Comparative multilocus phylogeography of two Palaearctic spruce bark beetles: influence of contrasting ecological strategies on genetic variation.

While phylogeographic patterns of organisms are often interpreted through past environmental disturbances, mediated by climate changes, and geographic barriers, they may also be strongly influenced by species-specific traits. To investigate the impact of such traits, we focused on two Eurasian spruce bark beetles that share a similar geographic distribution, but differ in their ecology and reproduction. Ips typographus is an aggressive tree-killing species characterized by strong dispersal, whereas Dendroctonus micans is a discrete inbreeding species (sib mating is the rule), parasite of living trees and a poor disperser. We compared genetic variation between the two species over both beetles' entire range in Eurasia with five independent gene fragments, to evaluate whether their intrinsic differences could have an influence over their phylogeographic patterns. We highlighted widely divergent patterns of genetic variation for the two species and argue that the difference is indeed largely compatible with their contrasting dispersal strategies and modes of reproduction. In addition, genetic structure in I. typographus divides European populations in a northern and a southern group, as was previously observed for its host plant, and suggests past allopatric divergence. A long divergence time was estimated between East Asian and other populations of both species, indicating their long-standing presence in Eurasia, prior to the last glacial maximum. Finally, the strong population structure observed in D. micans for the mitochondrial locus provides insights into the recent colonization history of this species, from its native European range to regions where it was recently introduced.

Bayesian species delimitation reveals generalist and specialist parasitic wasps on Galerucella beetles (Chrysomelidae): sorting by herbivore or plant host.

BACKGROUND: To understand the ecological and evolutionary consequences of species interactions in food webs necessitates that interactions are properly identified. Genetic analyses suggest that many supposedly generalist parasitoid species should rather be defined as multiple species with a more narrow diet, reducing the probability that such species may mediate indirect interactions such as apparent competition among hosts. Recent studies showed that the parasitoid Asecodes lucens mediate apparent competition between two hosts, Galerucella tenella and G. calmariensis, affecting both interaction strengths and evolutionary feedbacks. The same parasitoid was also recorded from other species in the genus Galerucella, suggesting that similar indirect effects may also occur for other species pairs. METHODS: To explore the possibility of such interactions, we sequenced mitochondrial and nuclear genetic markers to resolve the phylogeny of both host and parasitoid and to test the number of parasitoid species involved. We thus collected 139 Galerucella larvae from 8 host plant species and sequenced 31 adult beetle and 108 parasitoid individuals. RESULTS: The analysis of the Galerucella data, that also included sequences from previous studies, verified the five species previously documented as reciprocally monophyletic, but the Bayesian species delimitation for A. lucens suggested 3-4 cryptic taxa with a more specialised host use than previously suggested. The gene data analyzed under the multispecies coalescent model allowed us to reconstruct the species tree phylogeny for both host and parasitoid and we found a fully congruent coevolutionary pattern suggesting that parasitoid speciation followed upon host speciation. CONCLUSION: Using multilocus sequence data in a Bayesian species delimitation analysis we propose that hymenopteran parasitoids of the genus Asecodes that infest Galerucella larvae constitute at least three species with narrow diet breath. The evolution of parasitoid Asecodes and host Galerucella show a fully congruent coevolutionary pattern. This finding strengthens the hypothesis that the parasitoid in host search uses cues of the host rather than more general cues of both host and plant.

Stepping-stone expansion and habitat loss explain a peculiar genetic structure and distribution of a forest insect.

It is challenging to unravel the history of organisms with highly scattered populations. Such species may have fragmented distributions because extant populations are remnants of a previously more continuous range, or because the species has narrow habitat requirements in combination with good dispersal capacity (naturally or vector borne). The northern pine processionary moth Thaumetopoea pinivora has a scattered distribution with fragmented populations in two separate regions, northern and south-western Europe. The aims of this study were to explore the glacial and postglacial history of T. pinivora, and add to the understanding of its current distribution and level of contemporary gene flow. We surveyed published records of its occurrence and analysed individuals from a representative subset of populations across the range. A 633 bp long fragment of the mtDNA COI gene was sequenced and nine polymorphic microsatellite loci were genotyped. Only nine nucleotide sites were polymorphic in the COI gene and 90% of the individuals from across its whole range shared the same haplotype. The microsatellite diversity gradually declined towards the north, and unique alleles were found in only three of the northern and three of southern sites. Genetic structuring did not indicate complete isolation among regions, but an increase of genetic isolation by geographic distance. Approximate Bayesian model choice suggested recent divergence during the postglacial period, but glacial refugia remain unidentified. The progressive reduction of suitable habitats is suggested to explain the genetic structure of the populations and we suggest that T. pinivora is a cold-tolerant relict species, with situation-dependent dispersal.

Post-glacial colonization of the Fennoscandian coast by a plant parasitic insect with an unusual life history.

