Using DNA barcoding to identify the early life history stages of demersal coastal fishes in shallow nearshore and estuarine benthic habitats of Algoa Bay

Comparative phylogenetic and sequence identity analysis of internal transcribed spacer 2 and cytochrome c oxidase subunit I as DNA barcode markers for the most common equine Strongylidae species.

DNA barcoding based on a fragment of the cytochrome c oxidase subunit I (COI) gene is widely applied in species identification and biodiversity studies. The aim of this study was to establish a comprehensive barcoding database of coastal ray-finned fishes in Vietnam. A total of 3,638 specimens were collected from fish landing sites in northern, central and southern Vietnam. Seven hundred and sixty-five COI sequences of ray-finned fishes were generated, belonging to 458 species, 273 genera, 113 families and 43 orders. A total of 59 species were newly recorded in Vietnam and sequences of six species were new to the Genbank and BOLD online databases. Only 32 species cannot be annotated to species level because difficulty in morphological identifications and their Kimura-2-Parameter (K2P) genetic distances to most similar sequences were more than 2%. Moreover, intra-specific genetic distances in some species are also higher than 2%, implying the existence of putative cryptic species. The mean K2P genetic distances within species, genera, families, orders and classes were 0.34%, 12.14%, 17.39%, 21.42%, and 24.80, respectively. Species compositions are quite different with only 16 common species among northern, central and southern Vietnam. This may attribute to multiple habitats and environmental factors across the 3,260 km Vietnamese coastline. Our results confirmed that DNA barcoding is an efficient and reliable tool for coastal fish identification in Vietnam, and also established a reliable DNA barcode reference library for these fishes. DNA barcodes will contribute to future efforts to achieve better monitoring, conservation, and management of fisheries in Vietnam.

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 >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.

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 241:201-213 (2002) - doi:10.3354/meps241201 Trace elements in otoliths indicate the use of open-coast versus bay nursery habitats by juvenile California halibut Graham E. Forrester1,*, Stephen E. Swearer2,** 1Department of Biological Sciences, University of Rhode Island, Kingston, Rhode Island 02881, USA 2Department of Ecology, Evolution, and Marine Biology, University of California, Santa Barbara, California 93106, USA *E-mail: gforrester@uri.edu **Present address: Department of Zoology, University of Melbourne, Victoria 3010, Australia ABSTRACT: Many coastal fishes use inshore nursery habitats as juveniles, but it is often difficult to define which nursery areas supply most recruits to adult populations. We tested whether trace element concentrations in otoliths can be used to identify which of 2 nursery habitats (bays or shallow open coast) were occupied by juvenile California halibut. Juveniles from bays in 1998 had concentrations of Cu and Pb in their otoliths that were higher than those in open coast juveniles of the same year. This broad-scale difference between bay and open coast juveniles remained intact when bay juveniles from 1994 to 1997 were added to the comparison, and juvenile halibut could be assigned to their nursery habitat of origin quite accurately (83%) using otolith concentrations of Cu and Pb. At a finer spatial scale, otolith concentrations of Cu and Pb differed among individual bays, and fish from the same bay could differ among years, precluding their use as markers of nursery habitat use at these scales. Like halibut otoliths, sediments from bays had higher concentrations of Cu and Pb than open coast nursery sites, and this difference was consistent over 11 yr. Otoliths and sediments from individual bays, however, showed no correlation in Cu and Pb concentrations. The concentration of Cu and Pb in sediments and their deposition in otoliths were thus loosely matched at a broad scale, though the underlying cause of this link is not known. A discriminant model, parameterized using Cu and Pb levels in juvenile otoliths, was used to classify prior nursery habitat use by 19 larger halibut (of unknown origin). Eleven of these halibut had high levels of Cu and Pb in the part of the otolith deposited as a juvenile, and were classified as of bay origin. The other 8 halibut had low otolith Cu and Pb levels in the juvenile portion of their otoliths and were classified as having used open coast nurseries. Overall, our results suggest that this approach has the potential to allow identification of nursery habitat use by California halibut at a broad scale (bay vs open coast) but not at a fine scale (individual bays). KEY WORDS: Estuaries · Nursery habitat · Otoliths · Trace elements · Halibut Full text in pdf format PreviousNextExport citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 241. Online publication date: October 04, 2002 Print ISSN: 0171-8630; Online ISSN: 1616-1599 Copyright © 2002 Inter-Research.

