Metazoa-level USCOs as markers in species delimitation and classification.

Metazoa-level universal single-copy orthologs (mzl-USCOs) are universally applicable markers for DNA taxonomy in animals that can replace or supplement single-gene barcodes. Previously, mzl-USCOs from target enrichment data were shown to reliably distinguish species. Here, we tested whether USCOs are an evenly distributed, representative sample of a given metazoan genome and therefore able to cope with past hybridization events and incomplete lineage sorting. This is relevant for coalescent-based species delimitation approaches, which critically depend on the assumption that the investigated loci do not exhibit autocorrelation due to physical linkage. Based on 239 chromosome-level assembled genomes, we confirmed that mzl-USCOs are genetically unlinked for practical purposes and a representative sample of a genome in terms of reciprocal distances between USCOs on a chromosome and of distribution across chromosomes. We tested the suitability of mzl-USCOs extracted from genomes for species delimitation and phylogeny in four case studies: Anopheles mosquitos, Drosophila fruit flies, Heliconius butterflies and Darwin's finches. In almost all instances, USCOs allowed delineating species and yielded phylogenies that corresponded to those generated from whole genome data. Our phylogenetic analyses demonstrate that USCOs may complement single-gene DNA barcodes and provide more accurate taxonomic inferences. Combining USCOs from sources that used different versions of ortholog reference libraries to infer marker orthology may be challenging and, at times, impact taxonomic conclusions. However, we expect this problem to become less severe as the rapidly growing number of reference genomes provides a better representation of the number and diversity of organismal lineages.

Local stochastics and ecoclimatic situation shape phytophagous chafer assemblage composition.

Very little is known about factors determining the assemblage structure of megadiverse polyphagous-herbivore scarab chafers in the tropics (Coleoptera: Scarabaeidae). Here, we examined the composition of Sri Lankan chafer assemblages and investigated whether it is influenced more by the general ecoclimatic situation, macrohabitat, or indetermined stochastic biotic and abiotic factors of each locality. We also explored the influence of the latter on separate lineages and general body size. Based on dedicated field surveys conducted during the dry and wet seasons, we examined 4847 chafer individuals of 105 species sampled using multiple UV-light traps in 11 localities covering different forest types and altitudinal zones. Assemblages were assessed for compositional similarity, species diversity, and abundance within four major eco-spatial partitions: forest types, elevational zones, localities, and macrohabitats. Our results revealed that assemblages were shaped mainly by locality stochastics (i.e., multi-factor ensemble of all biotic and abiotic environmental conditions at local scale), and to a minor extent by ecoclimatic conditions. Macrohabitat had little effect on the assemblage composition. This was true for the entire chafer assemblage as well as for all single lineages or different body size classes. However, in medium and large species the contrasts between localities were less pronounced, which was not the case for individual lineages of the assemblage. Contrasts of assemblage similarity between localities were much more evident than those for forest types and elevation zones. Significant correlation between species composition and geographic distance was found only for the assemblage of small-bodied specimens. Seasonal change (dry-wet) in species composition was minor and only measurable in a few localities. The strong turnover between examined localities corroborates with the high degree of endemism in many phytophagous chafers, particularly in Sericini. Connected with their hypothetic poor habitat specificity and polyphagy, this might also explain why so many chafer crop pests in the Asian tropics are endemics.

From DNA barcodes to ecology: Meta-analysis of central European beetles reveal link with species ecology but also to data pattern and gaps.

DNA barcoding has been used worldwide to identify biological specimens and to delimit species. It represents a cost-effective, fast, and efficient way to assess biodiversity with help of the public Barcode of Life Database (BOLD) accounting for more than 236,000 animal species and more than 10 million barcode sequences. Here, we performed a meta-analysis of available barcode data of central European Coleoptera to detect intraspecific genetic patterns among ecological groups in relation to geographic distance with the aim to investigate a possible link between infraspecific variation and species ecology. We collected information regarding feeding style, body size, as well as habitat and biotope preferences. Mantel tests and two variants of Procrustes analysis, both involving the Principal Coordinates Neighborhood Matrices (PCNM) approach, were applied on genetic and geographic distance matrices. However, significance levels were too low to further use the outcome for further trait investigation: these were in mean for all ecological guilds only 7.5, 9.4, or 15.6% for PCNM + PCA, NMDS + PCA, and Mantel test, respectively, or at best 28% for a single guild. Our study confirmed that certain ecological traits were associated with higher species diversity and foster stronger genetic differentiation. Results suggest that increased numbers of species, sampling localities, and specimens for a chosen area of interest may give new insights to explore barcode data and species ecology for the scope of conservation on a larger scale. We performed a meta-analysis of available barcode data of central European beetles to detect intraspecific genetic patterns among ecological groups in relation to geographic distance, regarding feeding style, body size, as well as habitat and biotope preferences. Our study confirmed that certain ecological traits were associated with higher species diversity and foster stronger genetic differentiation. However, significance levels were too low to further use the outcome for further trait investigation.

