Rosemary E. Golding

Phylogenetic relationships and evolution of pulmonate gastropods (Mollusca): New insights from increased taxon sampling

Phylogenetic relationships among higher clades of pulmonate gastropods are reconstructed based on a data set including one nuclear marker (complete ribosomal 18S) and two mitochondrial markers (partial ribosomal 16S and Cytochrome oxidase I) for a total of 96 species. Sequences for 66 of these species are new to science, with a special emphasis on sampling the Ellobiidae, Onchidiidae, and Veronicellidae. Important results include the monophyly of Systellommatophora (Onchidiidae and Veronicellidae) as well as the monophyly of Ellobiidae (including Trimusculus, Otina, and Smeagol). Relationships within Ellobiidae, Onchidiidae, and Veronicellidae are evaluated here for the first time using molecular data. Present results are compared with those from the recent literature, and the current knowledge of phylogenetic relationships among pulmonate gastropods is reviewed: despite many efforts, deep nodes are still uncertain. Identification uncertainties about early fossils of pulmonates are reviewed. Impacts of those phylogenetic and fossil record uncertainties on our understanding of the macro-evolutionary history of pulmonates, especially transitions between aquatic and terrestrial habitats, are discussed.

Ten new complete mitochondrial genomes of pulmonates (Mollusca: Gastropoda) and their impact on phylogenetic relationships.

BackgroundReconstructing the higher relationships of pulmonate gastropods has been difficult. The use of morphology is problematic due to high homoplasy. Molecular studies have suffered from low taxon sampling. Forty-eight complete mitochondrial genomes are available for gastropods, ten of which are pulmonates. Here are presented the new complete mitochondrial genomes of the ten following species of pulmonates: Salinator rhamphidia (Amphiboloidea); Auriculinella bidentata, Myosotella myosotis, Ovatella vulcani, and Pedipes pedipes (Ellobiidae); Peronia peronii (Onchidiidae); Siphonaria gigas (Siphonariidae); Succinea putris (Stylommatophora); Trimusculus reticulatus (Trimusculidae); and Rhopalocaulis grandidieri (Veronicellidae). Also, 94 new pulmonate-specific primers across the entire mitochondrial genome are provided, which were designed for amplifying entire mitochondrial genomes through short reactions and closing gaps after shotgun sequencing.ResultsThe structural features of the 10 new mitochondrial genomes are provided. All genomes share similar gene orders. Phylogenetic analyses were performed including the 10 new genomes and 17 genomes from Genbank (outgroups, opisthobranchs, and other pulmonates). Bayesian Inference and Maximum Likelihood analyses, based on the concatenated amino-acid sequences of the 13 protein-coding genes, produced the same topology. The pulmonates are paraphyletic and basal to the opisthobranchs that are monophyletic at the tip of the tree. Siphonaria, traditionally regarded as a basal pulmonate, is nested within opisthobranchs. Pyramidella, traditionally regarded as a basal (non-euthyneuran) heterobranch, is nested within pulmonates. Several hypotheses are rejected, such as the Systellommatophora, Geophila, and Eupulmonata. The Ellobiidae is polyphyletic, but the false limpet Trimusculus reticulatus is closely related to some ellobiids.ConclusionsDespite recent efforts for increasing the taxon sampling in euthyneuran (opisthobranchs and pulmonates) molecular phylogenies, several of the deeper nodes are still uncertain, because of low support values as well as some incongruence between analyses based on complete mitochondrial genomes and those based on individual genes (18S, 28S, 16S, CO1). Additional complete genomes are needed for pulmonates (especially for Williamia, Otina, and Smeagol), as well as basal heterobranchs closely related to euthyneurans. Increasing the number of markers for gastropod (and more broadly mollusk) phylogenetics also is necessary in order to resolve some of the deeper nodes -although clearly not an easy task. Step by step, however, new relationships are being unveiled, such as the close relationships between the false limpet Trimusculus and ellobiids, the nesting of pyramidelloids within pulmonates, and the close relationships of Siphonaria to sacoglossan opisthobranchs. The additional genomes presented here show that some species share an identical mitochondrial gene order due to convergence.

