Characterizing DNA preservation in degraded specimens of Amara alpina (Carabidae: Coleoptera)

DNA preserved in degraded beetle (Coleoptera) specimens, including those derived from dry‐stored museum and ancient permafrost‐preserved environments, could provide a valuable resource for researchers interested in species and population histories over timescales from decades to millenia. However, the potential of these samples as genetic resources is currently unassessed. Here, using Sanger and Illumina shotgun sequence data, we explored DNA preservation in specimens of the ground beetle Amara alpina, from both museum and ancient environments. Nearly all museum specimens had amplifiable DNA, with the maximum amplifiable fragment length decreasing with age. Amplification of DNA was only possible in 45% of ancient specimens. Preserved mitochondrial DNA fragments were significantly longer than those of nuclear DNA in both museum and ancient specimens. Metagenomic characterization of extracted DNA demonstrated that parasite‐derived sequences, including Wolbachia and Spiroplasma, are recoverable from museum beetle specimens. Ancient DNA extracts contained beetle DNA in amounts comparable to museum specimens. Overall, our data demonstrate that there is great potential for both museum and ancient specimens of beetles in future genetic studies, and we see no reason why this would not be the case for other orders of insect.     

Five microsatellite DNA markers for the ground beetle Abax parallelepipedus (Coleoptera,Carabidae)

We characterized five microsatellite loci from Abax parallelepipedus, one of the most common forest ground beetles in central Europe. The polymorphism of the loci was assessed in 30 individuals from one population. The number of alleles ranged between two and 19 and the observed heterozygosities between 0.10 and 0.97. No deviations from Hardy–Weinberg equilibrium were detected for any of the markers.     

Physical mapping of rDNA genes in the ground beetle Carabus and related genera (Carabidae: Coleoptera)

The major ribosomal DNA (rDNA) loci were localized on meiotic and mitotic chromosomes and in interphase nuclei of 18 ground-beetle species belonging to three tribes of the supertribe Carabitae by fluorescence in situ hybridization (FISH), using a PCR-amplified 18S rDNA as a probe. Meiotic observations indicate that the 18S rDNA sequences are located on the largest autosomal bivalent in 12 species of Carabus , two species of Calosoma (both genera belonging to the tribe Carabini), and three sibling species of Ceroglossus chilensis (tribe Ceroglossini). The data suggest the occurrence of a conservative pattern in these three genera despite the chromosomal rearrangements that have led to karyotypes with higher chromosome numbers in Ceroglossus . A different result is found in Cychrus caraboides (tribe Cychrini), where ribosomal cistrons are located in two medium-sized autosomal pairs. Further species of Cychrini should be studied for corroborating the occurrence of molecular and karyotypical apomorphies in Cychrus with regard to the genera Carabus, Calosoma and Ceroglossus .     

Isolation and characterization of microsatellite loci in the ground beetle Carabus problematicus (Coleoptera,Carabidae)

Six microsatellite loci were isolated from the ground beetle Carabus problematicus. The polymorphism ranged from two to 17 alleles and observed and expected heterozygosities ranged from 0.137 to 0.676 and 0.147 to 0.710, respectively.     

Isolation and characterization of microsatellite loci in the ground beetle Carabus nemoralis (Coleoptera,Carabidae)

We isolated and characterized 10 microsatellite loci in the ground beetle Carabus nemoralis (Coleoptera, Carabidae), an ubiquist species largely distributed in Europe. Polymorphism ranged from 3 to 12 alleles on the 30 individuals from the same locality examined.     

A rapid method for detecting mitochondrial DNA variation in the brown trout, Salmo trutta

A method is described for the amplification and restriction digestion of two variable regions of mitochondrial DNA (mtDNA) in the brown trout ( Salmo trutta ) as a non-invasive sampling and rapid screening technique for measuring genetic variation in populations.     

Chromosome evolution in ground beetles: localization of the rDNA loci in the tribe Harpalini (Coleoptera,Carabidae)

The pattern of localization of the ribosomal genes was studied by means of fluorescence in situ hybridization in 39 species of the tribe Harpalini. Most of them show one pair of autosomes carrying the ribosomal genes in a distal position of a single chromosome arm. This pattern is hypothesized to be ancestral for the whole tribe. Both, chromosome number and the number and localization of rDNA loci, show little variation and are therefore of little phylogenetic value. Only in the subtribe Ditomina is there enough variation to characterize phyletic relationships. The stability of rDNA loci is even higher than the constancy of chromosome number, as most species of Ditomina (genera Dixus, Eocarterus, Carterus, Odontocarus and Ditomus) have the usual pair of autosomes with rDNA loci, in spite of remarkable differences in the diploid number. Only Dixus sphaerocephalus and Dixus clypeatus have two autosomal pairs with a fluorescent signal. These results do not support the hypothesis that the high chromosome numbers found within Ditomina are the result of polyploid change from the ancestral 2n = 37 karyotype of the tribe Harpalini. Chromosomal translocations or the presence of mobile genetic elements are plausible sources of the few cases of intraspecific polymorphism in the rDNA loci found in species of Harpalus.     

