The Role of Passive Defense (Selfish Herd and Dilution Effect) in the Gregarious Nesting of Liostenogaster Wasps (Vespidae, Hymenoptera, Stenogastrinae)

The advantages of the selfish herd and encounter-dilution effects were studied in two congeneric gregarious-nesting species of Stenogastrine wasps, Liostenogaster vechti Turillazzi and L. flavolineata (Cameron), in Malaysia. Results show that nesting in clusters affords these wasps some degree of protection against their main invertebrate predators, parasites and parasitoids. This advantage is reflected in the geometry of the aggregation as well as in the behavior of the individuals. The costs and benefits of group living and the implications of the selection of this nesting strategy in the evolution of sociality in these wasps are discussed.     

Climatic correlates of temporal demographic variation in the tropical hover wasp <Emphasis Type="Italic">Liostenogaster flavolineata</Emphasis>

Environmental factors, and particularly climate, play an important role in influencing behaviour in many insects. In social species, climate is known to have a strong influence on social traits, but how this manifests itself in tropical ecosystems is poorly understood. In Peninsular Malaysia, the climate is characterised by relatively consistent annual temperatures with wet/dry cycles, and the tropical hover-wasp Liostenogaster flavolineata Cameron is active year-round. Newly emerged females can choose to remain at the natal nest and help, or disperse and found their own nest depending on a balance of ecological and demographic factors. We collated long-term adult and brood census data for populations of L. flavolineata in Peninsular Malaysia in three different years to investigate temporal variation in demographics (brood and adult numbers) and how this might be related to climatic factors. Our data indicate that there are multiple, temporally distinct peaks of brood production in this population. The number of newly eclosing females and number of mature brood were positively associated with temperature and negatively associated with the number of rain-days during the observation period. Furthermore, larger females were produced during the peaks of brood production. We speculate how these patterns may influence the staying or leaving decisions of newly emerged females in a primitively eusocial species such as L. flavolineata.     

Rank and colony defense against conspecifics in a facultatively eusocial hover wasp

An important benefit of social living is increased capacityfor defense. Highly eusocial species have often evolved a fightingcaste for this purpose, but many facultatively eusocial insectsand cooperatively breeding vertebrates lack morphological castesand the decision to defend or not can depend on costs and benefitsto each individual. Defense by subordinates in a social groupcan be regarded as a form of helping, and helping input oftenvaries among subordinates of different age or size. Severalhypotheses attempt to explain variation in helping effort, includingthe effects of relatedness and differences in the costs of helping.Evidence for these hypotheses is mixed and often lacks dataon the rank of subordinates, an important determinant of expectedfuture fitness. We examined individual variation in propensityto defend the nest against conspecifics in the tropical hairy-facedhover wasp Liostenogaster flavolineata. Prior to experimentation,we determined the positions of all wasps in the age-based queueto inherit the single egg-laying position in each L. flavolineatagroup. Two approaches were then used: observations of defenseagainst natural intrusions by conspecifics and experimentaltrials where wasps were presented attached to a wire. Higherranks were more likely to defend the nest than lower ranks,opposite to the pattern previously documented for another formof helping: foraging effort. Possible explanations for thisresult are that higher ranked females are better defenders andthat they suffer a larger decrease in expected future fitnesswhen an intruder usurps their position in the inheritance queue.     

Speed and accuracy in nest-mate recognition: a hover wasp prioritizes face recognition over colony odour cues to minimize intrusion by outsiders

Social insects have evolved sophisticated recognition systems enabling them to accept nest-mates but reject alien conspecifics. In the social wasp, Liostenogaster flavolineata (Stenogastrinae), individuals differ in their cuticular hydrocarbon profiles according to colony membership; each female also possesses a unique (visual) facial pattern. This species represents a unique model to understand how vision and olfaction are integrated and the extent to which wasps prioritize one channel over the other to discriminate aliens and nest-mates. Liostenogaster flavolineata females are able to discriminate between alien and nest-mate females using facial patterns or chemical cues in isolation. However, the two sensory modalities are not equally efficient in the discrimination of ‘friend’ from ‘foe’. Visual cues induce an increased number of erroneous attacks on nest-mates (false alarms), but such attacks are quickly aborted and never result in serious injury. Odour cues, presented in isolation, result in an increased number of misses: erroneous acceptances of outsiders. Interestingly, wasps take the relative efficiencies of the two sensory modalities into account when making rapid decisions about colony membership of an individual: chemical profiles are entirely ignored when the visual and chemical stimuli are presented together. Thus, wasps adopt a strategy to ‘err on the safe side’ by memorizing individual faces to recognize colony members, and disregarding odour cues to minimize the risk of intrusion from colony outsiders.     