Three-dimensional reconstruction of the odontophoral cartilages of Caenogastropoda (Mollusca: Gastropoda) using micro-CT: Morphology and phylogenetic significance

Odontophoral cartilages are located in the molluscan buccal mass and support the movement of the radula during feeding. The structural diversity of odontophoral cartilages is currently known only from limited taxa, but this information is important for interpreting phylogeny and for understanding the biomechanical operation of the buccal mass. Caenogastropods exhibit a wide variety of feeding strategies, but there is little comparative information on cartilage morphology within this group. The morphology of caenogastropod odontophoral cartilages is currently known only from dissection and histology, although preliminary results suggest that they may be structurally diverse. A comparative morphological survey of 18 caenogastropods and three noncaenogastropods has been conducted, sampling most major caenogastropod superfamilies. Three‐dimensional models of the odontophoral cartilages were generated using X‐ray microscopy (micro‐CT) and reconstruction by image segmentation. Considerable morphological diversity of the odontophoral cartilages was found within Caenogastropoda, including the presence of thin cartilaginous appendages, asymmetrically overlapping cartilages, and reflexed cartilage margins. Many basal caenogastropod taxa possess previously unidentified cartilaginous support structures below the radula (subradular cartilages), which may be homologous to the dorsal cartilages of other gastropods. As subradular cartilages were absent in carnivorous caenogastropods, adaptation to trophic specialization is likely. However, incongruence with specific feeding strategies or body size suggests that the morphology of odontophoral cartilages is constrained by phylogeny, representing a new source of morphological characters to improve the phylogenetic resolution of this group. J. Morphol. 2009.

Molecular phylogeny and systematics of Australian and East Timorese Stenothyridae (Caenogastropoda: Truncatelloidea)

The Australian and East Timorese species belonging to the truncatelloid family Stenothyridae are revised using molecular data and morphological characters from the shell, operculum, radula and external and reproductive anatomy. The Australian species Stenothyra australis is redescribed and two previously recognised subspecies are shown to be synonyms. The New Guinean species Stenothyra paludicola van Benthem Jutting, 1963 is redescribed and recorded from the Torres Strait region of northern Australia, and two new subspecies of S. paludicola are described from the Northern Territory and East Timor; S. paludicola topendensis n. subsp. and S. paludicola timorensis n. subsp. respectively. Stenothyra gelasinosa n. sp. is described from Australia, comprising three allopatric subspecies; S. gelasinosa gelasinosa n. sp. and n. subsp. from the eastern seaboard, S. gelasinosa phrixa n. subsp. from northern Australia and S. gelasinosa apiosa n. subsp. from the Pilbara region of Western Australia. Stenothyra frustillum is considered a nomen dubium. Molecular phylogenetic analysis of these taxa and other Asian stenothyrids supports these systematic decisions and provides a preliminary interpretation of relationships within Stenothyridae.

The evolutionary and biomechanical implications of snout and proboscis morphology in Caenogastropoda (Mollusca: Gastropoda)

The caenogastropod proboscis is a complex morphological adaptation to a carnivorous diet. This comparative morphological investigation of proboscis and snout anatomy in Caenogastropoda shows that there is undescribed diversity in both snout/proboscis wall composition and introversion/retraction musculature. There is morphological evidence which suggests that a proboscis evolved separately in at least four separate caenogastropod groups, each characterized by the presence of novel retractor muscles and different modifications of plesiomorphic “aortic muscles”. The biomechanical operation of the proboscis and snout utilizes a hydrostatic skeleton, but several higher caenogastropods have evolved a complex muscular hydrostat in the snout/proboscis wall which may mitigate the need to isolate the proboscis haemocoel during proboscis eversion.

Molecular phylogenetic analysis of mudflat snails (Gastropoda: Euthyneura: Amphiboloidea) supports an Australasian centre of origin.

