Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae)

Detalhes bibliográficos
Autor(a) principal: Barateli, Luciana Oliveira
Data de Publicação: 2018
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFG
dARK ID: ark:/38995/0013000007397
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/9034
Resumo: Stryphnodendron adstringens presents a wide geographical distribution, being predominant in regions of Cerrado sensu stricto and is popularly known as "barbatimão". It is a species widely used in herbal medicine because of its anti-inflammatory, antibacterial and antiulcerogenic potential. The Cerrado’s removal of vegetation cover reduces significantly the number of several native species, among them S. adstringens. Thus, it is important that efficient strategies for the use and conservation of this species are drawn. In order to provide molecular tools for studies of genetic diversity and conservation of S. adstringens, the present work had the objective of developing microsatellite markers for the species. Genomic DNA was obtained from leaf tissue using the CTAB protocol. The identification of the microsatellite regions and the design of the primers were performed using the QDD program modules. From the identified microsatellite regions, 20 pairs of primers were designed, 14 of which flank microsatellite regions composed of dinucleotides, four by tetranucleotides and two by pentanucleotides. Initially, four individuals were used for the standardization tests of the PCR protocol and annealing temperatures. Subsequently, 48 individuals were selected, distributed in three populations, to evaluate polymorphism via 6% polyacrylamide gel. Of the 20 pairs of primers evaluated, 16 presented polymorphic amplification products and four monomorphic amplification products. Considering the 16 polymorphic markers, the number of alleles varied between two (SadH19) and 13 (SadH13), with a mean of seven alleles per locus. The observed heterozygosity (Ho) and expected (He) and PIC values were 0.506, 0.543, 0.635, respectively. The mean Hmax value founded (65,519) indicates values of genetic diversity that can be considered medians for this set of loci evaluated in three populations of S. adstringens. On the other hand, although genetic diversity is median, this set of 16 polymorphic markers exhibited a ombined probability of paternity exclusion high (0.9999983) and combined probability of genetic identity low (3,49x10-15). The analysis of variance of allelic frequencies presented significant values for two of the three estimated statistics with f not significant 0.050, significant θ equal to 0,329 and F in the overall value also significant 0.360. Thus, it can be concluded that the panel of polymorphic markers developed for S. adstringens is highly informative and indicated for population genetic studies for the species. Another important factor is that these markers can be tested in other evolutionarily close species for the availability of microsatellite markers, without the need to develop new primers.
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spelling Telles, Mariana Pires de Camposhttp://lattes.cnpq.br/4648436798023532Brito, Cíntia Pelegrineti Targueta de AzevedoSoares, Thannya NascimentoTelles, Mariana Pires de Camposhttp://lattes.cnpq.br/0489837542400486Barateli, Luciana Oliveira2018-11-05T14:49:58Z2018-08-27BARATELI, L. O. Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae). 2018. 59 f. Dissertação (Mestrado em Genética e Melhoramento de Plantas) - Universidade Federal de Goiás, Goiânia, 2018.http://repositorio.bc.ufg.br/tede/handle/tede/9034ark:/38995/0013000007397Stryphnodendron adstringens presents a wide geographical distribution, being predominant in regions of Cerrado sensu stricto and is popularly known as "barbatimão". It is a species widely used in herbal medicine because of its anti-inflammatory, antibacterial and antiulcerogenic potential. The Cerrado’s removal of vegetation cover reduces significantly the number of several native species, among them S. adstringens. Thus, it is important that efficient strategies for the use and conservation of this species are drawn. In order to provide molecular tools for studies of genetic diversity and conservation of S. adstringens, the present work had the objective of developing microsatellite markers for the species. Genomic DNA was obtained from leaf tissue using the CTAB protocol. The identification of the microsatellite regions and the design of the primers were performed using the QDD program modules. From the identified microsatellite regions, 20 pairs of primers were designed, 14 of which flank microsatellite regions composed of dinucleotides, four by tetranucleotides and two by pentanucleotides. Initially, four individuals were used for the standardization tests of the PCR protocol and annealing temperatures. Subsequently, 48 individuals were selected, distributed in three populations, to evaluate polymorphism via 6% polyacrylamide gel. Of the 20 pairs of primers evaluated, 16 presented polymorphic amplification products and four monomorphic amplification products. Considering the 16 polymorphic markers, the number of alleles varied between two (SadH19) and 13 (SadH13), with a mean of seven alleles per locus. The observed heterozygosity (Ho) and expected (He) and PIC values were 0.506, 0.543, 