Multiple species delimitation approaches with COI barcodes poorly fit each other and morphospecies - An integrative taxonomy case of Sri Lankan Sericini chafers (Coleoptera: Scarabaeidae).

DNA taxonomy including barcoding and metabarcoding is widely used to explore the diversity in biodiversity hotspots. In most of these hotspot areas, chafers are represented by a multitude of species, which are well defined by the complex shape of male genitalia. Here, we explore how well COI barcode data reflect morphological species entities and thus their usability for accelerated species inventorization. We conducted dedicated field surveys in Sri Lanka to collect the species-rich and highly endemic Sericini chafers (Coleoptera: Scarabaeidae). Congruence among results of a series of protocols for de novo species delimitation and with morphology-based species identifications was investigated. Different delimitation methods, such as the Poisson tree processes (PTP) model, Statistical Parsimony Analysis (TCS), Automatic Barcode Gap Discovery (ABGD), Assemble Species by Automatic Partitioning (ASAP), and Barcode Index Number (BIN) assignments, resulted in different numbers of molecular operational taxonomic units (MOTUs). All methods showed both over-splitting and lumping of morphologically identified species. Only 18 of the observed 45 morphospecies perfectly matched MOTUs from all methods. The congruence of delimitation between MOTUs and morphospecies expressed by the match ratio was low, ranging from 0.57 to 0.67. TCS and multirate PTP (mPTP) showed the highest match ratio, while (BIN) assignment resulted in the lowest match ratio and most splitting events. mPTP lumped more species than any other method. Principal coordinate analysis (PCoA) on a match ratio-based distance matrix revealed incongruent outcomes of multiple DNA delimitation methods, although applied to the same data. Our results confirm that COI barcode data alone are unlikely to correctly delimit all species, in particular, when using only a single delimitation approach. We encourage the integration of various approaches and data, particularly morphology, to validate species boundaries.

Dietary preference of cheetahs (Acinonyx jubatus) in south-eastern Kenya.

The conversion of natural ecosystems due to anthropogenic activities has led to the destruction of natural habitats and to the deterioration of habitat quality. Top predators particularly respond sensitively to changes in habitat structures, including the availability of prey. The cheetah Acinonyx jubatus prefers small-medium-sized, wild ungulate prey due to the cheetah's morphological adaptations. However, the majority of the species' population is found beyond protected areas, where habitat structures, species abundances, and community composition are highly influenced by human activities. Only few studies have analyzed the diet preference of cheetahs in relation to prey availability and abundance for rangelands beyond protected areas in Eastern Africa. The study aimed to determine cheetah prey preference in the rangelands of south-eastern Kenya based on scat analyses. We compared dietary preference of cheetah with prey availability. For this purpose, we conducted standardized game counts. We analyzed 27 cheetah scat samples collected across the same study area where we also conducted game counts. We found that Grant's gazelle Gazella granti contributed the highest portion of cheetah's diet, although Thomson's gazelle Gazella thomsonii was the most abundant medium-sized ungulate prey in the study areas. We also recorded two primate species, yellow baboon Papio cynocephalus and vervet monkey Chlorocebus pygerythrus, as well as the rock hyrax Procavia capensis in the cheetah diet. These species have never been documented as cheetah prey before. Furthermore, our results document livestock as potential prey for cheetahs. These observations underline that cheetah use diverse prey in rangelands outside protected areas, and that the abundance of specific prey does not influence cheetah prey preference.

Image-Based Automated Species Identification: Can Virtual Data Augmentation Overcome Problems of Insufficient Sampling?