Amphiboloidea is a small but widespread group of snails found exclusively, and often abundantly, in mudflat and associated salt marsh or mangrove habitat. This study uses molecular data from three loci (COI, 16S and 28S) to infer phylogenetic relationships in Amphiboloidea and examine its position in Euthyneura. All but two of the named extant species of Amphiboloidea and additional undescribed taxa from across Southeast Asia and the Arabian Gulf were sampled. In contrast to the current morphology-based classification dividing Amphiboloidea into three families, analysis of molecular data supports revision of the classification to comprise two families. Maningrididae is a monotypic family basal to Amphibolidae, which is revised to comprise three subfamilies: Amphibolinae, Phallomedusinae and Salinatorinae. Sequence divergence between Asian populations of Naranjia is relatively large and possibly indicative of species complexes divergent across the Strait of Malacca. Salinatorrosacea and Salinator burmana do not cluster with other Salinator species, and require generic reassignment. In addition, sequences were obtained from an undescribed species of Lactiforis from the Malay Peninsula. Reconstruction of ancestral distributions indicates a plesiomorphic distribution and centre of origin in Australasia, with two genera subsequently diversifying throughout Asia. Increasing the sampling density of amphiboloid taxa in a phylogenetic analysis of Euthyneura did not resolve the identity of the sister taxon to Amphibolidae, but confirmed its inclusion in Pulmonata/Panpulmonata.

Homology and morphology of the neogastropod valve of Leiblein (Gastropoda: Caenogastropoda)

The valve of Leiblein is a morphological synapomorphy defining Neogastropoda, but is also a significant adaptation in the evolution of carnivorous, predatory feeding and homologues have not been recognised amongst non-neogastropods to date. This study uses histology to examine the valve of Leiblein and associated oesophageal features of a buccinoidean (Euplica scripta—Columbellidae) and two muricoideans (Morula marginalba—Muricidae, and Columbarium pagodoides—Turbinellidae), and comparisons are made with the oesophagus of four non-neogastropods; the tonnoidean out-group Cabestana spengleri and three other littorinimorph caenogastropods. Several morphological details conflict with earlier descriptions of different neogastropod species, such as the presence of a diverticulum in the muricoidean valve and torsion in the buccinoidean valve. This supports recent findings that the valve of Leiblein is morphologically heterogeneous, but, unlike some other studies, does not dispute the homology of the valve in different neogastropod groups. Structure and histology support the homology of the valve of Leiblein in Muricoidea and Buccinoidea and its derivation from elements of the anterior oesophagus of higher littorinimorph caenogastropods (Tonnoidea). A thick, secretory, pseudostratified epithelium lining the ventral anterior oesophagus of C. spengleri is comparable with the tissue lining the glandular chamber of the valve of Leiblein. A modified theory regarding the evolutionary origin of the valve of Leiblein is proposed in which it is homologous to the anterior oesophagus of Tonnoidea.

Molecular phylogeny and systematics of Australian ‘Iravadiidae’ (Caenogastropoda: Truncatelloidea)

The family Iravadiidae is found to be polyphyletic in a molecular phylogenetic analysis using a subset of Australian taxa. Taxa previously assigned to Iravadia form a monophyletic clade, but Nozeba topaziaca clusters with Auricorona queenslandica n. gen. and n. sp. in an unnamed family related to Tornidae. Aenigmula criscionei n. gen. and n. sp., an iravadiid-like species from the Northern Territory, belongs to another unnamed family related to Caecidae, Calopiidae and Clenchiellidae. A systematic revision of some Australian ‘iravadiids’ raises the subgenera Fluviocingula and Pseudomerelina to full generic rank and reinstates two former synonyms of Iravadia (Fairbankia), Pellamora and Wakauraia, as genera. The species formerly identified in Australia as Iravadia quadrasi is recognised as three allopatric species; Iravadia pilbara n. sp. and the reinstated species Iravadia goliath and Iravadia quadrina. Pellamora splendida n. sp., from Western Australia, is recognised as distinct from Pellamora australis, and Fluviocingula superficialis n. sp. from Fluviocingula resima. Wakauraia fukudai n. sp. is recorded from central Queensland. http://zoobank.org/urn:lsid:zoobank.org:pub:1B9917F6-48B2-4597-85C1-F90BA9093475