A genomic perspective on an old question: Salmo trouts or Salmo trutta (Teleostei: Salmonidae)?

There are particular challenges in defining the taxonomic status of recently radiated groups due to the low level of phylogenetic signal. Members of the Salmo trutta species-complex, which mostly evolved during and following the Pleistocene, show high morphological and ecological diversity that, along with their very wide geographic distribution, have led to morphological description of 47 extant nominal species. However, many of these species have not been supported by previous phylogenetic studies, which could be partly due to lack of significant genetic differences among them, the limited resolution offered by molecular methods previously used, as well as the often local scale of these studies. The development of next-generation sequencing (NGS) and related analytical tools have enhanced our ability to address such challenging questions. In this study, Genotyping-by-Sequencing (GBS) of 15,169 filtered SNPs and mitochondrial DNA (mtDNA) D-loop sequences were combined to assess the phylogenetic relationships among 166 brown trouts representing 21 described species and three undescribed groups collected from 84 localities throughout their natural distribution in Europe, west Asia, and North Africa. The data were analysed using different clustering algorithms (admixture analysis and discriminant analysis of principal components-DAPC), a Bayes Factor Delimitation (BFD) test, species tree reconstruction, gene flow tests (three- and four-population tests), and Rogue taxa identification tests. Genomic contributions of the Atlantic lineage brown trout were found in all major sea basins excluding the North African and Aral Sea basins, suggesting introgressive hybridization of native brown trouts driven by stocking using strains of the Atlantic lineage. After removing the phylogenetic noise caused by the Atlantic brown trout, admixture clusters and DAPC clustering based on GBS data, respectively, resolved 11 and 13 clusters among the previously described brown trout species, which were also supported by BFD test results. Our results suggest that natural hybridization between different brown trout lineages has probably played an important role in the origin of several of the putative species, including S. marmoratus, S. carpio, S. farioides, S. pellegrini, S. caspius (in the Kura River drainage) and Salmo sp. in the Danube River basin. Overall, our results support a multi-species taxonomy for brown trouts. They also resolve some species in the Adriatic-Mediterranean and Black Sea drainages as members of very closely related genomic clusters that may need taxonomic revision. However, any final conclusions pertaining to the taxonomy of the brown trout complex should be based on an integrative approach combining genomic, morphological, and ecological data. To avoid challenges in taxonomy and conservation of species complexes like brown trouts, it is suggested to describe species based on genomic clusters of populations instead of describing species based only on morphologically differentiated single type populations.

Testing the devil's impact on southern Baltic and North Sea basins whitefish (Coregonus spp.) diversity.

BACKGROUND: The diversity and phylogeny of whitefish of the genus Coregonus is complex, and includes many endemic species of high conservation concern. However, because of commercial importance of whitefish fisheries, stockings and translocations have occurred repeatedly, which challenges the identification of local populations as conservation units. This study analyses the phylogenetic relationships of 15 contemporary and two historical populations of lake-resident and anadromous whitefish (Coregonus spp.) from the southern Baltic and North Sea basins. We elucidated the complex history of Lake Schaal (northern Germany) whitefish, for which a local tale suggests that the devil threw whitefish from the Central European Lake Constance into this lake. Studies from the early twentieth century indeed suggested numerous stocking events for Lake Schaal from Lake Constance, from Estonian/Russian Lake Peipsi and from the anadromous whitefish of the Baltic Sea. RESULTS: Analyses of 13 microsatellite markers showed that Lake Constance whitefish are unrelated to any northern Germany whitefish population, including the contemporary whitefish population from Lake Schaal. Comparison with four historical specimens further showed that the native Lake Schaal whitefish (C. holsatus) vanished from the lake, but has survived as a non-native population in the north German Lake Drewitz. The whitefish currently occurring in Lake Schaal and three adjacent lakes are identified as C. maraenoides, introduced from Lake Peipsi. The contemporary anadromous whitefish populations from the Baltic (German and Finnish coast) and the German River Treene (North Sea basin, stocked from Danish River Vida) grouped together, but showed significant genetic differentiation. The 14 historical specimens of C. oxyrinchus from Rivers Rhine and Schelde were assigned to several contemporary whitefish populations, but among them only one specimen was assigned to the contemporary River Treene population. Therefore, we do not support the view that the whitefish from River Vida/Treene are identical with the historical C. oxyrinchus. CONCLUSIONS: Our study demonstrates that lake and anadromous whitefish in the Baltic and North Sea basins reflect a complex phylogeography, which is further blurred by the effects of repeated stocking and translocations. To identify conservation units, the genetic identity of each population has to be scrutinized.

