Transferibilidade e desenvolvimento de sistema multiplex de genotipagem de marcadores microssatélites para Pterodon emarginatus Vogel (Fabaceae)
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tede/4501 |
Resumo: | Microsatellite markers are widely used to assess the genetic variability in natural populations. These markers can be developed from derived Expressed sequence tag (ESTs) or genomic libraries. A feasible and economical alternative for obtaining of these markers for species that do not already have them is the transferability from primers designed for species evolutionarily close. The species Pterodon emarginatus Vogel (sucupira) belongs to the Fabaceae family and shows great potential for use timber and medicinal, which makes them a target of exploitation. Thus, studies that contribute to their use and conservation are needed, including the genetic population. Therefore, the aim of this study was to evaluate the potential for cross-amplification primers developed for the species Phaseolus vulgaris to P. emarginatus and to analyze the polymorphism of the loci transferred from multiplex genotyping in three populations of this species. For this, were evaluated 539 primers developed for Phaseolus vulgaris, with 345 derived from ESTs libraries and 194 genomics. Amplification tests were performed using three individuals, while the assessment initial of the polymorphism with eight. Finally, genetic diversity parameters were estimated using 88 plants from three natural populations of P. emarginatus. Primers that showed amplification products visualized on agarose gel were then evaluated in 6% acrylamide gel stained with silver nitrate. Polymorphic loci were synthesized with fluorescence for evaluation and genotyping by capillary electrophoresis in ABI3500 automatic DNA analyzer. The loci were transferred to P. emarginatus 23 (4 %) being 7 polymorphic. Polymorphic, 6 electropherograms profiles. These 6 loci were evaluated in populations along with three polymorphic genomic loci previously standardized sucupira, totaling nine loci suitable for genotyping. It was possible to develop two multiplex systems, a compound of 5primers and another for 4. The number of alleles per locus ranged from two to 13, with an average of five alleles per loci. The mean values of He and Ho for both populations were 0,563 and 0,463 respectively. The value of f was not significant for the total population and equal to 0,144. The values FIT and FST were significant and equal to 0,06 and 0,195 respectively. Microsatellite markers transferred and polymorphic for P. emarginatus showed polymorphism and can be used in studies genetic population more depth to the species. |
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Soares, Thannya Nascimentohttp://lattes.cnpq.br/5590256762396056Vianello, Rosana PereiraRodrigues, Flávia MeloSilva, Daniela de Melo eSoares, Thannya Nascimentohttp://lattes.cnpq.br/2986817803295152Santana, Ariane Rolins de2015-05-08T15:37:12Z2014-04-10SANTANA, A. R. Transferibilidade e desenvolvimento de sistema multiplex de genotipagem de marcadores microssatélites para Pterodon emarginatus Vogel (Fabaceae). 2014. 61 f. Dissertação (Mestrado em Genetica e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2014.http://repositorio.bc.ufg.br/tede/handle/tede/4501ark:/38995/001300000bwh3Microsatellite markers are widely used to assess the genetic variability in natural populations. These markers can be developed from derived Expressed sequence tag (ESTs) or genomic libraries. A feasible and economical alternative for obtaining of these markers for species that do not already have them is the transferability from primers designed for species evolutionarily close. The species Pterodon emarginatus Vogel (sucupira) belongs to the Fabaceae family and shows great potential for use timber and medicinal, which makes them a target of exploitation. Thus, studies that contribute to their use and conservation are needed, including the genetic population. Therefore, the aim of this study was to evaluate the potential for cross-amplification primers developed for the species Phaseolus vulgaris to P. emarginatus and to analyze the polymorphism of the loci transferred from multiplex genotyping in three populations of this species. For this, were evaluated 539 primers developed for Phaseolus vulgaris, with 345 derived from ESTs libraries and 194 genomics. Amplification tests were performed using three individuals, while the assessment initial of the polymorphism with eight. Finally, genetic diversity parameters were estimated using 88 plants from three natural populations of P. emarginatus. Primers that showed amplification products visualized on agarose gel were then evaluated in 6% acrylamide gel stained with silver nitrate. Polymorphic loci were synthesized with fluorescence for evaluation and genotyping by capillary electrophoresis in ABI3500 automatic DNA analyzer. The loci were transferred to P. emarginatus 23 (4 %) being 7 polymorphic. Polymorphic, 6 electropherograms profiles. These 6 loci were evaluated in populations along with three polymorphic genomic loci previously standardized sucupira, totaling nine loci suitable for genotyping. It was possible to develop two multiplex systems, a compound of 5primers and another for 4. The number of alleles per locus ranged from two to 13, with an average of five