Patterns of herbivory on Passiflora leaf tissues and species by generalized and specialized feeding insects

Abstract.

• 1 The feeding patterns of specialized (Heliconius butterflies) and generalized (the grasshopper, Osmilia flavolineata) herbivores on leaf age classes and species of neotropical vines in the genus Passiflora are compared by offering them leaf discs in choice experiments.
• 2 Heliconius larvae selected young meristematic leaves over medium aged and mature leaves of Passiflora auriculata on which to feed.
• 3 Adults of O. flavolineata consumed more young leaves of P. pittieri from an array of different ages of leaf tissue, but showed no preference for young over mature leaves of P. tetrastylus or P. data.
• 4 In choice tests conducted with medium aged leaves of four species of Passiflora, O. flavolineata selected P. tetrastylus over P. pittieri and P. vitifolia, which were consumed more than P. alata.
• 5 P. alata, which was avoided by O. flavolineata in feeding trials, was the only abundant Passiflora in neotropical habitats where generalized herbivores (grasshoppers and katydids) were common.
• 6 Our results suggest that Passiflora species in forest habitats where some generalized herbivores are rare have fewer chemical defences, but retain physical defences (tough leaves and hooked trichomes) rendering them unpalatable to specialized herbivores like the larvae of Heliconius butterflies.

     

Next‐generation sequencing to inventory taxonomic diversity in eukaryotic communities: a test for freshwater diatoms

The recent emergence of barcoding approaches coupled to those of next‐generation sequencing (NGS) has raised new perspectives for studying environmental communities. In this framework, we tested the possibility to derive accurate inventories of diatom communities from pyrosequencing outputs with an available DNA reference library. We used three molecular markers targeting the nuclear, chloroplast and mitochondrial genomes (SSU rDNA, rbcL and cox1) and three samples of a mock community composed of 30 known diatom strains belonging to 21 species. In the goal to detect methodological biases, one sample was constituted directly from pooled cultures, whereas the others consisted of pooled PCR products. The NGS reads obtained by pyrosequencing (Roche 454) were compared first to a DNA reference library including the sequences of all the species used to constitute the mock community, and second to a complete DNA reference library with a larger taxonomic coverage. A stringent taxonomic assignation gave inventories that were compared to the real one. We detected biases due to DNA extraction and PCR amplification that resulted in false‐negative detection. Conversely, pyrosequencing errors appeared to generate false positives, especially in case of closely allied species. The taxonomic coverage of DNA reference libraries appears to be the most crucial factor, together with marker polymorphism which is essential to identify taxa at the species level. RbcL offers a high resolving power together with a large DNA reference library. Although needing further optimization, pyrosequencing is suitable for identifying diatom assemblages and may find applications in the field of freshwater biomonitoring.     

Characterization of microsatellite markers in peach [Prunus persica (L.) Batsch]

Microsatellites have emerged as an important system of molecular markers. We evaluated the potential of microsatellites for use in genetic studies of peach [Prunus persica (L.) Batsch]. Microsatellite loci in peach were identified by screening a pUC8 genomic library, a λZAPII leaf cDNA library, as well as through database searches. Primer sequences for the microsatellite loci were tested from the related Rosaceae species apple (Malus×domestica) and sour cherry (Prunus cerasus L.). The genomic library was screened for CT, CA and AGG repeats, while the cDNA library was screened for (CT)_n- and (CA)_n-containing clones. Estimates of microsatellite frequencies were determined from the genomic library screening, and indicate that CT repeats occur every 100 kb, CA repeats every 420 kb, and AGG repeats every 700 kb in the peach genome. Microsatellite- containing clones were sequenced, and specific PCR primers were designed to amplify the microsatellite- containing regions from genomic DNA. The level of microsatellite polymorphism was evaluated among 28 scion peach cultivars which displayed one to four alleles per primer pair. Five microsatellites were found to segregate in intraspecific peach-mapping crosses. In addition, these microsatellite markers were tested for their utility in cross-species amplification for use in comparative mapping both within the Rosaceae, and with the un- related species Arabidopsis thaliana L. Received: 18 June 1999 / Accepted: 6 December 1999     

