European Species Research Articles

Macro- and micro-geographic genetic variation in early fitness traits in populations of maritime pine (Pinus pinaster Ait.).

Assessing adaptive genetic variation and its spatial distribution is crucial to conserve forest genetic resources and manage species' adaptive potential. Macro-environmental gradients commonly exert divergent selective pressures that enhance adaptive genetic divergence among populations. Steep micro-environmental variation might also result in adaptive divergence at finer spatial scales, even under high gene flow, but it is unclear how often this is the case. Here, we assess genetic variation in early fitness traits among distant and nearby maritime pine (Pinus pinaster Ait.) populations, to investigate climatic factors associated with trait divergence, and to examine trait integration during seedling establishment. Open pollinated seeds were collected from seven population pairs across the European species distribution, with paired populations spatially close (between <1 km up to 21 km) but environmentally divergent. Seeds were sown in semi-natural conditions at three environmentally contrasting sites, where we monitored seedling emergence, growth and survival. At large spatial scales, we found significant genetic divergence among populations in all studied traits, with certain traits exhibiting association with temperature and precipitation gradients. Significant trait divergence was also detected between pairs of nearby populations. Besides, we found consistent trait correlations across experimental sites, notably heavier seeds and earlier seedling emergence were both associated with higher seedling survival and fitness over two years in all experimental conditions. We identified mean annual temperature and precipitation seasonality as potential drivers of P. pinaster population divergence in the studied early-life traits. Populations genetically diverge also at local spatial scales, potentially suggesting that divergent natural selection can override gene flow along local-scale ecological gradients. These results suggest the species exhibits substantial adaptive potential that has allowed it to survive and evolve under contrasting environmental conditions.

Wandering the taxonomic mine-field: the Podocotyle species complex (Digenea: Opecoelidae).

This study focuses on the opecoelid genus Podocotyle, particularly the species in the northern European seas (White, Barents and Pechora), with additional data from few isolates from the Far East. The research combines molecular analyses (28S, 5.8S+ITS2 rDNA, and cox1 mtDNA) and morphological examinations. Molecular analyses suggested that there are most likely four species of Podocotyle in the northern European seas and three more in the Far East. Morphological redescriptions are provided for three northern European species: P. atomon, P. reflexa, and P. odhneri, with the latter recognized as a valid species. The study also verified the life cycle of P. atomon and identified Lacuna vincta (Littorinidae) as the first intermediate host for P. reflexa. In the Sea of Okhotsk, L. turrita is the first intermediate host for two apparently undescribed Podocotyle species. The results also highlight the morphological variability of P. atomon, emphasizing the need for an integrative approach in the taxonomy and biodiversity studies.

Persistence Over Millennia Through Extreme Clonal Longevity: Phylogenomic Insight Into History of One of the World's Rarest Plant Species

ABSTRACTAimThe evolutionary history of European alpine plant species with medium to large geographical ranges is relatively well explored. Here, we investigate the genetic structure and diversity of an extremely narrow endemic and one of the world's rarest plants.LocationEastern Carpathians, Romania.TaxonAndryala laevitomentosa (Asteraceae), an evolutionarily isolated herb species with a worldwide range limited to five micropopulations distributed along a 1.8 km long mountain ridge.MethodsWe used three plastid loci, nuclear ribosomal ITS and genome‐wide, mostly nuclear 26,272 Single‐Nucleotide Polymorphism (SNPs) obtained from RAD‐seq data. We assessed haplotype and genotype diversity, dated the resulting phylogeographic structure, quantified seed production and inferred vegetative propagation.ResultsMaternally inherited plastid markers and nuclear genomic data revealed a concordant pattern: (i) limited genetic diversity, with seven cpDNA haplotypes and 11 RAD‐seq multilocus genotypes; (ii) a strong geographic structure corresponding to spatially isolated genets (clones). The species is likely of early Pleistocene origin (c. 2 Mya), and the estimated age of individual clones varied from c. 24 to 64 Kya. The average seed set assessed over 3 years was only 0.4%. However, the species reproduces veg by axillary and adventitious rosettes formed on rhizomes and roots, respectively.Main ConclusionsThe strong trade‐off between sexual and vegetative reproduction explains not only a deep and ancient phylogeographic structure but also the rarity of the species. Its survival depends almost entirely on vegetative reproduction. The genets of A. laevitomentosa are amongst the oldest clones ever documented in angiosperms. The persistence of these clones in situ for tens of thousands of years suggests an exceptional ability of this species to adapt to major climatic oscillations throughout the Pleistocene and Holocene and challenge our perception of the extent of resilience in plants.