alleles per loci. The mean values of He and Ho for both populations were 0,563 and 0,463 respectively. The value of f was not significant for the total population and equal to 0,144. The values FIT and FST were significant and equal to 0,06 and 0,195 respectively. Microsatellite markers transferred and polymorphic for P. emarginatus showed polymorphism and can be used in studies genetic population more depth to the species.Os marcadores microssatélites são amplamente utilizados para acessar a variabilidade genética em populações naturais. Estes marcadores podem ser desenvolvidos a partir de bibliotecas derivadas de Sequências Expressas Transcritas (ESTs) ou genômicas. Uma alternativa possível e econômica para a obtenção destes marcadores para espécies que ainda não os possuem, é a transferibilidade a partir de primers desenhados para espécies evolutivamente próximas. A espécie Pterodon emarginatus Vogel (sucupira) pertence à família Fabaceae e apresenta grande potencial de uso medicinal e madeireiro, o que faz dela alvo da exploração extrativista. Assim, estudos que contribuam com o seu melhor uso e conservação são necessários, dentre eles os genético-populacionais. Diante disso, o objetivo deste estudo foi avaliar o potencial de amplificação cruzada de primers desenvolvidos para a espécie Phaseolus vulgaris para P. emarginatus e analisar o polimorfismo dos locos transferidos a partir de sistema multiplex de genotipagem em três populações desta espécie. Para tanto, foram avaliados 539 primers desenvolvidos para Phaseolus vulgaris, sendo 345 derivados de bibliotecas de ESTs e 194 genômicos. Os testes de amplificação foram realizados utilizando três indivíduos, enquanto a avaliação inicial do polimorfismo com oito. Por fim, os parâmetros genéticos de diversidade foram estimados utilizando 88 plantas, provenientes de três populações naturais de P. emarginatus. Os primers que apresentaram produtos de amplificação visualizados em gel de agarose foram, posteriormente, avaliados em gel de acrilamida 6% corado com nitrato de prata. Os locos polimórficos foram sintetizados com fluorescência para a avaliação e genotipagem via eletroforese capilar, no analisador automático de DNA ABI3500. Os locos transferidos para P. emarginatus foram 23 (4%) sendo 7 polimórficos. Dos polimórficos, 6 apresentaram bons perfis de eletroferogramas. Estes 6 locos foram avaliados nas populações juntamente com três locos genômicos polimórficos previamente padronizados para sucupira, totalizando nove locos adequados para genotipagem. Foi possível o desenvolvimento de dois sistemas multiplex, um composto de 5 primers e outro por 4. O número de alelos por loco variou de dois a 13, com uma média de cinco alelos por loco. Os valores médios de He e Ho para o conjunto das populações foram de 0,563 e 0,463 respectivamente. O valor de f não foi significativo para o total das populações e igual a 0,144. Os valores de FST e FIT foram significativos e iguais a 0,06 e 0,195 respectivamente. Os marcadores microssatélites transferidos e polimórficos para P. emarginatus apresentaram bom polimorfismo e podem ser utilizados em estudos genético-populacionais mais aprofundados com a espécie.Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2015-05-08T15:19:29Z No. of bitstreams: 2 Dissertação - Ariane Rolins de Santana - 2014.pdf: 1616629 bytes, checksum: 564437916d584de9d3769067e398b29b (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2015-05-08T15:37:12Z (GMT) No. of bitstreams: 2 Dissertação - Ariane Rolins de Santana - 2014.pdf: 1616629 bytes, checksum: 564437916d584de9d3769067e398b29b (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5)Made available in DSpace on 2015-05-08T15:37:12Z (GMT). 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dc.title.por.fl_str_mv |
Transferibilidade e desenvolvimento de sistema multiplex de genotipagem de marcadores microssatélites para Pterodon emarginatus Vogel (Fabaceae) |
dc.title.alternative.eng.fl_str_mv |
Transferability and development of multiplex system of genotyping microsatellite markers for Pterodon emarginatus vogel (Fabaceae) |
title |
Transferibilidade e desenvolvimento de sistema multiplex de genotipagem de marcadores microssatélites para Pterodon emarginatus Vogel (Fabaceae) |
spellingShingle |
Transferibilidade e desenvolvimento de sistema multiplex de genotipagem de marcadores microssatélites para Pterodon emarginatus Vogel (Fabaceae) Santana, Ariane Rolins de Amplificação cruzada Cerrado Sucupira Diversidade genética SSRs Cross-amplification Cerrado Sucupira Genetic diversity SSRs CIENCIAS BIOLOGICAS |
title_short |
Transferibilidade e desenvolvimento de sistema multiplex de genotipagem de marcadores microssatélites para Pterodon emarginatus Vogel (Fabaceae) |
title_full |
Transferibilidade e desenvolvimento de sistema multiplex de genotipagem de marcadores microssatélites para Pterodon emarginatus Vogel (Fabaceae) |
title_fullStr |
Transferibilidade e desenvolvimento de sistema multiplex de genotipagem de marcadores microssatélites para Pterodon emarginatus Vogel (Fabaceae) |
title_full_unstemmed |
Transferibilidade e desenvolvimento de sistema multiplex de genotipagem de marcadores microssatélites para Pterodon emarginatus Vogel (Fabaceae) |
title_sort |
Transferibilidade e desenvolvimento de sistema multiplex de genotipagem de marcadores microssatélites para Pterodon emarginatus Vogel (Fabaceae) |
