Barcode Of Life Data System Research Articles

State of biodiversity documentation in the Philippines: Metadata gaps, taxonomic biases, and spatial biases in the DNA barcode data of animal and plant taxa in the context of species occurrence data.

Anthropogenic changes in the natural environment have led to alarming rates of biodiversity loss, resulting in a more urgent need for conservation. Although there is an increasing cognizance of the importance of incorporating biodiversity data into conservation, the accuracy of the inferences generated from these records can be highly impacted by gaps and biases in the data. Because of the Philippines’ status as a biodiversity hotspot, the assessment of potential gaps and biases in biodiversity documentation in the country can be a critical step in the identification of priority research areas for conservation applications. In this study, we systematically assessed biodiversity data on animal and plant taxa found in the Philippines by examining the extent of metadata gaps, taxonomic biases, and spatial biases in DNA barcode data while using species occurrence data as a backdrop of the ‘Philippines’ biodiversity. These barcode and species occurrence datasets were obtained from public databases, namely: GenBank, Barcode of Life Data System and Global Biodiversity Information Facility. We found that much of the barcode data had missing information on either records and publishing, geolocation, or taxonomic metadata, which consequently, can limit the usability of barcode data for further analyses. We also observed that the amount of barcode data can be directly associated with the amount of species occurrence data available for a particular taxonomic group and location–highlighting the potential sampling biases in the barcode data. While the majority of barcode data came from foreign institutions, there has been an increase in local efforts in recent decades. However, much of the contribution to biodiversity documentation only come from institutions based in Luzon.

Gap analysis for DNA-based biomonitoring of aquatic ecosystems in China

DNA-based taxon identification is improving the assessment and management of biodiversity in rivers. However, the lack of comprehensive DNA barcode reference libraries and globally highly unequal coverage are still hindering the application prospects of this method worldwide. Here, we analyzed the COI barcode gap in two reference libraries, Barcode of Life Data Systems (BOLD) and NCBI GenBank, with a focus on three aquatic animal groups (freshwater fish, aquatic insects and molluscs) in Chinese rivers. Our data show gaps in barcode coverage (e.g., organisms without barcodes) of ca. 40–70% of taxa in these groups in the BOLD or NCBI GenBank database, respectively. These gaps can rise even further if the barcode thresholds are set to contain at least five reference sequences per taxon. Furthermore, most barcodes are from non-local samples, and only 14.4% (BOLD) and 28.8% (NCBI GenBank) of reference sequences were from organisms sampled in China, respectively. The pairwise genetic distance of local barcodes is 3 to 5 times lower than non-local barcodes, indicating that the latter may not be a good substitute. When looking at individual catchments, ca. 60% of the potentially occurring aquatic species have one or more barcodes, yet the barcode coverage varies slightly across ten major river catchments, ranging from 54.3% (Liao River basin) to 68.2% (Huai River basin). The taxa Salmoniformes and Perciformes in freshwater fish, Odonata and Diptera in aquatic insects, and Bivalvia in molluscs have the best barcode coverage in most catchments (mean coverage >70%). This study gives the first overview and current status of barcode reference libraries of three major aquatic animal groups in Chinese rivers. Our results will help to better interpret current metabarcoding studies from China, and also provide a basis to develop a strategy of filling the gaps in the reference libraries of aquatic species in China.

Trophic markers and biometric measurements in Southern Ocean sea stars (1985-2017).