Outbreak of Contagious Ecthyma in Free-Ranging Iberian Ibex (Capra pyrenaica) in the Montgrí Massif Natural Park, Catalonia, Northeastern Spain.

Contagious ecthyma is a common, worldwide, and highly transmissible viral zoonotic skin disease caused by the orf virus (ORFV). It mainly affects farmed small ruminants, but it has also been described in a broad range of wild and domestic mammals, with Caprinae species most susceptible. Between November 2019 and January 2020, adults, juveniles, yearlings, and kids from an Iberian ibex (Capra pyrenaica) population established in the Montgrí Massif Natural Park (Catalonia, northeastern Spain) were observed with clinical signs and lesions compatible with ORFV infection. The carcass of an adult male with severe disease enabled sample collection for histopathology and molecular studies, confirming ORFV DNA in the skin lesions. Sequence analyses indicated that the ORFV strain detected had high homology (>98%) with strains previously obtained from other European wild ruminant species. The outbreak peaked in December 2019, with an estimated prevalence of 68.97% (95% confidence interval, 53.35-84.59). From February 2020 forward, no individuals with lesions were observed. The yearly counts for population monitoring corroborated the apparently negligible ORFV impact on the ibex population of the Montgrí Natural Park. There are no previous reports of contagious ecthyma in a free-ranging Iberian ibex population.

Oxidative state is associated with migration distance, but not traits linked to flight energetics

Flight can be highly energy demanding, but its efficiency depends largely on flight style, wing shape and wing loading, and a range of morphological and lifestyle adaptations that can modify the cost of sustained flight. Such behavioural and morphological adaptations can also influence the physiological costs associated with migration. For instance, during intense flight and catabolism of reserves, lipid damage induced by pro‐oxidants increases, and to keep oxidative physiological homeostasis under control, the antioxidant machinery can be upregulated. Studies on the oxidative physiology of endurance flight have produced contradictory results, making generalization difficult, especially because multispecies studies are missing. Therefore, to explore the oxidative cost of flight and migration, we used samples collected during the breeding season from 113 European bird species and explored the associations of measures of antioxidant capacity (total antioxidant status, uric acid and glutathione concentration) and oxidative damage of lipids (malondialdehyde) with variables reflecting flight energetics (year‐round or specifically during migration) using a phylogenetic framework. We found that none of the traits predicting year‐round flight energy expenditure (flight style, wing morphology and flight muscle morphology) explained any measures of oxidative state. Our results suggest that birds endure their everyday flight exercise without or with low oxidative cost. However, oxidative damage to lipids and one component of the endogenous antioxidant system (uric acid), measured after the end of spring migration on breeding adult birds, increased with migration distance. Our results suggest that migration could have oxidative consequences that might be carried over to subsequent life‐history stages (breeding).

High phenotypic diversity correlated with genomic variation across the European Batrachochytrium salamandrivorans epizootic.

Recognizing the influence of pathogen diversity on infection dynamics is crucial for mitigating emerging infectious diseases. Characterising such diversity is often complex, for instance when multiple pathogen variants exist that interact differently with the environment and host. Here, we explore genotypic and phenotypic variation of Batrachochytrium salamandrivorans (Bsal), an emerging fungal pathogen that is driving declines among an increasing number of European amphibian species. For thirteen isolates, spanning most of the known temporal and geographical Bsal range in Europe, we mapped phenotypic diversity through numerous measurements that describe varying reproductive rates in vitro across a range of temperatures. Bsal isolates are revealed to have different thermal optima and tolerances, with phenotypic variation correlating with genomic diversity. Using a mechanistic niche model of the fire salamander (Salamandra salamandra) as an example, we illustrate how host steady-state body temperature and Bsal thermal range variation may influence pathogen growth through space and time across Europe. Our combined findings show how the identity of emergent pathogen variants may strongly influence when and which host populations are most at risk.