Accelerating plant DNA barcode reference library construction using herbarium specimens: improved experimental techniques

A well‐covered reference library is crucial for successful identification of species by DNA barcoding. The biggest difficulty in building such a reference library is the lack of materials of organisms. Herbarium collections are potentially an enormous resource of materials. In this study, we demonstrate that it is likely to build such reference libraries using the reconstructed (self‐primed PCR amplified) DNA from the herbarium specimens. We used 179 rosaceous specimens to test the effects of DNA reconstruction, 420 randomly sampled specimens to estimate the usable percentage and another 223 specimens of true cherries (Cerasus, Rosaceae) to test the coverage of usable specimens to the species. The barcode rbcLb (the central four‐sevenths of rbcL gene) and matK was each amplified in two halves and sequenced on Roche GS 454 FLX+. DNA from the herbarium specimens was typically shorter than 300 bp. DNA reconstruction enabled amplification fragments of 400–500 bp without bringing or inducing any sequence errors. About one‐third of specimens in the national herbarium of China (PE) were proven usable after DNA reconstruction. The specimens in PE cover all Chinese true cherry species and 91.5% of vascular species listed in Flora of China. It is very possible to build well‐covered reference libraries for DNA barcoding of vascular species in China. As exemplified in this study, DNA reconstruction and DNA‐labelled next‐generation sequencing can accelerate the construction of local reference libraries. By putting the local reference libraries together, a global library for DNA barcoding becomes closer to reality.     

Chromosomal Mapping of Repetitive DNAs in the Grasshopper Abracris flavolineata Reveal Possible Ancestry of the B Chromosome and H3 Histone Spreading

Supernumerary chromosomes (B chromosomes) occur in approximately 15% of eukaryote species. Although these chromosomes have been extensively studied, knowledge concerning their specific molecular composition is lacking in most cases. The accumulation of repetitive DNAs is one remarkable characteristic of B chromosomes, and the occurrence of distinct types of multigene families, satellite DNAs and some transposable elements have been reported. Here, we describe the organization of repetitive DNAs in the A complement and B chromosome system in the grasshopper species Abracris flavolineata using classical cytogenetic techniques and FISH analysis using probes for five multigene families, telomeric repeats and repetitive C₀t-1 DNA fractions. The 18S rRNA and H3 histone multigene families are highly variable and well distributed in A. flavolineata chromosomes, which contrasts with the conservation of U snRNA genes and less variable distribution of 5S rDNA sequences. The H3 histone gene was an extensively distributed with clusters occurring in all chromosomes. Repetitive DNAs were concentrated in C-positive regions, including the pericentromeric region and small chromosomal arms, with some occurrence in C-negative regions, but abundance was low in the B chromosome. Finally, the first demonstration of the U2 snRNA gene in B chromosomes in A. flavolineata may shed light on its possible origin. These results provide new information regarding chromosomal variability for repetitive DNAs in grasshoppers and the specific molecular composition of B chromosomes.     

Influence of method of presentation on results of plant-host preference tests with two species of grasshopper

Leaf material of two Piper spp. (Piperaceae) was presented to two different generalist herbivores (Orthoptera: Acrididae) to determine whether method of leaf presentation in preference trials affects choice. For the grasshopper Leptomerinthoprora brevipennis, the method of presentation had no effect on preference ranking: more of Piper sancti-felicis was eaten than of P. arieianum, regardless of whether leaf material was presented as leaf disks, whole leaves supplemented with water, or whole leaves without water. In contrast, preference of the grasshopper Abracis flavolineata depended on presentation method. In that case, whole leaves of P. scancti-felicis supplemented with water were preferred over the same of P. arieianum, while dry weight consumption of leaf disks of P. arieianum was greater than that for disks of P. sancti-felicis. When presented with leaves not supplemented with water, A. flavolineata showed no significant preference for one plant species over another.

