Speciation and mitochondrial DNA diversification of the diving beetles Agabus bipustulatus and A. wollastoni (Coleoptera, Dytiscidae) within Macaronesia

Diversification of populations of two morphologically similar diving beetles within the Agabus tristis group, A. wollastoni and A. bipustulatus, was investigated, with partial mtDNA cytochrome b (Cyt b) sequences, allozymes and landmark‐based morphometrics. The Madeiran endemic A. wollastoni was collected from 11 localities. Population genetic and morphological variation was compared to Scandinavian localities of the widespread west Palearctic A. bipustulatus, recorded also from the Azores. Agabus wollastoni and European A. bipustulatus specimens representing eight and 13 localities respectively, were used in evaluating their phylogenetic relationship. Maximum parsimony analysis of the Cyt b sequences showed that both the A. bipustulatus and A. wollastoni specimens form well‐supported monophyletic groups. Three lines of evidence suggest that Agabus wollastoni has speciated through a few founders: (1) a well‐supported mtDNA line; (2) the mean heterozygosity of A. wollastoni is lower when compared to A. bipustulatus on the mainland; and (3) several uncommon alleles of A. bipustulatus are missing in A. wollastoni. The Azorean A. bipustulatus population was drastically affected by the colonization event, since several loci have become fixed with a resulting lower mean heterozygosity. The colonization was relatively recent, as the mtDNA lineage represented in the Azores is deeply nested within the A. bipustulatus clade. Population structure shows moderate inbreeding of A. wollastoni, and extensive substructuring at all localities with moderate gene‐flow between them. Morphological variation in A. wollastoni showed significant differentiation among several populations. Island colonizations, population structure of A. wollastoni, and an observed pattern of variation of the α‐glycerophosphate dehydrogenase locus are discussed. © 2003 The Linnean Society of London, Biological Journal of the Linnean Society, 2003, 79, 653–666.     

Morphological and molecular species delimitation within the Holarctic Ilybius angustior complex with a focus on Beringia (Coleoptera: Dytiscidae)

A fourth species of the Holarctic Ilybius angustior complex, Ilybius minakawai n. sp., is described from the Island of Sakhalin in the Russian far east. Male genitalia are diagnostic within this species complex, although from body size and shape the new species cannot be separated from I. churchillensis Wallis from north Alaska and the Canadian low arctic. The small and narrow body characteristic of these two species represents an extreme of the cold climate form of the widespread I. angustior (Gyllenhal). Low variation within the studied large ribosomal and cytochrome c oxidase subunit I mitochondrial genes suggest that speciation events within the I. angustior-complex are fairly recent. In combination with known geographical ranges, the low genetic variation within this species complex suggest speciation within Pleistocene refugia including Beringia.     

Ecotype differentiation in the face of gene flow within the diving beetle Agabus bipustulatus (Linnaeus, 1767) in northern Scandinavia

The repeated occurrence of habitat-specific polyphyletic evolved ecotypes throughout the ranges of widely distributed species implies that multiple, independent and parallel selection events have taken place. Ecological transitions across altitudinal gradients over short geographical distances are often associated with variation in habitat-related fitness, these patterns suggest the action of strong selective forces. Genetic markers will therefore contribute differently to differences between ecotypes in local hybrid zones. Here we have studied the adaptive divergence between ecotypes of the water beetle Agabus bipustulatus along several parallel altitudinal gradients in northern Scandinavia. This water beetle is well known for its remarkable morphological variation associated with mountain regions throughout the western Palaearctic. Two morphological ecotypes are recognised: a montane type with reduced flight muscles and a lowland type with fully developed muscles. Using a multilocus survey of allozyme variation and a morphological analysis with landmark-based morphometrics, across thirty-three populations and seven altitudinal gradients, we studied the local adaptive process of gene flow and selection in detail. Populations were sampled at three different elevations: below, at and above the tree line. The results indicate that the levels of divergence observed between ecotypes in morphology and allele frequencies at α-Glycerophosphate dehydrogenase relative to those shown by neutral molecular markers reflects local diversifying selection in situ. Four main lines of evidence are shown here: (1) A repeated morphological pattern of differentiation is observed across all altitudinal transects, with high reclassification probabilities. (2) Allele and genotype frequencies at the α-Gpdh locus are strongly correlated with altitude, in sharp contrast to the presumable neutral markers. (3) Genetic differentiation is two to three times higher among populations across the tree line than among populations at or below. (4) Genetic differentiation between ecotypes within independent mountain areas is reflected by different sets of allozymes.     