The development of L. major, L. donovani and L. martiniquensis, Leishmania currently emerging in Europe, in the sand fly species Phlebotomus perniciosus and P. tobbi.

Several new species of Leishmania have recently emerged in Europe, probably as the result of global changes and increased human migration from endemic areas. In this study, we tested whether two sand fly species, the Western Mediterranean Phlebotomus perniciosus and the Eastern Mediterranean P. tobbi, are competent vectors of L. donovani, L. major and L. martiniquensis. Sand flies were infected through the chick skin membrane using Leishmania species and strains of various geographical origins. Leishmania infections were evaluated by light microscopy and qPCR, and the representation of morphological forms was assessed from Giemsa-stained gut smears. Neither P. perniciosus nor P. tobbi supported the development of L. martiniquensis, but L. major and L. donovani in both species survived defecation of blood meal remnants, colonized the stomodeal valve and produced metacyclic stages. The results with L donovani have shown that infection rates in sand flies can be strain-specific; therefore, to determine vector competence or refractoriness, it is optimal to test at least two strains of Leishmania. Both sand fly species tested are potential vectors of L. donovani and L. major in Mediterranean area. However, further studies will be needed to identify European vectors of L. martiniquensis and to test the ability of other European sand fly species to transmit L. major, L. donovani, L. tropica and L. infantum.

Resilience of genetic diversity in forest trees over the Quaternary

The effect of past environmental changes on the demography and genetic diversity of natural populations remains a contentious issue and has rarely been investigated across multiple, phylogenetically distant species. Here, we perform comparative population genomic analyses and demographic inferences for seven widely distributed and ecologically contrasting European forest tree species based on concerted sampling of 164 populations across their natural ranges. For all seven species, the effective population size, Ne, increased or remained stable over many glacial cycles and up to 15 million years in the most extreme cases. Surprisingly, the drastic environmental changes associated with the Pleistocene glacial cycles have had little impact on the level of genetic diversity of dominant forest tree species, despite major shifts in their geographic ranges. Based on their trajectories of Ne over time, the seven tree species can be divided into three major groups, highlighting the importance of life history and range size in determining synchronous variation in genetic diversity over time. Altogether, our results indicate that forest trees have been able to retain their evolutionary potential over very long periods of time despite strong environmental changes.

Automated long‐term monitoring of RFID‐tagged individuals reveals high hibernaculum site fidelity in Daubenton's bats and Natterer's bats

AbstractIn the temperate zone, many bat species depend on underground sites during the autumn mating period and as hibernacula in winter. Anthropogenic or natural alteration or destruction of a hibernaculum may severely threaten the bats that rely on the site, especially if they are not aware of suitable alternatives. Currently, the rate at which bats visit other hibernacula, or disperse between them, is poorly understood. Here, we studied hibernation site fidelity in two European bat species, Daubenton's bat (Myotis daubentonii) and Natterer's bat (Myotis nattereri), that share hibernacula but differ in their hibernation phenology and mating systems. The activity of over 2500 RFID‐tagged individuals was monitored using an automated recording system at the entrances of three hibernacula over eight full years. Hibernation site fidelity was high in both species, with only 1.6% of Daubenton's bats and 4.1% of Natterer's bats moving between the monitored sites during their lifetimes. Juvenile bats moved more frequently than adults and contrary to our expectations, in Natterer's bats, more females moved between hibernacula than males. Movements almost always occurred during the autumn mating season, and more frequently between neighboring hibernacula than between distant sites. Finally, we also found that individuals rarely explored more than one of the three entrances to a site, despite their close proximity. Taken together, our results suggest that some juvenile bats may explore multiple sites, but that they rarely disperse to other hibernacula as adults. As a result, most individuals are either not aware of alternative sites or do not regularly update this information. We therefore argue that the creation of new underground objects as a mitigation measure may not function as an immediate compensation for the disturbance or the loss of an existing hibernaculum. Instead, the proactive identification and protection of existing hibernacula should be prioritized.

