Frank Köhler

A comprehensive DNA barcode database for Central European beetles with a focus on Germany: adding more than 3500 identified species to BOLD

Beetles are the most diverse group of animals and are crucial for ecosystem functioning. In many countries, they are well established for environmental impact assessment, but even in the well‐studied Central European fauna, species identification can be very difficult. A comprehensive and taxonomically well‐curated DNA barcode library could remedy this deficit and could also link hundreds of years of traditional knowledge with next generation sequencing technology. However, such a beetle library is missing to date. This study provides the globally largest DNA barcode reference library for Coleoptera for 15 948 individuals belonging to 3514 well‐identified species (53% of the German fauna) with representatives from 97 of 103 families (94%). This study is the first comprehensive regional test of the efficiency of DNA barcoding for beetles with a focus on Germany. Sequences ≥500 bp were recovered from 63% of the specimens analysed (15 948 of 25 294) with short sequences from another 997 specimens. Whereas most specimens (92.2%) could be unambiguously assigned to a single known species by sequence diversity at CO1, 1089 specimens (6.8%) were assigned to more than one Barcode Index Number (BIN), creating 395 BINs which need further study to ascertain if they represent cryptic species, mitochondrial introgression, or simply regional variation in widespread species. We found 409 specimens (2.6%) that shared a BIN assignment with another species, most involving a pair of closely allied species as 43 BINs were involved. Most of these taxa were separated by barcodes although sequence divergences were low. Only 155 specimens (0.97%) show identical or overlapping clusters.

MULTIPLE ORIGIN OF VIVIPARITY IN SOUTHEAST ASIAN GASTROPODS (CERITHIOIDEA: PACHYCHILIDAE) AND ITS EVOLUTIONARY IMPLICATIONS

Abstract This study aims at a better understanding of the evolutionary significance of viviparity in some freshwater gastropods. We use a phylogeny based on partial sequences of the mitochondrial 16S gene of representatives of the limnetic and pantropical Pachychilidae to infer the relationships within this particular group of cerithioideans and the evolution of reproductive strategies. The phylogeny presented herein implies a new systematization and suggests that viviparity has appeared three times among the Pachychilidae. This is supported by the finding of very distinct reproductive morphologies in different lineages of viviparous taxa that are exclusively found in Southeast Asia. Based on the observation that oviparity is the ancestral character state in this freshwater family, we conclude that viviparity has evolved subsequent to the exploration of freshwater. We present data showing that all Pachychilidae produce considerably larger but fewer egg capsules compared to most marine snails. In other studies on freshwater gastropods, this has been discussed as an adaptation to freshwater environments. In this context we hypothesize that the increased parental investment involved in the enlargement of eggs in concert with the reduction of clutch sizes was the driving factor that ultimately lead to the evolution of viviparity in the Asian taxa. Consequently, although not directly correlated with the colonization of the new adaptive zone, viviparity is strongly favored by other consequences of this step. Hence, we hypothesize that the production of large eggs, which is necessitated by the exploration of freshwater, represents a preadaptation existing in those ancestors from which viviparous pachychilid lineages eventually evolved in Southeast Asia.

The Camaenid Species of the Kimberley Islands, Western Australia (Stylommatophora: Helicoidea)