Larval consumption rates,interspecific predation,and local guild composition of egg-overwintering Agabus (Coleoptera,Dytiscidae) species in vernal ponds

Summary In vernal ponds in the boreal region, egg-over-wintering Agabus species form a guild that feeds mainly on larvae and pupae of aedine mosquitoes. The regular co-existence of very similar Agabus species indicates local communities not structured by interspecific competition. However, the lower number of species in local guilds than in the regional species pool poses a problem of limited membership. We suggest that the species of this guild display habitat differences mainly with respect to water temperature, pond size and prey density. In this view, habitat selection reflects body size and thermal growth response of the species, mainly in connection with larval development. We present field data from two northern Swedish vernal ponds. Based on these data, feeding experiments were performed to test the hypothesis outlined above. At a high prey density, larvae of all instars of the larger species A. erichsoni Gemm. & Har. had a significantly higher consumption rate than those of the smaller species A. opacus Aubé. At a low prey density the differences were smaller, and only the third instar larvae differed significantly. At 2° C, larvae of A. opacus had a significantly higher consumption rate than those of A. congener (Thunberg). At 15° C, no significant difference was observed. In studies of within-guild interspecific predation, always the larger larvae consumed the smaller ones. Field data show that egg hatching is spread out in time, and show interspecific differences. Consequently, the effects of unexpected droughts differ with species.     

Sexual dimorphism and sexual conflict in the diving beetle Agabus uliginosus (L.) (Coleoptera: Dytiscidae)

Sexual conflict can drive intersexual arms races, with female resistance and male persistence traits coevolving antagonistically. Such arms races are well documented in some diving beetles, although the extent of sexual conflict in this family remains unclear. The European dytiscid Agabus uliginosus has a strikingly dimorphic female; individuals from most regions are smooth and male‐like, whereas those from some populations have a strongly roughened dorsum, a trait that has attracted the name dispar. We demonstrate that rough and smooth females differ consistently in the development of dorsal surface microreticulation, and that these females are associated with males that differ in the development of their persistence traits. These findings extend the occurrence of pre‐insemination sexual conflict and associated intrasexual dimorphism in Dytiscidae, and suggest that such mating systems are relatively widespread in these beetles.     

Species delimitation in the Aloysia gratissima complex (Verbenaceae) following the phylogenetic species concept

The Aloysia gratissima complex is a group of 22 accepted, specific and infraspecific taxa, the boundaries of which are controversial. Various different authors have chosen different characters as criteria for delimiting taxa in this complex, which has been reflected in the variable number of accepted species. To resolve the taxonomy of this group, a modified population aggregation analysis, based on the phylogenetic species concept, was employed. As a result, the Aloysia gratissima complex is here circumscribed as compromising only four species, A. decipiens, A. gratissima s.s., A. oblanceolata and A. schulziana. These species are sustained by a different combination of four characters, of the 16 qualitative attributes analysed. A new approach based on cluster analysis is proposed here delimiting varieties. As a consequence two varieties, supported by significant differences in quantitative characters, are recognized here: A. gratissima var. gratissima and A. gratissima var. sellowii. A complete taxonomic revision of the accepted taxa is presented and six new synonyms are proposed. A key, illustrations and geographical distribution maps are included. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2016, 180 , 193–212.     

Thermal tolerance and geographical range size in the Agabus brunneus group of European diving beetles (Coleoptera: Dytiscidae)

Aim Within clades, most taxa are rare, whilst few are common, a general pattern for which the causes remain poorly understood. Here we investigate the relationship between thermal performance (tolerance and acclimation ability) and the size of a species’ geographical range for an assemblage of four ecologically similar European diving beetles (the Agabus brunneus group) to examine whether thermal physiology relates to latitudinal range extent, and whether Brown’s hypothesis and the environmental variability hypothesis apply to these taxa. Location Europe. Methods In order to determine the species tolerances to either low or high temperatures we measured the lethal thermal limits of adults, previously acclimated at one of two temperatures, by means of thermal ramping experiments (± 1°C min^?1). These measures of upper and lower thermal tolerances (UTT and LTT respectively) were then used to estimate each species’ thermal tolerance range, as total thermal tolerance polygons and marginal UTT and LTT thermal polygons. Results Overall, widespread species have higher UTTs and lower LTTs than restricted ones. Mean upper lethal limits of the Agabus brunneus group (43 to 46°C), are similar to those of insects living at similar latitudes, whilst mean lower lethal limits (?6 to ?9°C) are relatively high, suggesting that this group is not particularly cold‐hardy compared with other mid‐temperate‐latitude insects. Widespread species possess the largest thermal tolerance ranges and have a relatively symmetrical tolerance to both high and low temperatures, when compared with range‐restricted relatives. Over the temperature range employed, adults did not acclimate to either high or low temperatures, contrasting with many insect groups, and suggesting that physiological plasticity has a limited role in shaping distribution. Main conclusions Absolute thermal niche appears to be a good predictor of latitudinal range, supporting both Brown’s hypothesis and the environmental variability hypothesis. Restricted‐range species may be more susceptible to the direct effect of climate change than widespread species, notwithstanding the possibility that even ‘thermally‐hardy’, widespread species may be influenced by the indirect effects of climate change such as reduction in habitat availability in Mediterranean areas.     

