Desenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae)
Autor(a) principal: | |
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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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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). No. of bitstreams: 2 Dissertação - Luciana Oliveira Barateli - 2018.pdf: 3641421 bytes, checksum: a96d12f5fce8d5c02a9793dec50ebfbf (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-08-27Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESapplication/pdfporUniversidade Federal de GoiásPrograma de Pós-graduação em Genética e Melhoramento de Plantas (EA)UFGBrasilEscola de Agronomia - EA (RG)http://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessBarbatimãoCerradoPlanta medicinalSSRMedicinal plantCIENCIAS AGRARIAS::AGRONOMIADesenvolvimento de marcadores microssatélites para Stryphnodendron adstringens (barbatimão - Fabaceae)Development of microssatellite markers for Stryphnodendron adstringens (barbatimão - Fabaceae)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis-6265679607231828330600600600600-6046953723502374070-30911387149076039072075167498588264571reponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGLICENSElicense.txtlicense.txttext/plain; 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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 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.program.fl_str_mv |
-6265679607231828330 |
dc.relation.confidence.fl_str_mv |
600 600 600 600 |
dc.relation.department.fl_str_mv |
-6046953723502374070 |
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/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
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 |
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Repositório Institucional da UFG |
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Repositório Institucional da UFG |
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Repositório Institucional da UFG - Universidade Federal de Goiás (UFG) |
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tasesdissertacoes.bc@ufg.br |
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