Molecular phylogeny and biogeography of the West‐Palaearctic Velia (Heteroptera: Gerromorpha: Veliidae)

We investigated the systematics and biogeography of the West‐Palaearctic water cricket genus Velia Latreille based on a phylogenetic analysis of five molecular markers obtained from 79% of all known taxa of the subgenera Velia (s.s.) and Velia (Plesiovelia ) Tamanini. The results revealed a sister‐group relationship between Velia (Plesiovelia ) and the monotypic subgenus Velia (s.s.), and showed that the former is divided into three major clades. All taxa of Velia (Plesiovelia) were recovered as monophyletic, except for V . (P.) serbica Tamanini, which was paraphyletic with respect to V . (P.) mancinii mancinii Tamanini. Our results also indicate the existence of several unrecognized species. Molecular dating based on fossil data and COI rates indicates that the split between Velia (s.s.) and Velia (Plesiovelia ) occurred between 40 and 22 Ma. An ancestral area reconstruction suggests that the latter originated in southeastern Europe, from where it radiated to the west and east, along the Neogene archipelagos of Europe and Asia Minor. Northwestern Africa served as the second most important diversification centre of the subgenus. The low genetic variability in the widespread V . (P.) caprai caprai Tamanini and V . (P.) saulii Tamanini implies a rapid postglacial colonization of Europe, whereas high diversity within the lineages of V . (P.) serbica indicates survival of Pleistocene glaciations in microrefugia throughout southeastern Europe. These results serve as a useful framework for future studies ranging from the systematics of the group to historical biogeography, ecology and biodiversity conservation.     

Comparative analysis of overwintering physiology in nine species of semi‐aquatic bugs (Heteroptera: Gerromorpha)

In semi‐aquatic bugs (Heteroptera: Gerromorpha), the strategies of overwintering in a cryothermic state (i.e. at body temperatures below the equilibrium freezing point) remain largely unexplored. The present study provides an analysis of the ecophysiological aspects of overwintering in nine gerromorphan species. All nine species avoid ice formation by means of a more or less extensive supercooling of their body fluids. There is a tight correlation between the supercooling point (SCP) and the lower lethal temperature. Different species use different physiological adjustments to increase the likelihood of survival in a supercooled state. These include stabilization of the supercooled state by active antifreeze factors that cause thermal hysteresis between equilibrium melting and freezing points, the accumulation of low‐molecular weight sugars and polyols with putative cryoprotective functions, or by having a relatively high body fluid osmolality, combined with a low level of hydration. The majority of species under study overwinter only as adults, whereas Velia caprai Tamanini can overwinter either as an adult or in the egg stage. The supercooling capacity of V. caprai adults is insufficient to prevent the risk of lethal freezing. The adults therefore survive only opportunistically in suitable microhabitats, and/or during mild winters. The survival of V. caprai in winter is assured by extensive supercooling and having overwintering eggs that are highly cold tolerant.     

Two new species in the Chaetoceros socialis complex (Bacillariophyta): C. sporotruncatus and C. dichatoensis,and characterization of its relatives,C. radicans and C. cinctus

The diatom genus Chaetoceros is one of the most abundant and diverse phytoplankton in marine and brackish waters worldwide. Within this genus, Chaetoceros socialis has been cited as one of the most common species. However, recent studies from different geographic areas have shown the presence of pseudo‐cryptic diversity within the C. socialis complex. Members of this complex are characterized by curved chains (primary colonies) aggregating into globular clusters, where one of the four setae of each cell curves toward the center of the cluster and the other three orient outwards. New light and electron microscopy observations as well as molecular data on marine planktonic diatoms from the coastal waters off Chile revealed the presence of two new species, Chaetoceros sporotruncatus sp. nov. and C. dichatoensis. sp. nov. belonging to the C. socialis complex. The two new species are similar to other members of the complex (i.e., C. socialis and C. gelidus) in the primary and secondary structure of the colony, the orientation pattern of the setae, and the valve ultrastructure. The only morphological characters that can be used to differentiate the species of this complex are aspects related to resting spore morphology. The two newly described species are closely related to each other and form a sister clade to C. gelidus in molecular phylogenies. We also provide a phylogenetic status along with the morphological characterization of C. radicans and C. cintus, which are genetically related to the C. socialis complex.     

