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Aim Membranous nephropathy (MN) and primary biliary cholangitis (PBC) are autoimmune diseases that coexist in some cases and might share a common pathogenesis. In 75 - 80% of MN patients, PLA2R1 or THSD7A is the target antigen responsible for disease development, while in the remaining cases, MN pathogenesis is not clear. Our aim was to identify potential antigens playing an overlapping pathogenic role for development of both PBC and MN. Materials and methods Serum from a patient with PBC-associated MN was analyzed for MN and PBC-specific autoantibodies and kidney biopsy tissue was stained for the respective antigens. A review of the literature for published PBC-associated MN cases was performed. Results A 39-year-old male patient was diagnosed with PBC-associated MN. Serology tests revealed negativity for PLA2R1-ab and THSD7A-ab, but positivity for two PBC-specific antibodies: M2-ab and gp210-ab. Kidney biopsy was stained for both PBC-specific antigens, PDC-E2 and gp210, as well as PLA2R1 and THSD7A, showing no MN-specific positivity. Human glomerular extracts also did not contain PDC-E2 or gp210. A review of all 17 published cases of PBC-associated MN showed that 71% of patients suffered from at least one additional autoimmune disease, and different IgG-subclasses were found in the renal immune deposits of these patients. Conclusion These results indicate that both PBC-antigens are not the putative antigen(s) leading to MN development in this patient. PBC-antigens might not be directly responsible for MN development. Both diseases seem to present as autoimmune phenomena triggered by interaction between unknown factors.

The International Code of Nomenclature (ICN) for algae, fungi, and plants calls for indexing of names in nomenclatural repositories (Turland 2018, Art. 42). Scientific names, new combinations, validations, and typifications of algae are novelties tracked by PhycoBank, the registration system for algae. PhycoBank was established and institutionalized at the Botanic Garden Berlin as the repository for nomenclatural acts concerning algae. Since June 2018, PhycoBank staff have been operating the registration system permanently. All data entered into the system undergo a curatorial process to assure a high level of data quality. PhycoBank’s three main components comprise a user-friendly data entry web application available for all registered submitters (self-registration allowed) and curators, a public data access portal, and a search engine that integrates most of the larger online repositories for algae names. The latter is not only a prerequisite for reliable data curation during the registration process but also a valuable, publicly available online tool for algae names. A fourth component still under development is handling the tight integration of the registration system with the workflow of digital publishers. The resulting data are expected to be of high quality and to have been intensively checked against existing nomenclatural acts worldwide. A crucial requirement for the data entry application is thus an intensive support for data validation and curation. Besides the search index giving access to a huge number of existing external names, the data entry application offers a set of tools for quality assurance. It supports and requires the creation of mostly fully atomized data that strictly follow the rules of the ICN. Additionally, completeness for core data is mandatory. Both, atomization and completeness are a precondition to achieve the required uniqueness in the dataset. Completeness also implies that new combinations can only be registered after or together with their original name. The same applies to bi- or trinomials and the respective uni- and binomials they are built on. The publicly available data portal allows accessing published registrations by searching for scientific names, nomenclatural authors, higher ranks and for each part of a bibliographic reference (such as bibliographic author, journal, title, year) stored in PhycoBank. Being a name centric application PhycoBank is neutral with respect to taxonomic opinions. Therefore, only nomenclatural synonymies (basionym or replaced synonym relationships) are stored in the system and names are not attached to a unique classification. Instead, to facilitate search via higher ranks as required by users, classification information is stored as a directed graph of higher taxa with registered names linking to this graph. By this, searches can be performed on multiple classifications simultaneously and can return all possible matches. PhycoBank uses http-based persistent identifiers (e.g. http://phycobank.org/102170), which makes them resolvable and actionable. These identifiers link to nomenclatural acts only and not to the information in the act. That is, a PhycoBank identifier should not be used to refer to a scientific name even if this name was established in the nomenclatural act. PhycoBank makes use of the application stack offered by the EDIT Platform for Cybertaxonomy (Kohlbecker 2017). However, as registration workflows essentially differ from other taxonomic workflows, a completely new user interface for editing has been developed, which provides the users with an intuitive and fluent user experience despite the high complexity of the data. The Common Data Model (Müller 2017) is the core data model for the Platform and already covered the vast majority of data types and fields required by the registration in terms of completeness and degree of atomization. For PhycoBank, it only needed to be extended by a single data type representing the registration/nomenclatural act itself. As of July 2022, PhycoBank includes 4,332 registrations, of which 4,202 are name novelties (1,523 in preparation or ready, 239 under curation, 2,407 published, and 33 rejected); 130 registrations refer to lectotypes or epitypes of existing names. PhycoBank will apply for recognition as a repository in 2022. This is a prerequisite for a proposal to make registration of nomenclatural acts for algae mandatory. This is possible before, at or after the 20th International Botanical Congress 2024.