Freshwater Megafauna: Flagships for Freshwater Biodiversity under Threat.

Freshwater biodiversity is highly threatened and is decreasing more rapidly than its terrestrial or marine counterparts; however, freshwaters receive less attention and conservation investment than other ecosystems do. The diverse group of freshwater megafauna, including iconic species such as sturgeons, river dolphins, and turtles, could, if promoted, provide a valuable tool to raise awareness and funding for conservation. We found that freshwater megafauna inhabit every continent except Antarctica, with South America, Central Africa, and South and Southeast Asia being particularly species rich. Freshwater megafauna co-occur with up to 93% of mapped overall freshwater biodiversity. Fifty-eight percent of the 132 megafauna species included in the study are threatened, with 84% of their collective range falling outside of protected areas. Of all threatened freshwater species, 83% are found within the megafauna range, revealing the megafauna's capacity as flagship and umbrella species for fostering freshwater conservation.

Phylogeny and polyploidy: resolving the classification of cyprinine fishes (Teleostei: Cypriniformes).

Cyprininae is the largest subfamily (>1300 species) of the family Cyprinidae and contains more polyploid species (∼400) than any other group of fishes. We examined the phylogenetic relationships of the Cyprininae based on extensive taxon, geographical, and genomic sampling of the taxa, using both mitochondrial and nuclear genes to address the phylogenetic challenges posed by polyploidy. Four datasets were analyzed in this study: two mitochondrial gene datasets (465 and 791 taxa, 5604bp), a mitogenome dataset (85 taxa, 14,771bp), and a cloned nuclear RAG1 dataset (97 taxa, 1497bp). Based on resulting trees, the subfamily Cyprininae was subdivided into 11 tribes: Probarbini (new; Probarbus+Catlocarpio), Labeonini Bleeker, 1859 (Labeo & allies), Torini Karaman, 1971 (Tor, Labeobarbus & allies), Smiliogastrini Bleeker, 1863 (Puntius, Enteromius & allies), Poropuntiini (Poropuntius & allies), Cyprinini Rafinesque, 1815 (Cyprinus & allies), Acrossocheilini (new; Acrossocheilus & allies), Spinibarbini (new; Spinibarbus), Schizothoracini McClelland, 1842 (Schizothorax & allies), Schizopygopsini Mirza, 1991 (Schizopygopsis & allies), and Barbini Bleeker, 1859 (Barbus & allies). Phylogenetic relationships within each tribe were discussed. Two or three distinct RAG1 lineages were identified for each of the following tribes Torini, Cyprinini, Spinibarbini, and Barbini, indicating their hybrid origin. The hexaploid African Labeobarbus & allies and Western Asian Capoeta are likely derived from two independent hybridization events between their respective maternal tetraploid ancestors and Cyprinion.

Genome differentiation in a species pair of coregonine fishes: an extremely rapid speciation driven by stress-activated retrotransposons mediating extensive ribosomal DNA multiplications.