Sea stars (Echinodermata: Asteroidea) are a key component of Southern Ocean benthos, with 16% of the known sea star species living there. In temperate marine environments, sea stars commonly play an important role in food webs, acting as keystone species. However, trophic ecology and functional role of Southern Ocean sea stars are still poorly known, notably due to the scarcity of large-scale studies. Here, we report 24,332 trophic marker (stable isotopes and elemental contents of C, N, and S of tegument and/or tube feet) and biometric (arm length, disk radius, arm to disk ratio) measurements in 2,456 specimens of sea stars. Samples were collected between 12 January 1985 and 8 October 2017 in numerous locations along the Antarctic littoral and subantarctic islands. The spatial scope of the data set covers a significant portion of the Southern Ocean (47.717° S to 86.273° S; 127.767° W to 162.201° E; depth, 6-5,338 m). The data set contains 133 distinct taxa, including 72 currently accepted species spanning 51 genera, 20 families, and multiple feeding guilds/functional groups (suspension feeders, sediment feeders, omnivores, predators of mobile or sessile prey). For 505 specimens, mitochondrial CO1 genes were sequenced to confirm and/or refine taxonomic identifications, and those sequences are already publicly available through the Barcode of Life Data System. This number will grow in the future, as molecular analyses are still in progress. Overall, thanks to its large taxonomic, spatial, and temporal extent, as well as its integrative nature (combining genetic, morphological, and ecological data), this data set can be of wide interest to Southern Ocean ecologists, invertebrate zoologists, benthic ecologists, and environmental managers dealing with associated areas. Please cite this data paper in research products derived from the data set, which is freely available without copyright restrictions.

An Incomplete European Barcode Library Has a Strong Impact on the Identification Success of Lepidoptera from Greece

Species identification by means of DNA barcodes depends essentially on the scope and quality of a relevant reference library. The first analysis of a large number (about 600 morphospecies) of southern European Lepidoptera (Greece: Peloponnese) shows both the advantages and disadvantages with regard to a reliable identification of Mediterranean species. We determined 946 DNA barcode sequences from 47 families, of which 929 sequences from 46 families were successfully assigned to a Barcode Index Number (BIN) in the global Barcode of Life Data Systems (BOLD) database. A species level identification for 485 BINs representing 477 Linnaean names was successful. These taxa include 34 new records for Greece. However, 128 BINs (c. 20% of the inventory) could not be attached to a Linnaean name from referenced sequences available in BOLD. Of these BINs, 99 are new and hence represent unique records for BOLD. Intra- and inter-BIN divergences are presented and discussed. An initial and preliminary in-depth analysis of randomly selected species indicates an incomplete DNA barcode library in terms of Linnean taxa, in addition to a considerable number of probably undescribed species. It is therefore strongly recommended that the already advanced European barcode library of Lepidoptera should be supplemented with not-yet-sequenced taxa from the Mediterranean.

MetaCOXI: an integrated collection of metazoan mitochondrial cytochrome oxidase subunit-I DNA sequences.

Nucleotide sequences reference collections or databases are fundamental components in DNA barcoding and metabarcoding data analyses pipelines. In such analyses, the accurate taxonomic assignment is a crucial aspect, relying directly on the availability of comprehensive and curated reference sequence collection and its taxonomy information. The currently wide use of the mitochondrial cytochrome oxidase subunit-I (COXI) as a standard DNA barcode marker in metazoan biodiversity studies highlights the need to shed light on the availability of the related relevant information from different data sources and their eventual integration. To adequately address data integration process, many aspects should be markedly considered starting from DNA sequence curation followed by taxonomy alignment with solid reference backbone and metadata harmonization according to universal standards. Here, we present MetaCOXI, an integrated collection of curated metazoan COXI DNA sequences with their associated harmonized taxonomy and metadata. This collection was built on the two most extensive available data resources, namely the European Nucleotide Archive (ENA) and the Barcode of Life Data System (BOLD). The current release contains more than 5.6 million entries (39.1% unique to BOLD, 3.6% unique to ENA, and 57.2% shared between both), their related taxonomic classification based on NCBI reference taxonomy, and their available main metadata relevant to environmental DNA studies, such as geographical coordinates, sampling country and host species. MetaCOXI is available in standard universal formats (‘fasta’ for sequences & ‘tsv’ for taxonomy and metadata), which can be easily incorporated in standard or specific DNA barcoding and/or metabarcoding data analysis pipelines. Database URL: https://github.com/bachob5/MetaCOXI

