Ecomorphological and genetic divergence between lowland and montane forms of the Pieris napi species complex (Pieridae, Lepidoptera)

The present study aimed to investigate the morphological and genetic differentiation of lowland and montane populations of the Pieris napi species complex in Norway and to make inferences about the subspecific status of Pieris napi adalwinda and Pieris napi napi . We measured 22 morphological characters on 228 individuals from eight populations in Central Norway (20–1100 m a.s.l.). In addition, lowland and mountain animals were reared at a high altitude locality. Half the animals from either locality were reared on mountain plant, and the other half on a lowland plant. Finally, a fragment of the mitochondrial cytochrome b gene was sequenced for individuals from Central and South-eastern Norway and Germany. Principal component analysis of morphological characters showed a zone of abrupt change from lowland to mountain morphological character states between populations at 506 m and 730 m a.s.l., respectively. The transplant experiment showed that the morphological differences have a genetic basis and that food plants have no impact on morphology. Limited, but significant, molecular genetic differentiation was found between lowland and mountain animals, but the phylogenetic analysis, however, showed that the lowland form ( P. n. napi ) is paraphyletic and the montane form ( P. n. adalwinda ) is monophyletic. Further study is required before taxonomic recognition can be applied.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 727–745.     

Comparative morphology of seeds of four closely related species of Aristolochia subgenus Siphisia (Aristolochiaceae, Piperales)

The primary purpose of this study was to determine whether morphological divergence has occurred in seeds of species in a subclade of Aristolochia subgenus Siphisia . Thus, we compared seed mass, surface area, and wings (presence or absence), embryo shape and state, and embryo length to seed length (E:S) ratio in A. californica Torr., endemic to California, A. macrophylla Lamk. and A. tomentosa Sims of eastern USA, and A. manshuriensis Komarov of eastern Asia. We also compared capsule length and number of seeds per capsule in the three US species. All four species have linear, underdeveloped embryos, but for the other morphological features statistically significant differences occurred among the taxa. However, none of the seed morphological features differed between the eastern North American species A. macrophylla and its eastern Asian sister species A. manshuriensis.   © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 148 , 433–436.     

A revision of Cerdia (Caryophyllaceae)

Cerdia is a poorly understood genus of minute cushiony plants endemic to Mexico that is traditionally placed in subfamily Paronychioideae, tribe Polycarpeae (Caryophyllaceae). Morphological observations and phylogenetic analysis of ITS DNA sequences were carried out to determine the number of species, heretofore controversial, that constitute the genus as well as its position within Caryophyllaceae. Samples of Cerdia were collected through its entire range of distribution and together with previous specimens were used in morphological analyses. Nineteen terminal taxa were considered in a phylogenetic analysis with representatives of the different lineages in Caryophyllaceae. Results indicated that Cerdia is a monotypic genus, including only one variable species, C .  virescens , but its phylogenetic affinities remain doubtful.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 152 , 1–13.     

A new divergent lineage of Daphnia (Cladocera: Anomopoda) and its morphological and genetical differentiation from Daphnia curvirostris Eylmann, 1887

The systematic biology of the subgenus Daphnia s . s . remains confused. Prior attempts at resolution used chiefly postabdominal claw morphology, chromosome numbers and rRNA gene sequences as characters for higher-level relations. Still, several taxa, such as Daphnia curvirostris Eylmann, 1878, have unclear affiliations. We addressed the position of D. curvirostris in this genus by estimating phylogenetic trees from a rapidly evolving protein coding gene (ND2), conducting broad geographical comparisons and carrying out detailed morphological comparisons. The Japanese ' curvirostris' was found to be a new divergent lineage in the subgenus Daphnia , and to possess distinctive morphological characteristics from D. curvirostris . We described this new species as Daphnia tanakai sp. nov. , and redescribed D. curvirostris . The polymorphic postabdominal claw morphology and the distinctive chromosome number of D. tanakai sp. nov. provided evidence for rapid evolution of these traits. Our new morphological, chromosomal and genetic assessment of Daphnia weakened the argument for division of the subgenus Daphnia ( Daphnia ) O. F. Müller, 1785 sensu Johnson, 1952, into two further subgenera.  © 2006 The Linnean Society of London, Zoological Journal of the Linnean Society , 2006, 146 , 385–405.     