Expansion of the area of the trematodes &lt;i&gt;Paracoenogonimus ovatus&lt;/i&gt; (Katsurada, 1914) in Western Siberia

The distribution of trematodes Paracoenogonimus ovatus and its first intermediate hosts, viviparous Viviparus viviparus and V. contectus in the Ob–Irtysh basin were analyzed. Until the 1990s the spreading of P. ovatus in this region was associated with habitats of V. contectus, which is considered a native species in Western Siberia. Since the mid-1990s European species V. viviparus began to spread in the Ob basin. Before this, only one source of infection with P. ovatus was known associated with finding V. viviparus in the Lower Irtysh. This paper describes another such outbreak in the Tura River near the city of Tyumen. In fish from the Tura River, a generally high extent and intensity of invasion were noted. Further expansion of the range is predicted P. ovatus associated with expansion V. viviparus.

Kinematics of elongate harvestmen chelicerae: Comparative range of motion analyses in extant Ischyropsalis (Dyspnoi, Opiliones)

Chelicerae, the mouthparts of chelicerates, are essential for food processing. Particularly within harvestmen (Opiliones), some species have greatly elongated their tripartite chelicerae and utilize them for mating behavior, defense, and primarily for predation. We investigated two European species, Ischyropsalis muellneri and Ischyropsalis hellwigii, which occupy different niches (caves, forests), exhibit different feeding ecologies (opportunist, specialist), and first and foremost possess different chelicerae morphologies (long and thin, short and robust). We scanned the specimens using state-of-the-art micro-CT, generated surface reconstructions, and equipped one chelicera of each specimen with artificial joints to explore their Range of Motion in a 3D kinematic approach. For a size-corrected comparison of the two species, we analyzed the Range of Motion in addition to three different settings (original body size, body scaled to 5 mm, chelicerae scaled to 5 mm). Ischyropsalis muellneri reached a higher maximum excursion angle (= single Range of Motion) in all three joints, also exhibiting a greater total Range of Motion in the original body length setting, as well as the scaled body length setting. Only in the third setting, the total Range of Motion of Ischyropsalis hellwigii was slightly higher, although Ischyropsalis muellneri still extended further ventrally. Our results suggest that the sturdier, more massive chelicerae of Ischyropsalis hellwigii, attributable to strong specialization on snails as prey, are associated with reduced Range of Motion. The less food-specialized species Ischyropsalis muellneri apparently requires higher flexibility of its chelicerae for prey capture, likely due to its restriction to cave ecosystems, where food availability is relatively scarce. We could show that virtual Range of Motion analyses in harvestmen chelicerae can play a pivotal role in understanding the theoretical feeding ecology and functional morphology of this group. This approach can be verified by in-vivo observations and measurements or extended to other arachnid taxa and other body parts.

Pre-emptive augmentative biological control of Spodoptera frugiperda in Europe using Trichogramma spp.

BackgroundFall armyworm, Spodoptera frugiperda, is a highly invasive pest of maize and other crops worldwide. It has recently been detected in Europe, and it is urgent to test and develop new sustainable control methods adapted to the European context and market. Trichogramma spp. are egg parasitoids that are sometimes used as biological control agent against S. frugiperda in other continents. However, a major issue using Trichogramma spp. against this pest is that females cannot reach all eggs in an egg mass, which is usually composed of one to three layers of eggs, often covered with scales and hair. Three European Trichogramma species were tested for their ability to parasitize egg masses with one to three layers and with or without hair and scale cover.MethodsTrichogramma brassicae, T. dendrolimi and T. cacoeciae were offered five types of S. frugiperda egg masses: one-layer without hair; one-layer with hair; two-layers without hair; two-layers with hair; three-layers with hair. For each treatment, an egg mass laid on paper was placed in a small vial saturated with females, to be sure that all reachable eggs would be parasitized. For each egg mass, the number of eggs in each layer was counted. Fifteen replicates were made per treatment, as well as 10 control vials without Trichogramma sp. Parasitism rates were calculated for each egg mass.ResultsThere were significant differences in the ability of parasitoid species to oviposit through hair and scales and to reach the lower egg layers. Trichogramma dendrolimi was the most efficient species and T. cacoeciae the least. Depending on the number of egg layers and hair and scales, parasitism rates by T. brassicae, T. dendrolimi and T. cacoeciae varied between 99 and 41%, 100 and 43% and 100 and 28%, respectively.ConclusionsThe assays confirmed that Trichogramma females cannot easily oviposit through thick layers of hair and scales but overall parasitism rates were higher than found previously. Important variations between species were found, and more Trichogramma sp. and other local natural enemies should be tested pre-emptively before S. frugiperda has invaded Europe.