Phylogenetic signal of sub‐arctic beetle communities

Postglacial dispersal and colonization processes have shaped community patterns in sub‐Arctic regions such as Churchill, Manitoba, and Canada. This study investigates evolutionary community structure within the beetle (Coleoptera) families of Churchill and tests whether biological traits have played a role in governing colonization patterns from refugial and southerly geographic regions. This study quantifies sub‐Arctic beetle phylogenetic community structure for each family using the net relatedness index (NRI) and nearest taxon index (NTI), calculated using publicly available data from the Barcode of Life Data Systems (BOLD); compares patterns across families with different traits (habitat, diet) using standard statistical analysis (ANOVA) as well as phylogenetic generalized least squares (PGLS) using a family‐level beetle phylogeny obtained from the literature; and compares community structure in Churchill with a region in southern Canada (Guelph, Ontario). These analyses were also repeated at a genus level. The dominant pattern detected in our study was that aquatic families were much better represented in Churchill compared to terrestrial families, when compared against richness sampled from across Canada and Alaska. Individually, most families showed significant phylogenetic clustering in Churchill, likely due to the strong environmental filtering present in Arctic environments. There was no significant difference in phylogenetic structure between Churchill and Guelph but with a trend toward stronger clustering in the North. Fungivores were significantly more overdispersed than other feeding modes, predators were significantly more clustered, and aquatic families showed significantly stronger clustering compared to terrestrial. This study contributes to our understanding of the traits and processes structuring insect biodiversity and macroecological trends in the sub‐Arctic.

Metabarcoding of canopy arthropods reveals negative impacts of forestry insecticides on community structure across multiple taxa

Abstract Insecticides used to combat outbreaks of forest defoliators can adversely affect non‐target arthropods. Forestry insecticides typically suppress Lepidoptera larvae which are the cornerstone of the canopy community of deciduous oak forests. The abrupt removal of this dominant component of the food web could have far‐reaching implications for forest ecosystems, yet it is rarely investigated in practice owing to several methodological shortcomings. The taxonomic impediment and the biased nature of arthropod sampling techniques particularly impede the assessment of insecticide impacts on diverse communities. To tackle this issue, we propose an experimental approach combining sampling by pyrethrum knockdown and species determination via DNA metabarcoding, using community subsampling to derive estimates of species abundances. We applied this protocol to investigate the short‐term effects of the insecticides diflubenzuron (DFB) and Bacillus thuringiensis var. kurstaki (BTK) on canopy‐dwelling arthropod communities in German oak woodlands. Our approach allowed us to detect most of the diversity and integrate species abundances in our analyses. By classifying arthropod species into assemblages based on their expected sensitivity rather than coarse taxonomic groupings, we could unveil substantial effects of DFB across multiple taxa 5 weeks after application. Although strong effects on single species appear related to direct toxicity, substantial impacts of DFB on parasitoids and xylophagous beetles suggest that anti‐defoliator treatments can have previously unsuspected indirect effects on some components of forest arthropod communities. The impacts of BTK on community structure were consistent with that of DFB though much weaker. Synthesis and applications. Comparing diversity patterns in the arthropod communities of sprayed and unsprayed oak canopies, our results show that selective insecticides can alter species diversity in presumably non‐sensitive taxa. Even though the ecological significance of these impacts has yet to be assessed in an operational setting, their existence calls for increased regulatory scrutiny on indirect effects. As community approaches become more attainable with the rapid development of DNA metabarcoding, we suggest the inclusion of community‐level end points as regulatory requirements for the approval of forest‐use insecticides.

Population genetics and molecular identification of Crocodylus acutus and C. moreletii (Crocodilia: Crocodylidae) in captive and wildlife populations

