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      Development of microsatellite markers in Pterodon pubescens and transferability to Pterodon emarginatus, two Brazilian plant species with medicinal potential

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          Abstract

          Abstract Pterodon pubescens and P. emarginatus (Leguminosae) are native medicinal plants of Brazil. Extractivism due to its therapeutic properties threatens populations of both species. Studies of genetic diversity is a way to reason the use and promote conservation. We developed microsatellite markers for P. pubescens and transferred them to P. emarginatus to further genetic diversity investigation of these species. From genomic sequences of P. pubescens, obtained via the Illumina MiSeq platform, it was possible to identify 6,514 microsatellite regions, to design 5,419 primer pairs, and to test 30 markers amplification. We provide 26 polymorphic microsatellite markers, 10 of which were genotyped in 48 individuals per species. The number of alleles per locus range from 3 to 16, with high average genetic diversity ( P. pubescens HE = 0.753; P. emarginatus HE = 0.691). The genotyped markers have a high paternity exclusion probability (Q values greater than 0.99) and low probability of identity, indicating that set of loci is capable of individual discriminating in P. pubescens and P. emarginatus. Microsatellite markers provided in this study are a tool for population genetics studies and conservation of the two species and can be applied to closely related non-model species.

          Translated abstract

          Resumo Pterodon pubescens e P. emarginatus (Leguminosae) são duas espécies de plantas nativas medicinais do Brasil. As populações têm sido ameaçadas pelo extrativismo para uso de suas propriedades terapêuticas. Estudos de diversidade genética ajudam a racionalizar o uso e a promover a conservação dessas espécies. Nós desenvolvemos marcadores microssatélites para P. pubescens e transferimos para P. emarginatus com intuito de permitir investigações futuras da diversidade genética das espécies. A partir de sequências genômicas de P. pubescens, obtidas via plataforma Illumina MiSeq, foi possível identificar 6.514 regiões microssatélites, desenhar 5.419 pares de primers, e testar a amplificação de 30 marcadores. Nós fornecemos 26 marcadores moleculares, 10 dos quais foram usados para genotipar 48 indivíduos de cada espécie. O número de alelos por locus variou de 3 a 16, com alta diversidade genética média para ambas as espécies ( P. pubescens HE = 0,753; P. emarginatus HE = 0,691). Os dez marcadores apresentaram boa probabilidade de exclusão de falsa paternidade (com valores acima de 0,99) e baixos valores de probabilidade de identidade, indicando que esse conjunto é adequado para discriminar indivíduos em P. pubescens e P. emarginatus. Os marcadores microssatélites desenvolvidos neste trabalho representam uma ferramenta promissora para estudos de genética de populações e conservação das duas espécies e podem, eventualmente, ser aplicados a espécies não-modelos filogeneticamente próximas.

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          Trimmomatic: a flexible trimmer for Illumina sequence data

          Motivation: Although many next-generation sequencing (NGS) read preprocessing tools already existed, we could not find any tool or combination of tools that met our requirements in terms of flexibility, correct handling of paired-end data and high performance. We have developed Trimmomatic as a more flexible and efficient preprocessing tool, which could correctly handle paired-end data. Results: The value of NGS read preprocessing is demonstrated for both reference-based and reference-free tasks. Trimmomatic is shown to produce output that is at least competitive with, and in many cases superior to, that produced by other tools, in all scenarios tested. Availability and implementation: Trimmomatic is licensed under GPL V3. It is cross-platform (Java 1.5+ required) and available at http://www.usadellab.org/cms/index.php?page=trimmomatic Contact: usadel@bio1.rwth-aachen.de Supplementary information: Supplementary data are available at Bioinformatics online.
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            Sequencing technologies - the next generation.

            Demand has never been greater for revolutionary technologies that deliver fast, inexpensive and accurate genome information. This challenge has catalysed the development of next-generation sequencing (NGS) technologies. The inexpensive production of large volumes of sequence data is the primary advantage over conventional methods. Here, I present a technical review of template preparation, sequencing and imaging, genome alignment and assembly approaches, and recent advances in current and near-term commercially available NGS instruments. I also outline the broad range of applications for NGS technologies, in addition to providing guidelines for platform selection to address biological questions of interest.
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              Genotyping errors: causes, consequences and solutions.

              Although genotyping errors affect most data and can markedly influence the biological conclusions of a study, they are too often neglected. Errors have various causes, but their occurrence and effect can be limited by considering these causes in the production and analysis of the data. Procedures that have been developed for dealing with errors in linkage studies, forensic analyses and non-invasive genotyping should be applied more broadly to any genetic study. We propose a protocol for estimating error rates and recommend that these measures be systemically reported to attest the reliability of published genotyping studies.
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                Author and article information

                Journal
                rod
                Rodriguésia
                Rodriguésia
                Instituto de Pesquisas Jardim Botânico do Rio de Janeiro (Rio de Janeiro, RJ, Brazil )
                0370-6583
                2175-7860
                2022
                : 73
                : e01112021
                Affiliations
                [1] Goiânia Goiás orgnameUniversidade Federal de Goiás orgdiv1Inst. Ciências Biológicas orgdiv2Lab. Genética & Biodiversidade Brazil
                [3] Goiânia Goiás orgnameUniversidade Federal de Goiás orgdiv1Rede Pró-Centro-Oeste orgdiv2Prog. Pós-graduação em Biotecnologia e Biodiversidade Brazil
                [2] Goiânia Goiás orgnameUniversidade Federal de Goiás orgdiv1Prog. Pós-graduação em Genética e Melhoramento de Plantas - Escola de Agronomia Brazil
                Article
                S2175-78602022000100256 S2175-7860(22)07300000256
                10.1590/2175-7860202273091
                56c28f66-a27f-4b25-bd30-2aed3903e865

                This work is licensed under a Creative Commons Attribution 4.0 International License.

                History
                : 19 July 2021
                : 18 April 2022
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 41, Pages: 0
                Product

                SciELO Brazil

                Categories
                Original Papers

                Illumina MiSeq,molecular markers,diversidade genética,“sucupira-branca”,neotropical tree,genetic diversity,árvores neotropicais,marcadores moleculares

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