Molecular markers locate genes for resistance to the Colorado potato beetle,Leptinotarsa decemlineata,in hybrid Solanum tuberosum x S. berthaultii potato progenies

The wild Bolivian potato, Solanum berthaultii Hawkes, has been used as a source of resistance to the Colorado potato beetle (CPB), Leptinotarsa decemlineata Say, one of the most significant pests of potato. In this study, two reciprocal backcross S. tuberosum x S. berthaultii potato progenies, BCB and BCT, were mapped with RFLP markers and screened for resistance to CPB consumption, oviposition and defoliation. The genotypic and phenotypic data were combined and analysed to locate quantitative trait loci (QTLs) for resistance to CPB. Three QTLs on three chromosomes in BCB, and two QTLs on two chromosomes in BCT influenced resistance. The QTLs were generally additive but one instance of epistasis was noted. Each QTL accounted for 4–12% of the phenotypic variation observed in resistance. In the more resistant BCB population, a three QTL model explained ca. 20% of the variation in CPB oviposition. When alleles at the three QTLs were homozygous S. berthaultii, oviposition was reduced ca. 60% compared to the heterozygotes. The QTLs for resistance to CPB were compared to those previously identified for the type A and B glandular trichomes, which have been implicated in resistance in the same progenies. Generally, the QTLs for resistance to CPB coincided with loci associated with the glandular trichomes confirming the importance of the glandular trichomes in mediating resistance. However, a relatively strong and consistent QTL for insect resistance in both BCB and BCT on chromosome 1 was observed that was not associated with any trichome traits, suggesting the trichomes may not account for all of the resistance observed in these progenies.     

Tackling the taxonomic impediment: a global assessment for ant‐nest beetle diversity (Coleoptera: Carabidae: Paussini)

We evaluated the completeness and historical trends of the taxonomic knowledge of the myrmecophilous ground beetle tribe Paussini (Coleoptera, Carabidae, Paussinae). Species accumulation curves were modelled using a logistic function. Similar analyses were conducted for genera and subgenera. Although not all biogeographical regions have been equally explored, accumulation curves reached a plateau in all cases. Our models predict that about 96% of the world fauna has been already described. However, the asymptotes calculated for the Australian and Oriental species should be interpreted as false plateaus because of the lack of recent research. Similarly, patterns of genera accumulation indicate that a plateau has been reached. As a result of continued debate on the use and validity of Paussini subgenera, the accumulation curves of subgenera showed stepped patterns, with no evidence of plateaus. Thus, although relatively few species are expected to be described in the future, the species accumulation curves indicate that the taxonomic inventory is not yet complete. Differences in accumulation patterns among biogeographical regions can be used to highlight the areas where more species are expected, and hence where taxonomical efforts should be concentrated. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 330–339.     

Restriction fragment analysis of the secalin loci of rye

Analyses of wheat/rye addition lines by Southern blotting confirmed the presence of sequences related to theSec 1, Sec 2, andSec 3 loci on chromosomes 1R and 2R. Comparison of the 1R and 2R addition lines allowed the identification of -secalin genes atSec 1 andSec 2, respectively, while -secalin and -secalin genes atSec 1 were discriminated by comparative hybridization with three probes: -secalin, total -secalin, and 3 -secalin. The high molecular weight (HMW) secalin genes atSec 3 were identified using a homologous HMW subunit probe from wheat. Gene copy numbers were estimated as about 40–60 for -secalins, 5–10 for -secalins, and 2 for HMW secalins. Comparison of individual plants of cv. Gazelle showed a high degree of polymorphism, particularly for sequences related to -secalins and HMW secalins.     