DNA barcoding based on a fragment of the cytochrome c oxidase subunit I (COI) gene from the mitochondrial genome is widely applied in species identification, species discovery and biodiversity studies. The aim of this study was to establish a barcoding reference database of sponges collected from South Africa, and evaluate the applicability of the COI gene for aiding in the identification of sponges in combination with tentative morphological identifications. A total of 317 mitochondrial COI barcode sequences, with an additional 21 extended COI fragments and 24 nuclear ITS sequences, were obtained from 11 orders, 38 families, 58 genera and 124 species of spiculated sponges. A Neighbour Joining (NJ) trees that were reconstructed using these sequences in most cases clustered species in accordance with their current taxonomic identification, and we conclude that COI sequencing can be used to aid in the identification of sponge species. We further demonstrate that DNA barcoding analysis has potential to uncover cryptic sponge species, and to reveal dubious morphological identifications. We recommend that future taxonomic studies of South African sponges incorporate multiple sources of information for species identification or discovery.

BackgroundArctic ecosystems, especially those near transition zones, are expected to be strongly impacted by climate change. Because it is positioned on the ecotone between tundra and boreal forest, the Churchill area is a strategic locality for the analysis of shifts in faunal composition. This fact has motivated the effort to develop a comprehensive biodiversity inventory for the Churchill region by coupling DNA barcoding with morphological studies. The present study represents one element of this effort; it focuses on analysis of the spider fauna at Churchill.Results198 species were detected among 2704 spiders analyzed, tripling the count for the Churchill region. Estimates of overall diversity suggest that another 10–20 species await detection. Most species displayed little intraspecific sequence variation (maximum <1%) in the barcode region of the cytochrome c oxidase subunit I (COI) gene, but four species showed considerably higher values (maximum = 4.1-6.2%), suggesting cryptic species. All recognized species possessed a distinct haplotype array at COI with nearest-neighbour interspecific distances averaging 8.57%. Three species new to Canada were detected: Robertus lyrifer (Theridiidae), Baryphyma trifrons (Linyphiidae), and Satilatlas monticola (Linyphiidae). The first two species may represent human-mediated introductions linked to the port in Churchill, but the other species represents a range extension from the USA. The first description of the female of S. monticola was also presented. As well, one probable new species of Alopecosa (Lycosidae) was recognized.ConclusionsThis study provides the first comprehensive DNA barcode reference library for the spider fauna of any region. Few cryptic species of spiders were detected, a result contrasting with the prevalence of undescribed species in several other terrestrial arthropod groups at Churchill. Because most (97.5%) sequence clusters at COI corresponded with a named taxon, DNA barcoding reliably identifies spiders in the Churchill fauna. The capacity of DNA barcoding to enable the identification of otherwise taxonomically ambiguous specimens (juveniles, females) also represents a major advance for future monitoring efforts on this group.

Sphaeroma terebrans, a wood-boring isopoda, is distributed worldwide in tropical and subtropical mangroves. The taxonomy of S. terebrans is usually based on morphological characteristics, with its molecular identification still poorly understood. The number of teeth on the uropodal exopod and the length of the propodus of the seventh pereopod are considered as the major morphological characteristics in S. terebrans, which can cause difficulty in regards to accurate identification. In this study, we identified S. terebrans via molecular and morphological data. Furthermore, the validity of the mitochondrial cytochrome c oxidase subunit I (COI) gene as a DNA barcode for the identification of genus Sphaeroma, including species S. terebrans, S. retrolaeve, and S. serratum, was examined. The mitochondrial COI gene sequences of all specimens were sequenced and analysed. The interspecific Kimura 2-parameter distances were higher than intraspecific distances and no intraspecific-interspecific distance overlaps were observed. In addition, genetic distance and nucleotide diversity (π) exhibited no differences within S. terebrans. Our results revealed that the mitochondrial COI gene can serve as a valid DNA barcode for the identification of S. terebrans. Furthermore, the number of teeth on the uropodal exopod and the length of the propodus of the seventh pereopod were found to be unreliable taxonomic characteristics for S. terebrans.