Temperature and salinity are some of the most influential abiotic parameters shaping biota in aquatic ecosystems. In recent decades, climate change has had a crucial impact on both factors—especially around the Antarctic Peninsula—with increasing air and water temperature leading to glacial melting and the accompanying freshwater increase in coastal areas. Antarctic soft and hard bottoms are typically inhabited by microphytobenthic communities, which are often dominated by benthic diatoms. Their physiology and primary production are assumed to be negatively affected by increased temperatures and lower salinity. In this study, six representative benthic diatom strains were isolated from different aquatic habitats at King George Island, Antarctic Peninsula, and comprehensively identified based on molecular markers and morphological traits. Photosynthesis, respiration, and growth response patterns were investigated as functions of varying light availability, temperature, and salinity. Photosynthesis–irradiance curve measurements pointed to low light requirements, as light-saturated photosynthesis was reached at

The diversity of centric diatoms is documented for the transitional zone of the lower part of the Southern Bug River (Ukraine) just before entering the Dnipro-Bug Estuary and compared to earlier results from the upstream sampling sites of the same river system. Benthic samples of the following sites were investigated: north of Mykolaiv City (approximately 5 km), in Mykolaiv City (near Varvarivskyi Bridge), and 5 km south of the city. Twenty-four centric diatom taxa belonging to 11 genera were identified, analysed, and documented by scanning electron microscopy (SEM) and light microscopy (LM). Among them, Aulacoseira nivalis is the first report for Ukraine, A. islandica and is the first confirmed record for the studied area since the 1930s. The maximum number of centric diatom taxa found in one station was 21, the minimum 10. Melosira subglobosa was the most common (documented in 57–80% of sites with centric diatoms) and abundant species 7.3–15.7% in relative abundance to all diatom taxa. The discovered diversity of taxa and its comparison with previous results is discussed with regard to the relevance of estuary zones in the research of diatoms.

In order to understand the taxonomy and phylogeny of the genus Gomphonema, the type of the name of the genus, G. acuminatum, and related species complexes are studied with an integrative taxonomic ...

Five taxa in the genus Neidium, N. iridis, N. beatyi sp. nov., N. vandusenense sp. nov., N. collare sp. nov. and N. lavoieanum sp. nov. are documented from a pond and stream system in the VanDusen Botanical Garden, Vancouver, Canada. Neidium beatyi is a large linear species with multiple longitudinal canals and sagittate apices. The areolae are occluded by finger-like silica extensions on the external surface. This taxon is distinguished from Neidium iridis by the number of longitudinal canals (>5), shape of the valve apices, and smaller size. Neidium vandusenense is broadly linear with distinct rostrate apices. Two-three longitudinal canals are present along each margin. Plastid rbcL sequence data associates this taxon with N. amphigomphus. Neidium collare is an elliptic lanceolate taxon with one longitudinal canal. This taxon is genetically related to N. bisculatum sensu lato, but with a different shape form. Neidium lavoieanum has a valve shape form similar to Neidium potapovae, but is larger and genetically similar to N. productum sensu lato. The five Neidium taxa were observed in a small stream next to Lake Victoria (pond) in the VanDusen Botanical Garden Vancouver, Canada. The water was mildly alkaline with a pH of 7.86, a conductance of 163 µS/cm, higher nutrient loads and low metal content.

Diatoms are among the most commonly used bioindicators. Correct taxonomic identifications are critical to their use as bioindicators because closely related diatom species can respond diffe...

In central Mexico, streams flow from mountain protected areas at high elevations, i.e. abive 2,400 m a.s.l., being exposed to different human activities along their course. The main human threats of these aquatic ecosystems include the presence of hydraulic infrastructure (e.g. canals and dams), the construction of bridges or roads, fish farming, agriculture, livestock, unregulated tourism or irregular human settlements, resulting in nutrient enrichment, organic pollution and low hydromorphological quality. Diatoms, widely recognized as valuable indicators of environmental conditions, offer a unique opportunity to assess the ecological quality of the streams in and around this megacity, as they respond directly and sensitively to specific physical, chemical and biological changes. To conduct ecological assessment, the composition of diatom communities has traditionally been characterized by morphological methods and most recently by environmental DNA (eDNA) metabarcoding. However, the diatom communities of only a few of these streams in the Basin of Mexico have been characterized morphologically at low resolution light microscopy (LM), and no eDNA-based studies have been conducted yet. In this context, our aims are to 1) implement morphological and eDNA metabarcoding methods for the identification and quantification of epilithic diatoms and to assess their seasonal and spatial response in three peri-urban mountain streams from the Basin of Mexico, and 2) develop a metric to assess and monitor the ecological quality in these streams. For this purpose, we characterized a) the physical, environmental and hydro-morphological conditions related to ecosystem quality, and b) the diatom diversity and community structure of nine sites per stream (27 sites). The sites include headwaters, as well as middle and lower reaches with different land-use and anthropologic pressures, and were investigated over three hydrological periods, i.e. post-rainy, dry-cold and dry-warm. Species identification was first conducted under LM and scanning electron microscopy (SEM), and is currently been done by eDNA metabarcoding of the V4 region of the 18S rRNA gene and a fragment of the chloroplast marker rbcL. Our study has resulted in 520 infrageneric taxa identified by LM and SEM, the largest diversity ever found for freshwater diatoms of Mexico. From this data, we found a clear environmental gradient from almost pristine conditions towards heavily polluted states as the streams enter the city. These changes are well reflected by variations in the diatom community structure, as revealed by the Diatom Ecological Quality Index (DEQI), newly developed to evaluate the ecological quality of the streams in and around this megacity.