Natural enemies of Lemna minuta in its native range and their potential as biological control agents for Europe

ABSTRACT Lemna minuta is native to North America but an invasive alien plant in Europe, where it poses significant threats to freshwater ecosystems. Explorations for biological control agents were conducted in two regions of the United States (Louisiana and California), revealing three candidate species. Subsequent laboratory investigations focused on the weevil Tanysphyrus lemnae and the fly Lemnaphila scotlandae as the most promising agents and confirmed that both the herbivorous insects attack L. minuta plants. To evaluate the host specificity of these two potential agents, insects were exposed to five species of duckweed commonly occurring in North America and Europe. Preliminary host-range testing indicated that T. lemnae develops on all evaluated duckweed species, including those from different genera (Lemna, Spirodela, Landoltia). Conversely, data revealed that feeding and development of the dipteran L. scotlandae are limited to species in the genus Lemna, but this includes L. minor, a native European species threatened in Europe due to the spread of L. minuta. No-choice and choice tests confirmed that neither larvae nor adults of L. scotlandae discriminate between the two Lemna species, except for pupation, which occurs more frequently in L. minor under choice conditions. We conclude that the broad host-range of T. lemnae and L. scotlandae render them unsuitable as biological control agents of L. minuta in Europe.

An Update on the Conservation Status Assessment of two Endangered Freshwater Mussel Species in Bavaria, Germany

ABSTRACTThe two highly endangered European mussel species Margaritifera margaritifera and Unio crassus are target species of conservation. Based on a recently completed systematic state‐wide monitoring of each 22 M. margaritifera and 22 U. crassus streams in Bavaria, Germany, we present an update on population trends, conservation status, habitat quality and threats for both species. Populations status and habitat quality varied strongly between M. margaritifera and U. crassus streams, but there was also great variability within each of those groups. The population decline of M. margaritifera has continued, albeit higher proportions of juveniles originating from artificial breeding programmes have been established in some streams. Habitat quality often did not match known requirements as evident from poor stream bed quality, lack of hosts and elevated nutrient levels. In contrast, U. crassus populations showed a better status, with an increase in population size over all sampled streams. Successful recruitment was indicated by high proportions of juveniles. However, no mussels older than 16 years were found, probably due to predation and structural stream maintenance measures. Climate change effects, such as extreme droughts, affected both species. This study demonstrates different needs in conservation management for both species. Although mitigation of drought effects is commonly needed for both species, tackling host fish management and direct threats such as predation should be prioritized in U. crassus, whereas restoration of prime habitat quality and intact catchments is key to enable natural recruitment and sustainable populations of M. margaritifera.

Stable Isotope Labelling Reveals Water and Carbon Fluxes in Temperate Tree Saplings Before Budbreak.

Despite considerable experimental effort, the physiological mechanisms governing temperate tree species' water and carbon dynamics before the onset of the growing period remain poorly understood. We applied 2H-enriched water during winter dormancy to the soil of four potted European tree species. After 8 weeks of chilling, hydrogen isotopes in stem, twig and bud water were measured six times during 2 consecutive weeks of forcing conditions (Experiment 1). Additionally, we pulse-labelled above-ground plant tissues using 2H-enriched water vapour and 13C-enriched CO2 7 days after exposure to forcing conditions to trace atmospheric water and carbon uptake (Experiment 2). Experiment 1 revealed soil water incorporation into the above-ground organs of all species during the chilling phase and significant species-specific differences in water allocation during the forcing conditions, which we attributed to differences in structural traits. Experiment 2 illustrated water vapour incorporation into all above-ground tissue of all species. However, the incorporation of carbon was found for evergreen saplings only. Our results suggest that temperate trees take up and reallocate soil water and absorb atmospheric water to maintain sufficient above-ground tissue hydration during winter. Therefore, our findings provide new insights into the water allocation dynamics of temperate trees during early spring.