Introduction: There is low evidence of genetic diversity and hybridization processes within Crocodylus acutus and C. moreletii populations. Objetive: To evaluate genetic diversity and some phylogenetic relationships in wild and captive populations of C. acutus and C. moreletii using the Barcode of Life Data System (COX1, cytochrome C oxidase subunit 1 gene). Methods: 28 individuals phenotypically like C. acutus located in the state of Guerrero, Oaxaca and Quintana Roo were sampled, as well as animals belonging to C. moreletii located in the states of Tabasco, Campeche, and Quintana Roo. 641 base pairs of nucleotide sequence from COX1 were used to obtain the haplotype and nucleotide diversity per population, and a phylogenetic and network analysis was performed. Results: Evidence of hybridization was found by observing C. moreletti haplotypes in animals phenotypically determined as C. acutus, as well as C. acutus haplotypes in animals classified as C. moreletti. Low haplotypic diversity was observed for C. acutus (0.455 ± 0.123) and for C. moreletii (0.505 ± 0.158). A phylogenetic tree was obtained in which the sequences of C. acutus and C. moreletii were grouped into two well-defined clades. Organisms identified phenotypically as C. acutus but with C. moreletii genes were separated into a different clade within the clade of C. moreletii. Conclusions: There are reproductive individuals with haplotypes different from those of the species. This study provides a small but significant advance in the genetic knowledge of both crocodile species and the use of mitochondrial markers, which in this case, the COX1 gene allowed the detection of hybrid organisms in wild and captive populations. Conservation efforts for both species of crocodiles should prevent the crossing of both threatened species and should require the genetic identification of pure populations, to design effective conservation strategies considering the possibility of natural hybridization in areas of sympatry.
 Palabras clave: ADN mitocondrial; citocromo C oxidasa; diversidad haplotípica; diversidad de nucleótidos; hibridación, Sistema de código de barras de la vida.

In memory of Gary Bauchan: Integrated taxonomy of soil mites in farming systems

Current and future legislation requiring the reduction of pesticides use, coupled with global initiatives for the promotion of soil health and conservation of soil biodiversity are creating opportunities for studies aimed at highlighting ecosystem services provided by functioning soil food webs in agricultural systems, including soil predatory mites. However, the key personnel for performing such studies are expert taxonomists, who are already spread very thin. To meet this demand, we propose an integrative approach where scientists (without expertise in taxonomy) play a significant role in supporting expert taxonomists. Soil samples were collected at the USDA ARS Farming Systems Project (FSP) at the Beltsville Agricultural Research Center, either incubated or not, followed by extraction of mites and nematodes. Improved modified Berlese funnels and an extraction protocol were utilized to improve sequencing success. Incubation dramatically enhanced the number of extracted individuals per sample whilst the daily freezing of extracted mites substantially improved the sequencing success rate compared to previous studies. Taken together, this led to the addition of records for eight Mesostigmata and ten Oribatida BINs to BOLD (Barcode of Life Datasystem). Fifteen species of Mesostigmata species were found, with three dominant species, Cycetogamasus diviortus (Athias-Henriot, 1967) (Parasitidae), Lasioseius youcefi Athias-Henriot 1959 (Blattisociidae) and a new species of Gamasellodes (Ascidae). LTSEM imaging followed by molecular identification contributed further details to the published descriptions of C. diviortus and L. youcefi. In line with our general aim, collecting, extracting, identification to morpho-species, sample preparation for DNA barcoding and uploading relevant information to BOLD was performed by trained personnel, but without taxonomic expertise. Whereas our skilled taxonomists focused on the morphological identifications using light microscopy, expanding on existing descriptions using LTSEM images and in a subsequent manuscript the description of a new species. We believe this division of tasks and labor will set the stage for further collaborative integrated studies between ecologists, biocontrol specialists and expert taxonomists for the identification, evaluation, and description of known and novel soil acarine biological control agents (BCAs).

Contribution to a reference library of DNA barcodes of Colombian freshwater fishes.

BackgroundThe Barcode of Life initiative was originally motivated by the large number of species, taxonomic difficulties and the limited number of expert taxonomists. Colombia has 1,610 freshwater fish species and comprises the second largest diversity of this group in the world. As genetic information continues to be limited, we constructed a reference collection of DNA sequences of Colombian freshwater fishes deposited in the Ichthyology Collection of the University of Antioquia (CIUA), thus joining the multiple efforts that have been made in the country to contribute to the knowledge of genetic diversity in order to strengthen the inventories of biological collections and facilitate the solution of taxonomic issues in the future.New informationThis study contributes to the knowledge on the DNA barcodes and occurrence records of 96 species of Colombian freshwater fishes. Fifty-seven of the species represented in this dataset were already available in the Barcode Of Life Data System (BOLD System), while 39 correspond to new species to the BOLD System. Forty-nine specimens were collected in the Atrato River Basin and 708 in the Magdalena-Cauca asin during the period 2010-2020. Two species (Loricariichthysbrunneus (Hancock, 1828) and Poeciliasphenops Valenciennes, 1846) are considered exotic to the Atrato, Cauca and Magdalena Basins and four species (Oncorhynchusmykiss (Walbaum, 1792), Oreochromisniloticus (Linnaeus, 1758), Parachromisfriedrichsthalii (Heckel, 1840) and Xiphophorushelleri Heckel, 1848) are exotic to the Colombian hydrogeographic regions. All specimens are deposited in CIUA and have their DNA barcodes made publicly available in the BOLD online database. The geographical distribution dataset can be freely accessed through the Global Biodiversity Information Facility (GBIF).