ABSTRACT The present paper provides a complete overview of the camaenid land snail fauna of the 22 largest islands along the Kimberley coast, northwesternmost Western Australia, which were surveyed between 2007 and 2010. These islands were found to harbour a total of 104 species (83 new) in 15 genera (5 new), including taxa described recently. Species were essentially delimited by means of comparative analyses of shells (size, shape, sculpture, colouration), genital anatomy (in particular penial anatomy), and genetic differentiation in a mitochondrial marker (partial 16S rRNA sequences). Additionally, the radular and jaw morphology of selected species was studied but found to provide little useful information for taxonomic purposes. Species of the following genera are treated: Amplirhagada Iredale, 1933 (22 species), Carinotrachia Solem, 1985 (2 subspecies), Kimberieymelon Köhler, 2010 (1 species), Kimberleydiscus Köhler, 2010 (1 species), Australocosmica Köhler, 2011 (6 species), Kimboraga Iredale, 1939 (4 species), Xanthomelon Martens, 1860 (1 species), Globorhagada Iredale, 1933 (8 species; the name Globorhagada being removed from synonymy of Xanthomelon), Rhagada Albers, 1860 (8 species), Retroterra Solem, 1985 (3 species), Baudinella Thiele, 1931 (5 species), Setobaudinia Iredale, 1933 (14 species), Torresitrachia Iredale, 1939 (17 species or subspecies). In addition, two new genera, Molema, n. gen. (one species) and Kimberleytrachia, n. gen. (12 species), are described. The taxa Torresitrachia umbonis Solem, 1979, T. deflecta Solem, 1979, and T. crawfordi Solem, 1979, are transferred to the new genus Kimberleytrachia. Species previously assigned to Damochlora are transferred to Setobaudinia; the former genus name likely being a junior synonym of the latter. The following species are delimited differently compared to the latest available revisions: “Xanthomelon” (now: Globorhagada) prudhoensis (E. A. Smith, 1894); “Damochlora” (now: Setobaudinia) rectilabrum (E. A. Smith, 1894); “Torresitrachia” (now: Kimberleytrachia) umbonis (Solem, 1979); Torresitrachia amaxensis Solem, 1979; T. bathurstensis (E. A. Smith, 1894). Most species were found to be island endemics. The work increases the number of known camaenids in northwestern Australia considerably and highlights the region as significant hotspot of land snail diversity.

Morphology, reproductive biology and molecular genetics of ovoviviparous freshwater gastropods (Cerithioidea, Pachychilidae) from the Philippines, with description of a new genus Jagora

The new genus Jagora of the family Pachychilidae Troschel, 1857 is described for the type species Melania asperata Lamarck, 1822 from the Philippines. In addition to J. asperata, a second endemic species of the new genus, J. dactylus (I. Lea & H.C. Lea, 1850), is recognized on the basis of shell morphometry and molecular genetic data (cytochrome C oxidase I and 16S rRNA). The taxonomic history of J. asperata and J. dactylus is revised and the recent distribution documented on basis of available museum material and the authors’ own field collections. J. asperata occurs on Luzon and its satellite islands Leyte and Samar, while J. dactylus is restricted to the Visayan islands Bohol, Cebu, and Guimeras. The morphology of the two species is presented and illustrated in detail, and compared to taxa of the closely related genus Brotia H. Adams, 1866 to which they were previously assigned. Among the South‐east Asian freshwater Cerithioidea of the family Pachychilidae, which were previously subsumed under Brotia for their more or less similar shell morphology and operculum, three distinct lineages can be distinguished, in particular by means of distinct reproductive anatomy: (1) the species of Brotia sensu stricto from mainland South‐east Asia, Sumatra, Borneo and Java, which all exhibit a subhaemocoelic brood pouch; (2) the pachychilid species endemic to Sulawesi, currently assigned to the genera Brotia and Tylomelania, as well as both species of Pseudopotamis endemic to the Torres Strait Islands, which possess a uterine brood pouch; (3) the females of the Philippine Jagora, which carry egg capsules, embryos and advanced juvenile stages within the mantle cavity — a unique reproductive feature. Associated with this mode of ovoviviparity, Jagora is characterized by additional unique properties of the reproductive system including a deeply incised and long sperm gutter in the medial lamina, a very short and posteriorly positioned spermatophore bursa formed by the medial lamina, and a prominent lateral ridge functioning as a seminal receptacle. These characteristics are exclusive to Jagora and are consequently considered to represent autapomorphies of this clade which is endemic only to the Philippines. The zoogeographical implications are discussed in connection with a recently developed palaeogeographical reconstruction.

Tethyan relicts on continental coastlines of the northwestern Pacific Ocean and Australasia: molecular phylogeny and fossil record of batillariid gastropods (Caenogastropoda, Cerithioidea)