During the past decade genetic approaches have been developed to monitor biodiversity in aquatic ecosystems. These enable access to taxonomic and genetic information from biological communities using DNA from environmental samples (e.g. water, biofilm, soil) and methods based on high-throughput sequencing technologies, such as DNA metabarcoding. Within the context of the Water Framework Directive (WFD), such approaches could be applied to assess Biological Quality Elements (BQE). These are used as indicators of the ecological status of aquatic ecosystems as part of national monitoring programs of the european network of 110,000 surface water monitoring sites with 79.5% rivers and 11% lake sites (Charles et al. 2020). A high-throughput method has the potential to increase our spatio-temporal monitoring capacity and to accelerate the transfer of information to water managers with the aim to increase protection of aquatic ecosystems. Good progress has been made with developing DNA metabarcoding approaches for benthic diatom assemblages. Technological innovation and protocol optimization have allowed robust taxonomic (species) and genetic (OTU, ESV) information to be obtained from which diatom quality indices can be calculated to infer ecological status to rivers and lakes. Diatom DNA metabarcoding has been successfully applied for biomonitoring at the scale of national river monitoring networks in several countries around the world and can now be considered technically ready for routine application (e.g. Apothéloz-Perret-Gentil et al. 2017, Bailet et al. 2019, Mortágua et al. 2019, Vasselon et al. 2019, Kelly et al. 2020, Pérez-Burillo et al. 2020, Pissaridou et al. 2021). However, protocols and methods used by each laboratory still vary between and within countries, limiting their operational transferability and the ability to compare results. Thus, routine use of DNA metabarcoding for diatom biomonitoring requires standardization of all steps of the metabarcoding procedure, from the sampling to the final ecological status assessment in order to define good practices and standards. Following previous initiatives which resulted in a CEN technical report for biofilm sampling and preservation (CEN 2018), a set of experiments was initiated during the DNAqua-Net WG2 diatom workshop (Cyprus, 2019) to focus on DNA extraction and PCR amplification steps in order to evaluate: i) the transferability and reproducibility of a protocol between different laboratories; ii) the variability introduced by different protocols currently applied by the scientific community. 19 participants from 14 countries performed DNA extraction and PCR amplification in parallel, using i) the same fixed protocol and ii) their own protocol. Experiments were performed by each participant on a set of standardized DNA and biofilm samples (river, lake, mock community). In order to specifically test the variability of DNA extraction and PCR amplification steps, all other steps of the metabarcoding process were fixed and the preparation of the Miseq sequencing was performed by only one laboratory. The variability within and between participants will be evaluated on DNA extracts quantity, taxonomic (genus, species) and genetic richness, community structure comparison and diatom quality index scores (IPS). We will also evaluate the variability introduced by different DNA extraction and PCR amplification protocols on diatom quality index scores and the final ecological status assessment. The results from this collaborative work will not serve to define “one protocol to rule them all”, but will provide valuable information to define guidelines and minimum requirements that should be considered when performing diatom metabarcoding for biomonitoring.