Two Palaearctic species of Orthonevra (Diptera: Syrphidae) under the name O. brevicornis

During studying European material of Orthonevra Macquart, 1829 the presence of two widespread species so far interpreted as Orthonevra brevicornis (Loew, 1843) has been discovered. One species occurs from Central Europe into Transcaucasia and agrees with the taxon O. brevicornis of Loew (1843). The second species, O. atlantica Żóralski &amp; Van de Meutter sp. nov., has its main distribution range in the Atlantic parts of Europe and is described here. We provide a redescription of O. brevicornis and designate a neotype from the type locality (Poland, Posen [= Poznań] vicinity). The most important characters for distinguishing males of these two closely related species are in the structures of the terminalia. A diagnosis and identification key to European black-legged Orthonevra species is given as well as new sequences of the cytochrome c oxidase subunit I (COI) of mitochondrial DNA (mtDNA), also known as DNA barcodes, for most of the species of this group. Some new information and changes to the known ranges of Orthonevra species are provided as a result of this revisionary work: O. frontalis (Loew, 1843) is removed from the list of species occurring in Poland; O. gemmula Violovitsh, 1979 is confirmed from Hungary; O. incisa (Loew, 1843) is confirmed from its locus typicus (Poland, Posen vicinity); and O. montana Vujić, 1999 is reported from Poland and Austria for the first time and confirmed for Germany. An explanation for some previous records published as O. plumbago (Loew, 1840) is also given.

The genus Kelliella (Bivalvia) in the Atlantic Ocean

ABSTRACT Kelliella M. Sars, 1870 is a genus of small epibenthic bivalves inhabiting muddy bottoms of the outer shelf and slope worldwide, commonly in habitats that are oxygen-poor or enriched in organic carbon. Currently, there are five accepted species of Kelliella with an Atlantic type locality, and six more Atlantic species, including Vesicomya atlantica (Smith, 1885), the type species of Vesicomya Dall, 1886, were at some time placed in Kelliella. In this work, we revise the genus Kelliella from the North Atlantic waters, based on large amounts of new material from both sides, and describe a new species from Martinique (Kelliella madininae n. sp.). We here propose, after revision of the shell morphology, the synonymy of K. biscayensis Allen, 2001 with K. miliaris (Philippi, 1844), and consider a single West European and Mediterranean species of Kelliella. The fossil “Kelliella” barbara is also excluded from the genus Kelliella and reallocated as Coralliophaga barbara (Studencka, 1987) n. comb. In the Atlantic margin of America, we consider that Diplodonta pilula Dall, 1881 remains a nomen dubium. The type material of K. concentrica Allen, 2001 is figured, and this species considered a member of the Vesicomyinae. The new Caribbean species, K. madininae, resemble K. brasuca Passos, Machado &amp; Fantinatti, 2017 from southeastern Brazil. We recorded new localities for Kelliella goesi Odhner, 1960, so far known only from its original description. Diagnostic characters discriminating Vesicomya from Kelliella, stated by Krylova et al. (2018), Journal of Molluscan Studies, 84: 69–91, are discussed, and we consider the hinge architecture as the most robust.

Effects of wind turbine dimensions on the collision risk of raptors: A simulation approach based on flight height distributions