BACKGROUND: Sympatric species pairs are particularly common in freshwater fishes associated with postglacial lakes in northern temperate environments. The nature of divergences between co-occurring sympatric species, factors contributing to reproductive isolation and modes of genome evolution is a much debated topic in evolutionary biology addressed by various experimental tools. To the best of our knowledge, nobody approached this field using molecular cytogenetics. We examined chromosomes and genomes of one postglacial species pair, sympatric European winter-spawning Coregonus albula and the local endemic dwarf-sized spring-spawning C. fontanae, both originating in Lake Stechlin. We have employed molecular cytogenetic tools to identify the genomic differences between the two species of the sympatric pair on the sub-chromosomal level of resolution. RESULTS: Fluorescence in situ hybridization (FISH) experiments consistently revealed a distinct variation in the copy number of loci of the major ribosomal DNA (the 45S unit) between C. albula and C. fontanae genomes. In C. fontanae, up to 40 chromosomes were identified to bear a part of the major ribosomal DNA, while in C. albula only 8-10 chromosomes possessed these genes. To determine mechanisms how such extensive genome alternation might have arisen, a PCR screening for retrotransposons from genomic DNA of both species was performed. The amplified retrotransposon Rex1 was used as a probe for FISH mapping onto chromosomes of both species. These experiments showed a clear co-localization of the ribosomal DNA and the retrotransposon Rex1 in a pericentromeric region of one or two acrocentric chromosomes in both species. CONCLUSION: We demonstrated genomic consequences of a rapid ecological speciation on the level undetectable by neither sequence nor karyotype analysis. We provide indirect evidence that ribosomal DNA probably utilized the spreading mechanism of retrotransposons subsequently affecting recombination rates in both genomes, thus, leading to a rapid genome divergence. We attribute these extensive genome re-arrangements associated with speciation event to stress-induced retrotransposons (re)activation. Such causal interplay between genome differentiation, retrotransposons (re)activation and environmental conditions may become a topic to be explored in a broader genomic context in future evolutionary studies.

Atlantor, a new generic name for Barbus reinii Günther, 1874 from Morocco(Teleostei: Cyprinidae).

Atlantor, new genus, is proposed for Barbus reinii from the Atlantic basin in Morocco. The sole species of the genus is clearly separated from related genera based on its phylogenetic position, and from Carasobarbus, Mesopotamichthys and Pterocapoeta in its morphological characters. Atlantor was previously included in Labeobarbus, making this genus paraphyletic with respect to several genera from Western Asia and North Africa. With the description of Atlantor, all these genera become monophyletic. Within Torinae, Atlantor can be diagnosed by having 5½ branched anal-fin rays, modally 8½ branched dorsal-fin rays, two pairs of long barbels, and by its distinct phylogenetic position.

Mystus misrai Anuradha, 1986, a valid species from the Orontes drainage (Teleostei: Bagridae).

Mystus misrai from the northern Orontes drainage (Mediterranean Sea basin) is re-examined and recognised as a valid species. It is distinguished from M. pelusius from the Gulf basin by the lack of stripes on the flank, shorter fins, the eye situated below the dorsal head profile, and a K2P distance of 7.3% in its COI barcoding gene. Mystus misrai is likely Critically Endangered: only a single, spring-fed lake in Türkiye is known as its habitat. The biogeographic connection between the Orontes and the Gulf is discussed based on molecular data of 27 species native to the region.

Glyptothorax vatandousti, a new species of torrent catfish from the upper Karkheh drainage in Iran (Teleostei: Sisoridae).

Glyptothorax vatandousti, new species, from the upper Karkheh drainage, a tributary of the Iranian Tigris, is distinguished from its congeners in the Persian Gulf basin by having the flank with a fine, pale-brown mottled pattern overlaid by small and large, blackish or dark-brown blotches, deep caudal-peduncle (its depth 1.1-1.3 times in length), and without, or with a pale-brown triangle-shaped blotch in front of dorsal-fin origin.

Glyptothorax sardashtensis, a new species of torrent catfish from the upper Lesser Zab drainage in Iran (Teleostei: Sisoridae).