DNA barcoding by sequencing a standard region of cytochrome c oxidase subunit I (COI) provides an accurate, rapid method for identifying different species. In this study, we provide a molecular taxonomic assessment of demersal fishes in the Bering Sea and Chukchi Sea based on DNA barcoding, and a total of 123 mitochondrial COI partial fragments with a length of 652 bp were obtained. The consensus among all sequences was determined by alignment via a BLAST search in GenBank. Phylogenetic relationships were reconstructed based on neighbor-joining trees and barcoding gaps. The 39 species investigated in this analysis were distributed among 10 families. Five families within Scorpaeniformes including 19 species accounted for almost half of the species. The next largest group was Perciformes, with 9 species, followed by Pleuronectiformes and Gadiformes, with 5 species each, and the smallest number of species belonged to Rajiformes. At the family level, Cottidae was the largest family, followed by Zoarcidae, accounting for 8 species. The other eight families—Gadidae, Pleuronectidae, Psychrolutidae, Agonidae, Liparidae, Ammodytidae, Hexagrammidae, and Rajidae—accounted for a smaller proportion of species. In brief, our study shows that DNA barcodes are an effective tool for studying fish diversity and taxonomy in the Bering Sea and Chukchi Sea. The contribution of DNA barcoding to identifying Arctic fish species may benefit further Arctic fish studies on biodiversity, biogeography and conservation in the future.

Current-mediated downstream dispersal by the early developmental stages of fish in rivers is a common phenomenon. Knowledge of patterns and processes in the dispersal, or ‘drift’, of young fishes provides important information on spawning location and spawning success, habitat use, movement paths and flow-ecology relationships more generally, all of which are critical for effective river conservation and management. But despite the importance of such information, our understanding of the patterns and processes of the drift of the early life stages of riverine fishes is limited. Furthermore, riverine fish drift research has tended to occur in isolation from movement studies of other organisms, limiting its integration with higher level concepts and theory. This manuscript reviews the literature on the dispersal of young fishes in running waters. Relevant studies from all climatic zones and geographical regions are investigated, with particular attention given to the types and life history stages of fishes that drift and the seasonal and diel patterns of drifting. We then consider how fish enter the drift and their mode of drifting, attempting to reconcile a long-running discussion, under what we call the ‘active–passive conundrum’. We argue that, aside from eggs, the early stages of fish are not exclusively either passive or active drifters, but usually a mixture of the two, which we term ‘actipassive’ drift. Finally, we evaluate existing knowledge in the context of a general conceptual framework for movement ecology, identifying gaps in our understanding of the roles of internal state, navigation capacity, motion capacity, external factors and internal factors in influencing the dispersal process.

Objective To make up the limitations of traditional morphological classification methods, we identified vector fleas by DNA barcoding in Qinghai Province. Methods The mt DNA cytochrome c oxidase subunit Ⅰ(COⅠ) gene was amplified by PCR from 36 muscle tissues of fleas in 3 states, 2 cities and 5 counties of Qinghai Province, and the obtained COⅠ gene fragments were sequenced and aligned. The intra- and inter-species genetic distances were calculated with Mega 6 software using K2-P model and a phylogenetic tree was constructed with neighbor-joining (NJ) method. Results Totally 36 COⅠ gene sequences of 2 superfamilies, 4 genera and 6 kinds of vector fleas were measured, the average genetic distance was 0.119, and the intraspecific distance was 0.002 - 0.027, the interspecific distance was 0.039 - 0.207, and the interspecific genetic distance was significantly greater than the intraspecific genetic distance. NJ tree showed the same species had formed a single line with high support rate and interspecific branch was clear. Conclusion DNA barcoding is suitable for identification of vector fleas in Qinghai Province, may make up the limitations of traditional morphological classification methods. Key words: Siphonaptera; Cytochrome c oxidase subunit Ⅰ; DNA barcoding

