.8d-F, prr). Buccal mass slightly shorter than retracted proboscis (Fig.8C, bm), attached to its walls by multiple odontophoral retractors (Fig.8C, odr). Radula lying in middle of buccal mass and attached to proboscis walls by median retractor (Fig.8C, mrr). Anterior oesophagus straight, following along ventral side of proboscis (Fig.8E, aoe). Valve of Leiblein (Fig.8d, E, vl) medium large, coniform, situated immediately before nerve ring (Fig.8d, nr). Salivary glands medium-large (about 0.3proboscis length), fused ventrally beneath nerve ring (Fig.8E, F, sg), with very thin strongly convoluted salivary ducts following along anterior oesophagus. Gland of Leiblein medium in size (Fig.8d, E, gl), following along posterior oesophagus and anterior aorta. Stomach rather large, spanning about 0.4whorl (Fig.8G). Posterior mixing area (pma) twice larger than in Phaenomenella samadiae, posterior oesophagus and intestine medium wide. reMarks The new species is highly variable in shell shape. Some of the specimens are much more slender (Fig.7d, I, G) and the axial ribs are either very weak or obsolete. The specimen with no axial ribs was collected at a maximal depth (1634-1683m), but there is not clear correlation with depth, since syntopic specimens can have strong or weak axial ribs. Nevertheless the molecular data clearly indicates the conspecifity of “typical” angulated specimens with well-developed ribs and smooth ones. The specimens collected at greater depth have the protoconch and upper teleoconch whorls more eroded or missing. In general shape the new species has some resemblance to P.mokenorum Fraussen, 2008 from the Andaman Sea, differing in better pronounced axial ribs in later teleoconch whorls and more attenuated narrow siphonal canal. Also P.nicoi n. sp. has smaller size (maximal shell length 42mm versus 55.6mm in P.mokenorum). Genus Siphonalia A.Adams, 1863 type species. — Buccinum cassidariaeforme Reeve, 1846 (Subsequent designation by Cossmann 1889). Siphonalia cassidariaeformis (Reeve, 1846) (Figs9A-C; 10; 11A) Buccinum cassidariaeformis Reeve, 1846: pl. 2, sp. 11.. M aterial exaMined .— Japan • 2lots, 3specimens; Off Hashima, Miyazaki Prefecture, Kyushu; 10.V.1996 (no.1, Fig.9A); Off Atsumi Peninsula, Aichi Prefecture; 30m; 13.V.2001 (nos. 2, 3, Figs9B, C). coMpleMent to description Radula Radula studied in three specimens. Rather similar in all specimens (Fig.11A, B); central tooth with rectangular basal part and weakly arcuate anterior margin and three medium long triangular broad cusps, central one equal in length but slightly narrower than lateral ones. Lateral teeth tricuspate with weakly curved, nearly straight basal side, attached to membrane. Outermost cusp recurved, medium long, inner cusp weakly recurved, about 2/3 of outer cusp length. Intermediate cusp shortest, situated closer to inner cusp. 49 Taxonomic position of Phaenomenella ZOOSYSTEMA • 2020 • 42 (3) Fig. 9. — Shells of studied Siphonalia spp.: A-C, S. cassidariaeformis (Reeve, 1846): specimen no. 1 (A), specimen no. 2 (B), specimen no. 3 (C); D, E, S. pfefferi G. B. Sowerby III, 1900; D, specimen no. 1; E, specimen no. 2. Scale bar: 10 mm. A B C D E 50 ZOOSYSTEMA • 2020 • 42 (3) Kantor Y. et al. A A-C E, F D G H I B C E F D G I H dg kd ft pr s sg prr aoe poe lfl adg int poe oeo gl prr n nr pr vl pr sg sd dg st pma pma gl poe nr aoe adg int poe oeo pdg poe op ft st dg m m hd eyes cep.t hd s s pr sp ft eye Fig. 10 . — Anatomy of Siphonalia cassidariaeformis (Reeve, 1846): A, B; ventral (A) and dorsal (B) views of body of specimen no. 1, removed from the shell; C, cephalopodium of no. 2, front view; D, penis of no. 3; E, F, foregut of no. 1, with right salivary gland in original position (E) and removed (F); G, stomach of no. 2, external view; H, stomach of no. 1, opened dorsally; I, stomach of no. 2, opened dorsally. Abbreviations: see material and methods. Scale bars: A-F, 10 mm; G, I, 5 mm; H, 1 mm 51 Taxonomic position of Phaenomenella ZOOSYSTEMA • 2020 • 42 (3) Anatomy Soft body (no.1, female, Fig.10A, B, E, F, no.2, female, Fig.10C) with approximately 3whorls. Head short and broad, with short contracted tentacles. Eyes small, situated at small lobes in the middle of tentacles (Fig.10B, C, eyes). Foot contracted, with very narrow propodium and large operculum with terminal nucleus. Penis of spm. no.3 (Fig.10d) medium long, flattened, contracted, with small (contracted) rounded seminal papilla at the top, surrounded by circular fold of skin. Mantle with very long siphon in dissected specimens (longer than half mantle width). digestive system. Proboscis half everted out of rhynchodaeum, thick, contracted (Fig.10E, F, pr). Proboscis retractors (prr) attached to rhynchodaeum along both sides of anterior oesophagus (mostly on its right side), connecting rhynchodaeum to lateral walls of body haemocoel. Anterior oesophagus short and wide, dorso-ventrally flattened, along ventral side of proboscis (Fig.10E, F, aoe) into relatively small rounded valve of Leiblein (Fig.10F, vl), situated immediately anterior to nerve ring (Fig.10F, nr). Salivary glands medium small (about 0.25proboscis length) (Fig.10E, F, sg), with very thin strongly convoluted salivary ducts (Fig.10F, sd) following along anterior oesophagus. Gland of Leiblein large (Fig.10E, F, gl), following along thick, round in section posterior oesophagus (Fig.10E, F, poe). Stomach spanning about 0.3whorl (Fig.10G). Posterior mixing area not large in spm. no.1, large in spm. no.2 (Fig.10G, I, pma). Posterior oesophagus and intestine medium wide in both specimens. Opening of posterior duct of digestive gland (found in spm. no.2) located near oesophageal opening (Fig.10I, pdg), opening of anterior duct (found in spms. nos. 1 and 2) located near beginning AB CD Fig. 11. — Radulae of Siphonalia spp. A, S. cassidariaeformis (Reeve, 1846) no. 1; B, S. cassidariaeformis no. 2; C, S. pfefferi G. B. Sowerby III, 1900 no. 1; D, S. pfefferi no. 2. Scale bars: 200 µm. 52 ZOOSYSTEMA • 2020 • 42 (3) Kantor Y. et al. of intestine. Inner stomach wall between two openings with longitudinal fold (Fig.10H, lfl), lined with low oblique folds, remaining part of inner and outer stomach walls lined with moderately high transverse folds. Siphonalia pfefferi G. B. Sowerby III, 1900 (Figs9d, E; 11C, d; 12) Siphonalia pfefferi G. B. Sowerby III, 1900: 440, pl. 11, fig. 3. Material exaMined.— Japan • 1lot, 2specimens; Off Hashima, Miyazaki Prefecture, Kyushu; 10.V.1996 (nos. 1, 2, figs8d, E). coMpleMent to description Radula Radula rather similar in both specimens (Fig.11C, d); central tooth with rectangular basal part and weakly arcuate anterior margin and three medium long triangular broad cusps, central one equal in length but slightly narrower than lateral ones. Lateral teeth tricuspate with weakly curved basal side, attached to membrane. Outermost cusp recurved, medium long, inner cusp weakly recurved, about 2/3of outer cusp length; inner cusp in right longitudinal row of specimen spm. no.2partially subdivided (Fig.11d). Intermediate cusp shortest, situated closer to inner cusp; inner cusp of spm. no.2partially subdivided in left longitudinal row. Anatomy (spm. no.1, male, Fig.12) Head very short and broad, tentacles short, contracted, with small eyes at lobes. Foot contracted, propodium moderately wide, operculum oval with terminal nucleus. Penis rather large (Fig.12C), flattened, with long narrow seminal papilla in deepening at the top. Mantle