author |
Santana, Ariane Rolins de |
author_facet |
Santana, Ariane Rolins de |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Soares, Thannya Nascimento |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/5590256762396056 |
dc.contributor.advisor-co1.fl_str_mv |
Vianello, Rosana Pereira |
dc.contributor.referee1.fl_str_mv |
Rodrigues, Flávia Melo |
dc.contributor.referee2.fl_str_mv |
Silva, Daniela de Melo e |
dc.contributor.referee3.fl_str_mv |
Soares, Thannya Nascimento |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/2986817803295152 |
dc.contributor.author.fl_str_mv |
Santana, Ariane Rolins de |
contributor_str_mv |
Soares, Thannya Nascimento Vianello, Rosana Pereira Rodrigues, Flávia Melo Silva, Daniela de Melo e Soares, Thannya Nascimento |
dc.subject.por.fl_str_mv |
Amplificação cruzada Cerrado Sucupira Diversidade genética SSRs |
topic |
Amplificação cruzada Cerrado Sucupira Diversidade genética SSRs Cross-amplification Cerrado Sucupira Genetic diversity SSRs CIENCIAS BIOLOGICAS |
dc.subject.eng.fl_str_mv |
Cross-amplification Cerrado Sucupira Genetic diversity SSRs |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS |
description |
Microsatellite markers are widely used to assess the genetic variability in natural populations. These markers can be developed from derived Expressed sequence tag (ESTs) or genomic libraries. A feasible and economical alternative for obtaining of these markers for species that do not already have them is the transferability from primers designed for species evolutionarily close. The species Pterodon emarginatus Vogel (sucupira) belongs to the Fabaceae family and shows great potential for use timber and medicinal, which makes them a target of exploitation. Thus, studies that contribute to their use and conservation are needed, including the genetic population. Therefore, the aim of this study was to evaluate the potential for cross-amplification primers developed for the species Phaseolus vulgaris to P. emarginatus and to analyze the polymorphism of the loci transferred from multiplex genotyping in three populations of this species. For this, were evaluated 539 primers developed for Phaseolus vulgaris, with 345 derived from ESTs libraries and 194 genomics. Amplification tests were performed using three individuals, while the assessment initial of the polymorphism with eight. Finally, genetic diversity parameters were estimated using 88 plants from three natural populations of P. emarginatus. Primers that showed amplification products visualized on agarose gel were then evaluated in 6% acrylamide gel stained with silver nitrate. Polymorphic loci were synthesized with fluorescence for evaluation and genotyping by capillary electrophoresis in ABI3500 automatic DNA analyzer. The loci were transferred to P. emarginatus 23 (4 %) being 7 polymorphic. Polymorphic, 6 electropherograms profiles. These 6 loci were evaluated in populations along with three polymorphic genomic loci previously standardized sucupira, totaling nine loci suitable for genotyping. It was possible to develop two multiplex systems, a compound of 5primers and another for 4. The number of alleles per locus ranged from two to 13, with an average of five alleles per loci. The mean values of He and Ho for both populations were 0,563 and 0,463 respectively. The value of f was not significant for the total population and equal to 0,144. The values FIT and FST were significant and equal to 0,06 and 0,195 respectively. Microsatellite markers transferred and polymorphic for P. emarginatus showed polymorphism and can be used in studies genetic population more depth to the species. |
publishDate |
2014 |
dc.date.issued.fl_str_mv |
2014-04-10 |
dc.date.accessioned.fl_str_mv |
2015-05-08T15:37:12Z |
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 |
SANTANA, A. R. Transferibilidade e desenvolvimento de sistema multiplex de genotipagem de marcadores microssatélites para Pterodon emarginatus Vogel (Fabaceae). 2014. 61 f. Dissertação (Mestrado em Genetica e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2014. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tede/4501 |
dc.identifier.dark.fl_str_mv |
ark:/38995/001300000bwh3 |
identifier_str_mv |
SANTANA, A. R. Transferibilidade e desenvolvimento de sistema multiplex de genotipagem de marcadores microssatélites para Pterodon emarginatus Vogel (Fabaceae). 2014. 61 f. Dissertação (Mestrado em Genetica e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2014. ark:/38995/001300000bwh3 |
url |
http://repositorio.bc.ufg.br/tede/handle/tede/4501 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.program.fl_str_mv |
-7235106986317239737 |
dc.relation.confidence.fl_str_mv |
600 600 600 600 600 |
dc.relation.department.fl_str_mv |
-3872772117827373404 |
dc.relation.cnpq.fl_str_mv |
-3439178843068202161 |
dc.relation.sponsorship.fl_str_mv |
-2555911436985713659 -961409807440757778 |
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 Genetica e Biologia Molecular |
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UFG |
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Brasil |
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Instituto de Ciências Biológicas - ICB (RG) |
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Universidade Federal de Goiás |
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