Ambiguities within species description and identification may compromise research validity. Species identification has typically been based upon morphological characteristics, yet recent technological advances have led to identifications achieved via DNA approaches, including DNA barcoding. DNA barcoding studies typically use cytochrome c oxidase subunit I (COI) as the proposed universal molecular marker for animals. Here, we test 12 mitochondrial protein coding genes for the presence of a clear barcoding gap allowing us to unequivocally define species. Using the African Great Apes as our model group, we assess this at the species (Pan troglodytes), genus (Pan) and family (Hominidae) level. Based on 279 complete mitochondrial genomes, sequences were partitioned by gene for analysis and pairwise distances were calculated. No barcoding gap was observed at the within species level, i.e., the four recognised chimpanzee taxa were not distinguishable through DNA barcoding. However, NADH dehydrogenase subunit 5 (ND5) and cytochrome c oxidase subunit II (COII) produce the largest barcoding gaps at the genus (ND5 2%, COII 0.5%) and family (ND5 1.5%, COII 0.5%) level. Rather than focusing on COI, our analysis suggests that these two genes may be more, or at least as, appropriate markers in primate species delineation, with uses in the identification of extinct and extant species. Further use may be beneficial to taxonomists, providing additional evidence and new insights for these morphologically similar species.

DNA barcoding is a novel concept for the taxonomic identification, in that it uses a specific short genetic marker in an organism’s DNA to discriminate species. In 2003, professor Paul D. N. Hebert, “the father of DNA barcoding”, of the University of Guelph, Ontario, Canada first proposed the idea to identify biological species using DNA barcode, where the mitochondrial gene cytochrome c oxidase subunit I (COI) was supposed to be the first candidate for animals (Hebert et al. 2003a). Their studies of COI profiling in both higher taxonomic categories and species-level assignment demonstrated that COI gene has significant resolutions across the animal kingdom except the phylum Cnidaria (Hebert et al. 2003b, Ward et al. 2005, Hajibabaei et al. 2006). From then on, a wide broad of taxonomic groups (i.e. birds, fish, butterflies, spiders, ants, etc) were examined by COI gene for its usability as the barcode (i.e. Hebert et al. 2004a, Hebert et al. 2004b, Greenstone et al. 2005, Smith et al. 2005, Barber and Boyce 2006, Meier et al. 2006, Kerr et al. 2007, Kumar et al. 2007, Pfenninger et al. 2007, Stahls and Savolainen 2008, Zhou et al. 2009). Meanwhile, other candidate genes, including Internal Transcribed Spacer (ITS), trnH-psbA intergenic spacer (trnH-psbA), Ribulose-bisphosphate carboxylase (rbcL) and Maturase K (matK) were analysed by different research groups (Jaklitsch et al. 2006, Evans et al. 2007, Ran et al. 2010, de Groot et al. 2011, Liu et al. 2011, Piredda et al. 2011, Yesson et al. 2011). Till recently, there are about 30 DNA barcode candidates are tested, and 4 to 8 of them are widely used for the identification of diversified taxonomic groups with a relatively good resolution. It has been estimated that there are 10 to 100 million species of living creatures in the earth, while what we know is very limited. Knowing the biodiversity is one of the crucial biological issues of ecology, evolutionary biology, bio-security, agro-biotechnology, bioresources and many other areas. For very long period, taxonomists have provided a nomenclatural hierarchy and key prerequisites for the society. However, the needs for species identification requested by non-taxonomists require the knowledge held by taxonomists. Therefore, a standardized, rapid and inexpensive species identification approach is needed to establish for the non-specialists. There had some attempts on the molecular identification systems based on polymerase chain reaction (PCR), especially in bacterial studies (Woese 1996, Zhou et al. 1997, Maiden et al. 1998, Wirth et al. 2006), but no successful solutions for broader scopes of eukaryotes (reviewed in Frezal and Leblois 2008). The DNA Barcode of Life project is another attempt to create a universal eukaryotic identification system based on molecular approaches. Following studies by Hebert et al.