Automated species identification and delimitation is challenging, particularly in rare and thus often scarcely sampled species, which do not allow sufficient discrimination of infraspecific versus interspecific variation. Typical problems arising from either low or exaggerated interspecific morphological differentiation are best met by automated methods of machine learning that learn efficient and effective species identification from training samples. However, limited infraspecific sampling remains a key challenge also in machine learning. In this study, we assessed whether a data augmentation approach may help to overcome the problem of scarce training data in automated visual species identification. The stepwise augmentation of data comprised image rotation as well as visual and virtual augmentation. The visual data augmentation applies classic approaches of data augmentation and generation of artificial images using a generative adversarial networks approach. Descriptive feature vectors are derived from bottleneck features of a VGG-16 convolutional neural network that are then stepwise reduced in dimensionality using Global Average Pooling and principal component analysis to prevent overfitting. Finally, data augmentation employs synthetic additional sampling in feature space by an oversampling algorithm in vector space. Applied on four different image data sets, which include scarab beetle genitalia (Pleophylla, Schizonycha) as well as wing patterns of bees (Osmia) and cattleheart butterflies (Parides), our augmentation approach outperformed a deep learning baseline approach by means of resulting identification accuracy with nonaugmented data as well as a traditional 2D morphometric approach (Procrustes analysis of scarab beetle genitalia). [Deep learning; image-based species identification; generative adversarial networks; limited infraspecific sampling; synthetic oversampling.].

Is it time to describe new species without diagnoses?A comment on Sharkey et al. (2021).

New methods in taxonomy and systematics can influence the overall practice of formally naming and describing biodiversity. DNA barcoding has been controversial since its emergence, but now, large scale species descriptions exclusively based on barcodes have created what can be called a 'new quality of performance. Its limitations are discussed from different perspectives: nomenclature, general pragmatism, and problems of DNA-based species delimitation in the light of the central aim of achieving a robust and stable nomenclature of organisms, essential for all applications of biodiversity research. This issue needs to be addressed to prevent restraining the progress of taxonomy and its ability to contribute to modern science.

Excluding spatial sampling bias does not eliminate oversplitting in DNA-based species delimitation analyses.

DNA barcoding and DNA-based species delimitation are major tools in DNA taxonomy. Sampling has been a central debate in this context, because the geographical composition of samples affects the accuracy and performance of DNA barcoding. Performance of complex DNA-based species delimitation is to be tested under simpler conditions in absence of geographic sampling bias. Here, we present an empirical dataset sampled from a single locality in a Southeast-Asian biodiversity hotspot (Laos: Phou Pan mountain). We investigate the performance of various species delimitation approaches on a megadiverse assemblage of herbivorous chafer beetles (Coleoptera: Scarabaeidae) to infer whether species delimitation suffers in the same way from exaggerate infraspecific variation despite the lack of geographic genetic variation that led to inconsistencies between entities from DNA-based and morphology-based species inference in previous studies. For this purpose, a 658 bp fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) was analyzed for a total of 186 individuals of 56 morphospecies. Tree-based and distance-based species delimitation methods were used. All approaches showed a rather limited match ratio (max. 77%) with morphospecies. Poisson tree process (PTP) and statistical parsimony network analysis (TCS) prevailingly over-splitted morphospecies, while 3% clustering and Automatic Barcode Gap Discovery (ABGD) also lumped several species into one entity. ABGD revealed the highest congruence between molecular operational taxonomic units (MOTUs) and morphospecies. Disagreements between morphospecies and MOTUs have to be explained by historically acquired geographic genetic differentiation, incomplete lineage sorting, and hybridization. The study once again highlights how important morphology still is in order to correctly interpret the results of molecular species delimitation.

A Plea for Standardized Nuclear Markers in Metazoan DNA Taxonomy.

The ease of sequencing DNA barcodes promoted a species identification system universally applicable across animal phyla. However, relying on a single mitochondrial DNA fragment has a number of drawbacks that can mislead species delimitation and identification. Implementation of multiple nuclear markers would mitigate the limits of the current barcoding system if these markers are universally applicable across species, carry sufficient information to discriminate between closely related species, and if sequencing and analyzing these markers can be automatized. As sequencing costs continue to fall, we believe that the time is right to extend DNA barcoding. Here we argue that nearly universal single-copy nuclear protein-coding genes deliver the desired characteristics and could be used to reliably delimit and identify animal species.