Importance of nonindigenous harpacticoids (Crustacea: Copepoda) decrease with depth in Lake Ontario

Harpacticoid copepods can be a substantial component of the meiobenthic community in lakes and serve an ecological role as detritivores. Here we present the first species-level lake-wide quantitative assessment of the harpacticoid assemblage of Lake Ontario with emphasis on the status of nonindigenous species. Additionally, we provide COI-5P sequences of harpacticoid taxa through Barcode of Life Data System (BOLD). Harpacticoids were collected at depths from 0.1 to 184 m and from a range of substrates from August to September 2018 as part of the Cooperative Science and Monitoring Initiative (CSMI) offshore benthic survey. Twenty-six meiobenthic samples were analyzed using microscopy for community composition analysis of harpacticoids. We found thirteen indigenous and three nonindigenous species of harpacticoid, with the introduced species dominating at shallow depths. The community transitioned from nonindigenous to indigenous species dominance as depth increased. Nonindigenous species accounted for 79% of the community (by abundance) at depths <20 m, 55% from 20 to 40 m, and only 24% at depths >40 m. The nonindigenous species encountered included the first detections of Schizopera borutzkyi (Monchenko, 1967) and Heteropsyllus nunni (Coull, 1975) from Lake Ontario. S. borutzkyi was the most abundant harpacticoid species in the lake, approaching a maximum density of 50,000/m2 and a lake-wide average density of 7,900/m2. Numerically important indigenous species included Bryocamptus nivalis (Willey, 1925), Canthocamptus robertcokeri (Wilson, 1958), Canthocamptus staphylinoides (Pearse, 1905), and Moraria cristata (Chappuis, 1929). The prevalence of nonindigenous harpacticoids in the meiobenthos of Lake Ontario suggests further investigations of Great Lakes meiofauna communities are warranted.

First insights on the diversity of the genus Psychoda Latreille (Diptera: Psychodidae) in Corsica using DNA barcoding

Among the 44 European Psychoda species identified to date, we expected to observe in Corsica species with large distribution ranges and/or that are frequent in adjacent territories (i.e. south mainland France, mainland Italy, and Sardinia). Each studied specimen was morphologically identified and then barcoded using cytochrome c oxidase subunit I (COI) and subunit II (COII) to confirm its identification, and to detect potential cryptic species. From 211 mounted specimens, 16 Psychoda sensu lato morphotypes were recorded in Corsica and mainland France. While 14 out of the 15 species recorded in Corsica are new records for the island, two species are also new records for France and Europe, P. (Psychodocha) uncinula Quate, 1954 and P. (Chodopsycha) divaricata Duckhouse, 1968. Moreover, our barcoding approach allowed us to detect 18 haplotypes in total, including three undescribed Corsican haplotype complexes, namely P. (Tinearia) alternata, P. (Logima) satchelli and P. (Psychodocha) cinerea. With only mitochondrial COI barcodes, it cannot be asserted whether these complexes represent the within-species coexistence of various haplotypes or the existence of distinct cryptic species. In BOLD (Barcode of Life Datasystems), only 30 out of the 58 barcode indice numéros (BINs) corresponding to the genus Psychoda have a species name. All of the COI barcodes generated in this study already exist in BOLD, suggesting that the barcodes of most of the temperate Psychoda diversity are already known, but not properly named.