Glyptothorax sardashtensis, new species, from the upper Lesser Zab in Iran, is distinguished from its congeners in the Persian Gulf basin by: a plain flank without black or brown blotches; a wide and round anterior margin of the medial pit in the thoracic adhesive apparatus; few, short median striae in the thoracic adhesive apparatus; three yellowish blotches arranged in a crescent-shaped arch on the nuchal plate in front of the dorsal-fin origin; no tubercles on the head and flank; and a short adipose fin. The new species is also distinguished by a minimum K2P sequence divergence of 2.16% in the mtDNA-COI barcode region from G. daemon and G. galaxias, its closest relatives. Glyptothorax kurdistanicus is re-discovered close to its type locality.

Barbus urmianus, a synonym of Barbus cyri (Teleostei: Cyprinidae).

Barbus urmianus, from the upper Mahabad River in Lake Urmia basin, was distinguished from B. cyri based on several morphological characters. Our analysis demonstrated very small molecular (COI) differences between both species and mostly overlapping or identic morphological character states. Therefore, Barbus urmianus is treated as a junior synonym of B. cyri.

Two new freshwater blennies from the Eastern Mediterranean basin (Teleostei: Blenniidae).

Two new species of Salariopsis are described from the Eastern Mediterranean basin. Salariopsis burcuae, new species, from the Bay of Antalya east to the Jordan, is characterised by having a short cirrus, usually not overlapping the 9th circum-orbital sensory pore, and many tiny black dots on the cheek not organised in rows or bands. The new species shows a 4.1% K2P sequence divergence on the cytochrome-c-oxidase subunit 1 (COI) barcoding region from its closest relative, S. fluviatilis. Salariopsis renatorum, new species, from the upper Ceyhan drainage and a coastal stream in Arsuz, is distinguished by having an unbranched supraocular tentacle, black lateral line pores, a short snout, and no black dots on the upper part of the flank and on the cheek. It is also distinguished from its geographically closest congener, S. burcuae, by a molecular distance of 8.8% K2P in its COI barcode region.

A multi-taxon analysis of European Red Lists reveals major threats to biodiversity.

Biodiversity loss is a major global challenge and minimizing extinction rates is the goal of several multilateral environmental agreements. Policy decisions require comprehensive, spatially explicit information on species' distributions and threats. We present an analysis of the conservation status of 14,669 European terrestrial, freshwater and marine species (ca. 10% of the continental fauna and flora), including all vertebrates and selected groups of invertebrates and plants. Our results reveal that 19% of European species are threatened with extinction, with higher extinction risks for plants (27%) and invertebrates (24%) compared to vertebrates (18%). These numbers exceed recent IPBES (Intergovernmental Platform on Biodiversity and Ecosystem Services) assumptions of extinction risk. Changes in agricultural practices and associated habitat loss, overharvesting, pollution and development are major threats to biodiversity. Maintaining and restoring sustainable land and water use practices is crucial to minimize future biodiversity declines.

Phylogenetic relationships of the algae scraping cyprinid genus Capoeta (Teleostei: Cyprinidae).

We reconstructed the matrilineal phylogeny of Asian algae-eating fishes of the genus Capoeta based on complete mitochondrial gene for cytochrome b sequences obtained from 20 species sampled from the majority of the range and 44 species of closely related barbs of the genera Barbus s. str. and Luciobarbus. The results of this study show that Capoeta forms a strongly supported monophyletic subclade nested within the Luciobarbus clade, suggesting that specialized scraping morphology appeared once in the evolutionary history of the genus. We detected three main groups of Capoeta: the Mesopotamian group, which includes three species from the Tigris-Euphrates system and adjacent water bodies, the Anatolian-Iranian group, which has the most diversified structure and encompasses many species distributed throughout Anatolian and Iranian inland waters, and the Aralo-Caspian group, which consists of species distributed in basins of the Caspian and Aral Seas, including many dead-end rivers in Central Asia and Northern Iran. The most probable origination pathway of the genus Capoeta is hypothesized to occur as a result of allopolyploidization. The origin of Capoeta was found around the Langhian-Serravallian boundary according to our molecular clock. The diversification within the genus occurred along Middle Miocene-Late Pliocene periods.

Molecular phylogeny of the cyprinid tribe Labeonini (Teleostei: Cypriniformes).