Trends in detoxification enzymes and heavy metal accumulation in ground beetles (Coleoptera: Carabidae) inhabiting a gradient of pollution

Non-specfic carboxylesterase and glutathione S-transferase activity was measured in the ground beetle, Pterosthicus oblongopunctatus (Coleoptera: Carabidae), from five sites along a gradient of heavy metal pollution. A previous study determined that beetles from the two most polluted sites (site codes OLK2 and OLK3) were more susceptible to additional stressors compared with beetles from the reference site (Stone et al., Environ. Pollut. 113, 239-244 2001), suggesting the possibility of physiological impairment. Metal body burdens in ground beetles from five sites along the gradient ranged from 79 to 201 microg/g Zn, 0.174 to 8.66 microg/g Pb and 1.14 to 10.8 microg/g Cd, whereas Cu seemed to be efficiently regulated regardless of metal levels in the soil. Beetle mid- and hindguts were homogenized and the soluble fraction containing glutathione S-transferase (GST) and carboxylesterase (CaE) was assayed using kinetic analyses. Significantly higher levels of GST were found only in female beetles from the most polluted sites (OLK2 and OLK3; P=0.049, P<0.001, respectively) compared with the reference site (OLK7). In addition, OLK3 females had significantly higher levels of CaE compared with the reference beetles (P=0.01). Male beetles did not differ in enzyme activity along the metal gradient. Overall, obvious trends in detoxification enzymes were not detected in ground beetles in association with metal body burdens.     

The ground beetle fauna of ancient and recent woodlands in the lowlands of north-west Germany (Coleoptera, Carabidae)

In north-west Germany the ground beetle fauna was investigated using pitfall traps at 79 sampling sites in ancient woodlands and recent woodlands (with and without direct contact to old stands). Two woodland types were considered: The Quercion robori-petraea-woodlands (oak-beech-type) on mainly sandy soils and the Stellario-Carpinetum-woodlands (hornbeam-type) on mainly loamy soils. The number of recorded ground beetle species inhabiting exclusively or predominantly woodlands in the investigation area is significantly higher in ancient stands of both woodland types than in recent ones. No statistically substantiated relation between habitat size (both about 1800 and in 1990) and the number of characteristic woodland ground beetle species could be ascertained. Carabus glabratus and Abax parallelus show a distinct focus in ancient woodlands. Significantly more records of Carabus violaceus and Abax parallelepipedus are known from ancient woodlands than from recent ones. Twelve of the 16 ground beetle species, for which no difference in the colonisation of ancient and recent woodlands was ascertained, are macropterous. Half of the eight brachypterous woodland species is exclusively or predominantly found in ancient woodlands, suggesting that power of dispersal is an important factor which determines the species number in woodland fragments of different age.     

A multivariate analysis of cropping effects on Irish ground beetle assemblages (Coleoptera: Carabidae) in mixed arable and grass farmland

The assemblage of carabid species trapped over a 3 yr period in eight separate but contiguous fields was analysed by Canonical Correspondence Analysis (CCA), Seven of the fields were subject to arable crop rotations involving a mixture of autumn and early spring‐sown cereals (wheat or barley), late spring‐sown crops (potatoes, sugar and fodder beet and maize) and short‐term uncultivated grass leys, The eighth field was an established ‘permanent’ grass pasture of at least 50 yrs standing, Pooled samples collected in the first half of the growing season (April‐June) showed clear evidence of soil cultivation effects on community structure, Systematic exclusion of samples and re‐analysis distinguished the fauna of firstly, old pasture samples, samples from ley pastures of increasing age and finally samples from fields with different times of soil cultivation, In the latter analysis, the main effects of soil cultivation were related to differences in ground cover over the winter and a direct effect of spring soil cultivation on autumn breeding populations, The ordination of pooled catches for the second half of the growing season (July‐September) could not be related to known year, field or crop cultivation variables, The underlying species ordination suggested that later in the summer the effect of crop cover on soil microclimate may mask cultivation effects by influencing the post‐emergence dispersal of autumn‐breeding populations and the reproductive success of spring‐breeders, The combination of earlier soil cultivation effects, and probably microclimatic influences later in the season, resulted in the strong distinction of whole season carabid catches from individual fields, It is concluded that the uniqueness of individual field histories may provide a mechanism to promote the co‐existence of ecologically similar species within the farmed landscape and enhance the abundance and bio‐diversity of species in the face of routine soil cultivation.     