Diatoms (Bacillariophyta) are ubiquitous microalgae, which present a huge taxonomic diversity, changing in correlation with differing environmental conditions. This makes them excellent ecological indicators for various ecosystems and ecological problematics (ecotoxicology, biomonitoring, paleo-environmental reconstruction …). Current standardized methodologies for diatoms are based on microscopic determinations, which is time consuming and prone to identification uncertainties. DNA metabarcoding has been proposed as a way to avoid these flaws, enabling the sequencing of a large quantity of barcodes from natural samples. A taxonomic identity is given to these barcodes by comparing their sequences to a barcoding reference library. However, to identify environmental sequences correctly, the reference database should contain a representative number of reference sequences to ensure a good coverage of diatom diversity. Moreover, the reference database needs to be carefully taxonomically curated by experts, as its content has an obvious impact on species detection. Diat.barcode is an open-access library for diatoms linking diatom taxonomic identities to rbcL barcode sequences (a chloroplast marker suitable for species-level identification of diatoms), which has been maintained since 2012. Data are accumulated from three sources: (1) the NCBI nucleotide database, (2) unpublished sequencing data of culture collections and more recently (3) environmental sequences. Since 2017, an international network of experts in diatom taxonomy curate this library. The last version of the database (version 9.2), includes 8066 entries that correspond to more than 280 different genera and 1490 different species. In addition to the taxonomic information, morphological features (e.g. biovolumes, chloroplasts, etc.), life-forms (mobility, colony-type) and ecological features (taxa preferences to pollution) are given. The database can be downloaded from the website (www6.inrae.fr/carrtel-collection/Barcoding-database/) or directly through the R package diatbarcode. Ready-to-use files for commonly used metabarcoding pipelines (Mothur and DADA2) are also available.

DNA barcoding and metabarcoding is increasingly used to effectively and precisely assess and monitor biodiversity in aquatic ecosystems. As these methods rely on data availability and quality of barcode reference libraries, it is important to develop and follow best practices to ensure optimal quality and traceability of the metadata associated with the reference barcodes used for identification. Sufficient metadata, as well as vouchers, corresponding to each reference barcode must be available to ensure reliable barcode library curation and, thereby, provide trustworthy baselines for downstream molecular species identification. This document (1) specifies the data and metadata required to ensure the relevance, the accessibility and traceability of DNA barcodes and (2) specifies the recommendations for DNA harvesting and for the storage of both voucher specimens/samples and barcode data, (undefined)

Ecological assessment of lakes and rivers using benthic diatom assemblages currently requires considerable taxonomic expertise to identify species using light microscopy. This traditional approach is also time-consuming. Diatom metabarcoding is a promising alternative and there is increasing interest in using this approach for routine assessment. However, until now, analysis protocols for diatom metabarcoding have been developed and optimised by research groups working in isolation. The diversity of existing bioinformatics methods highlights the need for an assessment of the performance and comparability of results of different methods. The aim of this study was to test the correspondence of outputs from six bioinformatics pipelines currently in use for diatom metabarcoding in different European countries. Raw sequence data from 29 biofilm samples were treated by each of the bioinformatics pipelines, five of them using the same curated reference database. The outputs of the pipelines were compared in terms of sequence unit assemblages, taxonomic assignment, biotic index score and ecological assessment outcomes. The three last components were also compared to outputs from traditional light microscopy, which is currently accepted for ecological assessment of phytobenthos, as required by the Water Framework Directive. We also tested the performance of the pipelines on the two DNA markers (rbcL and 185-V4) that are currently used by the working groups participating in this study. The sequence unit assemblages produced by different pipelines showed significant differences in terms of assigned and unassigned read numbers and sequence unit numbers. When comparing the taxonomic assignments at genus and species level, correspondence of the taxonomic assemblages between pipelines was weak. Most discrepancies were linked to differential detection or quantification of taxa, despite the use of the same reference database. Subsequent calculation of biotic index scores also showed significant differences between approaches, which were reflected in the final ecological assessment. Use of the rbcL marker always resulted in better correlation among molecular datasets and also in results closer to these generated using traditional microscopy. This study shows that decisions made in pipeline design have implications for the dataset's structure and the taxonomic assemblage, which in turn may affect biotic index calculation and ecological assessment. There is a need to define best-practice bioinformatics parameters in order to ensure the best representation of diatom assemblages. Only the use of similar parameters will ensure the compatibility of data from different working groups. The future of diatom metabarcoding for ecological assessment may also lie in the development of new metrics using, for example, presence/absence instead of relative abundance data. (C) 2020 The Authors. Published by Elsevier B.V.

Colonies of a new, stalked diatom species Gomphoneis tegelensis sp. nov. were sampled in September 2017 from Lake Tegel (Berlin, Germany) where this large species was first observed in 2009. Cultiv...

Molecular data (rbcL and 18S) of 36 Planothidium strains were analysed. Twenty-seven strains were also studied morphologically by light microscopy and scanning electron microscopy: six strains from Berlin, two strains from the Faroe Islands, four strains from Lake Baikal, seven strains from Korea and eight strains from Mexico. The findings were compared to International Nucleotide Sequence Database Collaboration data of strains from New Zealand, Germany, France and the USA. The molecular and morphological data differentiated eight species, and the molecular trees underlined the clear differentiation between the two clades, with taxa possessing an asymmetrical central area on the sternum valve which is devoid of striae, either in the form of a sinus or of a cavum clustering together, versus the one taxon lacking a sinus or a cavum. In addition to Planothidium lanceolatum and P. cf. subantarcticum, the clade with a sinus contains two new species, P. cryptolanceolatum and P. taeansa, whereas the other clade ...