Mitochondrial Cox-1 Barcoding For The Identification and Phylogenetic Analysis of Sarcophaga (Liosarcophaga) dux Recorded From Jeddah City of Saudi Arabia

This study was aimed to identify and phylogenetically analyze the sarcophagid flies from Jeddah (Saudi Arabia) on the basis of mitochondrial cytochrome-C-oxidase subunit-1 (COX-1) gene. Five flesh fly samples were collected from different locations of Jeddah region. All the samples were first morphologically identified as Sarcophaga dux and confirmed on the basis of COX-1 gene amplification and sequencing techniques using universal primers LCO1490 and HCO2198. For phylogenetic analysis, 5 sequences were checked by COX-1 service on barcode of life data system (BOLD) database. To determine the phylogenetic relationships and genetic variability between 5 collected and BOLD samples, large Neighbor-joining tree comprising of the collected samples and all the available S. dux BOLD samples were constructed. The 32 most related BOLD sequences were used to construct maximum likelihood (ML) and Neighbor-joining trees. The phylogenetic analysis of COX-1 gene sequences revealed significant match with S. dux sequences from BOLD database: GQ254446 (Australia), MG780139 (India), AY879255 (China), JN869983 (Malaysia) and KC249664 (Egypt). This is the first report of the flesh fly species recorded in the Western regions of Saudi Arabia. Results indicated that the genetic diversity and phylogenetic relationship analysis of the collected S. dux samples have an important role in understanding the evolutionary processes of Sarcophaga cryptic species in Western regions of Saudi Arabia.

Assessing the effectiveness of DNA barcoding for exploring hidden genetic diversity in deep-sea fishes

The numbers of deep-sea fish species and their genetic diversities are poorly understood because of taxonomic confusion and the lack of robust diagnostic features. However, DNA barcoding using mitochondrial DNA sequences may offer an effective approach to identifying cryptic species and characterizing their genetic diversities. To validate the genetic differentiation identified by DNA mitochondrial barcoding, it is necessary to show that these reflect variations present in nuclear genomic markers. Here, we performed DNA barcoding using cytochrome c oxidase subunit I (COI) sequences and also carried out multiplexed intersimple sequence repeat genotyping by sequencing (MIG-seq) for mesopelagic and demersal fish species from the continental shelf and upper slope of the northwestern Pacific Ocean. We obtained the COI sequences of 115 species from 48 families; the species were identified using the Barcode of Life Data System. Phylogenetic analyses using COI sequences showed high levels of intraspecific genetic differentiation (Kimura 2-parameter distances &gt;2%) in 20 of 115 species, suggesting many cryptic species or intraspecific genetic differentiation previously unknown in these species. We performed phylogenetic and population genetic analyses using multiple single-nucleotide polymorphism loci obtained by MIG-seq of 3 species that showed high levels of intraspecific genetic differentiation in COI sequences. The nuclear markers confirmed the genetic differentiation in all 3 species identified by the COI sequences. The high concordance between these different genetic markers indicates the effectiveness of DNA barcoding for identifying cryptic deep-sea species and characterizing genetic differentiation in these species.

DNA Barcoding of Key Storage Pests Using Mitochondrial Cytochrome Oxidase I

Stored grain insect pests are of economic importance, and they spread rapidly through the grain trade. Hence, it is important to identify these pests accurately. Although several morphological methods exist, it is tedious and time-consuming. DNA barcoding using mitochondrial COI is an alternative approach that aids in precise species identification. In this study, 13 stored grain insect pests belonging to the order Coleoptera and Lepidoptera were collected from different storage structures in Coimbatore. A fragment of mt-COI was amplified and sequenced. Sequence analyses were carried out with BOLD (Barcode of Life Data System) and ABGD (Automatic Barcode Gap Discovery) tools. The barcoding gap analysis revealed that the inter-species genetic distance is greater than the intra-species genetic distance. The ABGD analysis for species delimitation partitioned the Coleopteran and Lepidopteran datasets into 9 and 4 putative species, respectively. The barcode gap was absent in more closely related species. However, analysis of their sequences revealed significant variations. Our results showed the ability of the mt-COI to discriminate between the species, thus provide a complementary technique for the diagnosis of stored grain insect pests.