The Batillariidae are a family of cerithioidean gastropods consisting of 14 living species, classified in six to eight genera. They are abundant on sandy mudflats and sometimes on rocky shores, on continental margins in the warm‐temperate to tropical regions of the northwestern Pacific Ocean, Australasia and the Americas. Using samples from all 14 nominal batillariid species, we present a molecular phylogenetic hypothesis constructed from two genes, mitochondrial 16S rRNA and nuclear 28S rRNA, with taxa from four marine cerithioidean families as outgroups (Cerithiidae, Modulidae, Planaxidae, Potamididae). The Batillariidae as traditionally conceived are not monophyletic, because the Neotropical ‘batillariids’Lampanella and Rhinocoryne are sister to the Planaxidae. The monophyletic Batillariidae sensu stricto are restricted to the northwestern Pacific and Australasia; we suggest a revised generic classification consistent with our phylogenetic hypothesis and recognize the four genera Batillaria, Pyrazus, Velacumantus and Zeacumantus. We propose a definition of the family using shell characters, as a basis for a review of the extensive fossil record. Batillariids appeared in the Late Cretaceous or Palaeocene, and the extinct genera Pyrazopsis, Vicinocerithium and Granulolabium became diverse in the Tethyan realm before the group disappeared from Europe at the end of the Miocene. The Batillariidae s. s. reached Australia and New Zealand by the Late Oligocene, and the genera Pyrazus, Velacumantus and Zeacumantus still survive in this refugium of Tethyan fauna. Two lineages, Batillaria and the extinct Tateiwaia, migrated north to China and Japan in the Early Miocene, to establish the present disjunct distribution of this relictual group in southern Australasia and the Oriental region.

Conserved shell disguises diversity in Mesodontrachia land snails from the Australian Monsoon Tropics (Gastropoda: Camaenidae)

We comprehensively revise the taxonomy of the camaenid genus Mesodontrachia Solem, 1985, which is endemic to the Victoria River District and East Kimberley in the north‐western Australian Monsoon Tropics based on comparative analyses of key morphological features and mitochondrial DNA sequences. We examined newly collected samples from several collection sites spread over nearly 20 000 km2 of mostly inaccessible land, which represented all three currently known and three previously undescribed species. All species were initially identified as members of Mesodontrachia based on their similar, putatively typical shell. However, Mesodontrachia as so delimited was polyphyletic in a mitochondrial phylogeny with respect to several other camaenid genera from NW Australia. Contrary to the shell, we found considerable variation in the penial anatomy that was consistent with the mtDNA differentiation. To retain monophyletic taxa, we propose a revised taxonomy whereby Mesodontrachia is maintained as a monotypic taxon. In addition, four genera (Nodulabium, Ototrachia, Pseudomesodontrachia and Vincentrachia) and three species (P. gregoriana, O. compressa and N. solidum) are newly described. The shell of all these taxa is highly conserved and of little taxonomic utility. Shell similarity is attributed to a similar life style in a similar and harsh environment.

A molecular phylogeny of camaenid land snails from north-western Australia unravels widespread homoplasy in morphological characters (Gastropoda, Helicoidea)

North-western Australia harbours more than 300 species of camaenid land snail in 41 genera exhibiting considerable and well documented morphological diversity. We performed Bayesian and Maximum Likelihood analyses of concatenated nuclear (28S) and mitochondrial (COI, 16S) DNA sequences from 140 species of 37 of these genera plus an additional 27 extralimital species in order to resolve their phylogenetic relationships and to address the significance of morphological characters for the delineation of monophyletic taxa. While north-western Australian Camaenidae in their entirety are not monophyletic with respect to extralimital groups, they underwent extensive in situ-diversification in several independent phylogenetic radiations. A Maximum Likelihood-based character history reconstruction revealed extensive convergence in all studied shell characters and in three out of four genital features across this group. However, in some genera close morphological similarity is best explained by the retention of ancestral characters. We hypothesize that both morphological stasis in some groups and convergent character evolution in others can to a large extent be attributed to adaptive transformations in response to historically increasing aridity throughout north-western Australia in concert with structural constraints.

Australocosmica, A New Genus of Land Snails from the Kimberley, Western Australia (Eupulmonata, Camaenidae)

ABSTRACT The Western Australian Kimberley region harbours a diverse fauna of camaenid land snails characterised by marked patterns of narrow range endemism. A recent survey of previously poorly known areas along the Kimberley coast has resulted in the discovery of a number of further, yet undeschbed camaenid taxa. One of these, the genus Australocosmica, is newly described herein based on comparative studies of genital anatomy, shell and radular morphology, and analyses of partial sequences of the mitochondrial marker 16S rRNA. Australocosmica is characterised by a broadly conical to semi-globose shell with moderately elevated spire, well-rounded, convex whorls that are separated by a deeply incised suture, and a sculpture of dense, regularly spaced axial ribs. The most distinguishing feature of the inner penial anatomy is the presence of a furrowed, collar-like vergic papillum forming the entrance of the vas deferens into the lumen of the penial chamber. Three new species, Australocosmia augustae, A. sanctumpatriciusae, and A. vulcanice, are described based mainly on differences in penial anatomy. All species are well differentiated by uncorrected pair-wise p-distances of 15% to 20% in the 16S rRNA gene. Similar to most camaenids in the Kimberley region, species of Australocosmica are narrow-range endemics being restricted to single islands, sometimes including the immediately adjacent mainland coast.