Sex-Biased Dispersal Obscures Species Boundaries in Integrative Species Delimitation Approaches.

Accurate delimitation of species is crucial for a stable taxonomy, which provides the foundation for the study of evolutionary biology, ecology, and essentially all biological disciplines. Several approaches toward impartial and repeatable taxonomic practices are available but all existing methods have potentially unacceptable shortcomings. In particular, problems can arise when the underlying model assumptions are violated, for instance, in the presence of reduced gene flow. This is observed in the context of sex-biased dispersal, which is a common but underappreciated feature in many groups of organisms. Previously, simulations have indicated that sex-biased dispersal may lead to erroneous estimations of the true species numbers. However, this phenomenon has never been examined using empirical data. We evaluate the bias introduced by extreme female philopatry on a range of de novo [GMYC, PTP, ABGD, statistical parsimony, trinomial distribution of triplets model (tr2)] and validation (STACEY, iBPP) approaches to species delimitation in the scarab beetle genus Pachypus. Since female philopatry exhibited in this genus in particular can affect mitochondrial gene flow, we compared the results from analyses of single loci, mitochondrial loci, nuclear loci and combined data, as well as the performance of morphometric data as a secondary data source in a fully integrative Bayesian framework. Large overestimation of species numbers was observed across all analyses of combined and mitochondrial DNA data sets, suggesting specimens from nearly every sampling location as separate species. The use of nuclear data resulted in more reasonable estimations of species boundaries, which were largely supported by morphometrics of linear measurements, while geometric morphometrics of body outlines resulted in stronger splitting. Simulations of population divergence with migration, corresponding to the biology of Pachypus, showed that female philopatry strongly increases reciprocal monophyly of mitochondrial markers and may substantially contribute to over-splitting in species delimitation. Robust results recovered using nuclear DNA and morphological data nevertheless enabled us to reach novel conclusions about species boundaries in Pachypus. Our findings suggest that mitochondrial DNA will be less suited to species delimitation in many cases, in particular in the presence of sex-biased dispersal.

The phylogeny of pholcid spiders: a critical evaluation of relationships suggested by molecular data (Araneae, Pholcidae).

With almost 600 species, the latest molecular phylogeny of pholcid spiders (Eberle et al. 2018, BMC Evolutionary Biology) more than triples the largest previously available molecular phylogeny of the family. At the level of genera, the coverage is high (86%, i.e., 75 of the 87 named genera), and at the level of subfamilies it is complete. The present paper is an effort to critically evaluate the implications of this phylogeny for pholcid systematics. The analyses largely support the division of Pholcidae into five subfamilies: Ninetinae, Arteminae, Modisiminae, Smeringopinae, and Pholcinae. Their compositions are largely unchanged except that Chisosa Huber, 2000 is moved from Ninetinae to Arteminae. The positions of Artema Walckenaer, 1837 and Priscula Simon, 1893 in this system remain dubious. Relationships among subfamilies remain weakly supported, except for the sister group relationship between Smeringopinae and Pholcinae. Several major clades within subfamilies are separated from each other along geographical boundaries; for example within Modisiminae a South American clade and a Central + North American + Caribbean clade, and within Smeringopinae a Sub-Saharan clade and a clade ranging from the Mediterranean to Central Asia. Central + North American + Caribbean clades in both Ninetinae and Modisiminae may originate from South American ancestors. Many taxonomic changes are suggested by the data, some of which are formally implemented herein. Two new genera result from the splitting of Calapnita Simon, 1892 and Panjange Deeleman-Reinhold & Deeleman, 1983, respectively: Nipisa Huber, gen. n.; and Apokayana Huber, gen. n. Nine new genera result from splitting of Pholcus: Cantikus Huber, gen. n.; Kelabita Huber, gen. n.; Kintaqa Huber, gen. n.; Muruta Huber, gen. n.; Meraha Huber, gen. n.; Paiwana Huber, gen. n.; Pribumia Huber, gen. n.; Teranga Huber, gen. n.; and Tissahamia Huber, gen. n. Two genera are newly synonymized: Platnicknia Özdikmen & Demir, 2009 is synonymized with Modisimus Simon, 1893; Sihala Huber, 2011 is synonymized with Pholcus Walckenaer, 1805. Pholcusagadir Huber, 2011 is moved to Micropholcus Deeleman-Reinhold & Prinsen, 1987, resulting in the new combination Micropholcusagadir (Huber, 2011).