Revision and annotation of DNA barcode records for marine invertebrates: report of the 8th iBOL conference hackathon

The accuracy of specimen identification through DNA barcoding and metabarcoding relies on reference libraries containing records with reliable taxonomy and sequence quality. The considerable growth in barcode data requires stringent data curation, especially in taxonomically difficult groups such as marine invertebrates. A major effort in curating marine barcode data in the Barcode of Life Data Systems (BOLD) was undertaken during the 8th International Barcode of Life Conference (Trondheim, Norway, 2019). Major taxonomic groups (crustaceans, echinoderms, molluscs, and polychaetes) were reviewed to identify those which had disagreement between Linnaean names and Barcode Index Numbers (BINs). The records with disagreement were annotated with four tags: a) MIS-ID (misidentified, mislabeled, or contaminated records), b) AMBIG (ambiguous records unresolved with the existing data), c) COMPLEX (species names occurring in multiple BINs), and d) SHARE (barcodes shared between species). A total of 83,712 specimen records corresponding to 7,576 species were reviewed and 39% of the species were tagged (7% MIS-ID, 17% AMBIG, 14% COMPLEX, and 1% SHARE). High percentages (&amp;gt;50%) of AMBIG tags were recorded in gastropods, whereas COMPLEX tags dominated in crustaceans and polychaetes. The high proportion of tagged species reflects either flaws in the barcoding workflow (e.g., misidentification, cross-contamination) or taxonomic difficulties (e.g., synonyms, undescribed species). Although data curation is essential for barcode applications, such manual attempts to examine large datasets are unsustainable and automated solutions are extremely desirable.

Mitochondrial sequence diversity reveals the hybrid origin of invasive gibel carp (Carassius gibelio) populations in Hungary.

BackgroundInvasive gibel carp, Carassius gibelio (Bloch, 1782) has become well-established in the Hungarian waters and now are spreading in the European waters. On major concern now is the potential hybridization between gibel carp and the other invasive species in the Carassius auratus complex (CAC), which may further accelerate the spread of the whole invasive species complex. The identification of gibel carp and their hybrids is difficult because of its morphological similarity to the other species in CAC. Here we carry out a genomic assessment to understand the history of gibel carp invasion and its phylogenetic relationship with the other species in CAC. Three loci of the mitochondrial genome (D-loop, CoI, Cytb) were used to determine the phylogenetic origin of individuals and relarionship among six gibel carp populations and the other species in the CAC.MethodolgyA total of 132 gibel carp samples from six locations in Southern Transdanubia (Hungary) were collected after phenotypic identification to measure the genetic diversity within and among gibel carp populations of Southern Transdanubia (Hungary). The genetic background was examined by the sequences of the mitochondrial genome: D-loop, Cytochrome c oxidase I (CoI) and Cytochrome b (Cytb). Mitochondrial genetic markers are excellent tools for phylogenetic studies because they are maternally inherited. Successfully identified haplotypes were aligned and with reference sequences in nucleotide databases (i.e., NCBI-BLAST: National Centre for Biotechnology Information and BOLD: Barcode of Life Data System). The phylogenetic relationships among gibel carp populations were then analyzed together with the reference sequences to understand the relationship and the level of hybridization with the species in CAC.ResultsAmong the 132 aligned D-loop sequences 22 haplotypes were identified. Further examination of representative individuals of the 22 haplotypes, six Cytb and four CoI sequences were detected. The largest number of haplotypes of all three loci were found in Lake Balaton, the largest shallow lake in Central Europe. Based on the NCBI-BLAST alignment of the D-loop, haplotypes of Carassius auratus auratus and Carassius a. buergeri in CAC were identified in the C. gibelio samples. Further analysis of haplotypes with the other two mitochondrial markers confirmed the occurrence of intragenus hybridization of C. gibelio in the Hungarian waters.ConclusionBy using three mitochondrial markers (D-loop, Cytb, CoI), we genomically characterized a gibel carp-complex in Hungarian waters and assessed the C. gibelio phylogenetic status between them. Hybrid origin of locally invasive Carassius taxon was detected in Hungary. It points out that invasive species are not only present in Hungary but reproduce with each other in the waters, further accelerating their spread.