Species that exhibit very peculiar ecological traits combined with limited dispersal ability pose a challenge to our understanding of ecological and evolutionary mechanisms. This is especially true when they have managed to spread over long distances, overcome physical barriers, and colonize large areas. Climate and landscape changes, trophic web relations, as well as life history all interact to shape migration routes and present-day species distributions and their population genetic structures. Here we analyzed the post-glacial colonization of northern Europe by the gall midge Contarinia vincetoxici, which is a monophagous parasite on the perennial herb White swallowwort (Vincetoxicum hirundinaria). This insect not only has a narrow feeding niche but also limited dispersal ability and an exceptionally long dormancy. Gall midge larvae (n = 329) were collected from 16 sites along its distribution range in Denmark, Sweden, and Finland. Using microsatellite loci and knowledge of the species and the regions' history, we investigated the role of landscape change, host plant distribution, insect population dynamics, and life history in shaping the population genetic structure of the insect. We devoted particular interest to the role of the insect's presumed poor dispersal capacity in combination with its exceptionally extended diapause. We found significant levels of local inbreeding (95% highest posterior density interval = 0.42-0.47), low-level within-population heterozygosity (mean H E = 0.45, range 0.20-0.61) with private alleles in all populations except two. We also found significant (p < .001) regional isolation-by-distance patterns, suggesting regularly recurring mainly short-distance dispersal. According to approximate Bayesian computations, C. vincetoxici appears to have colonized the study area via wind-aided flights from remote areas approximately 4600-700 years before present when the land has gradually risen above the sea level. Extremely long dormancy periods have allowed the species to "disperse in time", thereby aiding population persistence despite generally low census population sizes.

Limited emigration from an outbreak of a forest pest insect.

Population density and individual dispersal behaviour affect species' distribution dynamics. Population densities vary over time, and some species occasionally increase to very high numbers, for example during outbreaks. In such situations, populations are expected to expand into new areas as a result of density-dependent dispersal which sometimes even results in range expansion. A local population of the northern pine processionary moth Thaumetopoea pinivora has recently reached outbreak densities at the edge of its northern range at the southern tip of Gotland Island in the Baltic Sea. We first investigated whether the outbreak had resulted in establishment of populations in suitable habitats on Gotland Island outside the outbreak area. Six small populations were found that could potentially have originated from the outbreak area. However, data from 12 microsatellite markers strongly suggest that these populations did not originate from the recent outbreak. Genetic variability was not reduced in these small, isolated populations, and there were several unique alleles, indicating instead a different population history and that there has been no recent range expansion. In addition, there was apparent genetic isolation by geographic distance, implying that despite the high density of the outbreak population, significant gene flow has not occurred.

Gene flow relates to evolutionary divergence among populations at the range margin.

BACKGROUND: Morphological differentiation between populations resulting from local adaptations to environmental conditions is likely to be more pronounced in populations with increasing genetic isolation. In a previous study a positive clinal variation in body size was observed in isolated Roesel's bush-cricket, Metrioptera roeselii, populations, but were absent from populations within a continuous distribution at the same latitudinal range. This observational study inferred that there was a phenotypic effect of gene flow on climate-induced selection in this species. METHODS: To disentangle genetic versus environmental drivers of population differences in morphology, we measured the size of four different body traits in wild-caught individuals from the two most distinct latitudinally-matched pairs of populations occurring at about 60°N latitude in northern Europe, characterised by either restricted or continuous gene flow, and corresponding individuals raised under laboratory conditions. RESULTS: Individuals that originated from the genetically isolated populations were always bigger (femur, pronotum and genital appendages) when compared to individuals from latitudinally-matched areas characterised by continuous gene flow between populations. The magnitude of this effect was similar for wild-caught and laboratory-reared individuals. We found that previously observed size cline variation in both male and female crickets was likely to be the result of local genetic adaptation rather than phenotypic plasticity. CONCLUSIONS: This strongly suggests that restricted gene flow is of major importance for frequencies of alleles that participate in climate-induced selection acting to favour larger phenotypes in isolated populations towards colder latitudes.

Genetic diversity of a successful colonizer: isolated populations of Metrioptera roeselii regain variation at an unusually rapid rate.

Newly founded isolated populations need to overcome detrimental effects of low genetic diversity. The establishment success of a population may therefore depend on various mechanisms such as assortative mating, purging of deleterious alleles, creation of new mutations and/or repeated inflow of new genotypes to reduce the effects of inbreeding and further loss of genetic variation. We compared the level of genetic variation in introduced populations of an insect species (Metrioptera roeselii) far beyond its natural distribution with levels found in their respective founder populations and coupled the data with timing since establishment. This allowed us to analyze if the introduced populations showed signs of temporal changes in genetic variation and have made it possible to evaluate underlying mechanisms. For this, we used neutral genetic markers, seven microsatellite loci and a 676-bp-long sequence of the mtDNA COI gene. All tested indices (allelic richness, unbiased expected heterozygosity, effective size, haplotype diversity, and nucleotide diversity) except inbreeding coefficient had significantly higher values in populations within the founding populations inside the continuous area of the species distribution compared with the introduced populations. A logarithmic model showed a significant correlation of both allelic richness and unbiased expected heterozygosity with age of the isolated populations. Considering the species' inferred colonization history and likely introduction pathways, we suggest that multiple introductions are the main mechanism behind the temporal pattern observed. However, we argue that influences of assortative mating, directional selection, and effects of an exceptional high intrapopulation mutation rate may have impacts. The ability to regain genetic diversity at this level may be one of the main reasons why M. roeselii successfully continue to colonize northern Europe.