An apposition‐like compound eye with a layered rhabdom in the small diving beetle Agabus japonicus (Coleoptera,Dytiscidae)

The fine structure of the compound eyes of the adult diving beetle Agabus japonicus is described with light, scanning, and transmission electron microscopy. The eye of A. japonicus is mango‐shaped and consists of about 985 ommatidia. Each ommatidium is composed of a corneal facet lens, an eucone type of crystalline cone, a fused layered rhabdom with a basal rhabdomere, seven retinula cells (including six distal cells and one basal cell), two primary pigment cells and an undetermined number of secondary pigment cells that are restricted to the distalmost region of the eye. A clear‐zone, separating dioptric apparatus from photoreceptive structures, is not developed and the eye thus resembles an apposition eye. The cross‐sectional areas of the rhabdoms are relatively large indicative of enhanced light‐sensitivity. The distal and central region of the rhabdom is layered with interdigitating microvilli suggesting polarization sensitivity. According to the features mentioned above, we suggest that 1) the eye, seemingly of the apposition type, occurs in a taxon for which the clear‐zone (superposition) eye is characteristic; 2) the eye possesses adaptations to function in a dim‐light environment; 3) the eye may be sensitive to underwater polarized light or linearly water‐reflected polarized light. J. Morphol. 275:1273–1283, 2014. © 2014 Wiley Periodicals, Inc.     

The species delimitation problem in the Simulium damnosum complex, blackfly vectors of onchocerciasis

The Simulium damnosum Theobald complex (Diptera: Simuliidae) comprises 57 cytoforms grouped into six subcomplexes. Previous phylogenetic studies using gene sequences have not completely resolved the evolutionary relationships of the cytoforms. The present study investigated the systematics of the complex using a phylogeographic approach. The differentiation between eastern and western forms observed in the phylogenetic studies is confirmed in the estimated haplotype networks. However, haplotypes tend to group in geographical clades and not according to cytoforms. Spatial analyses of the molecular variance also resulted in optimal groupings of sequences that did not correspond to cytoform boundaries. Moreover, Mantel tests showed significant correlations, although not strong, between genetic and geographical distances. This suggests an isolation-by-distance model of differentiation. Furthermore, there are instances in which genetic differentiation between cytoforms is low and not significant. These results indicate a lack of clear genetic differentiation between the cytoforms, which may be explained either by a separation of the taxa recent enough to allow the accumulation of few genetic differences or by recombination between the genomes of the cytoforms, which may be the result of hybridization with introgression or of non-independent evolutionary lineages. The results also emphasize the need for further sampling and for the use of more variable markers in order to clarify the evolutionary history of the group.     

Selective predation by larval Agabus (Coleoptera: Dytiscidae) on mosquitoes: support for conservation-based mosquito suppression in constructed wetlands

1. Wetland insect predators can structure aquatic prey communities via selective predation, but receive considerably less attention than vertebrate predators. We conducted laboratory experiments to test selective predation by two species of larval dytiscid beetles ( Agabus ; Coleoptera: Dytiscidae) and the potential contribution of these beetles to suppression of mosquito populations in constructed wetlands.
2.  Agabus consumed copepods, ostracods and mosquito larvae in no-choice tests. When offered a choice, 76% of all prey consumed were mosquito larvae, indicating selective predation. Subsequent experiments revealed this preference was due to ease of capture of mosquito larvae over alternative prey.
3. Cannibalism and intraguild predation were common within and between species of Agabus , which may reduce the overall impact of the observed selective predation.
4.  Agabus larvae selectively preyed on mosquito larvae over alternative prey, which is not characteristic of some fish used as biological control agents for mosquitoes. Predator exclusion or similar experiments in the field could document how these results translate into a natural setting.
5. The findings of this study suggest developing mosquito suppression strategies focused on conservation of native wetland predators. These strategies are preferable to introducing non-native generalist predators, or applying pesticides.     