Ligulate Desmarestia (Desmarestiales,Phaeophyceae) revisited: D. japonica sp. nov. and D. dudresnayi differ from D. ligulata

The phylogeny of ligulate and sulfuric‐acid containing species of Desmarestia, occurring worldwide from polar to temperate regions, was revised using a multigenic and polyphasic approach. Sequence data, gametophyte characteristics, and sporophyte morphology support reducing a total of 16 taxa to four different species. (1) D. herbacea, containing broad‐bladed and highly branched forms, has dioecious gametophytes. The three other species have monoecious gametophytes: (2) D. ligulata which is profusely branched and, except for one subspecies, narrow‐bladed, (3) Japanese ligulate Desmarestia, here described as D. japonica sp. nov., which is morphologically similar to D. ligulata but genetically distant from all other ligulate taxa. This species may have conserved the morphology of original ligulate Desmarestia. (4) D. dudresnayi, including unbranched or little branched broad‐bladed taxa. A figure of the holotype of D. dudresnayi, which was lost for decades, was relocated. The taxonomy is complemented by a comparison of internal transcribed spacer and cytochrome c oxidase subunit I (cox1) as potential barcode loci, with cox1 offering good resolution, reflecting species delimitations within the genus Desmarestia.     

No evidence of contemporary interploidy gene flow between the closely related European woodland violets Viola reichenbachiana and V. riviniana (sect. Viola,Violaceae)

• Viola reichenbachiana (2n = 4x = 20) and V. riviniana (2n = 8x = 40) are closely related species widely distributed in Europe, often sharing the same habitat throughout their overlapping ranges. It has been suggested in numerous studies that their high intraspecific morphological variability and plasticity might have been further increased by interspecific hybridisation in contact zones, given the sympatry of the species and the incomplete sterility of their hybrid. The aims of this study were to: (i) confirm that V. reichenbachiana and V. riviniana have one 4x genome in common, and (ii) determine the impact of hybridisation and introgression on genetic variation of these two species in selected European populations.
• For our study, we used 31 Viola populations from four European countries, which were analysed using AFLP and sequencing of a variable plastid intergenic spacer, trnH‐psbA.
• Our analysis revealed that V. reichenbachiana exhibited larger haplotype diversity, having three species‐specific haplotypes versus one in V. riviniana. The relationships among haplotypes suggest transfer of common haplotypes into V. riviniana from both V. reichenbachiana and hypothetically the other, now extinct, parental species. AFLP analysis showed low overall genetic diversity of both species, with V. riviniana showing higher among‐population diversity. None of the morphologically designated hybrid populations had additive AFLP polymorphisms that would have indicated recent hybridisation. Also, kinship coefficients between both species did not indicate gene flow. V. riviniana showed significant population subdivision and significant isolation by distance, in contrast to V. reichenbachiana.
• The results indicate lack of gene flow between species, high influence of selfing on genetic variability, as well as probably only localised introgression toward V. riviniana.

     

Complete mitochondrial genome of the black‐tailed hornet,Vespa ducalis (Hymenoptera: Vespidae): Genomic comparisons in Vespoidea

We sequenced the complete mitochondrial genome (mitogenome) of the black‐tailed hornet, Vespa ducalis (Hymenoptera: Vespidae). The genome was 15,779‐bp long and contained typical sets of genes [13 protein‐coding genes (PCGs), 22 tRNAs, and 2 rRNAs]. The V. ducalis A + T‐rich region was 166‐bp long and was the shortest of all sequenced Vespoidea genomes, including Vespa. The genome was highly biased toward A/T nucleotides—80.1 % in the whole genome, 77.8 % in PCGs, 83.4–85.6 % in RNAs, and 92.8 % in the A + T‐rich region. These values are well within the typical range for genes and regions of Vespoidea mitogenomes. Start and stop codons in several Vespa species—including V. ducalis—were diversified, despite these species belonging to the same genus. In comparison with the ancestral mitogenomes, Vespa mitogenomes—including that of V. ducalis—showed substantial gene rearrangement; however, we detected no gene rearrangement among Vespa species. We conducted phylogenetic reconstruction based on concatenated sequences of 13 PCGs and two rRNAs (12,755 bp ) in available species of Vespoidea—21 species in six subfamilies in two families (Vespidae and Formicidae). The Bayesian inference and maximum likelihood (ML) methods revealed that each family formed strong monophyletic groups [Bayesian posterior probability (BPP) = 1; ML, 100 %]. Moreover, V. ducalis and V. mandarinia formed a strong sister group (BPP = 1; ML, 94 %).     