Diatoms (Bacillariophyta) are ubiquitous microalgae which produce a siliceous exoskeleton and which make a major contribution to the productivity of oceans and freshwaters. They display a huge diversity, which makes them excellent ecological indicators of aquatic ecosystems. Usually, diatoms are identified using characteristics of their exoskeleton morphology. DNA-barcoding is an alternative to this and the use of High-Throughput-Sequencing enables the rapid analysis of many environmental samples at a lower cost than analyses under microscope. However, to identify environmental sequences correctly, an expertly curated reference library is needed. Several curated libraries for protists exists; none, however are dedicated to diatoms. Diat.barcode is an open-access library dedicated to diatoms which has been maintained since 2012. Data come from two sources (1) the NCBI nucleotide database and (2) unpublished sequencing data of culture collections. Since 2017, several experts have collaborated to curate this library for rbcL, a chloroplast marker suitable for species-level identification of diatoms. For the latest version of the database (version 7), 605 of the 3482 taxonomical names originally assigned by the authors of the rbcL sequences were modified after curation. The database is accessible at https://www6.inra.fr/carrtel-collection_eng/Barcoding-database.

Background and aims – Within the project “German Barcode of Life – Diatoms” common diatoms of German waters were routinely isolated and cultivated. In order to understand the taxonomy and phylogeny of the genus Gomphonema, one of the most common taxa of Central Europe, known currently either under the name Gomphonema olivaceum in Europe or Gomphoneis olivacea in America, was studied. Methods – Twenty unialgal strains were established from five different water bodies in Germany and one from Lake Balaton, Hungary, which supplied molecular data (18S V4 and rbcL) besides morphometric and ultrastructural data. In addition, on eight populations from different water bodies including the type from Denmark, morphometric and micromorphological studies by light and scanning electron microscopy were performed. Key results – Molecular and micromorphological data show that the target taxon neither belongs to Gomphonema Ehrenb. nor to Gomphoneis Cleve. By reinstating the genus name Gomphonella Rabenh., the nomenclatural and taxonomic enigma of this taxon is solved, and with the presentation of the type by Hornemann the authorship of the epithet is clarified. Molecular data for the unialgal strains and several environmental clones show that there is more diversity in the Gomphonella olivacea clade than can be identified morphologically. In addition, the establishment of the new species Gomphonella coxiae and Gomphonella acsiae is supported. The molecular data classified Gomphonella species as belonging to the Cymbellales but not to the Gomphonemataceae. In addition, molecular data put Gomphoneis tegelensis R.Jahn & N.Abarca also into Gomphonella. In order to make the genera Gomphoneis and Gomphonema monophyletic, their astigmate members are transferred to Gomphonella. Conclusions – The results clarify that the gomphonemoid outline is not restricted to the family Gomphonemataceae but seem to be distributed across the entire order Cymbellales. This is shown in this paper for the revived genus Gomphonella, which contains the astigmate group of Gomphoneis and Gomphonema besides the longly disputed G. olivacea. Only a polyphasic approach, combining molecular and micromorphological data for taxonomy, nomenclatural evaluation, and observations from clonal cultures can reveal the full intricacies of evolutionary relations.

Aquatic biomonitoring has become an essential task in Europe and many other regions as a consequence of strong anthropogenic pressures affecting the health of lakes, rivers, oceans and groundwater. A typical assessment of the environmental quality status, such as it is required by European but also North American and other legislation, relies on matching the composition of assemblages of organisms identified using morphological criteria present in aquatic ecosystems to those expected in the absence of anthropogenic pressures. Through decade-long and difficult intercalibration exercises among networks of regulators and scientists in European countries, a pragmatic biomonitoring approach was developed and adopted, which now produces invaluable information. Nonetheless, this approach is based on several hundred different protocols, making it susceptible to issues with comparability, scale and resolution. Furthermore, data acquisition is often slow due to a lack of taxonomic experts for many taxa and regions and time-consuming morphological identification of organisms. High-throughput genetic screening methods such as (e)DNA metabarcoding have been proposed as a possible solution to these shortcomings. Such "next-generation biomonitoring", also termed "biomonitoring 2.0", has many advantages over the traditional approach in terms of speed, comparability and costs. It also creates the potential to include new bioindicators and thereby further improves the assessment of aquatic ecosystem health. However, several major conceptual and technological challenges still hinder its implementation into legal and regulatory frameworks. Academic scientists sometimes tend to overlook legal or socioeconomic constraints, which regulators have to consider on a regular basis. Moreover, quantification of species abundance or biomass remains a significant bottleneck to releasing the full potential of these approaches. Here, we highlight the main challenges for next-generation aquatic biomonitoring and outline principles and good practic, COST - European Cooperation in Science and Technology(CA15219). COST Action DNAqua-Net (CA15219), supported by the COST (European Cooperation in Science and Technology) programme

