DNA barcoding the Lepidoptera inventory of a large complex tropical conserved wildland, Area de Conservacion Guanacaste, northwestern Costa Rica

Genome ◽  
2016 ◽  
Vol 59 (9) ◽  
pp. 641-660 ◽  
Author(s):  
Daniel H. Janzen ◽  
Winnie Hallwachs

The 37-year ongoing inventory of the estimated 15 000 species of Lepidoptera living in the 125 000 terrestrial hectares of Area de Conservacion Guanacaste, northwestern Costa Rica, has DNA barcode documented 11 000+ species, and the simultaneous inventory of at least 6000+ species of wild-caught caterpillars, plus 2700+ species of parasitoids. The inventory began with Victorian methodologies and species-level perceptions, but it was transformed in 2004 by the full application of DNA barcoding for specimen identification and species discovery. This tropical inventory of an extraordinarily species-rich and complex multidimensional trophic web has relied upon the sequencing services provided by the Canadian Centre for DNA Barcoding, and the informatics support from BOLD, the Barcode of Life Data Systems, major tools developed by the Centre for Biodiversity Genomics at the Biodiversity Institute of Ontario, and available to all through couriers and the internet. As biodiversity information flows from these many thousands of undescribed and often look-alike species through their transformations to usable product, we see that DNA barcoding, firmly married to our centuries-old morphology-, ecology-, microgeography-, and behavior-based ways of taxonomizing the wild world, has made possible what was impossible before 2004. We can now work with all the species that we find, as recognizable species-level units of biology. In this essay, we touch on some of the details of the mechanics of actually using DNA barcoding in an inventory.

Author(s):  
Takeru Nakazato

DNA barcoding technology has become employed widely for biodiversity and molecular biology researchers to identify species and analyze their phylogeny. Recently, DNA metabarcoding and environmental DNA (eDNA) technology have developed by expanding the concept of DNA barcoding. These techniques analyze the diversity and quantity of organisms within an environment by detecting biogenic DNA in water and soil. It is particularly popular for monitoring fish species living in rivers and lakes (Takahara et al. 2012). BOLD Systems (Barcode of Life Database systems, Ratnasingham and Hebert 2007) is a database for DNA barcoding, archiving 8.5 million of barcodes (as of August 2020) along with the voucher specimen, from which the DNA barcode sequence is derived, including taxonomy, collected country, and museum vouchered as metadata (e.g. https://www.boldsystems.org/index.php/Public_RecordView?processid=TRIBS054-16). Also, many barcoding data are submitted to GenBank (Sayers et al. 2020), which is a database for DNA sequences managed by NCBI (National Center for Biotechnology Information, US). The number of the records of DNA barcodes, i.e. COI (cytochrome c oxidase I) gene for animal, has grown significantly (Porter and Hajibabaei 2018). BOLD imports DNA barcoding data from GenBank, and lots of DNA barcoding data in GenBank are also assigned BOLD IDs. However, we have to refer to both BOLD and GenBank data when performing DNA barcoding. I have previously investigated the registration of DNA barcoding data in GenBank, especially the association with BOLD, using insects and flowering plants as examples (Nakazato 2019). Here, I surveyed the number of species covered by BOLD and GenBank. I used fish data as an example because eDNA research is particularly focused on fish. I downloaded all GenBank files for vertebrates from NCBI FTP (File Transfer Protocol) sites (as of November 2019). Of the GenBank fish entries, 86,958 (7.3%) were assigned BOLD identifiers (IDs). The NCBI taxonomy database has registrations for 39,127 species of fish, and 20,987 scientific names at the species level (i.e., excluding names that included sp., cf. or aff.). GenBank entries with BOLD IDs covered 11,784 species (30.1%) and 8,665 species-level names (41.3%). I also obtained whole "specimens and sequences combined data" for fish from BOLD systems (as of November 2019). In the BOLD, there are 273,426 entries that are registered as fish. Of these entries, 211,589 BOLD entries were assigned GenBank IDs, i.e. with values in “genbank_accession” column, and 121,748 entries were imported from GenBank, i.e. with "Mined from GenBank, NCBI" description in "institution_storing" column. The BOLD data covered 18,952 fish species and 15,063 species-level names, but 35,500 entries were assigned no species-level names and 22,123 entries were not even filled with family-level names. At the species level, 8,067 names co-occurred in GenBank and BOLD, with 6,997 BOLD-specific names and 599 GenBank-specific names. GenBank has 425,732 fish entries with voucher IDs, of which 340,386 were not assigned a BOLD ID. Of these 340,386 entries, 43,872 entries are registrations for COI genes, which could be candidates for DNA barcodes. These candidates include 4,201 species that are not included in BOLD, thus adding these data will enable us to identify 19,863 fish to the species level. For researchers, it would be very useful if both BOLD and GenBank DNA barcoding data could be searched in one place. For this purpose, it is necessary to integrate data from the two databases. A lot of biodiversity data are recorded based on the Darwin Core standard while DNA sequencing data are sometimes integrated or cross-linked by RDF (Resource Description Framework). It may not be technically difficult to integrate these data, but the species data referenced differ from the EoL (The Encyclopedia of Life) for BOLD and the NCBI taxonomy for GenBank, and the differences in taxonomic systems make it difficult to match by scientific name description. GenBank has fields for the latitude and longitude of the specimens sampled, and Porter and Hajibabaei 2018 argue that this information should be enhanced. However, this information may be better described in the specimen and occurrence databases. The integration of barcoding data with the specimen and occurrence data will solve these problems. Most importantly, it will save the researcher from having to register the same information in multiple databases. In the field of biodiversity, only DNA barcode sequences may have been focused on and used as gene sequences. The museomics community regards museum-preserved specimens as rich resources for DNA studies because their biodiversity information can accompany the extraction and analysis of their DNA (Nakazato 2018). GenBank is useful for biodiversity studies due to its low rate of mislabelling (Leray et al. 2019). In the future, we will be working with a variety of DNA, including genomes from museum specimens as well as DNA barcoding. This will require more integrated use of biodiversity information and DNA sequence data. This integration is also of interest to molecular biologists and bioinformaticians.