Taxonomic notes on Trechini and description of a new hypogean species from the Iberian Peninsula (Coleoptera: Carabidae: Trechinae)

In this contribution, a new pubescent Trechus species for the Iberian Peninsula is described, and the use of some morphological traits in the systematic of Trechini are discussed. Micropubescence is common in endogean and troglobite species which are closely related with the “terrestrial-phreatic habitat”. It probably enhances survival in swampy environments through the retention of air bubbles in immersion conditions and promoting flotation. Neither this trait, anophthalmy nor inner sac of aedeagus without sclerotized pieces can be used to support supraspecific taxa. Thus, we propose the formal synonymy of Hydrotrechus with Trechus [Hydrotrechus syn. nov. of Trechus]. To avoid secondary homonymy, we propose the name Trechus triamicorum nom. nov. for Hydrotrechus cantabricus Carabajal, García & Rodríguez 1999 [already occupied by Trechus cantabricus Jeannel 1927, synonym of Trechus distigma Kiesenwetter 1851]. We also propose the synonymization of the subgenus Trichapoduvalius with Apoduvalius [Trichapoduvalius syn. nov. of Apoduvalius]. The new hypogean species with pubescent elytra, Trechus cifrianae sp. nov., is described. The morphology of the species suggests that it probably inhabits the Mesovoid Shallow Substratum (MSS) or the fissure network (mesocaverns and microcaverns) of the cave where the only individual was found. The structure of internal female genitalia suggests a close relationship between T. cifrianae sp. nov. and T. carrilloi Toribio & Rodríguez 1997.     

GIS analysis of the biogeography of beetles of the subgenus Anisodactylus (Insecta: Coleoptera: Carabidae: genus Anisodactylus)

The advent of GIS technology and the World Wide Web, respectively, facilitate analysing geographical relationships and electronically storing and exchanging biogeographic data. This paper illustrates GIS technology with a study of the subgenus Anisodactylus Dejean (Insecta: Coleoptera: Carabidae: genus Anisodactylus). Species are concentrated in three centres of biodiversity in North America and in four in lands near the western Mediterranean. These centres largely correspond to current areas of wetlands. Eurasia has fewer species than expected based on its area, probably because large portions have habitats unfavourable for the subgenus and/or are poorly collected for Carabidae. Members of the subgenus are primarily adapted to areas with January temperatures between ?10 and 10 °C, July temperatures from 10 to 30 °C and mean annual precipitation from 20 to 200 mm. Cold is apparently a major limiting factor because it typically occurs during several consecutive months of winter and is difficult to escape except by hibernation. Heat is less of a stress when moisture is sufficient. The size of geographical ranges is often larger in the North than in the South and correlates with the latitude of the centre of ranges at r =0.42 (level of significance=0.05). Geographic ranges are often smaller in western North America and in the western Mediterranean than elsewhere in the Northern Hemisphere. Explanations for the smaller sizes include portions of western North America having unfavourable desert or montane habitats, the Rocky Mountains and deserts barring eastward dispersal of species, and the smaller size and more patchy distribution of climatic zones and habitats. In North America geographical ranges west of the Rocky Mountains are north–south elongated because they track primarily north–south orientated climatic zones and because mountains and deserts bar eastward extension. Ranges in north-eastern and north-central North America tend to extend east–west along temperature isotherms. In Eurasia many ranges are stretched east–west because of the shape of the continent and because many northern and southern areas lack suitable habitats. Species with relatively high numbers of apomorphic character states cluster in western Eurasia and to a lesser extent in western North America. The North American centres of biodiversity are post-Wisconsin phenomena while those near the western Mediterranean probably date from the Oligocene or Miocene.     

Amplified fragment length polymorphism (AFLP) suggests old and recent immigration into the Alps by the arctic-alpine annual Comastoma tenellum (Gentianaceae)

Aim This study aims to elucidate the phylogeography of the arctic‐alpine annual Comastoma tenellum (Rottb.) Toyok. (Gentianaceae) and to unravel the history of its immigration into the Alps. Location Although samples from Alaska and Central Asia were also included, our study focusses on Europe, especially on the Alps. Methods We applied amplified fragment length polymorphism (AFLP) fingerprinting on 37 populations (162 individuals) of C. tenellum and analysed the results phenetically. Results As C. tenellum is mainly inbreeding, there is typically little to no intrapopulational genetic variation. Two populations from Alaska and Altai are strongly separated from all other accessions. The majority of the populations from the Alps group together with high bootstrap support. They fall into an unsupported Alps I group (northwards of Gran Paradiso) and a well‐supported Alps II group (south‐western Alps). The remaining European populations form a weakly‐supported branch constituting accessions from the Carpathians, Scandinavia and two populations from the Eastern Alps. Main conclusions Comastoma tenellum reached the Alps at least twice. The first immigration event resulted in a lineage that is clearly separated from the other European accessions. The immigration must have occurred well before the last glaciation because this lineage shows further phylogeographical structuring into two groups (Alps II in the south‐western Alps and Alps I in the rest of the Alps). This pattern is presumably due to isolation in different glacial refugia. In addition to the old immigration event, the species reached the Alps in recent times either from Scandinavia or from the Carpathians via long‐distance dispersal. These immigrations resulted in (at least) two populations that are spatially small and poor in individuals.     