Phylogeny of South American Bufo (Anura: Bufonidae) inferred from combined evidence

Despite the considerable research that has focused on the evolutionary relationships and biogeography of the genus Bufo, an evolutionary synthesis of the entire group has not yet emerged. In the present study, almost 4 kb of DNA sequence data from mitochondrial (12S, tRNA^Val, and 16S) and nuclear (POMC; Rag-1) genes, and 83 characters from morphology were analysed to infer a phylogeny of South American toads. Phylogenies were reconstructed with parsimony and maximum likelihood and Bayesian model-based methods. The results of the analysis of morphological data support the hypothesis that within Bufo , some skull characters (e.g. frontoparietal width), correlated with the amount of cranial ossification, are prone to homoplasy. Unique and unreversed morphological synapomorphies are presented that can be used to diagnose recognized species groups of South American toads. The results of all phylogenetic analyses support the monophyly of most species groups of South American Bufo . In most DNA-only and combined analyses, the South American (minus the B. guttatus and part of the ' B. spinulosus ' groups), North American, Central American, and African lineages form generally well-supported clades: ((((((((South America) (North America + Central America)) Eurasia) Africa) Eurasia) South America) West Indies) South America). This result confirms and extends prior studies recovering South American Bufo as polyphyletic. The biogeographical results indicate that: (1) The origin of Bufo predates the fragmentation of Gondwana; (2) Central and North American species compose the sister group to a large, 'derived' clade of South American Bufo ; and (3) Eurasian species form the sister group to the New World clade.  © 2006 The Linnean Society of London, Zoological Journal of the Linnean Society , 2006, 146 , 407–452.     

Phenotypic and genetic differentiation of two major phylogeographical lineages of arctic grayling Thymallus arcticus in the Lena River, and surrounding Arctic drainages

Two phylogeographical lineages of arctic grayling, Thymallus arcticus , in Siberia are extensively characterized based on both molecular genetic (mtDNA control region sequences) and phenotypic (12 meristic characters) data. One lineage, occurring in the delta region of the Lena River as well as all other Arctic draining rivers sampled, corresponds to the subspecific taxon Thymallus arcticus pallasii , whose type locality is the Kolyma River. This taxon is proposed to be a postglacial colonizer of the Lena delta. The second lineage occurs throughout the rest of the Lena basin and is proposed to have survived in a glacial refugium in the middle reaches of the Lena. These lineages form reciprocally monophyletic groups based on mtDNA sequences (net divergence 3.2%), a relationship that is concordant with phenotypic data, and thus reflects distinct taxa. The upper Lena taxon is given the preliminary name of Thymallus arcticus lenensis . Phylogenetic analysis, together with previously published data from North America, reveals that mtDNA sequences from North American populations group within the diverse clade corresponding to T. a. pallasii in Siberia. Despite the relatively close genetic relationship of most North American haplotypes with those in northern Siberia, inferences of fragmentation between the continents are supported, but bidirectional movements between the two continents are seen as likely. Despite inclusion in the clade representing T. a. pallasii in Siberia, the source of the relatively divergent Nahanni refuge haplotypes in North America is not resolved. Otherwise, inferences of postglacial expansion across several thousand kilometres are well supported within North America, but only smaller-scale colonization events among drainages are supported in Siberia.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 88 , 511–525.     

Comparative osteology of the Danio (Cyprinidae: Ostariophysi) axial skeleton with comments on Danio relationships based on molecules and morphology

Danio is a diverse group of small, colourful and easily bred freshwater fishes native to Southern Asia. Biological interest in danios has increased in recent years because the zebrafish, Danio rerio , has become an important model organism, particularly for studies of vertebrate developmental biology and genetics. Though several phylogenetic studies of Danio have been done on a subset of Danio species, the resulting phylogenies conflict in detail. To examine the utility of osteology for systematics of this group at the species level, we studied the axial skeleton for 11 species of Danio. We analyse our morphological data alone and combined with DNA sequence data for five gene sequences generated in earlier phylogenetic studies. The axial skeleton of Danio exhibits 14 characters that prove useful in phylogenetic analysis. Both molecular and morphological data support the monophyly of the danios included in our analysis and both data sets support the monophyly of two subclades: a deep-bodied group and a slender-bodied group. Separate analysis of molecular and morphological data sets show that the molecular data resolves relationships within the slender subclade whereas the topology of the deep-bodied subclade is determined by morphological data.  © 2002 The Linnean Society of London. Zoological Journal of the Linnean Society , 2002, 135 , 529–546.     