Phylogenetic relationships and biogeography of Ptilophora (Gelidiales,Rhodophyta) with descriptions of P. aureolusa,P. malagasya,and P. spongiophila from Madagascar

The genus Ptilophora currently includes 16 species occurring mostly in subtidal habitats of the Indo‐Pacific Ocean, but its global diversity and biogeography are poorly understood. We analyzed mitochondrial cox1, plastid rbcL and plastid psbA sequences from specimens collected in southern Madagascar during the 2010 Atimo Vatae expedition and studied their morphologies. Both morphological and molecular data sets demonstrated the presence of five species in southern Madagascar: Ptilophora hildebrandtii, P. pterocladioides, and three new species described here, P. aureolusa, P. malagasya, and P. spongiophila. Ptilophora aureolusa is distinguished by its compound pinnae with uniformly spaced pinnules. Ptilophora malagasya has an indistinct midrib and irregularly spaced pinnules. Ptilophora spongiophila, heavily coated with sponges, has cylindrical to flattened main axes, lateral and surface proliferations, and spatulate tetrasporangial sori. The species of Ptilophora found in Madagascar are endemic, except P. hildebrandtii, which also occurs in eastern Africa. Ptilophora comprises four phylogenetic groups that map to eastern Australia, Japan, western Australia/Southeast Asia/Madagascar/eastern Africa, and Madagascar/eastern Africa/Aegean Sea. Biogeographical analysis revealed that the ancestor of Ptilophora originated in Australia, but most of the species radiated from Madagascar.     

Hybridization of cultivated Vitis vinifera with wild V. californica and V. girdiana in California

Hybridization of introduced domesticates and closely related natives is well documented in annual crops. The widespread introduction of the domesticated grapevine, Vitis vinifera, into California where it overlaps with two native congenerics, with which it is interfertile, provides opportunity to investigate hybridization between woody perennials. Although geographically widespread, the introduction over the past two centuries has been limited to a few elite clonal cultivars, providing a unique opportunity to study the effects of hybridization on the native species. The amount of hybridization with V. vinifera and the genetic diversity of wild‐growing Vitis californica and Vitis girdiana were examined using nineteen microsatellite markers. STRUCTURE analysis was used to define hybrid and introgressed individuals and to analyze genetic structure of the native species. FAMOZ software was used to identify which V. vinifera cultivars served as parents of F₁ hybrids. The three species were clearly distinguished by STRUCTURE analysis. Thirty percent of 119 V. californica vines were hybrids. The domesticated parent was identified for 16 F₁ hybrid vines; the original California cultivar, ‘Mission’, was the parent of eight. Backcrosses were also found, showing introgression into subsequent generations. Similar results were obtained for a small sample of V. girdiana. Removing hybrids greatly reduced the genetic variation of the presumed pure species, among which there was essentially no genetic structure. Limited genetic variability indicates the California natives may be threatened by genetic erosion. The discovery of F₁ hybrids of ‘Mission’, a cultivar not grown in the areas for ~100 years, suggests long generation times for wild vines that, often, grow into expansive liana and propagate by layering, all factors that limit recruitment in populations already disjunct by habitat lose. Hermaphroditic flowers and fruit that is more attractive to birds may favor the production of backcross seed and establishment of introgressed individuals.     