Adaptive latitudinal variation of the duration and thermal requirements for development in the ground beetle Amara communis (Panz.) (Coleoptera, Carabidae)

The data are obtained on development time at six constant temperatures (12, 14, 16, 18, 20, 22°C) and thermal requirements for preimaginal development in a ground beetle Amara communis from Arkhangelsk (64°34′N) and St. Petersburg (59°53′N). The larval and pupal development times were found to be significantly shorter in the Arkhangelsk than in the St. Petersburg population under all temperatures. As a result, total preimaginal development appeared to be shorter by 6.2–6.6% in the Arkhangelsk population. The regression lines of the larval, pupal and total (egg-to-adult) development rate on temperature for the Arkhangelsk population run above and steeper than the respective lines for the St. Petersburg population. Both populations share the similar values of the thermal thresholds (7.2–8.2°C). This explains faster preimaginal development in the northern population under all temperatures above the threshold. Thus, the slope of the regression lines increases, i.e., the sum of degree-days decreases, whereas the thermal threshold for development exhibited no distinctive changes from south to north in this species. Adults from Arkhangelsk reared in the experiments appeared heavier on the average in comparison with those from St. Petersburg, especially at 18–22°C. Temperature did not significantly affect adult weight, except the fact that the beetles were slightly heavier at 20 and 22°C. Consequently, the well-known “temperature-size rule” is violated in this species. Relative growth rate in larvae of A. communis increased considerably with temperature rise from 14 to 22°C. It was significantly higher in the beetles from Arkhangelsk at 18–22°C. There were no differences in larval growth rate between the two populations at 14 and 16°C.     

Organellar genetic diversity in Penstemonhaydenii (Scrophulariaceae): an endangered plantspecies

Genetic variation was analyzed in the endangered prairie forbPenstemon haydenii. Individuals from nine populations weresurveyed for chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA)variation. No cpDNA polymorphism was found. Analysis of mtDNA revealedvariation within and among populations. A minimum of eight haplotypeswere identified. The distribution of these genotypes suggests that thereis some correlation between geographic and genetic distances, whichreflects the minimal gene flow in thisspecies.     

Geographically congruent large‐scale patterns of plastid haplotype variation in the European herbs Silene dioica and S. latifolia (Caryophyllaceae)

The closely related dioecious herbs Silene latifolia and Silene dioica are widespread and predominantly sympatric in Europe. The species are interfertile, but morphologically and ecologically distinct. A study of large‐scale patterns of plastid DNA (polymerase chain reaction–restriction fragment length polymorphism) haplotypes in a sample of 198 populations from most of the European ranges of both species revealed extensive interspecific haplotype sharing. Four of the 28 detected haplotypes were frequent (found in > 40 populations) and widespread. Three of these frequent haplotypes occurred in both species and the geographic distribution of each haplotype was broadly congruent in both species. Each of these three, shared and widespread haplotypes is likely to have colonized central and/or northern Europe after the last glaciation from one or more of refugial areas in southern Europe. Interspecific hybridization and plastid introgression within refugial regions and/or during the early stages of postglacial expansion is the most plausible explanation for the broadly similar distribution patterns of the shared, frequent chloroplast haplotypes in the two species. The fourth frequent, widespread haplotype was absent from S. latifolia and almost entirely restricted to Nordic S. dioica. It is most likely that this haplotype spread into the Nordic countries from a central or northern European source or from a refugial area in Russia. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 161 , 153–170.     

Inventory of the carabid beetle fauna of the Gaoligong Mountains,western Yunnan Province,China: species of the tribe Zabrini (Coleoptera,Carabidae)

A ten-year multidisciplinary, multi-national and multi-institutional biodiversity inventory project in the Gaoligong Shan region of western Yunnan Province, China generated more than 35,000 specimens of the beetle (Coleoptera) family Carabidae. In this report, first of a planned series, we focus on diversity in tribe Zabrini. Our study of just over 1300 specimens of zabrine carabids from the project, all in genus Amara Bonelli, found a total of 13 species, all previously described, to occur in the study area, with none of them strictly endemic. We present a key for identification of adults of these species, as well as nomenclatural data, diagnoses, illustrations of dorsal habitus and male genitalia, and information about geographical, altitudinal and habitat distributions within the study area and overall geographical distribution for each species. Distributions of the species within the study area are compared, and broader geographical range patterns are characterized. We also discuss a possible role of the Gaoligong Shan region as one source area for the present-day fauna of the Himalaya and southern edge of the Qinghai-Xizang (Tibetan) Plateau.