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      A DNA barcode survey of insect biodiversity in Pakistan

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          Abstract

          Although Pakistan has rich biodiversity, many groups are poorly known, particularly insects. To address this gap, we employed DNA barcoding to survey its insect diversity. Specimens obtained through diverse collecting methods at 1,858 sites across Pakistan from 2010–2019 were examined for sequence variation in the 658 bp barcode region of the cytochrome c oxidase 1 (COI) gene. Sequences from nearly 49,000 specimens were assigned to 6,590 Barcode Index Numbers (BINs), a proxy for species, and most (88%) also possessed a representative image on the Barcode of Life Data System (BOLD). By coupling morphological inspections with barcode matches on BOLD, every BIN was assigned to an order (19) and most (99.8%) were placed to a family (362). However, just 40% of the BINs were assigned to a genus (1,375) and 21% to a species (1,364). Five orders (Coleoptera, Diptera, Hemiptera, Hymenoptera, Lepidoptera) accounted for 92% of the specimens and BINs. More than half of the BINs (59%) are so far only known from Pakistan, but others have also been reported from Bangladesh (13%), India (12%), and China (8%). Representing the first DNA barcode survey of the insect fauna in any South Asian country, this study provides the foundation for a complete inventory of the insect fauna in Pakistan while also contributing to the global DNA barcode reference library.

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          Biological identifications through DNA barcodes.

          Although much biological research depends upon species diagnoses, taxonomic expertise is collapsing. We are convinced that the sole prospect for a sustainable identification capability lies in the construction of systems that employ DNA sequences as taxon 'barcodes'. We establish that the mitochondrial gene cytochrome c oxidase I (COI) can serve as the core of a global bioidentification system for animals. First, we demonstrate that COI profiles, derived from the low-density sampling of higher taxonomic categories, ordinarily assign newly analysed taxa to the appropriate phylum or order. Second, we demonstrate that species-level assignments can be obtained by creating comprehensive COI profiles. A model COI profile, based upon the analysis of a single individual from each of 200 closely allied species of lepidopterans, was 100% successful in correctly identifying subsequent specimens. When fully developed, a COI identification system will provide a reliable, cost-effective and accessible solution to the current problem of species identification. Its assembly will also generate important new insights into the diversification of life and the rules of molecular evolution.
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            bold: The Barcode of Life Data System (http://www.barcodinglife.org)

            The Barcode of Life Data System (bold) is an informatics workbench aiding the acquisition, storage, analysis and publication of DNA barcode records. By assembling molecular, morphological and distributional data, it bridges a traditional bioinformatics chasm. bold is freely available to any researcher with interests in DNA barcoding. By providing specialized services, it aids the assembly of records that meet the standards needed to gain BARCODE designation in the global sequence databases. Because of its web-based delivery and flexible data security model, it is also well positioned to support projects that involve broad research alliances. This paper provides a brief introduction to the key elements of bold, discusses their functional capabilities, and concludes by examining computational resources and future prospects.
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              iNEXT: an R package for rarefaction and extrapolation of species diversity (Hill numbers)

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                Author and article information

                Contributors
                Journal
                PeerJ
                PeerJ
                PeerJ
                PeerJ
                PeerJ Inc. (San Diego, USA )
                2167-8359
                25 April 2022
                2022
                : 10
                : e13267
                Affiliations
                [1 ]Centre for Biodiversity Genomics & Department of Integrative Biology, University of Guelph , Guelph, Canada
                [2 ]Department of Biotechnology, University of Sargodha , Sargodha, Pakistan
                [3 ]Centre for Animal Sciences and Fisheries, University of Swat , Mingora, Pakistan
                [4 ]Directorate of Entomology, Ayub Agricultural Research Institute , Faisalabad, Pakistan
                [5 ]Department of Entomology, University of Poonch , Rawalakot, Azad Kashmir, Pakistan
                [6 ]Faculty of Life Sciences and Informatics, Balochistan University of Information Technology, Engineering and Management Sciences , Quetta, Pakistan
                [7 ]Department of Zoology, Lahore College for Women University , Lahore, Pakistan
                [8 ]Centre for Biodiversity Genomics, University of Guelph , Guelph, Canada
                [9 ]Institute of Zoology, University of the Punjab , Lahore, Pakistan
                [10 ]College of Agriculture, Bahauddin Zakariya University Bahadur Campus , Layyah, Pakistan
                [11 ]Pulses Research Institute, Ayub Agricultural Research Institute , Faisalabad, Pakistan
                [12 ]Faculty of Agriculture and Environmental Sciences, MNS University of Agriculture , Multan, Pakistan
                [13 ]Department of Zoology, University of Sindh , Jamshoro, Pakistan
                [14 ]Department of Zoology, Government College University Lahore , Lahore, Pakistan
                [15 ]National Insect Museum, National Agricultural Research Center , Islamabad, Pakistan
                [16 ]Department of Plant Pathology, Washington State University , Pullman, WA, United States
                [17 ]Department of Biology, University of Copenhagen , Copenhagen, Denmark
                [18 ]Government College Women University Faisalabad , Faisalabad, Pakistan
                [19 ]Ghazi University , Dera Ghazi Khan, Pakistan
                [20 ]Department of Zoology, Cholistan University of Veterinary and Animal Sciences , Bahawalpur, Pakistan
                [21 ]Department of Zoology, University of Narowal , Narowal, Pakistan
                [22 ]National Institute for Biotechnology and Genetic Engineering , Faisalabad, Pakistan
                Author information
                http://orcid.org/0000-0002-3081-6700
                Article
                13267
                10.7717/peerj.13267
                9048642
                35497186
                04087fd7-fd3c-4f38-90cc-ec149f0cf30c
                © 2022 Ashfaq et al.

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.

                History
                : 30 December 2021
                : 23 March 2022
                Funding
                Funded by: International Development Research Centre (IDRC) in Canada
                Award ID: 106106-001
                Funded by: Higher Education Commission of Pakistan
                Award ID: 20-1403/R& D/09
                Funded by: Government of Canada
                Funded by: Canada First Research Excellence Fund
                Funded by: New Frontiers in Research Fund
                This study was enabled by grant 106106-001 “Engaging Developing Nations in iBOL” from the International Development Research Centre (IDRC) in Canada and by grant HEC No. 20-1403/R& D/09 “Sequencing DNA Barcodes of Economically Important Insect Species from Pakistan” from the Higher Education Commission of Pakistan. Sequence analysis was made possible by a grant from the Government of Canada through Genome Canada and Ontario Genomics in support of the International Barcode of Life (iBOL) project. This is a contribution to the ‘Food from Thought’ project supported by the Canada First Research Excellence Fund. Paul Hebert and Nazeer Ahmed were supported by the New Frontiers in Research Fund through its award to BIOSCAN. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Ecology
                Entomology
                Genetics
                Taxonomy
                Population Biology

                dna barcoding,cytochrome c oxidase i,barcode index number,biodiversity overlap,bold

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