Phylogenetic relationships of thorny catfishes (Siluriformes: Doradidae) inferred from molecular and morphological data

Doradidae is a putatively monophyletic group of South American freshwater catfishes containing 30 extant genera and 72 valid species. Only one study to date has attempted to estimate phylogenetic relationships among doradids. This morphological analysis partitioned species into two basal genera ( Wertheimeria and Francisodoras ) and a crown group of three subfamilies (Platydoradinae, Astrodoradinae and Doradinae) whose relationships were unresolved. No subsequent work has been done to resolve the subfamilial trichotomy or to assess whether postulated intergeneric relationships are accurate. We address this problem with complete sequences (2.5 kilobases, kb) of mitochondrial 12S and 16S rRNA genes and partial (1.3 kb) sequences of the nuclear elongation factor-1 alpha (EF1α) gene from representatives of 23 doradid genera (43 species) and 13 outgroups from additional siluriform families. Phylogenetic analysis of these data yields strong support for the monophyly of Doradidae and Astrodoradinae (as well as other relationships), but otherwise shows significant conflict with morphological results. A partial re-examination of published morphological data indicates that many characters may have been incorrectly polarized and many taxa have incorrect state assignments. Our results provide a framework for ongoing efforts to describe the species-level diversity of this poorly understood neotropical family.  © 2004 The Linnean Society of London, Zoological Journal of the Linnean Society , 2004, 140 , 551–575.     

Reproductive character displacement by inversion of coiling in clausiliid snails (Gastropoda, Pulmonata)

In land snails, a change in the direction of coiling, being associated with a shift in the position of the genital apparatus, may act as a barrier against hybridization between sympatric species. Putative reproductive character displacement by an inversion in chirality has been reported in only a few land snails, based on observations in the field and interbreeding experiments. In this study, we present a new case of possible reproductive character displacement in the direction of coiling, in the clausiliid snail Isabellaria dextrorsa . This species is dextral, in contrast with its nearest relatives, including I. torifera and I. lophauchena , which share plesiomorphic sinistral coiling. Whereas I. dextrorsa occurs in sympatry and even syntopically with I. lophauchena throughout most of its range, the sinistral species have a mosaic distribution. Phylogenetic analyses of mitochondrial cytochrome c oxidase subunit I (COI) sequences demonstrated that I. dextrorsa constitutes a clade with I. torifera . In this clade, a shift in coiling direction occurred at least twice, maybe triggered by the presence of a sympatric congeneric sinistral species. The analyses separated the sequences of all I. dextrorsa samples from those of sympatric and syntopic I. lophauchena samples. The failure to demonstrate gene flow between these species is consistent with the hypothesis of genetic isolation by reproductive character displacement.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 88 , 155–164.     

Phylogenetic analysis of Russian Permian dicynodonts (Therapsida: Anomodontia): implications for Permian biostratigraphy and Pangaean biogeography

Dicynodont therapsids have been known from the Upper Permian of Eastern Europe since the beginning of the twentieth century, but the phylogenetic relationships of these taxa have not been examined cladistically. Here we present the results of a phylogenetic analysis that includes eight Permian dicynodonts from Russia as well as 18 taxa best known from southern Africa. Our results do not conflict with much of the established picture of Permian dicynodont phylogeny, but are consistent with several novel hypotheses. Most importantly, our analysis suggests that the genus Dicynodon is paraphyletic, and we question its use in correlating widely separated basins. However, we cannot strongly reject a monophyletic Dicynodon . Our results also indicate that the closest Permian relatives of Kannemeyeria lived in Russia, suggesting a Laurasian origin for the lineage that includes this important Triassic taxon. The phylogeny presented here also suggests a Laurasian origin for several other dicynodont clades, but a Gondwanan origin is equally likely given the data at hand. Regardless of where these groups originated, there appears to be some endemism among Late Permian dicynodont faunas. Although our understanding of dicynodont phylogeny is improving, this study emphasizes the disparity in sampling of the dicynodont record between Gondwana and Laurasia and the need for a large scale phylogenetic analysis of Permian and Triassic dicynodonts.  © 2003 The Linnean Society of London, Zoological Journal of the Linnean Society, 2003, 139 , 157−212.     