Microhabitat change drives diversification in pholcid spiders.

BACKGROUND: Microhabitat changes are thought to be among the main drivers of diversification. However, this conclusion is mostly based on studies on vertebrates. Here, we investigate the influence of microhabitat on diversification rates in pholcid spiders (Araneae, Pholcidae). Diversification analyses were conducted in the framework of the largest molecular phylogeny of pholcid spiders to date based on three nuclear and three mitochondrial loci from 600 species representing more than 85% of the currently described pholcid genera. RESULTS: Assessments of ancestral microhabitat revealed frequent evolutionary change. In particular, within the largest subfamily Pholcinae, numerous changes from near-ground habitats towards leaves and back were found. In general, taxa occupying leaves and large sheltered spaces had higher diversification rates than ground-dwelling taxa. Shifts in speciation rate were found in leaf- and space-dwelling taxa. CONCLUSIONS: Our analyses result in one of the most comprehensive phylogenies available for a major spider family and provide a framework for any subsequent studies of pholcid spider biology. Diversification analyses strongly suggest that microhabitat is an important factor influencing diversification patterns in pholcid spiders.

Landscape genetics indicate recently increased habitat fragmentation in African forest-associated chafers.

Today, indigenous forests cover less than 0.6% of South Africa's land surface and are highly fragmented. Most forest relicts are very small and typically occur in fire-protected gorges along the eastern Great Escarpment. Yet, they hold a unique and valuable fauna with high endemism and ancient phylogenetic lineages, fostered by long-term climatic stability and complex microclimates. Despite numerous studies on southern African vegetation cover, the current state of knowledge about the natural extension of indigenous forests is rather fragmentary. We use an integrated approach of population-level phylogeography and climatic niche modeling of forest-associated chafer species to assess connectivity and extent of forest habitats since the last glacial maximum. Current and past species distribution models ascertained potential fluctuations of forest distribution and supported a much wider potential current extension of forests based on climatic data. Considerable genetic admixture of mitochondrial and nuclear DNA among many populations and an increase in mean population mutation rate in Extended Bayesian Skyline Plots of all species indicated more extended or better connected forests in the recent past (<5 kya). Genetic isolation of certain populations, as revealed by population differentiation statistics (GST'), as well as landscape connectivity statistics and habitat succession scenarios suggests considerable loss of habitat connectivity. As major anthropogenic influence is likely, conservational actions need to be considered.

A historical biogeography of megadiverse Sericini-another story "out of Africa"?

Megadiverse insect groups present special difficulties for biogeographers because poor classification, incomplete knowledge of taxonomy, and many undescribed species can introduce a priori sampling bias to any analysis. The historical biogeography of Sericini, a tribe of melolonthine scarabs comprising about 4000 species, was investigated using the most comprehensive and time-calibrated molecular phylogeny available today. Problems arising through nomenclatural confusion were overcome by extensive sampling (665 species) from all major lineages of the tribe. A West Gondwanan origin of Sericini (c. 112 Ma) was reconstructed using maximum parsimony, maximum-likelihood and model-based ancestral area estimation. Vicariance in the tribe's earliest history separated Neotropical and Old World Sericini, whereas subsequent lower Cretaceous biogeography of the tribe was characterized by repeated migrations out of Africa, resulting in the colonization of Eurasia and Madagascar. North America was colonized from Asia during the Cenozoic and a lineage of "Modern Sericini" reinvaded Africa. Diversification dynamics revealed three independent shifts to increased speciation rates: in African ant-adapted Trochalus, Oriental Tetraserica, and Asian and African Sericina. Southern Africa is proposed as both cradle and refuge of Sericini. This area has retained many old lineages that portray the evolution of the African Sericini fauna as a series of taxon pulses.

A molecular phylogeny of rose chafers (Coleoptera: Scarabaeidae: Cetoniinae) reveals a complex and concerted morphological evolution related to their flight mode.