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Résumé Des feuilles provenant de deux espèces de Piper (Piperacae) ont été proposées à deux Orthoptères, Acrididae, herbivores généralistes, pour déterminer si la méthode de présentation des feuilles modifiait le choix. Pour Leptomerinthoprora brevipennis, la méthode de présentation est sans influence sur l'ordre de préférence; la consommation de Piper sancti-felicis est toujours supérieure à celle de P. arieianum, quelque soit le mode de présentation: disques de feuilles, feuilles entières humidifiées, ou feuilles entières sans addition d'eau. A l'opposé, l'ordre de préférence de Abracis flavolineata dépend du type de présentation; les feuilles entières humidifiées de P. sancti-felicis sont préférées à celles de P. arieianum proposées de la même façon, tandis que la consommation de disques secs de P. arieianum est plus importante que celle des disques secs de P. sancti-felicis. En présence de feuilles entières humidifiées, A. flavolineata ne manifeste aucune préférence.

     

Plant DNA barcode library for native flowering plants in the arid region of northwestern China

DNA barcoding is a well-established tool for rapid species identification and biodiversity monitoring. A reliable and traceable DNA barcode reference library with extensive coverage is necessary but unavailable for many geographical regions. The arid region in northwestern China, a vast area of about 2.5 million km², is ecologically fragile and often overlooked in biodiversity studies. In particular, DNA barcode data from the arid region in China are lacking. We develop and evaluate the efficacy of an extensive DNA barcode library for native flowering plants in the arid region of northwestern China. Plant specimens were collected, identified and vouchered for this purpose. The database utilized four DNA barcode markers, namely rbcL, matK, ITS and ITS2, for 1816 accessions (representing 890 species from 385 genera and 72 families), and consisted of 5196 barcode sequences. Individual barcodes varied in resolution rates: species- and genus-level rates for rbcL, matK, ITS and ITS2 were 79.9%–51.1%/76.1%, 79.9%–67.2%/88.9%, 85.0%–72.0%/88.2% and 81.0%–67.4%/84.9%, respectively. The three-barcode combination of rbcL + matK + ITS (RMI) revealed a higher species- and genus-level resolution (75.5%/92.1%, respectively). A total of 110 plastomes were newly generated as super-barcodes to increase species resolution for seven species-rich genera, namely Astragalus, Caragana, Lactuca, Lappula, Lepidium, Silene and Zygophyllum. Plastomes revealed higher species resolution compared to standard DNA barcodes and their combination. We suggest future databases include super-barcodes, especially for species-rich and complex genera. The plant DNA barcode library in the current study provides a valuable resource for future biological investigations in the arid regions of China.     

Ecological constraints on independent nesting in facultatively eusocial hover wasps

Recent field experiments suggest that cooperative breeding in vertebrates can be driven by a shortage of breeding territories. We did analogous experiments on facultatively eusocial hover wasps (Stenogastrinae: Liostenogaster flavolineata). We provided nesting opportunities by removing residents from 39 nests within a large aggregation (1995), and by glueing 20 nests obtained from a distant site into a second aggregation (1996). We prevented nest-less floaters from competing for these opportunities in 1995 but not in 1996. In both years, helpers in unmanipulated groups were given opportunities to nest independently without having to incur nest-building costs and with a reduced wait before potential helpers emerged. Helpers visited the nests we provided, but adopted only a small proportion (5% of 111 vacancies created in 1995). Others were adopted by floaters, but a significant proportion of nests were never adopted (nine out of 20 in 1995, seven out of 20 in 1996). Helpers that visited nests did not originate from particular kinds of social group. Nests containing older brood were more likely to be adopted, and adopting females rarely destroyed older brood. A general feature of social insect but not vertebrate life-histories, the long period of offspring dependency relative to the short life expectancy of adult carers, may be a key factor constraining independent nesting.     

DNA barcoding as a tool for early warning and monitoring alien duckweeds (Lemna sp.pl.): the case of Central Italy

Aquatic habitats are vulnerable to the invasion of alien species, so early warning protocols are necessary for eradication. The presence in Italy of two alien duckweeds in freshwaters has been documented: Lemna minuta, that showed high invasivity, and L. valdiviana, still confined to south Lazio. These two species may be mistaken for each other and for the domestic L. minor and L. gibba due to morphological variation. Here, we assess the applicability of DNA barcoding as a complement to morphological analysis for monitoring the spread of alien Lemna. We chose two chloroplast genome sequences for their ability to discriminate all Lemna species: the 5’ intron of the trnK gene and the matK gene. Among 48 samples of Lemna collected at 20 sites in Central Italy, 20 were identified as L. minor, 19 as L. minuta, five as L. trisulca and four as L. gibba. L. minuta was present at most sampling sites; in particular, at six locations of Lake Trasimeno, eight L. minuta samples were found. We demonstrate that DNA sequence analyses with cost-effective barcoding techniques can effectively support expert efforts in species determination for an early alert system of invasive Lemna species.     