2020 ◽  
Vol 8 ◽  
Author(s):  
Sonia Ferreira ◽  
Rui Andrade ◽  
Ana Gonçalves ◽  
Pedro Sousa ◽  
Joana Paupério ◽  
...  

The InBIO Barcoding Initiative (IBI) Diptera 01 dataset contains records of 203 specimens of Diptera. All specimens have been morphologically identified to species level, and belong to 154 species in total. The species represented in this dataset correspond to about 10% of continental Portugal dipteran species diversity. All specimens were collected north of the Tagus river in Portugal. Sampling took place from 2014 to 2018, and specimens are deposited in the IBI collection at CIBIO, Research Center in Biodiversity and Genetic Resources. This dataset contributes to the knowledge on the DNA barcodes and distribution of 154 species of Diptera from Portugal and is the first of the planned IBI database public releases, which will make available genetic and distribution data for a series of taxa. All specimens have their DNA barcodes made publicly available in the Barcode of Life Data System (BOLD) online database and the distribution dataset can be freely accessed through the Global Biodiversity Information Facility (GBIF).


Genome ◽  
2018 ◽  
Vol 61 (1) ◽  
pp. 21-31 ◽  
Author(s):  
Jason Gibbs

There is an ongoing campaign to DNA barcode the world’s >20 000 bee species. Recent revisions of Lasioglossum (Dialictus) (Hymenoptera: Halictidae) for Canada and the eastern United States were completed using integrative taxonomy. DNA barcode data from 110 species of L. (Dialictus) are examined for their value in identification and discovering additional taxonomic diversity. Specimen identification success was estimated using the best close match method. Error rates were 20% relative to current taxonomic understanding. Barcode Index Numbers (BINs) assigned using Refined Single Linkage Analysis (RESL) and barcode gaps using the Automatic Barcode Gap Discovery (ABGD) method were also assessed. RESL was incongruent for 44.5% of species, although some cryptic diversity may exist. Forty-three of 110 species were part of merged BINs with multiple species. The barcode gap is non-existent for the data set as a whole and ABGD showed levels of discordance similar to the RESL. The viridatum species-group is particularly problematic, so that DNA barcodes alone would be misleading for species delimitation and specimen identification. Character-based methods using fixed nucleotide substitutions could improve specimen identification success in some cases. The use of DNA barcoding for species discovery for standard taxonomic practice in the absence of a well-defined barcode gap is discussed.


2021 ◽  
Vol 9 ◽  
Author(s):  
Parvin Aghayeva ◽  
Salvatore Cozzolino ◽  
Donata Cafasso ◽  
Valida Ali-zade ◽  
Silvia Fineschi ◽  
...  