Larval morphology of Agabus clypealis (Thomson, 1867) and A. pseudoclypealis Scholz, 1933 and notes on their distribution (Coleoptera: Dytiscidae)

The third instar larvae of Agabus clypealis (Thomson, 1867) and A. pseudoclypealis Scholz, 1933 are described. Important morphological structures (head, legs, and abdominal segment 8 with urogomphi) are illustrated. Both species share a completely sclerotized abdominal segment 6 with A. setulosus (J. Sahlberg, 1895). Morphological characters and species phenology are discussed in relation to the A. confinis-species group. All available literature records and some new and additional data on distribution of the two species are provided. Agabus clypealis is recorded for the first time from Estonia, and there is no reason to doubt the records of A. pseudoclypealis from Kazakhstan.     

A new species of Agaporomorphus Zimmermann, 1921 from Peru (Coleoptera,Dytiscidae, Copelatinae)

Agaporomorphus julianeae sp. n. is described from the Biological Field Station Panguana, in Huànuco province of central Peru. The new species belongs to the Agaporomorphus knischi-group sensu Miller 2005. Together with Agaporomorphus knischi Zimmermann, 1921 and Agaporomorphus colberti Miller & Wheeler, 2008 this is the third species of the genus with broadly enlarged male antennomeres. The new species can be separated from Agaporomorphus colberti and Agaporomorphus knischi by the smaller please expanded male antennomere VIII, and the form of the median lobe. Important species characters (median lobe, male antennae, metafemur, colour pattern) of the new species and Agaporomorphus knischi are figured, and the habitat, a temporary blackwater forest pond, and its species rich water beetle coenosis are illustrated and described in detail. The Brazilian Agaporomorphus mecolobus Miller, 2001, only known from the type material from Sao Paulo, is here recorded for Minas Gerais. Habitus photos of four additional Agaporomorphus species and Hydrodytes opalinus (Zimmermann, 1921) are provided. Altogether ten species of Agaporomorphus are now known.     

Species delimitation in the Aleochara fucicola species complex (Coleoptera: Staphylinidae: Aleocharinae) and its phylogenetic relationships

To assess the accuracy of species delimitation and phylogenetic relationships of the Aleochara fucicola species complex, we performed molecular phylogenetic analyses. Detailed micromorphological characters were also examined using scanning electron microscopy (SEM). The molecular data set included two mitochondrial (COI and COII) and three nuclear protein‐coding genes (CAD, EF1‐α and wg) for 19 populations of five species. Significant discordance was found between mitochondrial and nuclear gene trees. Two species, A. puetzi (Assing) and A. segregata Yamamoto and Maruyama, were not separated in the mitochondrial gene trees, but clearly were differentiated in the nuclear and concatenated gene trees. The SEM data also supported the separation of these two species, but an analysis of genetic divergence data did not. Adaptation to extremely harsh environments might reduce morphological variation in the A. fucicola species complex during the colonization of seashores. We present a multilocus phylogeny of the species complex. It suggests that the ancestor of the A. fucicola species complex occurred along the southern coasts of Northeast Asia, followed by dispersals to northern coasts.     

Genetic variation within and between Iliamna corei and I. remota (Malvaceae): implications for species delimitation

Iliamna corei and I. remota are classified as endangered species; however, their designation as separate species has been questioned. In order to address this problem, intersimple sequence repeat (ISSR) data were generated to examine patterns of genetic differentiation within and between these two taxa. ISSRs were used to screen individuals for genetic diversity of I. corei from the single known natural population and two garden populations, and individuals of I. remota from the six natural populations and four garden populations. Using ten primers, 140 informative markers were generated. Ninety-four percent of loci detected revealed polymorphisms. Cluster analysis (neighbour-joining, NJ) revealed that the two species are genetically distinct and that the Illinois populations of I. remota were genetically distinct from the Virginia populations of I. remota . Ordination by Principal Coordinate Analysis (PCoA) supported the findings of the NJ, with a separation of I. corei and I. remota . Analysis of Molecular Variance (AMOVA) revealed that the majority of variation detected was within populations, which is consistent with other self-incompatible plants. The results indicate a correlation between the geographical distributions of the species and gene flow.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 151 , 345–354.     