Pollen morphology of the Swertiinae (Gentianaceae): phylogenetic implications

Recent molecular surveys of the Swertiinae (Gentianaceae–Gentianeae) revealed unexpected phylogenetic relationships, including polyphyly of the genera Gentianella , Jaeschkea , Lomatogonium and Swertia . To find new non-molecular characters supporting the phylogeny, we examined the exine variation of 73 species of all major lineages of subtribe Swertiinae using environmental scanning electron miscroscopy supplementing older, mainly light microscopical, studies. In contrast to previous studies, we were able to pick out taxa from phylogenetic key positions with particular focus on Swertia . Many distantly related taxa such as parts of Frasera , Gentianopsis , Halenia , Gentianella , Megacodon and several lineages of Swertia share a striate–reticulate or reticulate exine pattern. This is interpreted as the plesiomorphic character state of Swertiinae. There is also considerable variation of derived patterns; for example, different types of microechinate or almost smooth pollen was repeatedly observed in distantly related groups. Another extreme was the ring-shaped reticulation found in a North American species of Gentianopsis . Unfortunately, major relationships as revealed by molecular analyses were rarely supported because of the abundance of the plesiomorphic type and homoplasy even on low taxonomic levels; for example, within Lomatogonium . Exine variation was particularly useful in characterizing independent lineages of Swertia . For example, according to pollen characters and in agreement with other data, the Asian Swertia cuneata is a sister group of a strongly diversified African lineage and Swertia yunnanensis , which is rather aberrant in flower morphology, seems close to parts of Lomatogonium .  © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society , 2008, 157 , 323–341.     

Evolutionary and ecological implications of genome size in the North American endemic sagebrushes and allies (Artemisia, Asteraceae)

The genome size of 51 populations of 20 species of the North American endemic sagebrushes (subgenus Tridentatae ), related species, and some hybrid taxa were assessed by flow cytometry, and were analysed in a phylogenetic framework. Results were similar for most Tridentatae species, with the exception of three taxonomically conflictive species:  Artemisia bigelovii Gray,  Artemisia pygmaea Gray, and  Artemisia rigida Gray. Genome size homogeneity (together with the high morphological, chemical, and karyological affinities, as well as low DNA sequence divergence) could support a recent diversification process in this geographically restricted group, thought to be built upon a reticulate evolutionary framework. The Tridentatae and the other North American endemic Artemisia show a significantly higher genome size compared with the other subgenera. Our comparative analyses including genome size results, together with different kinds of ecological and morphological traits, suggest an evolutionary change in lifestyle strategy linked to genome expansion, in which junk or selfish DNA accumulation might be involved. Conversely, weed or invasive behaviour in Artemisia is coupled with lower genome sizes. Data for both homoploid and polyploid hybrids were also assessed. Genome sizes are close to the expected mean of parental species for homoploid hybrids, but are lower than expected in the allopolyploids, a phenomenon previously documented to be related with polyploidy.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 631–649.     

What,if anything,is a ‘typical’ viper? Biological attributes of basal viperid snakes (genus Causus Wagler, 1830)

For many major phylogenetic radiations of organisms, the available ecological knowledge is disproportionately derived from a small minority of taxa, and sometimes from organisms that are highly atypical. Viperid snakes provide a good example of this situation; high-latitude cold-climate taxa in northern Europe (vipers) and North America (rattlesnakes) have been studied intensively, but more speciose radiations in tropical Africa, Asia, and Central America remain poorly known. We dissected > 500 specimens (six species) of night adders (genus Causus ), mostly from Cameroon (68%) in equatorial Africa, to quantify morphology, diets, and reproductive biology. By contrast to the 'slow' life-histories of cold-climate viperids, night adders feed frequently on frogs (rather than infrequently on mammals), and produce frequent large clutches of relatively small eggs (rather than infrequent small litters of relatively large live young). Thus, putatively 'typical' viperid attributes such as low fecundity, viviparity, and predation on mammals reflect adaptations to the invasion of cold environments by a small and perhaps atypical subset of viperid taxa. Our data on prey size suggest that one of the critical innovations of early viperids may have been an ability to subdue and ingest relatively large prey.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 575–588.     