The cyprinid tribe Labeonini (sensuRainboth, 1991) is a large group of freshwater fishes containing around 40 genera and 400 species. They are characterized by an amazing diversity of modifications to their lips and associated structures. In this study, a total of 34 genera and 142 species of putative members of this tribe, which represent most of the generic diversity and more than one third of the species diversity of the group, were sampled and sequenced for four nuclear genes and five mitochondrial genes (totaling 9465bp). Phylogenetic relationships and subdivision of this tribe were investigated and the placement and status of most genera are discussed. Partitioned maximum likelihood analyses were performed based on the nuclear dataset, mitochondrial dataset, combined dataset, and the dataset for each nuclear gene. Inclusion of the genera Paracrossochilus, Barbichthys, Thynnichthys, and Linichthys in the Labeonini was either confirmed or proposed for the first time. None of the genera Labeo, Garra, Bangana, Cirrhinus, and Crossocheilus are monophyletic. Taxonomic revisions of some genera were made: the generic names Gymnostomus Heckel, 1843, Ageneiogarra Garman, 1912 and Gonorhynchus McClelland, 1839 were revalidated; Akrokolioplax Zhang and Kottelat, 2006 becomes a junior synonym of Gonorhynchus; the species Osteochilus nashii was found to be a member of the barbin genus Osteochilichthys. Five historical hypotheses on the classification of the Labeonini were tested and rejected. We proposed to subdivide the tribe, which is strongly supported as monophyletic, into four subtribes: Labeoina, Garraina, Osteochilina, and Semilabeoina. The taxa included in each subtribe were listed and those taxa that need taxonomic revision were discussed.

DNA barcode data confirm the placement of subterranean Noemacheilus (Troglocobitis) starostini Parin 1983 in the genus Paracobitis (Teleostei, Nemacheilidae).

DNA barcodes (COI) of Troglocobitis starostini, endemic to a single site in Turkmenistan, were analysed and put into the taxonomic context of the large group of nemacheilid loaches known from Western and Central Asia. All applied phylogenetic tree-based analyses place the species into the genus Paracobitis. This finding supports previous morphological studies. While the exact position of Troglocobitis starostini within Paracobitis was not resolved unambiguously, it was constantly recovered within Paracobitis, irrespective of the tree reconstruction method applied. With a minimum interspecific K2P distance of 7.19% P. persa was the closest hit in our dataset, which comprised a total of ten species of Paracobitis, which showed an average interspecific K2P distance of 5.43% (range 2.78-9.44%).

Garra rezai, a new species from two widely disjunct areas in the Tigris drainage (Teleostei: Cyprinidae).

Garra rezai, new species, is described from the Chooman, a tributary of the Lesser Zab in Iran, and from headwaters of the Yanarsu, a tributary of the upper Tigris in Turkey. It is distinguished from its congeners in the Garra variabilis species group by having two pairs of barbels, a well-developed mental disc, 35-40 total scales along the lateral line, 15-19 scales along the predorsal midline, and 15-18 circumpeduncular scales. It is further characterised by having ten diagnostic nucleotide substitutions and the K2P genetic distances with the closest species i.e. G. klatti, G. kemali and G. variabilis as 11.9, 12.0, and 13.7%, respectively in the mtDNA COI barcode region.

Spatial priorities for freshwater biodiversity conservation in light of catchment protection and connectivity in Europe.