Morphological versus genetic diversity of Viola reichenbachiana and V. riviniana (sect. Viola,Violaceae) from soils differing in heavy metal content

Morphological characters, AFLP markers and flow cytometry were used to investigate the morphological and genetic variability and differentiation of Viola reichenbachiana and V. riviniana in non‐metallicolous (NM) and metallicolous (M) populations. The aims were to clarify the taxonomic status of plants occurring in ore‐bearing areas, to determine any relationship in V. reichenbachiana and V. riviniana from sites not polluted with heavy metals, and to examine the genetic variability and differentiation of M and NM populations of both species. Multivariate analyses based on morphological characters showed significant differences between V. reichenbachiana and V. riviniana from non‐polluted sites, high levels of intra‐ and inter‐population variability, and the occurrence of inter‐specific hybrids. Plants from M populations showed hybrid characters but also fell within the range of V. riviniana or V. reichenbachiana. There were no significant differences in relative genome size between plants from polluted areas and V. riviniana from NM populations. Bayesian analysis of population genetic structure based on AFLP markers distinguished two main groups: V. reichenbachiana and V. riviniana together with the M populations. That analysis also revealed the occurrence of populations of inter‐specific hybrids from non‐polluted areas. Further Bayesian analysis of V. riviniana including NM and M populations separated all the studied M populations from NM populations. We conclude that plants forming the M populations are well adapted to a metal‐polluted environment, and could be considered as stabilised introgressive forms resulting from unidirectional (asymmetric) introgression toward V. riviniana.     

Molecular systematics,species limits,and diversification of the genus Dendrocolaptes (Aves: Furnariidae): Insights on biotic exchanges between dry and humid forest types in the Neotropics

The true diversity and interspecific limits in the Neotropical endemic avian genus Dendrocolaptes (Furnariidae) remain a highly controversial subject, with previous genus‐wide assessments, based mostly on morphological characters, producing poorly resolved phylogenies. The lack of well‐resolved, robust, and taxonomically densely sampled phylogenies for Dendrocolaptes prevents reliable inferences on the genus’ actual species diversity and evolutionary history. Here, we analyzed 2,741 base pairs of mitochondrial and nuclear genes from 43 specimens belonging to all species and the majority of subspecies described for Dendrocolaptes to evaluate species limits and reconstruct its diversification through time. Our phylogenies recovered a monophyletic Dendrocolaptes, with two main highly supported internal clades corresponding to the D. certhia and D. picumnus species complexes. Also, our analyses supported the monophyly of most Dendrocolaptes species recognized today, except D. picumnus, which was consistently recovered as paraphyletic with respect to D. hoffmannsi. A coalescent‐based test supported a total of 15 different lineages in Dendrocolaptes and indicated that the number of currently accepted species within the genus may be greatly underestimated. Particularly relevant, when combined with previous analyses based on plumage characters, comparative high levels of genetic differentiation and coalescent analyses support the recognition of D. picumnus transfasciatus as a full species that is already under threat. Ancestral area reconstructions suggest that diversification in Dendrocolaptes was centered in lowland Amazonia, with several independent dispersal events leading to differentiation into different adjacent dry and high elevation forest types throughout the Neotropics, mainly during the Middle and Late Pleistocene.     

Natural hybridisation among Quercus glabrescens,Q. rugosa and Q. obtusata (Fagaceae): Microsatellites and secondary metabolites markers

• Natural hybridisation has significant ecological, genetic and evolutionary consequences altering morphological and chemical characters of individuals. Quercus glabrescens, Q. rugosa and Q. obtusata are white oak species well separated by their morphological characters when they occur in allopatry in Mexican temperate forests. However, in sympatry, individuals with atypical morphology have been observed, suggesting hybridisation events.
• In this study, we determined, with microsatellites and secondary metabolites, if interspecific gene flow occurs when these three oak species coexist in sympatry. In total, 180 individuals belonging to seven populations [three allopatric (one for each parental species) and four sympatric sites] were analysed.
• Allopatric populations represent well‐defined genetic groups and the sympatric populations showed genetic evidence of hybridisation between Q. glabrescens × Q. rugosa and Q. glabrescens × Q. obtusata. The hybridisation percentage varied between sites and combination of involved species. We registered the presence of unique flavonoid compounds for Q. glabrescens (caffeic acid and flavonol 2), Q. rugosa (flavonol 5) and Q. obtusata (flavonol 1). Three compounds (quercetin rhamnoside, flavonol 3 and alkyl coumarate) were expressed in all taxa. Finally, the hybrid genotypes identified in this study (Q. glabrescens × Q. rugosa and Q. glabrescens × Q. obtusata) showed specific chemical profiles, resulting from a combination of those of their parental species.
• These results show that hybridisation events between these oak species alter chemical expression of secondary metabolites, creating a mosaic of resources and conditions that provide the substrate for different combinations of foliar‐associated species such as herbivores, endophytic fungi or epiphyte plants.