Integrating a comprehensive DNA barcode reference library with a global map of yews (Taxus L.) for forensic identification

Rapid and accurate identification of endangered species is a critical component of biosurveillance and conservation management, and potentially policing illegal trades. However, this is often not possible using traditional taxonomy, especially where only small or preprocessed parts of plants are available. Reliable identification can be achieved via a comprehensive DNA barcode reference library, accompanied by precise distribution data. However, these require extensive sampling at spatial and taxonomic scales, which has rarely been achieved for cosmopolitan taxa. Here, we construct a comprehensive DNA barcode reference library and generate distribution maps using species distribution modelling (SDM), for all 15 Taxus species worldwide. We find that trnL‐trnF is the ideal barcode for Taxus: It can distinguish all Taxus species and in combination with ITS identify hybrids. Among five analysis methods tested, NJ was the most effective. Among 4,151 individuals screened for trnL‐trnF, 73 haplotypes were detected, all species‐specific and some population private. Taxonomical, geographical and genetic dimensions of sampling strategy were all found to affect the comprehensiveness of the resulting DNA barcode library. Maps from SDM showed that most species had allopatric distributions, except T. mairei in the Sino‐Himalayan region. Using the barcode library and distribution map data, two unknown forensic samples were identified to species (and in one case, population) level and another was determined as a putative interspecific hybrid. This integrated species identification system for Taxus can be used for biosurveillance, conservation management and to monitor and prosecute illegal trade. Similar identification systems are recommended for other IUCN‐ and CITES‐listed taxa.     

Plant DNA‐barcode library and community phylogeny for a semi‐arid East African savanna

Applications of DNA barcoding include identifying species, inferring ecological and evolutionary relationships between species, and DNA metabarcoding. These applications require reference libraries that are not yet available for many taxa and geographic regions. We collected, identified, and vouchered plant specimens from Mpala Research Center in Laikipia, Kenya, to develop an extensive DNA‐barcode library for a savanna ecosystem in equatorial East Africa. We amassed up to five DNA barcode markers (rbcL, matK, trnL‐F, trnH–psbA, and ITS) for 1,781 specimens representing up to 460 species (~92% of the known flora), increasing the number of plant DNA barcode records for Africa by ~9%. We evaluated the ability of these markers, singly and in combination, to delimit species by calculating intra‐ and interspecific genetic distances. We further estimated a plant community phylogeny and demonstrated its utility by testing if evolutionary relatedness could predict the tendency of members of the Mpala plant community to have or lack “barcode gaps”, defined as disparities between the maximum intra‐ and minimum interspecific genetic distances. We found barcode gaps for 72%–89% of taxa depending on the marker or markers used. With the exception of the markers rbcL and ITS, we found that evolutionary relatedness was an important predictor of barcode‐gap presence or absence for all of the markers in combination and for matK, trnL‐F, and trnH–psbA individually. This plant DNA barcode library and community phylogeny will be a valuable resource for future investigations.     

Phylogeny of tall fescue and related species using RFLPs

The wild species of tall fescue (Festuca arundinacea var.genuina Schreb.) represent a wide range of genetic variation and constitute potential germplasm for tall fescue improvement. Our objective was to evaluate genome specificity of the previously-identified DNA probes and to examine the phylogenetic relationship of tall fescue with six related species by using RFLP data. A total of 29 DNA probes from aPstI-genomic library of tall fescue were hybridized toEcoRI-orHindIII-digested DNA of 32 plants from sixFestuca species and fromLolium perenne L. Fifteen probes hybridized to all seven species. The remaining 14 probes showed differential hybridization patterns (i.e., ±), especially at the diploid and tetraploid levels. This hybridization pattern reflected genome divergence in these species. The DNA probes will be useful markers in breeding programs involving interspecific and intergeneric hybridization. Cluster analyses were performed using the average genetic distances calculated with the RFLP data from 53 probe-enzyme combinations. Generally, genotypes from the same species were grouped in the same cluster. These data indicated that tall fescue has a close relationship withF. pratensis Huds. (diploid),F. arundinacea var.glaucescens Boiss. (tetraploid), andL. perenne L. (diploid) and thatFestuca pratensis andL. perenne had the closest degree of relationship.This paper is a contribution of the Missouri Agricultural Experimental Station, Journal Series no. 11,798     