Species delimitation in Lippia section Goniostachyum (Verbenaceae) using the phylogenetic species concept

Lippia section Goniostachyum comprises plants distinguished by their numerous axillary florescences (three to six, sometimes up to nine) and tetrastichous floral bracts. Species of section Goniostachyum occur in the Neotropics, from Mexico to northern Argentina. Delimitation of the species grouped under Goniostachyum has remained unclear. Forty‐one names exist under this section, but only c. eight to ten names have been used frequently. To resolve the taxonomy of this group, a modified population aggregation analysis, based on the phylogenetic species concept, was employed. As a result, Goniostachyum is here circumscribed to only four species: L. grata, L. origanoides, L. sericea and L. stachyoides. These species are supported by different combinations of three characters of the 13 qualitative attributes analysed: canescent sericeous pubescence, frondose or frondose‐bracteose inflorescences and free or fused florescence apical bracts. Two varieties based on significant differences among quantitative characters are recognized: L. stachyoides var. stachyoides and L. stachyoides var. martiana comb. nov. Fifteen lectotypifications and four neotypifications are proposed. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 170 , 197–219.     

A (not-so-radical) solution to the species problem

What are species? One popular answer is that species are individuals. Here I develop another approach to thinking about species, an approach based on the notion of a lineage. A lineage is a sequence of reproducing entities, individuated in terms of its components. I argue that one can conceive of species as groups of lineages, either organism lineages or population lineages. Conceiving of species as groups of lineages resolves the problems that the individual conception of species is supposed to resolve. It has added the virtue of focusing attention on the characteristic of species that is most relevant to understanding their role in evolutionary processes, namely, the lineage structure of species.     

Larvae of four species of the Hyphydrus lyratus species-group (Coleoptera: Dytiscidae: Hydroporinae)

Abstract  The third instars of the Australian species Hyphydrus lyratus Swartz, H. contiguus Wehncke, H. elegans (Montrouzier) and H. decemmaculatus Wehncke (Coleoptera: Adephaga: Dytiscidae) are described, including a chaetotaxic analysis of the cephalic capsule, head appendages, legs, last abdominal segment and urogomphi. Larvae of these species morphologically resemble other species of Hyphydrus Illiger for which the larvae have been described. A key to identify larvae of the Australian species of Hyphydrus is provided. Larvae of H. effeminatus Watts appear identical to those of H. decemmaculatus . A 822 bp fragment of the CO1 gene of larvae and adults of these species showed very slight differences, suggesting the possibility that, in Australia at least, H. decemmaculatus is polymorphic.     

A chromosomal analysis of some water beetle species recently transferred from Agabus Leach to Ilybius Erichson, with particular reference to the variation in chromosome number shown by I. montanus Stephens (Coleoptera: Dytiscidae)

The karyotypes of seven Ilybius species are described and illustrated. All except I. wasastjernae have a basic karyotype of 34 autosomes plus sex chromosomes which are X0 ( male symbol ), XX ( female symbol ), with the X chromosome among the largest in the nucleus. This karyotype appears to be the norm for Ilybius and supports the transfer of the species concerned from Agabus to Ilybius. I. wasastjernae has 36 autosomes and the X chromosome is the smallest in the nucleus and its karyotype is unlike any other known karyotype in either Ilybius or Agabus. In most of the species studied no intraspecific variation has been detected. Exceptions are I. chalconatus, where there is one inversion polymorphism in one of the autosomes, and I. montanus whose autosome number has been found to vary from 29 to 34. Such variation is highly unusual among Coleoptera. The variation results from fusion-fission polymorphisms involving three different pairs of autosomes. In each case the fusions may be homozygous, heterozygous or absent. All populations investigated were polymorphic for some of the fusions, but only one (La Salceda, Spain) included individuals lacking all fusions. The frequencies of fused and unfused chromosomes were analysed in three English populations. In only one case was there a departure from the values expected from the Hardy-Weinberg equilibrium, and this population also showed a significant difference from the other two. Meiosis in males heterozygous for fusions involves the production of trivalents in first division, but results in the production of abundant sperm, with no evidence of chromosomal abnormalities in second metaphase, or of degenerating cells as a result of failed meiosis. The three fusions sites are consistent in all the populations studied, and it is concluded that these fusions represent unique historical events rather than current chromosomal instability.