     

Phylogeny and species delimitation of North European Lumbricillus (Clitellata,Enchytraeidae)

The enchytraeid genus Lumbricillus comprises about 80 described species of clitellate worms, which are up to a few centimetres long, and they mostly inhabit the littoral zone of non‐tropical marine and brackish waters world‐wide. The phylogeny of this genus is poorly studied, but previous work has suggested that Lumbricillus is a non‐monophyletic group. In this study, species boundaries and the phylogeny of this genus is re‐assessed using more than 300 DNA‐barcoded specimens (using COI mtDNA), part of which was also sequenced for two additional mitochondrial and four nuclear molecular markers. Statistical and coalescent based applications were used for the delimitation of a total of 24 species, of which 20 were identified as belonging to 17 described morphospecies; one morphospecies was found to be a complex of four delimited species, and another four delimited species could not be matched with any described species. Furthermore, gene trees, concatenation and multispecies coalescent based species trees were estimated using Bayesian inference. The estimated phylogenies confirm a non‐monophyletic Lumbricillus as L. semifuscus is clearly excluded from the genus. Furthermore, the placement of a monophyletic clade consisting of L. arenarius, L. dubius, and an unidentified species varies between analyses; they are either found as the sister‐group to the genus Grania or as sister‐group to the remaining Lumbricillus, where the latter relationship is supported by the multispecies coalescent, which we consider as the most reliable method.     

Pollen dispersal and fruit production in Vaccinium oxycoccos and comparison with its sympatric congener V. uliginosum

Investigating plant–pollinator interactions and pollen dispersal are particularly relevant for understanding processes ensuring long‐term viability of fragmented plant populations. Pollen dispersal patterns may vary strongly, even between similar congeneric species, depending on the mating system, pollinator assemblages and floral traits. We investigated pollen dispersal and fruit production in a population of Vaccinium oxycoccos, an insect‐pollinated shrub, and compared the pollen dispersal pattern with a co‐flowering, sympatric congener, V. uliginosum. We examined whether they share pollinators (through interspecific fluorescent dye transfers) and may differently attract pollinators, by comparing their floral colour as perceived by insects. Fluorescent dyes were mainly dispersed over short distances (80% within 40.4 m (max. 94.5 m) for V. oxycoccos and 3.0 m (max. 141.3 m) for V. uliginosum). Dye dispersal in V. oxycoccos was not significantly affected by plant area, floral display or the proximity to V. uliginosum plants. Interspecific dye transfers were observed, indicating pollinator sharing. The significantly lower dye deposition on V. oxycoccos stigmas suggests lower visitation rates by pollinators, despite higher flower density and local abundance. The spectral reflectance analysis indicates that bees are unlikely to be able to discriminate between the two species based on floral colour alone. Fruit production increased with increasing floral display, but was not affected by proximity to V. uliginosum plants. Our study highlights that fragmented populations of V. oxycoccos, when sympatric with co‐flowering V. uliginosum, might incur increased competition for the shared pollinators in the case of pollination disruption, which might then reduce outcrossed seed set.     