Consumption of sugars, hemicellulose, starch, pectin and cellulose by the grasshopper Arcris flavolineata

Adults (0.61 g, fresh-weight) of Abracris flavolineata De Geer (Orthoptera: Acrididae) feeding on Brassica oleracea acephala leaves ingest 21 mg dry-weight/day with an approximate digestibility of 42%. Chemical determinations performed on the leaves ingested and on the feces expelled led to the determination of the approximate digestibilities (%) of the major carbohydrates of leaves as follows: soluble carbohydrates, 91; pectin, 32.1; hemicellulose, 0; starch, 66; cellulose, 15. The results are not sufficient to disregard the possibility that digestible hemicellulose polymers contaminate the pectin and the cellulose fraction. Thus, it is possible that the digestibility of hemicellulose is different from zero, and that the digestibility of pectin and cellulose are somewhat lower than reported. The data are used to propose physiological roles of the enzyme activities previously found in the A. flavolineata midgut.     

Construction of tomato genomic DNA libraries in a binary-BAC (BIBAC) vector

This is the first report of large insert genomic DNA libraries constructed in a binary-BAC (BIBAC) vector. Genomic DNA libraries containing approximately 4.6 haploid nuclear genomic equivalents were constructed for Lycopersicon esculentum (cv. Mogeor) and Lycopersicon pennellii (LA716) in the BIBAC2 vector. The L. esculentum library has an average insert size of 125 kb and is comprised of 42 272 individual colonies stored as frozen cultures in a 384-well format (108 plates). The L. pennellii library has an average insert size of 90 kb and is comprised of 53 760 individual clones (140 384-well plates). In each of the libraries, it is estimated that 90% of the colonies contain genomic DNA inserts. The composition of the L. esculentum and L. pennellii libraries was determined by analyzing a series of randomly selected clones. The L. esculentum library was surveyed for clones containing chloroplast DNA (1.4%), mitochondrial DNA (0.012%) and repetitive DNA motifs. BIBAC clones that may contain a gene of interest can be identified from these libraries by colony hybridization with homologous or heterologous probes or by PCR pooling techniques. Once identified, BIBAC genomic DNA library clones are immediately suitable for Agrobacterium tumefaciens-mediated plant transformation.     

Construction of a yeast artificial chromosome library of pepper (Capsicum annuum L.) and identification of clones from the Bs2 resistance locus

A yeast artificial chromosome (YAC) library was constructed using high-molecular-weight DNA isolated from pepper (Capsicum annuum L.) leaf protoplasts. Insert DNA was prepared by partial digestion using EcoRI and subjected to electrophoretic fractionation before in-gel ligation to the pJS97/98 YAC vector. Prior to transformation of yeast spheroplasts, ligation products were subjected to a second electrophoretic size selection. The library consists of about 19 000 clones with an average insert size of 500 kb, thus representing approximately three haploid genome equivalents. Three PCR-based markers tightly linked to the pepper Bs2 resistance gene were used to assess the utility of this library for positional cloning. Three YAC clones containing pepper genomic DNA from the Bs2 resistance locus were isolated from the library. The clones ranged in size from 270 kb to 1.2 Mb and should prove useful for the cloning of the Bs2 gene. Received: 15 January 1999 / Accepted: 11 May 1999     

Low genetic diversity in Polish populations of sibling ant species: <Emphasis Type="Italic">Lasius niger</Emphasis> (L.) and <Emphasis Type="Italic">Lasius platythorax</Emphasis> Seifert (Hymenoptera,Formicidae)

We present preliminary data on mitochondrial DNA diversity within and among populations of the ants Lasius niger and Lasius platythorax in Poland. Phylogenetic analysis based on the mitochondrial DNA markers: cytochrome c oxidase subunit I (cox1) and 16S ribosomal RNA (16S rRNA) confirms the species status of L. niger and L. platythorax. Intraspecific variability is low in both species, which might be a result of severe bottlenecks and rapid postglacial expansion into Central Europe.