DNA barcoding has rapidly become a useful complementary tool in floristic investigations particularly for identifying specimens that lack diagnostic characters. Here, we assess the capability of three DNA barcode markers (chloroplast rpoB, accD and nuclear ITS) for correct species assignment in a floristic survey on the Caucasus. We focused on two herbal groups with potential for ornamental applications, namely orchids and asterids. On these two plant groups, we tested whether our selection of barcode markers allows identification of the “barcoding gap” in sequence identity and to distinguish between monophyletic species when employing distance-based methods. All markers successfully amplified most specimens, but we found that the rate of species-level resolution amongst selected markers largely varied in the two plant groups. Overall, for both lineages, plastid markers had a species-level assignment success rate lower than the nuclear ITS marker. The latter confirmed, in orchids, both the existence of a barcoding gap and that all accessions of the same species clustered together in monophyletic groups. Further, it also allowed the detection of a phylogeographic signal.The ITS marker resulted in its being the best performing barcode for asterids; however, none of the three tested markers showed high discriminatory ability. Even if ITS were revealed as the most promising plant barcode marker, we argue that the ability of this barcode for species assignment is strongly dependent on the evolutionary history of the investigated plant lineage.


2022 ◽  
Vol 2022 ◽  
pp. 1-19
Author(s):  
Sarfraz Ahmed ◽  
Muhammad Ibrahim ◽  
Chanin Nantasenamat ◽  
Muhammad Farrukh Nisar ◽  
Aijaz Ahmad Malik ◽  
...  

DNA barcodes are regarded as hereditary succession codes that serve as a recognition marker to address several queries relating to the identification, classification, community ecology, and evolution of certain functional traits in organisms. The mitochondrial cytochrome c oxidase 1 (CO1) gene as a DNA barcode is highly efficient for discriminating vertebrate and invertebrate animal species. Similarly, different specific markers are used for other organisms, including ribulose bisphosphate carboxylase (rbcL), maturase kinase (matK), transfer RNA-H and photosystem II D1-ApbsArabidopsis thaliana (trnH-psbA), and internal transcribed spacer (ITS) for plant species; 16S ribosomal RNA (16S rRNA), elongation factor Tu gene (Tuf gene), and chaperonin for bacterial strains; and nuclear ITS for fungal strains. Nevertheless, the taxon coverage of reference sequences is far from complete for genus or species-level identification. Applying the next-generation sequencing approach to the parallel acquisition of DNA barcode sequences could greatly expand the potential for library preparation or accurate identification in biodiversity research. Overall, this review articulates on the DNA barcoding technology as applied to different organisms, its universality, applicability, and innovative approach to handling DNA-based species identification.


Botany ◽  
2019 ◽  
Vol 97 (9) ◽  
pp. 503-512 ◽  
Author(s):  
Deniz Aygoren Uluer ◽  
Rahma Alshamrani

Aesculus L. is a small genus of horticulturally important trees and shrubs, comprising 13–19 species. Frequent hybridization among species, particularly in cultivation, has contributed to taxonomic confusion and difficulties in the identification of plants. In this study, we evaluated three widely employed plant DNA barcode loci, matK, and the entire ITS region (ITS1+5.8S+ITS2) as well as subunit ITS2 for 50 individuals representing 13 species of Aesculus, excluding only A. wangii (=A. assamica). In contrast to the plastid matK region, both the ITS and ITS2 loci displayed low levels of species discrimination, especially in our “first hit” BLASTn searches. We also presented the phylogeny of Aesculus based on matK and the entire ITS region, with additional matK and ITS sequences from GenBank. Our results show that Aesculus chinensis, A. flava, A. glabra, A. pavia, and A. sylvatica are probably not monophyletic. Furthermore, with the widest taxon coverage until now, the current study highlights the importance of sampling multiple individuals, not only for DNA barcoding, but also for phylogenetic studies.


2014 ◽  
Vol 59 (2) ◽  
Author(s):  
Bjørn Rukke ◽  
Symira Cholidis ◽  
Arild Johnsen ◽  
Preben Ottesen

AbstractDNA barcoding is a practical tool for species identification, when morphological classification of an organism is difficult. Herein we describe the utilisation of this technique in a case of ophthalmomyiasis interna. A 12-year-old boy was infested during a summer holiday in northern Norway, while visiting an area populated with reindeer. Following medical examination, a Diptera larva was surgically removed from the boy’s eye and tentatively identified from its morphological traits as Hypoderma tarandi (L.) (Diptera: Oestridae). Ultimately, DNA barcoding confirmed this impression. The larval cytochrome c oxidase subunit 1 (COI) DNA sequence was matched with both profiles of five adult H. tarandi from the same region where the boy was infested, and other established profiles of H. tarandi in the Barcode of Life Data Systems (BOLD) identification engine.