A molecular study of phylogenetic relationships and evolution of antipredator strategies in Australian Diplodactylus geckos, subgenus Strophurus

We present phylogenetic analyses of the lizard genus Diplodactylus subgenus Strophurus using 1646 aligned positions of mitochondrial DNA sequences containing 893 parsimony-informative characters for samples of 12 species of Strophurus and 19 additional Australian gecko species. Sequences from three protein-coding genes (ND1, ND2 and COI) and eight intervening transfer RNA genes were examined using parsimony, maximum-likelihood and Bayesian analyses. Species of Strophurus appeared to form a monophyletic group with the possible exception of S. taenicauda . Strophurus has evolved two distinct defence/display characteristics: caudal glands, which expel an unpalatable substance, and striking mouth colours. Caudal glands appeared to have arisen once in a common ancestor of Strophurus , with dermal augmentation of caudal glands characterizing a subclade within the subgenus. Evolution of yellow and dark-blue mouth colours in Strophurus occurred in the context of diurnal activity and may be interpreted as an augmentation of defensive behavioural displays. Molecular divergence suggests that arboreality evolved in a common ancestor of Oedura and Strophurus approximately 29 Mya and that the caudal glands of Strophurus arose approximately 25 Mya.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 82 , 123–138.     

Canarian land snail diversity: conflict between anatomical and molecular data on the phylogenetic placement of five new species of Napaeus (Gastropoda, Pulmonata, Enidae)

Five new species of land snail (family Enidae) are described from La Gomera (Canary Islands) of which the majority, on the basis of anatomy alone, could be incorporated within a new supraspecific taxon. In addition to the morphological study of these new species, a region of the 16S mitochondrial gene is sequenced from three of the new species and a range of species of Napaeus from within its two subgenera ( Napaeinus and Napaeus ) . There is a disparity between the morphological and preliminary molecular phylogenetic data. Possible explanations for this conflict are discussed, as well as the evolutionary relationships among these different taxa, and it is suggested that this group may be an excellent model for further studies of adaptation and diversification.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 169–187.     

Genetic relationships of the western Mediterranean painted frogs based on allozymes and mitochondrial markers: evolutionary and taxonomic inferences (Amphibia, Anura, Discoglossidae)

Allozymes and sequencing of mitochondrial cytochrome  b (cyt  b ) and 12S genes were used to reconstruct the genetic structure and phylogenetic relationships of all Discoglossus taxa described so far (except the probably extinct D. nigriventer ). This is the first time that a comprehensive study on the Discoglossus painted frogs has used nuclear and mitochondrial markers, evidencing a discordant pattern between the two datasets. Comparison of these discrepancies suggests a role of stochastic sorting of ancestral polymorphisms, possibly associated with male-biased dispersal and present or past secondary contact. The genetic relationships between taxa with intermediate levels of divergence were well defined by allozyme data, but showed short internodes and low bootstrap values for mitochondrial data, suggesting a rapid radiation of their lineages during the Messinian Lago Mare phase. The results provide information about the taxonomic status of D. galganoi and D. jeanneae , considered as subspecies, and indicate D. pictus as nonmonophyletic, confirming D. scovazzi as a distinct species.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 87 , 515–536.     

Species diversity and endemism in the Daphnia of Argentina: a genetic investigation

Although the temperate regions of South America are known to have a diverse daphniid fauna, there has been no genetic evaluation of the existing taxonomic system or of the affinities between the North and South American faunas. The present study analyses mitochondrial DNA sequences and allozyme variation to investigate species diversity in 176 Daphnia populations from Argentina. This work established the presence of at least 15 species in Argentina, six of which are either undescribed or are currently misidentified and two of which represent range extensions of North American taxa. Eleven of the Argentine species appear endemic to South America, while the remaining four also occur in North America. In the latter cases, the close genetic similarity between populations from North and South America indicates the recent exchange of propagules between the continents. While biological interactions and habitat availability have undoubtedly contributed to the observed species distributions, chance dispersal has apparently played a dominant role in structuring large-scale biogeographical patterns in this genus and probably in other passively-dispersed organisms.  © 2004 The Linnean Society of London, Zoological Journal of the Linnean Society , 2004, 140 , 171−205.