with medium long siphon. digestive system. Proboscis partly everted out of rhynchodaeum, with contracted walls. Multiple proboscis retractors attaching mostly along right side of anterior oesophagus (Fig.12d, E, prr), connecting rhynchodaeum and lateral walls of body haemocoel. Buccal mass slightly shorter than retracted proboscis (Fig.12F, bm), attaching to its walls by multiple odontophoral retractors (odr). Radula lying in middle of buccal mass and attached to proboscis walls by median retractor (Fig.12F, mrr). Salivary glands (Fig.12d, E, sg) medium large (0.4proboscis length), oval, with salivary ducts following on both sides of anterior oesophagus. Anterior oesophagus wide, dorso-ventrally flattened (Fig.12E, aoe), valve of Leiblein rounded, medium large. Posterior oesophagus (poe) relatively narrow. Gland of Leiblein large, folded beneath nerve ring (Fig.12E, gl). Stomach spanning about 0.3whorl (Fig.12G). Posterior mixing area not large (Fig.12G, H, pma). Intestine medium wide. Opening of posterior duct of digestive gland located near oesophageal opening (Fig.12I, pdg), opening of anterior duct located closer to beginning of intestine. Inner stomach wall between two openings contains longitudinal fold (Fig.12H, lfl), lined with low oblique folds, rest part of inner and outer stomach wall lined with moderately high transverse folds. dISCuSSION Results of the phylogenetic analysis suggest close affinities of Siphonalia and Phaenomenella that remained unnoticed previously. Fraussen& Hadorn (2006), while describing Phaenomenella, compared it to Manaria and Eosipho, but not to Siphonalia. The shell outline of some Phaenomenella (e.g. Phaenomenella insulapratasensis) is rather similar to Siphonalia: the shell is stout, with strongly convex whorls and a recurved siphonal canal. Species of Phaenomenella though have a much larger (about twice) protoconch in comparison with Siphonalia. The intrageneric variability of shell shape in Phaenomenella in its current definition is very high (Fraussen& Stahlschmidt 2013) and in its extremes there is no resemblance between the two genera. It should also be born in mind that some of the most diverging species of Phaenomenella were not yet sequenced and may fall into other lineages. Representatives of both genera are also anatomically similar, particularly in the digestive system characters. Both Phaenomenella and Siphonalia have a broad, medium long in the contracted state proboscis, medium large salivary glands and a large gland of Leiblein. It should be mentioned that despite these general similarities, there are no unique morphological characters uniting both genera. The radular morphology is very similar in both genera (radula of one more species, S.marybethi Parth, 1996 was illustrated in Zhang& Zhang 2018), however, as in the case with the body anatomy, it is of rather generalized buccinid appearance; similar radular morphology can be found in many unrelated genera – eg. Latisipho dall, 1916 (Kosyan 2006), Plicifusus dall, 1902 (Kosyan& Kantor 2012), Aulacofusus dall, 1918 (Kosyan& Kantor 2013). Our molecular analysis did not recover Phaenomenella as monophyletic. In both COI and combined COI+28S analyses the internal relationships within Phaenomenella – Siphonalia clade are not resolved. Siphonalia spadicea cluster without significant support with P.samadiae n.sp. We have only a single species of Siphonalia in our analyses so it is too preliminary to change the classification on the basis of the incomplete dataset. Therefore we presently retain the validity of Phaenomenella, although it is possible that Phaenomenella and Siphonalia can belong to a single genus. One of the distinctions between the genera is the depth range of