Freshwater ecosystems host disproportionately high numbers of species relative to their surface area yet are poorly protected globally. We used data on the distribution of 1631 species of aquatic plant, mollusc, odonate and fish in 18,816 river and lake catchments in Europe to establish spatial conservation priorities based on the occurrence of threatened, range-restricted and endemic species using the Marxan systematic conservation planning tool. We found that priorities were highest for rivers and ancient lakes in S Europe, large rivers and lakes in E and N Europe, smaller lakes in NW Europe and karst/limestone areas in the Balkans, S France and central Europe. The a priori inclusion of well-protected catchments resulted in geographically more balanced priorities and better coverage of threatened (critically endangered, endangered and vulnerable) species. The a priori exclusion of well-protected catchments showed that priority areas that need further conservation interventions are in S and E Europe. We developed three ways to evaluate the correspondence between conservation priority and current protection by assessing whether a cathment has more (or less) priority given its protection level relative to all other catchments. Each method found that priority relative to protection was high in S and E Europe and generally low in NW Europe. The inclusion of hydrological connectivity had little influence on these patterns but decreased the coverage of threatened species, indicating a trade-off between connectivity and conservation of threatened species. Our results suggest that catchments in S and E Europe need urgent conservation attention (protected areas, restoration, management, species protection) in the face of imminent threats such as river regulation, dam construction, hydropower development and climate change. Our study presents continental-scale conservation priorities for freshwater ecosystems in ecologically meaningful planning units and will thus be important in freshwater biodiversity conservation policy and practice, and water management in Europe.

Contributions to the biodiversity of Vietnam - Results of VIETBIO inventory work and field training in Cuc Phuong National Park.

VIETBIO [Innovative approaches to biodiversity discovery and characterisation in Vietnam] is a bilateral German-Vietnamese research and capacity building project focusing on the development and transfer of new methods and technology towards an integrated biodiversity discovery and monitoring system for Vietnam. Dedicated field training and testing of innovative methodologies were undertaken in Cuc Phuong National Park as part and with support of the project, which led to the new biodiversity data and records made available in this article collection. VIETBIO is a collaboration between the Museum für Naturkunde Berlin - Leibniz Institute for Evolution and Biodiversity Science (MfN), the Botanic Garden and Botanical Museum, Freie Universität Berlin (BGBM) and the Vietnam National Museum of Nature (VNMN), the Institute of Ecology and Biological Resources (IEBR), the Southern Institute of Ecology (SIE), as well as the Institute of Tropical Biology (ITB); all Vietnamese institutions belong to the Vietnam Academy of Science and Technology (VAST). The article collection "VIETBIO" (https://doi.org/10.3897/bdj.coll.63) reports original results of recent biodiversity recording and survey work undertaken in Cuc Phuong National Park, northern Vietnam, under the framework of the VIETBIO project. The collection consist of this "main" cover paper - characterising the study area, the general project approaches and activities, while also giving an extensive overview on previous studies from this area - followed by individual papers for higher taxa as studied during the project. The main purpose is to make primary biodiversity records openly available, including several new and interesting findings for this biodiversity-rich conservation area. All individual data papers with their respective primary records are expected to provide useful baselines for further taxonomic, phylogenetic, ecological and conservation-related studies on the respective taxa and, thus, will be maintained as separate datasets, including separate GUIDs also for further updating.

Oxynoemacheilus shehabi, a new nemacheilid loach from the upper Orontes in southern Syria (Teleostei: Nemacheilidae).

Oxynoemacheilus shehabi, new species, is described from the upper Orontes in southern Syria. It is distinguished from other Oxynoemacheilus species in the Levant by possession of a complete lateral line, a deeply emarginate caudal fin, a narrow caudal peduncle, a suborbital groove in male individuals, a well-developed pelvic axillary lobe, and 5-7 dark-brown bars on the flank. Molecular data suggest that the new species is characterised by a minimum K2P distance of 3.8% to O. "seyhanicola" in the COI mtDNA barcode region.

Oxynoemacheilus amanos, a new nemacheilid loach from the Orontes River drainage (Teleostei: Nemacheilidae).

Oxynoemacheilus amanos, new species, is described from Incesu spring in the upper Hupnik drainage, a northern tributary of the lower Orontes in Turkey. It is distinguished from the other Oxynoemacheilus species in the Eastern Mediterranean Sea basin by possession of an incomplete lateral line with 23-45 pores, terminating between the vertical through the dorsal fin origin and the anus, 10-13 pores in the infraorbital canal, a deeply emarginate caudal fin, no suborbital groove in the male, and a series of irregularly shaped and set dark-brown bars on the flank, not connected to saddles on the back.