DNA barcodes evidence the contact zone of eastern and western caddisfly lineages in the Western Carpathians

The region of the Western Carpathians is, among other aspects, very important for survival and diversity of European freshwater fauna due to the presence of a large number of (sub)mountain springs and streams. However, these ecologically and faunistically diversified habitats are still understudied in the context of genetic diversity and population structure of their inhabitants. This study focuses on genetic diversity and distribution patterns of the caddisfly Rhyacophila tristis, common and widespread representative of mountain freshwater fauna. Analysis of the COI mitochondrial marker revealed presence of the western and eastern lineages, with samples from both lineages being grouped in BOLD (Barcode of Life Data System) into separate BINs (Barcode Index Numbers). Our data indicates that eastern lineage (BIN_E) is more closely related to the Balkan populations than to co-occurring western lineage (BIN_W), and that the contact zone of the lineages passes through the W Carpathians. The study revealed phylogeographic and demographic differences between lineages, supporting hypothesis of their evolutionary independence and specific ecological preferences. The obtained genetic data of the R. tristis population from W Carpathians improved our knowledge about population genetics of this aquatic species and can contribute to understanding the state and evolution of biodiversity of freshwater ecosystems in Europe.

A Review on Application of DNA Barcoding Technology for Rapid Molecular Diagnostics of Adulterants in Herbal Medicine.

The rapid molecular diagnostics of adulterants in herbal medicine using DNA barcoding forms the core of this meticulously detailed review, based on two decades of data. With 80% of the world's population using some form of herbal medicine, authentication, quality control, and detection of adulterants warrant DNA barcoding. A combined group of keywords were used for literature review using the PubMed, the ISI Web of Knowledge, Web of Science (WoS), and Google Scholar databases. All the papers (N=210) returned by the search engines were downloaded and systematically analyzed. Detailed analysis of conventional DNA barcodes were based on retrieved sequences for internal transcribed spacer (ITS) (412,189), rbcL (251,598), matK (210,835), and trnH-psbA (141,846). The utility of databases such as The Barcode of Life Data System (BOLD), NCBI, GenBank, and Medicinal Materials DNA Barcode Database (MMDBD) has been critically examined for the identification of unknown species from known databases. The current review gives an overview of the ratio of adulterated to authentic drugs for some countries along with the state of the art technology currently being used in the identification of adulterated medicines. In this review, efforts were made to systematically analyze and arrange the research and reviews on the basis of technical progress. The review concludes with the future of DNA-based herbal medicine adulteration detection, forecasting the reliance on the metabarcoding technology. DNA barcoding technology for differentiating adulterated herbal medicine.

Global gap-analysis of amphipod barcode library.

In the age of global climate change and biodiversity loss there is an urgent need to provide effective and robust tools for diversity monitoring. One of the promising techniques for species identification is the use of DNA barcoding, that in Metazoa utilizes the so called ‘gold-standard’ gene of cytochrome c oxidase (COI). However, the success of this method relies on the existence of trustworthy barcode libraries of the species. The Barcode of Life Data System (BOLD) aims to provide barcodes for all existing organisms, and is complemented by the Barcode Index Number (BIN) system serving as a tool for potential species recognition. Here we provide an analysis of all public COI sequences available in BOLD of the diverse and ubiquitous crustacean order Amphipoda, to identify the barcode library gaps and provide recommendations for future barcoding studies. Our gap analysis of 25,702 records has shown that although 3,835 BINs (indicating putative species) were recognised by BOLD, only 10% of known amphipod species are represented by barcodes. We have identified almost equal contribution of both records (sequences) and BINs associated with freshwater and with marine realms. Three quarters of records have a complete species-level identification provided, while BINs have just 50%. Large disproportions between identification levels of BINs coming from freshwaters and the marine environment were observed, with three quarters of the former possessing a species name, and less than 40% for the latter. Moreover, the majority of BINs are represented by a very low number of sequences rendering them unreliable according to the quality control system. The geographical coverage is poor with vast areas of Africa, South America and the open ocean acting as “white gaps”. Several, of the most species rich and highly abundant families of Amphipoda (e.g., Phoxocephalidae, Ampeliscidae, Caprellidae), have very poor representation in the BOLD barcode library. As a result of our study we recommend stronger effort in identification of already recognised BINs, prioritising the studies of families that are known to be important and abundant components of particular communities, and targeted sampling programs for taxa coming from geographical regions with the least knowledge.