Evaluating range-expansion models for calculating nonnative species' expansion rate.

Species range shifts associated with environmental change or biological invasions are increasingly important study areas. However, quantifying range expansion rates may be heavily influenced by methodology and/or sampling bias. We compared expansion rate estimates of Roesel's bush-cricket (Metrioptera roeselii, Hagenbach 1822), a nonnative species currently expanding its range in south-central Sweden, from range statistic models based on distance measures (mean, median, 95(th) gamma quantile, marginal mean, maximum, and conditional maximum) and an area-based method (grid occupancy). We used sampling simulations to determine the sensitivity of the different methods to incomplete sampling across the species' range. For periods when we had comprehensive survey data, range expansion estimates clustered into two groups: (1) those calculated from range margin statistics (gamma, marginal mean, maximum, and conditional maximum: ˜3 km/year), and (2) those calculated from the central tendency (mean and median) and the area-based method of grid occupancy (˜1.5 km/year). Range statistic measures differed greatly in their sensitivity to sampling effort; the proportion of sampling required to achieve an estimate within 10% of the true value ranged from 0.17 to 0.9. Grid occupancy and median were most sensitive to sampling effort, and the maximum and gamma quantile the least. If periods with incomplete sampling were included in the range expansion calculations, this generally lowered the estimates (range 16-72%), with exception of the gamma quantile that was slightly higher (6%). Care should be taken when interpreting rate expansion estimates from data sampled from only a fraction of the full distribution. Methods based on the central tendency will give rates approximately half that of methods based on the range margin. The gamma quantile method appears to be the most robust to incomplete sampling bias and should be considered as the method of choice when sampling the entire distribution is not possible.

Permanent Genetic Resources added to Molecular Ecology Resources Database 1 October 2009-30 November 2009.

This article documents the addition of 411 microsatellite marker loci and 15 pairs of Single Nucleotide Polymorphism (SNP) sequencing primers to the Molecular Ecology Resources Database. Loci were developed for the following species: Acanthopagrus schlegeli, Anopheles lesteri, Aspergillus clavatus, Aspergillus flavus, Aspergillus fumigatus, Aspergillus oryzae, Aspergillus terreus, Branchiostoma japonicum, Branchiostoma belcheri, Colias behrii, Coryphopterus personatus, Cynogolssus semilaevis, Cynoglossus semilaevis, Dendrobium officinale, Dendrobium officinale, Dysoxylum malabaricum, Metrioptera roeselii, Myrmeciza exsul, Ochotona thibetana, Neosartorya fischeri, Nothofagus pumilio, Onychodactylus fischeri, Phoenicopterus roseus, Salvia officinalis L., Scylla paramamosain, Silene latifo, Sula sula, and Vulpes vulpes. These loci were cross-tested on the following species: Aspergillus giganteus, Colias pelidne, Colias interior, Colias meadii, Colias eurytheme, Coryphopterus lipernes, Coryphopterus glaucofrenum, Coryphopterus eidolon, Gnatholepis thompsoni, Elacatinus evelynae, Dendrobium loddigesii Dendrobium devonianum, Dysoxylum binectariferum, Nothofagus antarctica, Nothofagus dombeyii, Nothofagus nervosa, Nothofagus obliqua, Sula nebouxii, and Sula variegata. This article also documents the addition of 39 sequencing primer pairs and 15 allele specific primers or probes for Paralithodes camtschaticus.

Characterization and cross-amplification of 13 microsatellite loci in the northern pine processionary moth, Thaumetopoea pinivora (Lepidoptera: Notodontidae).

Thirteen polymorphic microsatellite loci were developed for the northern pine processionary moth (Thaumetopoea pinivora) and tested for cross-amplification in seven other species within the Thaumetopoea family. Number of alleles ranged from two to 10 when at least 28 individuals from one population were screened and one locus, Thapin06, appears to be sex linked. Expected heterozygosity ranged from 0.094 to 0.856 and observed heterozygosity ranged from 0.097 to 0.806. Amplification success varied between sister species, with two up to seven loci being successfully amplified. The described loci will be valuable for studying the population genetic structure and dispersal behaviour of this forest pest.

Species-specific primers for predation studies of the pollen beetle, Meligethes aeneus (Coleoptera, Nitidulidae).

Species-specific primers were developed for the pollen beetle (a pest in oilseed rape) for studies of predation by natural insect predators. Two forward and three reverse primers were designed within the mitochondrial COI gene and used in combination to amplify fragments in the size range of 163-290 bp. Remains of pollen beetle DNA were consistently detected in Pardosa spiders up to 24 h after ingestion but dropped drastically at 48 h. These primers will facilitate studies on biological control of this oilseed rape pest. Detection time was not correlated with fragment length as might be expected as the DNA gradually degrades into progressively shorter fragments over time.