known species. Generally, species of Siphonalia dwell at shallower depths – from 10 to 300m (Okutani 2000), while Phaenomenella is recorded at 190-1389m (Fraussen& Stahlschmidt 2013; herein). The new species are attributed to Phaenomenella based on stronger conchological similarity to other species of the genus rather than to species of Siphonalia. unfortunately the protoconch of P.samadiae n.sp. was decollated in all available specimens, but the protoconch of P.nicoi n.sp. is large globose, similar to other species of Phaenomenella. The analysis of a broader dataset of Buccinoidea rejected the monophyly of Siphonaliinae in its original scope. None of the Recent genera, originally included by Finlay (1928) in the subfamily, that are Penion, Aeneator and Glaphyrina, are closely related neither to each other, nor to Siphonalia. The system of Buccinidae and Buccinoidea in general is still far 53 Taxonomic position of Phaenomenella ZOOSYSTEMA • 2020 • 42 (3) Fig. 12 . — Anatomy of Siphonalia pfefferi G. B. Sowerby III, 1900, specimen no. 1: A, cephalopodium, ventral view; B, cephalopodium, dorsal view, mantle removed; C, penis; D, foregut, dorsal view; E, foregut, ventral view; F, proboscis opened dorsally; G, stomach, external view; H, stomach, opened dorsally. Abbreviations: see material and methods. Scale bars: A-F, 10 mm; G, H, 5 mm. A A, B D-F G H C kd ct os cm s op sp sp p ft eye prpg od nr gl poe sd odr r aoe pw bm rd mrr pdg lfl adg int poe oeo pma aoe prr dg pma st int poe rd pr sg cep.t hd DEF GH BC 54 ZOOSYSTEMA • 2020 • 42 (3) Kantor Y. et al. from being resolved, with many problematic buccinoidean lineages (see e.g. Couto etal. 2016; Harasewych 2018). Therefore the rank of the inferred clade Siphonalia + Phaenomenella can be resolved only after obtaining the robust phylogeny of the entire superfamily Buccinoidea. Acknowledgements The material was collected mostly in research cruises in the South China Sea and the Philippines Sea organized respectively by National taiwan university (dONGSHA 2014, NANHAI 2014, ZHONGSHA 2015; PI Wei-Jen Chen), and Muséum national d’Histoire naturelle, Paris (MNHN) with the Philippines Bureau of Fisheries and Aquatic Resources (BFAR) (AuRORA 2007; PI Philippe Bouchet) The material in this paper originates from several deep sea cruises, conducted by MNHN, Institut de Recherche pour le développement (IRd) and other partners as part of the Tropical Deep-Sea Benthos programme. The taiwan and South China Sea cruises were supported by bilateral cooperation research funding from the taiwan Ministry of Science and technology (MOSt 1022923–B–002–001–MY3, PI Wei-Jen Chen) and the French National Research Agency (ANR 12-ISV7-000501, PI Sarah Samadi). All expeditions operated under the regulations then in force in the countries in question and satisfy the conditions set by the Nagoya Protocol for access to genetic resources. The study was conducted using Joint usage Center “Instrumental methods in ecology” at the IEE RAS. We thank A. Neretina and A. Nekrasov for their friendly and helpful assistance with the SEM facilities. The studies were supported by the grant from the Russian Science Foundation RSF 16–14– 10118. This project was partly supported by the Service de Systématique moléculaire (uMS 2700 CNRS-MNHN) and the bilateral cooperation research funding from the Ministry of Science and technology, taiwan (grant number MOSt 102-2923-B-002001-MY3) and the French National Research Agency (grant number ANR 12-ISV7-0005-01). 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Journal of Oceanology and Limnology 36: 2333-2336. https://doi.org/10.1007/s00343-019-7218-x Submitted on28 January 2019; accepted on 8 April 2019; published on 4 February 2020.