Rose chafers (Cetoniinae) are a large group of flower visitors within the pleurostict Scarabaeidae that are characterized by their distinctive flight mode with nearly closed forewings. Despite their popularity, this is the first study to use molecular data to infer their phylogenetic relationships. We used partial gene sequences for 28S rRNA, cytochrome oxidase I (cox1) and 16S rRNA (rrnL) for 299 species, representing most recognized subfamilies of Scarabaeidae, including 125 species of Cetoniinae. Combined analyses using maximum parsimony, maximum likelihood and Bayesian inferences recovered Cetoniinae as monophyletic in all analyses, with the sister clade composed of Rutelinae and Dynastinae. Rutelinae was always recovered as paraphyletic with respect to Dynastinae. Trichiini sensu lato (s.l.) was recovered as a polyphyletic clade, while Cetoniini s.l. was recovered as paraphyletic. The inferred topologies were also supported by site bootstrapping of the ML trees. With the exception of Cremastochelini, most tribes of Cetoniinae were poly- or paraphyletic, indicating the critical need for a careful revision of rose chafer classification. Analysis of elytral base structure (including 11 scored characters) in the context of phylogeny, revealed a complex, concerted and rapid transformation of the single trait elements linked to a modified flight mode with closed elytra. This appears to be unlinked to the lateral sinuation of the elytra, which originated independently several times at later stages in the evolution of the group.

Bayesian species delimitation in Pleophylla chafers (Coleoptera) - the importance of prior choice and morphology.

BACKGROUND: Defining species units can be challenging, especially during the earliest stages of speciation, when phylogenetic inference and delimitation methods may be compromised by incomplete lineage sorting (ILS) or secondary gene flow. Integrative approaches to taxonomy, which combine molecular and morphological evidence, have the potential to be valuable in such cases. In this study we investigated the South African scarab beetle genus Pleophylla using data collected from 110 individuals of eight putative morphospecies. The dataset included four molecular markers (cox1, 16S, rrnL, ITS1) and morphometric data based on male genital morphology. We applied a suite of molecular and morphological approaches to species delimitation, and implemented a novel Bayesian approach in the software iBPP, which enables continuous morphological trait and molecular data to be combined. RESULTS: Traditional morphology-based species assignments were supported quantitatively by morphometric analyses of the male genitalia (eigenshape analysis, CVA, LDA). While the ITS1-based delineation was also broadly congruent with the morphospecies, the cox1 data resulted in over-splitting (GMYC modelling, haplotype networks, PTP, ABGD). In the most extreme case morphospecies shared identical haplotypes, which may be attributable to ILS based on statistical tests performed using the software JML. We found the strongest support for putative morphospecies based on phylogenetic evidence using the combined approach implemented in iBPP. However, support for putative species was sensitive to the use of alternative guide trees and alternative combinations of priors on the population size (θ) and rootage (τ 0 ) parameters, especially when the analysis was based on molecular or morphological data alone. CONCLUSIONS: We demonstrate that continuous morphological trait data can be extremely valuable in assessing competing hypotheses to species delimitation. In particular, we show that the inclusion of morphological data in an integrative Bayesian framework can improve the resolution of inferred species units. However, we also demonstrate that this approach is extremely sensitive to guide tree and prior parameter choice. These parameters should be chosen with caution - if possible - based on independent empirical evidence, or careful sensitivity analyses should be performed to assess the robustness of results. Young species provide exemplars for investigating the mechanisms of speciation and for assessing the performance of tools used to delimit species on the basis of molecular and/or morphological evidence.

Rarity and Incomplete Sampling in DNA-Based Species Delimitation.