2021 ◽  
Vol 9 ◽  
Author(s):  
Sónia Ferreira ◽  
Pjotr Oosterbroek ◽  
Jaroslav Starý ◽  
Pedro Sousa ◽  
Vanessa Mata ◽  
...  

The InBIO Barcoding Initiative (IBI) Diptera 02 dataset contains records of 412 crane fly specimens belonging to the Diptera families: Limoniidae, Pediciidae and Tipulidae. This dataset is the second release by IBI on Diptera and it greatly increases the knowledge on the DNA barcodes and distribution of crane flies from Portugal. All specimens were collected in Portugal, including six specimens from the Azores and Madeira archipelagos. Sampling took place from 2003 to 2019. Specimens have been morphologically identified to species level by taxonomists and belong to 83 species in total. The species, represented in this dataset, correspond to about 55% of all the crane fly species known from Portugal and 22% of crane fly species known from the Iberian Peninsula. All DNA extractions and most specimens are deposited in the IBI collection at CIBIO, Research Center in Biodiversity and Genetic Resources. Fifty-three species were new additions to the Barcode of Life Data System (BOLD), with another 18 species' barcodes added from under-represented species in BOLD. Furthermore, the submitted sequences were found to cluster in 88 BINs, 54 of which were new to BOLD. All specimens have their DNA barcodes publicly accessible through BOLD online database and its collection data can be accessed through the Global Biodiversity Information Facility (GBIF). One species, Gonomyia tenella (Limoniidae), is recorded for the first time from Portugal, raising the number of crane flies recorded in the country to 145 species.


2020 ◽  
Vol 21 (8) ◽  
Author(s):  
Viet The Ho ◽  
MINH PHUONG NGUYEN

Abstract. Ho VT, Nguyen MP. 2020. An in silico approach for evaluation of rbcL and matK loci for DNA barcoding of Cucurbitaceae family. Biodiversitas 21: 3879-3885. DNA barcodes have been used intensively to discriminate different species in Cucurbitaceae family. The main of this study is to evaluate the effectiveness of rbcL and matK loci for 16 species of Cucurbitaceae family by using in silico approach. For analysis, sequences were firstly retrieved from NCBI and then calculated for sequence parameters. Sequences were then aligned and constructed phylogenetic try and examined for species resolution ability. The obtained data show the variability of resolving capacity among species. rbcL region is suitable for distinguishing five species namely S. edule, M. cochinchinensis, L. aegyptiaca, C. melo, and C. pepo, whereas matK locus is more proper for different five species consisting of M. balsamina, M. cochinchinensis, M. charantia, S. edule, and C. sativus. The resolving power is improved sharply by analyzing the rbcL + matK combination with up to nine species consisting of C. lanatus, B. hispida, C. melo, C. sativus, C. pepo, C. agryrosperma, L. aegyptiaca, S. edule, and M. cochinchinensis. Therefore, the integration of rbcL and matK loci may improve the competence of assessing genetic relatedness at species level of members in Cucurbitaceae family. The obtained information could be important for choosing proper DNA barcode loci for phylogenetic study of this crop family.


Author(s):  
Mikhail V. Chesalin ◽  
Salem A. Al-Ghassani ◽  
Mohammed A. Balkhair

Oysters are commonly found on rocky shores along the Dhofar coast of Oman, but the species composition has not yet been studied. In an attempt to determine the species of these oysters, samples from four sites (Mirbat, Sadah, Hadbin, and Hasik) were collected and analyzed using DNA sequences that were compared with the database of GenBank/National Center for Biotechnology Information (NCBI) and Barcoding of Life Data Systems (BOLD). BLAST results of DNA sequences showed that there were no existing sequences available on NCBI/GenBank and BOLD. The DNA barcode sequences generated in this study can be employed as standards for this species and may be of use in taxonomic studies. Phylogenetic analysis demonstrated that the oysters from Oman exhibit homology with Saccostrea species cucullata or mordax. 


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