A number of same species of Cerithiidae are morphologically unlike, whereas most of species in the same genus are morphologically similar and just exhibit subtle differences. It is difficult to identify them by morphological methods alone. DNA barcoding is a modern molecular technique that can be used to identify species accurately, and is particularly helpful when distinguishing morphologically similar species. In order to identify species of Cerithiidae using DNA barcoding technology based on mitochondrial cytochrome oxidase subunit I (COI) and 16S ribosomal RNA (16S rRNA) genes, this study calculated intraspecific and interspecific genetic distance and constructed the phylogenetic trees. A total of 80 COI and 16S rRNA barcode sequences were obtained from 10 species and 3 genera. Some unknown specimens were further identified and a cryptic species may exist in Cerithium traillii, showing that DNA barcoding technology has the potential to discover new species and cryptic species. The phylogenetic trees revealed that all of the cerithiids could converge upon a monophyly with high support values and two genera (Cerithium and Clypeomorus) maybe support the reclassification. It is necessary for traditional morphological methods to combine with the DNA barcoding for classification and identification of Cerithiidae.

Oomycete species occupy many different environments and many ecological niches. The genera Phytophthora and Pythium for example, contain many plant pathogens which cause enormous damage to a wide range of plant species. Proper identification to the species level is a critical first step in any investigation of oomycetes, whether it is research driven or compelled by the need for rapid and accurate diagnostics during a pathogen outbreak. The use of DNA for oomycete species identification is well established, but DNA barcoding with cytochrome c oxidase subunit I (COI) is a relatively new approach that has yet to be assessed over a significant sample of oomycete genera. In this study we have sequenced COI, from 1205 isolates representing 23 genera. A comparison to internal transcribed spacer (ITS) sequences from the same isolates showed that COI identification is a practical option; complementary because it uses the mitochondrial genome instead of nuclear DNA. In some cases COI was more discriminative than ITS at the species level. This is in contrast to the large ribosomal subunit, which showed poor species resolution when sequenced from a subset of the isolates used in this study. The results described in this paper indicate that COI sequencing and the dataset generated are a valuable addition to the currently available oomycete taxonomy resources, and that both COI, the default DNA barcode supported by GenBank, and ITS, the de facto barcode accepted by the oomycete and mycology community, are acceptable and complementary DNA barcodes to be used for identification of oomycetes.

Cryptic species present a challenge for conservation, as species diversity may remain undetected. In zoological research, DNA barcoding of the mitochondrial cytochrome c oxidase subunit I (COI) has become a useful heuristic tool for aiding species resolution and informing species discovery. Despite concerted efforts to genetically barcode bats and birds, comprehensive assessments have yet to be undertaken across the Afrotropics. We retrieved available DNA barcodes of native breeding Afrotropical bat and bird species. Using Bayesian phylogenetic modelling, we assessed DNA barcode performance at species identification, and sought to detect notable intraspecific clade partitioning hinting at cryptic speciation. Available DNA barcodes represent only 42.3% and 23.6% of the relevant bat and bird species diversity, respectively, with only 18.7% of bat species and 7.2% of bird species having geographically spread records. DNA barcodes afforded greater taxonomic resolution of Afrotropical bird species than of bats (96.8% vs. 84.0%), with bats having a higher proportion of species non-monophyly (25.5% vs. 4.8%). Well-supported (≥ 95% posterior probability) clade partitioning was inferable from twenty-one bat species and fifteen bird species, and a further single under-sampled bat species and fifteen such bird species showed deep (> 2.0%) intraspecific divergences. These phylogenetic signatures allude to cryptic speciation within these volant taxa, and serve to prompt more comprehensive assessments of Afrotropical fauna. These findings also indirectly affirm the importance of paleoclimatic refugia to endemic vertebrate diversity. The current taxonomic status of birds is better supported by this molecular evidence than that of bats.