Genetic markers (DNA barcodes) are often used to support and confirm species identification. Barcode sequences can be generated in the field using portable systems based on the Oxford Nanopore Technologies (ONT) MinION sequencer. However, to achieve a broader application, current proof-of-principle workflows for on-site barcoding analysis must be standardized to ensure a reliable and robust performance under suboptimal field conditions without increasing costs. Here, we demonstrate the implementation of a new on-site workflow for DNA extraction, PCR-based barcoding, and the generation of consensus sequences. The portable laboratory features inexpensive instruments that can be carried as hand luggage and uses standard molecular biology protocols and reagents that tolerate adverse environmental conditions. Barcodes are sequenced using MinION technology and analyzed with ONTrack, an original de novo assembly pipeline that requires as few as 1000 reads per sample. ONTrack-derived consensus barcodes have a high accuracy, ranging from 99.8 to 100%, despite the presence of homopolymer runs. The ONTrack pipeline has a user-friendly interface and returns consensus sequences in minutes. The remarkable accuracy and low computational demand of the ONTrack pipeline, together with the inexpensive equipment and simple protocols, make the proposed workflow particularly suitable for tracking species under field conditions.

Urbanization transforms environments in ways that alter biological evolution. We examined whether urban environmental change drives parallel evolution by sampling 110,019 white clover plants from 6169 populations in 160 cities globally. Plants were assayed for a Mendelian antiherbivore defense that also affects tolerance to abiotic stressors. Urban-rural gradients were associated with the evolution of clines in defense in 47% of cities throughout the world. Variation in the strength of clines was explained by environmental changes in drought stress and vegetation cover that varied among cities. Sequencing 2074 genomes from 26 cities revealed that the evolution of urban-rural clines was best explained by adaptive evolution, but the degree of parallel adaptation varied among cities. Our results demonstrate that urbanization leads to adaptation at a global scale.

BackgroundWe coin the term "taxon expeditions" for citizen scientists' field courses to carry out publishable taxonomic work in close association with trained taxonomists.New informationDuring the first-ever taxon expedition, in Maliau Basin Studies Centre, Sabah, Malaysian Borneo, the participants sampled leaf litter beetles from lowland dipterocarp forest using the Winkler apparatus. The collected material proved to contain at least three undescribed species of small-bodied (ca. 1 mm long) hemispherical litter-dwelling Coleoptera. As part of the field course work, taxonomic descriptions were prepared for the chrysomelid Clavicornaltica sabahensis sp. n. and the leiodids Colenisia chungi sp. n. and Dermatohomoeus maliauensis sp. n.

The genus Anthroherpon Reitter, 1889 exhibits the most pronounced troglomorphic characters among Coleoptera, and represents one of the most spectacular radiations of subterranean beetles. However, radiation, diversification, and biogeography of this genus have never been studied in a phylogenetic context. This study provides a comprehensive evolutionary analysis of the Anthroherpon radiation, using a dated molecular phylogeny as a framework for understanding Anthroherpon diversification, reconstructing the ancestral range, and exploring troglomorphic diversity. Based on 16 species and 22 subspecies, i.e. the majority of Anthroherpon diversity, we reconstructed the phylogeny using Bayesian analysis of six loci, both mitochondrial and nuclear, comprising a total of 4143 nucleotides. In parallel, a morphometric analysis was carried out with 79 landmarks on the body that were subjected to geometric morphometrics. We optimized morphometric features to phylogeny, in order to recognize the way troglomorphy was expressed in different clades of the tree, and did character evolution analyses. Finally, we reconstructed the ancestral range of the genus using BioGeoBEARS. Besides further elucidating the suprageneric classification of the East-Mediterranean Leptodirini, our main findings also show that Anthroherpon dates back to the Early Miocene (ca. 22 MYA) and that the genus diversified entirely underground. Biogeographic reconstruction of the ancestral range shows the origin of the genus in the area comprising three high mountains in western Montenegro, which is in the accordance with the available data on the paleogeography of the Balkan Peninsula. Character evolution analysis indicates that troglomorphic morphometric traits in Anthroherpon mostly evolve neutrally but may diverge adaptively under syntopic competition.

Further results are presented of the first field course at Maliau Basin, Malaysian Borneo organized by Taxon Expeditions, an organization which enables citizen scientists to be directly involved in taxonomic discoveries. Three new species of the aquatic beetle genus Grouvellinus Champion, 1923, namely G. leonardodicaprioi sp. n., G. andrekuipersi sp. n., and G. quest sp. n. were collected jointly by the citizen scientists and taxonomists during the fieldwork in Maliau Basin. Material was mainly sampled from sandstone bottom rocks of blackwater streams at altitudes between 900 m and 1,000 m using fine-meshed hand-nets. The genus is widely distributed in the Oriental and Palearctic regions, but these are the first records from the island of Borneo.

The parasitic wasp genus Aphaereta Foerster, 1863, belongs to the large subfamily Alysiinae Leach, 1815 (Hymenoptera: Braconidae) and contains solitary or gregarious endoparasitoids of larvae of cyclorrhaphous Diptera living in decaying organic matter. A new species, Aphaereta vondelparkensis sp. n. (Braconidae, Alysiinae, Alysiini), from a city park in the centre of Amsterdam (Vondelpark) is described and illustrated. The species was discovered on a so-called taxon expedition, in which experts and citizen scientists collaborate on taxonomic projects. The finding highlights the fact that new animal species can still be discovered today, even in the busiest park of the capital of one of the best studied European countries.

Long-palped Water Beetles were collected during a taxon expedition in Montenegro which involved citizen scientists, students and taxonomists. The material was collected from springs, brooks, fens and the Tara River, at altitudes between 600 m and 1450 m above sea level, using fine-meshed hand-nets and by manual checking of submerged substrates. The morphological species delimitation was supplemented and congruent with mtDNA sequences mainly obtained in the field using the newly-developed MinION-based ONTrack pipeline. The new species Hydraena dinarica Freitag & de Vries, sp. n. from Durmitor Mt. is described, illustrated and compared in detail to closely-related congeners of the H. saga d'Orchymont, 1930/H. emarginata Rey, 1885 species complex. Five additional species and female specimens of two unidentified morphospecies of the genus were also recorded in the vicinity of Durmitor National Park. New records and the first DNA barcodes for Hydraena biltoni Jäch & Díaz, 2012 (endemic to Montenegro) and H. morio Kiesenwetter, 1849 are provided. Further records of H. nigrita Germar, 1824, H. minutissima Stephens, 1829, H. subintegra Ganglbauer, 1901 and females of two unidentified morphospecies are commented upon. The resulting inter- and intraspecific genetic distances and some observations of low or zero sequence divergence between recently-diverged species of Hydraena Kugelann, 1794 are briefly discussed.

Two new species of Anthroherpon belonging to the "ganglbaueri" species group are described from northern Montenegro: Anthroherpon sinjajevina n. sp. and Anthroherpon cecai n. sp. Based on morphological investigations of other species of this group, we elevate A. udrzali Giachino & Vailati from subspecies to species rank, and suggest the synonymy of A. brckoensis Giachino & Guéorguiev with A ganglbaueri alticola Knirsch. An identification key of the ganglbaueri species group (sensu Guéorguiev, 1990) is given and the distributions of species and subspecies are mapped and discussed.