Diatoms are used routinely to assess pollution level in rivers and lakes. Current methods are based on identification by light microscopy, which is laborious. An alternative is to identify species based on short DNA fragments and High-Throughput Sequencing (HTS). However a potential limitation is the incomplete coverage of species in reference barcode libraries. Usually these libraries are compiled by isolating cells, before culturing and sequencing them, which is tedious and often unsuccessful. Here we propose the use of rbcL sequences from environmental samples analysed by HTS. We set several criteria to ensure good sequence quality and correspondence with the target species observed in microscopy: the sequence needed to be abundant in the sample, and with no insertions nor deletions or stop codon, phylogenetic neighbour taxa had to correspond to neighbour taxonomic taxa expected from morphological observations. Four species from tropical rivers are given as examples, including one that is new to science. info:eu-repo/semantics/publishedVersion

The protection, preservation and restoration of aquatic ecosystems and their functions are of global importance. For European states it became legally binding mainly through the EU-Water Framework Directive (WFD). In order to assess the ecological status of a given water body, aquatic biodiversity data are obtained and compared to a reference water body. The quantified mismatch obtained determines the extent of potential management actions. The current approach to biodiversity assessment is based on morpho-taxonomy. This approach has many drawbacks such as being time consuming, limited in temporal and spatial resolution, and error-prone due to the varying individual taxonomic expertise of the analysts. Novel genomic tools can overcome many of the aforementioned problems and could complement or even replace traditional bioassessment. Yet, a plethora of approaches are independently developed in different institutions, thereby hampering any concerted routine application. The goal of this Action is to nucleate a group of researchers across disciplines with the task to identify gold-standard genomic tools and novel eco-genomic indices for routine application in biodiversity assessments of European fresh- and marine water bodies. Furthermore, DNAqua-Net will provide a platform for training of the next generation of European researchers preparing them for the new technologies. Jointly with water managers, politicians, and other stakeholders, the group will develop a conceptual framework for the standard application of eco-genomic tools as part of legally binding assessments.

Plant genome size evolution is a very dynamic process: the ancestral genome of angiosperms was initially most likely small, which led to a tendency towards genome increase during evolution. However, findings in several angiosperm lineages demonstrate mechanisms that also led to genome size contraction. Recent molecular investigations on the Asteraceae genus Crepis suggest that several genomic reduction events have occurred during the evolution of the genus. This study focuses on the Mediterranean Crepis sect. Neglectoides, which includes three species with some of the smallest genomes within the whole genus. Crepis neglecta has the largest genome in sect. Neglectoides, approximately twice the size of the two species Crepis cretica and Crepis hellenica. Whereas C. cretica and C. hellencia are more closely related to each other than to C. neglecta the karyotypes of the latter species and C. cretica are similar, while that of C. hellenica differs considerably. Here, the karyotypic organisation of the three species is investigated with fluorescence in-situ hybridisation and studied in a molecular phylogenetic framework based on the nuclear markers Actin, CHR12, CPN60B, GPCR1 and XTH23. Our findings further corroborate the occurrence of genome size contraction in Crepis, and suggest that the difference in genome size between C. neglecta and C. cretica is mostly due to elimination of dispersed repetitive elements, whereas chromosomal reorganisation was involved in the karyotype formation of C. hellenica.