0.635, respectively. The mean Hmax value founded (65,519) indicates values of genetic diversity that can be considered medians for this set of loci evaluated in three populations of S. adstringens. On the other hand, although genetic diversity is median, this set of 16 polymorphic markers exhibited a ombined probability of paternity exclusion high (0.9999983) and combined probability of genetic identity low (3,49x10-15). The analysis of variance of allelic frequencies presented significant values for two of the three estimated statistics with f not significant 0.050, significant θ equal to 0,329 and F in the overall value also significant 0.360. Thus, it can be concluded that the panel of polymorphic markers developed for S. adstringens is highly informative and indicated for population genetic studies for the species. Another important factor is that these markers can be tested in other evolutionarily close species for the availability of microsatellite markers, without the need to develop new primers.Stryphnodendron adstringens apresenta ampla distribuição geográfica, sendo predominante em regiões de Cerrado stricto sensu e é conhecida popularmente como “barbatimão”. É uma espécie utilizada na medicina fitoterápica por seu potencial anti-inflamatório, antibacteriano e antiulcerogênico. A retirada de cobertura vegetal do Cerrado diminui significativamente o número de diversas espécies nativas, dentre elas S. adstringens. Dessa forma, é importante que sejam traçadas estratégias eficientes para o uso e a conservação dessa espécie. A fim de disponibilizar ferramentas moleculares para estudos de diversidade genética e conservação de S. adstringens, o objetivo desse estudo foi desenvolver marcadores microssatélites e estimar o potencial informativo desses marcadores para subsidiar estudos de genética populacional da espécie. A identificação das regiões microssatélites e o desenho dos primers foram realizados usando o programa QDD. Inicialmente, foram utilizados quatro indivíduos para os testes de padronização do protocolo de PCR e temperaturas de anelamento. Posteriormente, foram selecionados 48 indivíduos, provenientes de três populações, para avaliação de polimorfismo via gel de poliacrilamida a 6%. A partir das regiões microssatélites identificadas, foram desenhados 20 pares de primers, sendo que 14 flanqueiam regiões microssatélites compostas por dinucleotídeos, quatro por tetranucleotídeos e dois por pentanucleotídeos. Dos 20 pares de primers avaliados, 16 apresentaram produtos de amplificação polimórficos e quatro monomórficos. Considerando os 16 marcadores polimórficos, o número de alelos variou entre dois (SadH19) e 13 (SadH13), com uma média igual a sete alelos por loco. Os valores das médias da heterozigosidade observada (Ho) e esperada (He) e do PIC foram iguais a 0,506, 0,543, 0,635 respectivamente. A proporção da diversidade máxima encontrada foi de 65%, o que indica valores de diversidade genética que podem ser considerados medianos para esse conjunto de locos avaliados em três populações de S. adstringens. Por outro lado, embora a diversidade genética seja mediana, esse conjunto de 16 marcadores polimórficos exibiu uma probabilidade de exclusão de paternidade combinada alta (0,999983) e uma probabilidade de identidade combinada baixa (3,49x10-15). A análise de variância das frequências alélicas apresentou valores significativos para duas das três estatísticas estimadas com f não significativo 0,050, θ significativo igual a 0,329 e F no valor global também significativo 0,360. Assim, pode-se concluir que o painel de marcadores polimórficos desenvolvidos para S. adstringens é altamente informativo e indicado para estudos genéticos populacionais para a espécie. Outro fator importante é que esses marcadores podem ser testados em outras espécies próximas evolutivamente para possível disponibilização de marcadores microssatélites, sem a necessidade de desenvolvimento de novos primers.Submitted by Franciele Moreira (francielemoreyra@gmail.com) on 2018-11-05T13:39:32Z No. of bitstreams: 2 Dissertação - Luciana Oliveira Barateli - 2018.pdf: 3641421 bytes, checksum: a96d12f5fce8d5c02a9793dec50ebfbf (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-11-05T14:49:58Z (GMT) No. of bitstreams: 2 Dissertação - Luciana Oliveira Barateli - 2018.pdf: 3641421 bytes, checksum: a96d12f5fce8d5c02a9793dec50ebfbf (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2018-11-05T14:49:58Z (GMT). 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dc.title.eng.fl_str_mv Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae)
dc.title.alternative.eng.fl_str_mv Development of microssatellite markers for Stryphnodendron adstringens (barbatimão - Fabaceae)
title Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae)
spellingShingle Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae)
Barateli, Luciana Oliveira
Barbatimão
Cerrado
Planta medicinal
SSR
Medicinal plant
CIENCIAS AGRARIAS::AGRONOMIA
title_short Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae)
title_full Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae)
title_fullStr Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae)
title_full_unstemmed Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae)
title_sort Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae)
author Barateli, Luciana Oliveira
author_facet Barateli, Luciana Oliveira