Wind energy development is a key component of climate change mitigation. However, birds collide with wind turbines, and this additional mortality may negatively impact populations. Collision risk could be reduced by informed selection of turbine dimensions, but the effects of turbine dimensions are still unknown for many species.As analyses of mortality data have several limitations, we applied a simulation approach based on flight height distributions of six European raptor species. To obtain accurate flight height data, we used high-frequency GPS tracking (GPS tags deployed on 275 individuals). The effects of ground clearance and rotor diameter of wind turbines on collision risk were studied using the Band collision risk model.Five species had a unimodal flight height distribution, with a mode below 25 m above ground level, while Short-toed Eagle showed a more uniform distribution with a weak mode between 120 and 260 m. The proportion of positions within 32–200 m ranged from 11 % in Marsh Harrier to 54 % in Red Kite.With increasing ground clearance (from 20 to 100 m), collision risk decreased in the species with low mode (−56 to −66 %), but increased in Short-toed Eagle (+38 %). With increasing rotor diameter (from 50 to 160 m) at fixed ground clearance, the collision risk per turbine increased in all species (+151 to +558 %), while the collision risk per MW decreased in the species with low mode (−50 % to −57 %).These results underpin that wind turbine dimensions can have substantial effects on the collision risk of raptors. As the effects varied between species, wind energy planning should consider the composition of the local bird community to optimise wind turbine dimensions. For species with a low mode of flight height, the collision risk for a given total power capacity could be reduced by increasing ground clearance, and using fewer turbines with larger diameter.

A DNA barcode reference library for the Tipulidae (Insecta, Diptera) of Germany.

Tipulidae, commonly known as true crane flies, represent one of the most species-rich dipteran families, boasting approximately 4,500 known species globally. Their larvae serve as vital decomposers across diverse ecosystems, prompting their frequent and close observation in biomonitoring programs. However, traditional morphological identification methods are laborious and time-consuming, underscoring the need for a comprehensive DNA barcode reference library to speed up species determination. In this study, we present the outcomes of the German Barcode of Life initiative focused on Tipulidae. Our DNA barcode library comprises 824 high-quality cytochrome c oxidase I (COI) barcodes encompassing 76 crane fly species, counting for ca. 54% of the German tipulid fauna. Our results significantly increased the number of European tipulid species available in the Barcode of Life Data System (BOLD) by 14%. Additionally, the number of barcodes from European tipulid specimens more than doubled, with an increase of 118%, bolstering the DNA resource for future identification inquiries. Employing diverse species delimitation algorithms - including the multi-rate Poisson tree processes model (mPTP), Barcode Index Number assignments (BIN), Assemble Species by Automatic Partitioning (ASAP), and the TaxCI R-script - we successfully match 76-86% of the morphologically identified species. Further validation through neighbor-joining tree topology analysis and comparison with 712 additional European tipulid barcodes yield a remarkable 89% success rate for the species identification of German tipulids based on COI barcodes. This comprehensive DNA barcode dataset not only enhances species identification accuracy but also serves as a pivotal resource for ecological and biomonitoring studies, fostering a deeper understanding of crane fly diversity and distribution across terrestrial landscapes.

Development of Cost-Effective SNP Markers for Genetic Variation Analysis and Variety Identification in Cultivated Pears (Pyrus spp.).

Pear (Pyrus spp.) is a major fruit crop in the Rosaceae family, and extensive efforts have been undertaken to develop elite varieties. With advances in genome sequencing technologies, single-nucleotide polymorphisms (SNPs) are commonly used as DNA markers in crop species. In this study, a large-scale discovery of SNPs was conducted using genotyping by sequencing in a collection of 48 cultivated pear accessions. A total of 256,538 confident SNPs were found on 17 chromosomes, and 288 SNPs were filtered based on polymorphic information content, heterozygosity rate, and genome distribution. This subset of SNPs was used to genotype an additional 144 accessions, consisting of P. pyrifolia (53), P. ussuriensis (27), P. bretschneideri (19), P. communis (26), interspecific hybrids (14), and others (5). The 232 SNPs with reliable polymorphisms revealed genetic variations between and within species in the 192 pear accessions. The Asian species (P. pyrifolia, P. ussuriensis, and P. bretschneideri) and interspecific hybrids were genetically differentiated from the European species (P. communis). Furthermore, the P. pyrifolia population showed higher genetic diversity relative to the other populations. The 232 SNPs and four subsets (192, 96, 48, and 24 SNPs) were assessed for variety identification. The 192 SNP subset identified 173 (90.1%) of 192 accessions, which was comparable to 175 (91.1%) from the 232 SNPs. The other three subsets showed 81.8% (24 SNPs) to 87.5% (96 SNPs) identification rates. The resulting SNPs will be a useful resource to investigate genetic variations and develop an efficient DNA barcoding system for variety identification in cultivated pears.