Water covers over 70% of our planet's surface and it is a key resource for the survival of all organisms, among them humans. Unfortunately, water resources face increasing pressures due to the exponential expansion of and resource exploitation by human beings. The consequences of this on water ecosystems represent hallmarks of the Anthropocene such as chemical pollution, warming, scarcity of clean drinking water, ocean acidification and a dramatic loss of biodiversity. As a consequence, the direct and indirect benefits humanity obtained from these ecosystems as cheap services, such as clean water, biomass production, climate regulation and matter fluxes, are increasingly at risk. Therefore, it is of utmost importance to assess the ecological state of aquatic ecosystems and to protect and manage them in a sustainable way. In order to assess the ecological status of a given water body, aquatic biodiversity data are collected by morphological identification of bioindicator species and a comparison of site-specific species lists to those of fairly natural reference water bodies. Quantifying the differences between the lists guides subsequent management actions. Examples of European standard bioasseessments (so far morphologically-based) are Marine Strategy Framework Directive (2008/56/EC) and the Water Framework Directive (2000/60/EC). While the implementation of biomonitoring programs is already a great success, there is room for improvement. In the field of molecular genetics, revolutionary high-throughput DNA-based analyses have been developed. These can be applied to assess taxon lists of hundreds to many thousands at once and greatly improve speed and accuracy of assessments. However, while these novel genetic tools have attracted a lot of interest, they are not implemented in any of the regular legal biomonitoring programs. In order to change this, the European Co-Operation in Science and Technology (COST) program's Action CA15219 'DNAqua-Net' was launched in November 2016 (Leese et al. 2016). The Action aims to gather existing knowledge and complement those standard procedures by developing and implementing these novel genomic DNA-based approaches for biomonitoring and bioassessment. The Action is comprised of five working groups (WGs): WG1: DNA Barcode References; WG2: Biotic Indices & Metrics; WG3: Lab & Field Protocols; WG4: Data Analysis & Storage and WG5: Implementation Strategies & Legal Issues. However, central to this is the standardisation of the various protocols, methods and biotic indices and integration of DNA-based datasets (e.g. resulting from DNA metabarcoding, mito- and metagenomics) with existing data standards. Here, the TDWG community will be of central importance and its participation in DNAqua-Net is highly desirable. Moreover, the innovative open access journal Metabarcoding & Metagenomics (MBMG) has been recently initiated to promote open science and enhance data exchange in this field as well as to connect the diverse actors and communities (Leese et al. 2017).

The Lerma-Chapala Basin, in Central Mexico, is geologically heterogeneous, climatically diverse and boasts high biodiversity, lying within two Biodiversity Hotspots, namely Mesoamerica and the Madrean Pine–Oak Woodlands. Epilithon and water samples were collected in the basin from 14 sampling sites three times each, two sampling campaigns during the rainy season and one in the dry season. A total of 274 infrageneric taxa in 48 genera were recorded. The taxonomic composition observed was dominated by taxa from the genera Nitzschia, Gomphonema, Pinnularia, Navicula, Sellaphora and Eunotia. About a third of the taxa found could not be identified to the species level. From those unidentified morphodemes, two are described as new species, namely Brachysira altepetlensis and Sellaphora queretana. Furthermore, Eolimna rhombica is transferred to Sellaphora. Canonical Correspondence Analysis (CCA) revealed that specific conductivity and pH were the main environmental factors driving the community composition observed. Three groups of samples were identified after the CCA: 1) characterized by acidic waters and low conductivity; 2) with circumneutral waters, low specific conductivity and high temperature and phosphorous concentrations; and 3) characterized by circumneutral waters, high conductivity and low nitrogen concentrations. The indicator value method (IndVal), based on the relative abundance and relative frequency of the most abundant taxa was calculated based on the groups observed in the CCA, identifying the characteristic taxa for each of the three groups.

Diatoms are frequently used for water quality assessments; however, identification to species level is difficult, time-consuming and needs in-depth knowledge of the organisms under investigation, as nonhomoplastic species-specific morphological characters are scarce. We here investigate how identification methods based on DNA (metabarcoding using NGS platforms) perform in comparison to morphological diatom identification and propose a workflow to optimize diatom fresh water quality assessments. Diatom diversity at seven different sites along the course of the river system Odra and Lusatian Neisse from the source to the mouth is analysed with DNA and morphological methods, which are compared. The NGS technology almost always leads to a higher number of identified taxa (270 via NGS vs. 103 by light microscopy LM), whose presence could subsequently be verified by LM. The sequence-based approach allows for a much more graduated insight into the taxonomic diversity of the environmental samples. Taxa retrieval varies considerably throughout the river system, depending on species occurrences and the taxonomic depth of the reference databases. Mostly rare taxa from oligotrophic parts of the river systems are less well represented in the reference database used. A workflow for DNA-based NGS diatom identification is presented. 28 000 diatom sequences were evaluated. Our findings provide evidence that metabarcoding of diatoms via NGS sequencing of the V4 region (18S) has a great potential for water quality assessments and could complement and maybe even improve the identification via light microscopy.