Sequencing type material resolves the identity and distribution of the generitype Lithophyllum incrustans,and related European species L. hibernicum and L. bathyporum (Corallinales,Rhodophyta)

DNA sequences from type material in the nongeniculate coralline genus Lithophyllum were used to unambiguously link some European species names to field‐collected specimens, thus providing a great advance over morpho‐anatomical identifi‐cation. In particular, sequence comparisons of rbcL, COI and psbA genes from field‐collected specimens allowed the following conclusion: the generitype species, L. incrustans, occurs mostly as subtidal rhodoliths and crusts on both Atlantic and Mediterranean coasts, and not as the common, NE Atlantic, epilithic, intertidal crust reported in the literature. The heterotypic type material of L. hibernicum was narrowed to one rhodolith belonging in Lithophyllum. As well as occurring as a subtidal rhodolith, L. hibernicum is a common, epilithic and epizoic crust in the intertidal zone from Ireland south to Mediterranean France. A set of four features distinguished L. incrustans from L. hibernicum, including epithallial cell diameter, pore canal shape of sporangial conceptacles and sporangium height and diameter. An rbcL sequence of the lectotype of Lithophyllum bathyporum, which was recently proposed to accommodate Atlantic intertidal collections of L. incrustans, corresponded to a distinct taxon hitherto known only from Brittany as the subtidal, bisporangial, lectotype, but also occurs intertidally in Atlantic Spain. Specimens from Ireland and France morpho‐anatomically identified as L. fasciculatum and a specimen from Cornwall likewise identified as L. duckerae were resolved as L. incrustans and L. hibernicum, respectively.     

Morphology and ontogenetic changes in otoliths of the mesopelagic fishes Ceratoscopelus maderensis (Myctophidae), Vinciguerria attenuata and V. poweriae (Phosichthyidae) from the Strait of Messina (Mediterranean Sea)

Fish otoliths morphology is an important aid in taxonomic, phylogenetic, palaeoichthyological and dietary studies. This study compares the morphology of the otoliths of the myctophid species, Ceratoscopelus maderensis, and two phosichthyid species, Vinciguerria attenuata and V. poweriae, collected from the Strait of Messina in the central Mediterranean Sea to identify the most appropriate taxonomical characters that separate these species. Ontogenetic changes in the otoliths of the studied fishes are evident. The sagitta of the juvenile C. maderensis is the most similar to the adult otoliths, while otoliths of the juvenile V. poweriae are the least similar to the adult otoliths. A discussion on possible geographical variations in otolith's morphology is provided by comparing our results with sagitta's images of C. maderensis, V. attenuata and V. poweriae in other studies.     

Molecular phylogeny of the deep‐sea penaeid shrimp genus Parapenaeus (Crustacea: Decapoda: Dendrobranchiata)

The commercial deep‐sea penaeid shrimp genus Parapenaeus contains 15 species, three subspecies and two forms in the Indo‐West Pacific and the Atlantic. Novel nucleotide sequence data from five different genes (COI, 16S, 12S, NaK and PEPCK) were collected to estimate phylogenetic relationships and taxonomic status amongst all but one subspecies in this genus. The phylogenetic results only support two of the four species groups previously proposed for this genus and indicate an evolution direction of the genital organs from simple to complex. The present results suggest that Parapenaeus originated in the shallow waters of the West Pacific with subsequent migration to the deep sea and the Atlantic. The molecular data reveal that there was probably misidentification of females between Parapenaeus australiensis and Parapenaeus ruberoculatus, with females previously assigned as P. australiensis likely being the females of P. ruberoculatus, while material identified as P. australiensis forma nodosa being the true P. australiensis females. On the other hand, Parapenaeus longipes forma denticulata truly represents a variation of the same species, while the subspecies Parapenaeus fissuroides indicus warrants a specific rank.     

Description of a second species of <Emphasis Type="Italic">Joblingia</Emphasis> Dybas &; Wenzel, 1947 (Diptera: Streblidae)

A new species of Joblingia Dybas & Wenzel, 1947, collected on the bat Myotis ruber (E. Geoffroy) in Southern Brazil, is described, and the type-species, J. schmidti Dybas & Wenzel, 1947, redescribed. The diagnosis of the genus is amended. The external morphology and postgonites of both species are figured. Geographical distribution and host-parasite relationships are discussed.     

Salix kaptarae sp. nov. (Salicaceae) from Crete

A new species of willow from Crete, named Salix kaptarae Cambria, C. Brullo & Brullo, is described and illustrated. Morphologically, it shows close relationships with species belonging to the S. apennina group, mainly with respect to the leaf shape, and the hairy ovary, capsule and base of stamina filaments. Considerations on its ecology, leaf micro‐morphology and conservation status are also provided.