Morphological and molecular analysis of ‘Melania’ jacqueti Dautzenberg and Fischer, 1906: from anonymous orphan to critical basal offshoot of the Semisulcospiridae (Gastropoda: Cerithioidea)

Morphological and molecular studies have been inconclusive in assessing monophyly of the Pleuroceridae s.l and the affinity of western North American Juga to Asian members of the family currently grouped in the Semisulcospirinae. In part, this has been hampered by the rarity of anatomical accounts for Asian pleurocerids (s.l.). The present study provides a comprehensive anatomical description of ‘Melania’jacqueti— a Vietnamese species of uncertain generic placement forgotten in the scientific literature for over 100 years. This investigation confirmed that ‘Melania’jacqueti and Juga possess a number of features that differentiate them from eastern North American species, including features of the kidney, prostate and pallial oviduct. However, comparative data remain inadequate to allow phylogenetic analysis based on morphological data alone. Consequently, a data set of partial mitochondrial 16S ribosomal RNA gene sequences for 51 pleurocerids (s.l.) and five outgroups (Melanopsidae, Thiaridae), was assembled. Parsimony, Bayesian and Maximum Likelihood analyses are largely congruent and support monophyly of the Pleuroceridae s.l., and of two large clades: (i) a clade of eastern North American species, and (ii) an Asian–American clade with ‘Melania’jacqueti as the critical basal offshoot, decisively anchoring Juga within the Semisulcospirinae as sister to all remaining Asian forms. Given the morphological disparity between these two clades, this result is consistent with restriction of the concept of Pleuroceridae s.s. to eastern North American species, and elevation of the Asian–American clade to the rank of family — the Semisulcospiridae. Examination of available genus‐group names for Oriental semisulcospirids supports placement of ‘Melania’jacqueti in the genus Hua. Molecular calibration of the basal split between Semisulcospiridae and Pleuroceridae suggests they diverged in the Cretaceous (approximately 90 million years ago). The origin and diversification of these clades are roughly coincident with the subdivision of Laurasia by epicontinental seas and the opening of the Atlantic basin, consistent with a primarily vicariant explanation for their modern biogeographical distributions modified by dispersal in Juga.

Speciation and Radiation in a River: Assessing the Morphological and Genetic Differentiation in a Species Flock of Viviparous Gastropods (Cerithioidea: Pachychilidae)

The Kaek River in central Thailand is unique in harbouring a diverse species assemblage of viviparous gastropods of the genus Brotia. A stretch of this river less than 100 km long is inhabited by seven, mostly endemic species that are essentially differentiated by their shell morphology. Earlier, it has been suggested that this species flock fulfils some basic requirements of a radiation (monophyly and phenotype–habitat correlation). However, the present study has shown that there is no strict correlation between radula and shell morphology and the utilisation of substrates, such as rock or sand, thereby refuting the hypothesis that ecological speciation may have played a significant role. Phylogenetic analyses based on mtDNA show that haplotypes cluster together in drainage-specific clades rather than according to the taxonomy. There are also strong indications that introgressive hybridisation has occurred, which may have resulted from secondary contact of previously isolated species due to dispersal or river captures during the Cenozoic. It is assumed that the high species diversity in the Kaek River results from two phenomena that interdigitate. Firstly, the Kaek River fauna may have originated from multiple species invasions from different source areas, while traces of these events may have been obscured by introgression of Kaek River-specific haplotypes. Secondly, waterfalls in the Kaek River seem to affect the directionality and amount of gene flow between local populations within the river and several smaller tributaries. Together with temporally changing water regimes, this highly structured environment may have conserved local genetic differentiation and triggered diversification and speciation in peripheral isolates within relatively short periods of time.