The planktonic freshwater diatom Cyclotella comensis Grunow is widely used as an indicator of oligotrophic lake conditions. Taxonomic confusion arose with the discrimination of the morphologically similar species Cyclotella pseudocomensis Scheffler and Cyclotella costei Druart & Straub. In this study, we used morphological and molecular data to investigate the relationship between C. comensis, C. pseudocomensis and C. costei using 11 strains from eight lakes in Germany and Austria. Morphological variability was high among the three morphospecies such that they could not be separated unambiguously using morphological traits. DNA sequence analysis revealed only minor differences in 18S V4 (

Diatoms are present in all types of water bodies and their species diversity is influenced greatly by environmental conditions. This means that diatom occurrence and abundances are suitable indicators of water quality. Furthermore, continuous screening of algal biodiversity can provide information about diversity changes in ecosystems. Thus, diatoms represent a desirable group for which to develop an easy to use, quick, efficient, and standardised organism identification tool to serve routine water quality assessments. Because conventional morphological identification of diatoms demands specialised in-depth knowledge, we have established standard laboratory procedures for DNA barcoding in diatoms. We (1) identified a short segment (about 400 bp) of the SSU (18S) rRNA gene which is applicable for the identification of diatom taxa, and (2) elaborated a routine protocol including standard primers for this group of microalgae. To test the universality of the primer binding sites and the discriminatory power of the proposed barcode region, 123 taxa, representing limnic diatom diversity, were included in the study and identified at species level. The effectiveness of the barcode was also scrutinised within a closely related species group, namely the Sellaphora pupula taxon complex and relatives.

The GGBN Data Portal (http://www.ggbn.org, Droege et al. 2014) has established standardised data flows for genomic DNA samples, including voucher specimens, tissue samples, DNA samples as well as resulting sequences and publications. Dealing with different types of DNA (aDNA, gDNA, eDNA) is essential and closely related to user-friendly search and display functionalities. GGBN aims both at preserving voucher specimens of all kinds of DNA as well as making these important data accessible on the Internet. In addition to genomic DNA, the development and use of high-throughput-/next-generation-sequencing (HTS formerly designated NGS) has outstripped current plans of SYNTHESYS and GGBN to join natural history collection data with DNA and tissue collection data. HTS libraries can be considered as a preparation of the genetic material of a single organism or of multiple organism (e.g. from an environmental mixed sample). From that point of view, they are the actual physical molecular representation of a specimen or sample. However, these libraries come with specific adaptors that limit their transferability to other sequencing systems. The libraries are prepared at great expense, but frequently are only used for a single project, not making use of additional useful information that could potentially be generated. To increase the potential of the HTS libraries to be used for multiple projects they have to be discoverable via published metadata. Optimally, HTS library metadata will include specific standardized keywords (by e.g. organism, HTS method etc.). Here we present our ideas and a prototype for eDNA samples and HTS libraries based on the GGBN Data Standard (Droege et al. 2016). A use case collection from animals, diatoms, fungi and plants has been developed and is available in the GGBN Sandbox (see more details at http://wiki.ggbn.org/ggbn/Use_Case_Collection). These examples will be improved upon and kept on-line at least until 2020, so progress can be observed.

24 páginas, 2 figuras, 1 tabla., The protection, preservation and restoration of aquatic ecosystems and their functions are of global importance. For European states it became legally binding mainly through the EUWater Framework Directive (WFD). In order to assess the ecological status of a given water body, aquatic biodiversity data are obtained and compared to a reference water body. The quantified mismatch obtained determines the extent of potential management actions. The current approach to biodiversity assessment is based on morpho-taxonomy. This approach has many drawbacks such as being time consuming, limited in temporal and spatial resolution, and error-prone due to the varying individual taxonomic expertise of the analysts. Novel genomic tools can overcome many of the aforementioned problems and could complement or even replace traditional bioassessment. Yet, a plethora of approaches are independently developed in different institutions, thereby hampering any concerted routine application. The goal of this Action is to nucleate a group of researchers across disciplines with the task to identify gold-standard genomic tools and novel ecogenomic indices for routine application in biodiversity assessments of European fresh- and marine water bodies. Furthermore, DNAqua-Net will provide a platform for training of the next generation of European researchers preparing them for the new technologies. Jointly with water managers, politicians, and other stakeholders, the group will develop a conceptual framework for the standard application of eco-genomic tools as part of legally binding assessments.

Diatom cultures of the G. parvulum species complex were established from seven different sites in the Faroe Islands, Sweden, Germany, Mexico and Korea, and were studied in detail. Eight morphodemes were identified which corresponded to the descriptions of the cosmopolitan taxon G. parvulum (Kützing) Kützing sensu lato: its nominate variety (var. parvulum), G. parvulum var. exilissimum Grunow and G. parvulum f. saprophilum Lange-Bertalot & Reichardt, G. [parvulum var.] lagenula Kützing plus four unidentifiable morphodemes. The concatenated analysis of the sequences of the markers 18SV4, rbcL, and ITS as well as morphological data resulted in a separation of four taxa based on their biogeography in Mexico, Korea, central Continental Europe and Northern Atlantic Europe. Mantel tests showed a significant correlation between molecular and geographical distances. The diagnoses of two taxa, G. parvulum sensu stricto, and G. lagenula, were emended, G. saprophilum elevated to species rank and epitypes designated. One species was newly described.