DNA-based species delimitation may be compromised by limited sampling effort and species rarity, including "singleton" representatives of species, which hampers estimates of intra- versus interspecies evolutionary processes. In a case study of southern African chafers (beetles in the family Scarabaeidae), many species and subclades were poorly represented and 48.5% of species were singletons. Using cox1 sequences from >500 specimens and ∼100 species, the Generalized Mixed Yule Coalescent (GMYC) analysis as well as various other approaches for DNA-based species delimitation (Automatic Barcode Gap Discovery (ABGD), Poisson tree processes (PTP), Species Identifier, Statistical Parsimony), frequently produced poor results if analyzing a narrow target group only, but the performance improved when several subclades were combined. Hence, low sampling may be compensated for by "clade addition" of lineages outside of the focal group. Similar findings were obtained in reanalysis of published data sets of taxonomically poorly known species assemblages of insects from Madagascar. The low performance of undersampled trees is not due to high proportions of singletons per se, as shown in simulations (with 13%, 40% and 52% singletons). However, the GMYC method was highly sensitive to variable effective population size ([Formula: see text]), which was exacerbated by variable species abundances in the simulations. Hence, low sampling success and rarity of species affect the power of the GMYC method only if they reflect great differences in [Formula: see text] among species. Potential negative effects of skewed species abundances and prevalence of singletons are ultimately an issue about the variation in [Formula: see text] and the degree to which this is correlated with the census population size and sampling success. Clade addition beyond a limited study group can overcome poor sampling for the GMYC method in particular under variable [Formula: see text] This effect was less pronounced for methods of species delimitation not based on coalescent models.

Gross changes in reconstructions of historic land cover/use for Europe between 1900 and 2010.

Historic land-cover/use change is important for studies on climate change, soil carbon, and biodiversity assessments. Available reconstructions focus on the net area difference between two time steps (net changes) instead of accounting for all area gains and losses (gross changes). This leads to a serious underestimation of land-cover/use dynamics with impacts on the biogeochemical and environmental assessments based on these reconstructions. In this study, we quantified to what extent land-cover/use reconstructions underestimate land-cover/use changes in Europe for the 1900-2010 period by accounting for net changes only. We empirically analyzed available historic land-change data, quantified their uncertainty, corrected for spatial-temporal effects and identified underlying processes causing differences between gross and net changes. Gross changes varied for different land classes (largest for forest and grassland) and led to two to four times the amount of net changes. We applied the empirical results of gross change quantities in a spatially explicit reconstruction of historic land change to reconstruct gross changes for the EU27 plus Switzerland at 1 km spatial resolution between 1950 and 2010. In addition, the reconstruction was extended back to 1900 to explore the effects of accounting for gross changes on longer time scales. We created a land-change reconstruction that only accounted for net changes for comparison. Our two model outputs were compared with five commonly used global reconstructions for the same period and area. In our reconstruction, gross changes led in total to a 56% area change (ca. 0.5% yr(-1)) between 1900 and 2010 and cover twice the area of net changes. All global reconstructions used for comparison estimated fewer changes than our gross change reconstruction. Main land-change processes were cropland/grassland dynamics and afforestation, and also deforestation and urbanization.

The evolution of morphospace in phytophagous scarab chafers: no competition--no divergence?

Body shape reflects species' evolution and mediates its role in the environment as it integrates gene expression, life style, and structural morphology. Its comparative analysis may reveal insight on what shapes shape, being a useful approach when other evidence is lacking. Here we investigated evolutionary patterns of body shape in the highly diverse phytophagous chafers (Scarabaeidae: Pleurosticti), a polyphagous group utilizing different parts of angiosperms. Because the reasons of their successful diversification are largely unknown, we used a phylogenetic tree and multivariate analysis on twenty linear measurements of body morphology including all major Pleurosticti lineages to infer patterns of morphospace covariation and divergence. The chafer's different feeding types resulted to be not distinguishable in the described morphospace which was largely attributed to large occupancy of the morphospace of some feeding types and to multiple convergences of feeding behavior (particularly of anthophagy). Low correlation between molecular and morphological rates of evolution, including significant rate shifts for some lineages, indicated directed selection within feeding types. This is supported by morphospace divergence within feeding types and convergent evolution in Australian Melolonthinae. Traits driving morphospace divergence were extremities and traits linked with locomotion behavior, but also body size. Being highly adaptive for burrowing and locomotion these traits showed major changes in the evolution of pleurostict scarabs. These activities also affected another trait, the metacoxal length, which is highly influenced by key innovations of the metacoxa (extended mesal process, secondary closure) particularly in one lineage, the Sericini. Significant shape divergence between major lineages and a lack of strong differentiation among closely related lineages indicated that the question about the presence or absence of competition-derived directed selection needs to be addressed for different time scales. Striking divergence between some sister lineages at their origin revealed strong driven selection towards morphospace divergence, possibly linked with resource partitioning.