author_role author
dc.contributor.advisor1.fl_str_mv Telles, Mariana Pires de Campos
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/4648436798023532
dc.contributor.referee1.fl_str_mv Brito, Cíntia Pelegrineti Targueta de Azevedo
dc.contributor.referee2.fl_str_mv Soares, Thannya Nascimento
dc.contributor.referee3.fl_str_mv Telles, Mariana Pires de Campos
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/0489837542400486
dc.contributor.author.fl_str_mv Barateli, Luciana Oliveira
contributor_str_mv Telles, Mariana Pires de Campos
Brito, Cíntia Pelegrineti Targueta de Azevedo
Soares, Thannya Nascimento
Telles, Mariana Pires de Campos
dc.subject.por.fl_str_mv Barbatimão
Cerrado
Planta medicinal
SSR
topic Barbatimão
Cerrado
Planta medicinal
SSR
Medicinal plant
CIENCIAS AGRARIAS::AGRONOMIA
dc.subject.eng.fl_str_mv Medicinal plant
dc.subject.cnpq.fl_str_mv CIENCIAS AGRARIAS::AGRONOMIA
description Stryphnodendron adstringens presents a wide geographical distribution, being predominant in regions of Cerrado sensu stricto and is popularly known as "barbatimão". It is a species widely used in herbal medicine because of its anti-inflammatory, antibacterial and antiulcerogenic potential. The Cerrado’s removal of vegetation cover reduces significantly the number of several native species, among them S. adstringens. Thus, it is important that efficient strategies for the use and conservation of this species are drawn. In order to provide molecular tools for studies of genetic diversity and conservation of S. adstringens, the present work had the objective of developing microsatellite markers for the species. Genomic DNA was obtained from leaf tissue using the CTAB protocol. The identification of the microsatellite regions and the design of the primers were performed using the QDD program modules. From the identified microsatellite regions, 20 pairs of primers were designed, 14 of which flank microsatellite regions composed of dinucleotides, four by tetranucleotides and two by pentanucleotides. Initially, four individuals were used for the standardization tests of the PCR protocol and annealing temperatures. Subsequently, 48 individuals were selected, distributed in three populations, to evaluate polymorphism via 6% polyacrylamide gel. Of the 20 pairs of primers evaluated, 16 presented polymorphic amplification products and four monomorphic amplification products. Considering the 16 polymorphic markers, the number of alleles varied between two (SadH19) and 13 (SadH13), with a mean of seven alleles per locus. The observed heterozygosity (Ho) and expected (He) and PIC values were 0.506, 0.543, 0.635, respectively. The mean Hmax value founded (65,519) indicates values of genetic diversity that can be considered medians for this set of loci evaluated in three populations of S. adstringens. On the other hand, although genetic diversity is median, this set of 16 polymorphic markers exhibited a ombined probability of paternity exclusion high (0.9999983) and combined probability of genetic identity low (3,49x10-15). The analysis of variance of allelic frequencies presented significant values for two of the three estimated statistics with f not significant 0.050, significant θ equal to 0,329 and F in the overall value also significant 0.360. Thus, it can be concluded that the panel of polymorphic markers developed for S. adstringens is highly informative and indicated for population genetic studies for the species. Another important factor is that these markers can be tested in other evolutionarily close species for the availability of microsatellite markers, without the need to develop new primers.
publishDate 2018
dc.date.accessioned.fl_str_mv 2018-11-05T14:49:58Z
dc.date.issued.fl_str_mv 2018-08-27
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv BARATELI, L. O. Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae). 2018. 59 f. Dissertação (Mestrado em Genética e Melhoramento de Plantas) - Universidade Federal de Goiás, Goiânia, 2018.
dc.identifier.uri.fl_str_mv http://repositorio.bc.ufg.br/tede/handle/tede/9034
dc.identifier.dark.fl_str_mv ark:/38995/0013000007397
identifier_str_mv BARATELI, L. O. Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae). 2018. 59 f. Dissertação (Mestrado em Genética e Melhoramento de Plantas) - Universidade Federal de Goiás, Goiânia, 2018.
ark:/38995/0013000007397
url http://repositorio.bc.ufg.br/tede/handle/tede/9034
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language por
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dc.relation.cnpq.fl_str_mv -3091138714907603907
dc.relation.sponsorship.fl_str_mv 2075167498588264571
dc.rights.driver.fl_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
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dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Goiás
dc.publisher.program.fl_str_mv Programa de Pós-graduação em Genética e Melhoramento de Plantas (EA)
dc.publisher.initials.fl_str_mv UFG
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Escola de Agronomia - EA (RG)
publisher.none.fl_str_mv Universidade Federal de Goiás
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFG
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http://repositorio.bc.ufg.br/tede/bitstreams/e8ab2045-0be3-4ef9-bef1-1942c91696d3/download
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