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      A molecular study of Neophyllaphisvaricolor (Hemiptera, Aphididae) in Costa Rica

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          The genus Neophyllaphis (Takahashi) ( Aphididae : Neophyllaphidinae ) is composed of 18 species; however, in the Americas only nine species have been reported previously. A new species, Neophyllaphis varicolor Miller & Halbert, was described in 2014 in USA. Colonies resembling those of this new species have been observed in Costa Rica on Podocarpus spp. In order to determine if N. varicolor is also present in Costa Rica, we sampled Neophyllaphis colonies from Podocarpus falcatus and P. chinensis . Additionally, we sampled individuals from Podocarpus sp. in Spain and Vietnam. DNA of each sample was extracted and used to amplify and sequence the cytochrome c oxidase subunit I (COI) and elongation factor I (EF-1α) partial regions. According to morphological characteristics, sequences comparisons done in GenBank and BOLD, and phylogenetic analyses, the colonies collected from Podocarpus spp. in Costa Rica and the colony from Vietnam corresponded to the species N. varicolor . To the best of our knowledge this is the first report of the presence of N. varicolor in Central America and Vietnam.

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            DNA barcodes distinguish species of tropical Lepidoptera.

            Although central to much biological research, the identification of species is often difficult. The use of DNA barcodes, short DNA sequences from a standardized region of the genome, has recently been proposed as a tool to facilitate species identification and discovery. However, the effectiveness of DNA barcoding for identifying specimens in species-rich tropical biotas is unknown. Here we show that cytochrome c oxidase I DNA barcodes effectively discriminate among species in three Lepidoptera families from Area de Conservación Guanacaste in northwestern Costa Rica. We found that 97.9% of the 521 species recognized by prior taxonomic work possess distinctive cytochrome c oxidase I barcodes and that the few instances of interspecific sequence overlap involve very similar species. We also found two or more barcode clusters within each of 13 supposedly single species. Covariation between these clusters and morphological and/or ecological traits indicates overlooked species complexes. If these results are general, DNA barcoding will significantly aid species identification and discovery in tropical settings.
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              Critical factors for assembling a high volume of DNA barcodes.

              Large-scale DNA barcoding projects are now moving toward activation while the creation of a comprehensive barcode library for eukaryotes will ultimately require the acquisition of some 100 million barcodes. To satisfy this need, analytical facilities must adopt protocols that can support the rapid, cost-effective assembly of barcodes. In this paper we discuss the prospects for establishing high volume DNA barcoding facilities by evaluating key steps in the analytical chain from specimens to barcodes. Alliances with members of the taxonomic community represent the most effective strategy for provisioning the analytical chain with specimens. The optimal protocols for DNA extraction and subsequent PCR amplification of the barcode region depend strongly on their condition, but production targets of 100K barcode records per year are now feasible for facilities working with compliant specimens. The analysis of museum collections is currently challenging, but PCR cocktails that combine polymerases with repair enzyme(s) promise future success. Barcode analysis is already a cost-effective option for species identification in some situations and this will increasingly be the case as reference libraries are assembled and analytical protocols are simplified.

                Author and article information

                Pensoft Publishers
                22 July 2019
                : 865
                : 123-135
                [1 ] Universidad de Costa Rica, Escuela de Agronomía, San José, 11501-2060, Costa Rica
                [2 ] Universidad de Costa Rica, Centro de Investigación en Biología Celular y Molecular, San José, 11501-2060, Costa Rica
                [3 ] Nong Lam University, Research Institute for Biotechnology and Environment, Ho Chi Minh City, 700000, Vietnam
                [4 ] Instituto de Biología Integrativa de Sistemas (I2SysBio), Centro Mixto Universidad de Valencia-CSIC, Valencia, 46980, España
                Author notes
                Corresponding author: Nicolás Pérez Hidalgo ( nipehi@ )

                Academic editor: Roger Blackman

                Adonay Zúñiga-Centeno, Izayana Sandoval-Carvajal, Mauricio Montero-Astúa, William Villalobos-Muller, Nguyễn Bảo Quốc, Nicolás Pérez Hidalgo

                This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                This research and the scientific visit of NPH to the CIBCM in 2014, were funded by University of Costa Rica (grants 801-B7-169 and 801-A1-801) and the investigation in Spain actually is conducted in the context of Project CGL2015-68188-P, funded by “Ministerio de Economia, Industria y Competitividad” (MIMECO) of Spain.
                Research Article
                Biodiversity & Conservation
                Molecular systematics
                Central America and the Caribbean


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