Análises moleculares em Melipona rufiventris e Melipona mondury (Hymenoptera: Apidae)

Detalhes bibliográficos
Autor(a) principal: Lopes, Denilce Meneses
Data de Publicação: 2008
Tipo de documento: Tese
Idioma: por
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: http://locus.ufv.br/handle/123456789/231
Resumo: Melipona mondury and M. rufiventris are two species of indigenous stingless bees which are very similar morphologically. Studies carried out with those species and different kinds of molecular markers revealed low genetic variability. Besides, populations of some regions revealed patterns which suggest that more than one species are receiving the same scientific name. Today, however, several methodologies make it possible to differentiate species and evaluate the genetic structure and the genic flow, besides supplying the basis for the understanding of other important aspects such as the genome evolution of different groups and organisms. So, in this work, it was tried to optimize tools that could help populational and evolutionary studies of the two mentioned species, which could also be used with other bee species. In order to achieve this, several methodologies were used: (1) the drawing of primers microsatellites for Melipona mondury and M. rufiventris and the evaluation of their amplification in other stingless bee species; (2) comparison between the polymorphism found with the specific primers and that of the heterologous primers; (3) analysis of the cytochrome B sequence and (4) standardization of the flow cytometry protocol, in order to determine the DNA content of bees and its use on M. rufiventris, M. mondury and Scaptotrigona xantrotricha. Using the ISSR technique, it was possible to achieve the drawing of 10 primers microsatellites for M. rufiventris and 11 for M. mondury, which presented the following genetic diversity values: H=0,47 and H=0,38, respectively. Those values were higher than those observed in amplifications in which it was used primers drawn for M. bicolor (H=0,16 and H=0,18 for M. mondury and M. rufiventris, respectively). In the analysis of cytochrome B sequences, eleven haplotypes were found which allowed the separation of three different groups, showing that could there are more than one species in these regions. Finally, the standardized methodology for the determination of the bee DNA content furnished good quality histograms, with variation coefficient values of 2,87-4,14%. It demonstrates the usefulness of this methodology in studies involving bees. The DNA 1C content values found for females of M. rufiventris, M. mondury and S. xantotricha were C = 0.93 pg, C = 0,95 pg and C = 0,44 pg, respectively, which reveals that the size of the genome of the two species of Melipona was about twice the size of the one belonging to S. xantotricha. This variation may show differences in the genomic structure of the species analyzed. However, further studies should be carried out to investigate the genome of other species of Melipona and other genders of stingless bees, in order to verify the processes involved in the expansion and contraction of their genomes.
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spelling Lopes, Denilce Meneseshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4707735U1Campos, Lúcio Antonio de Oliveirahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783908P9Salomão, Tânia Maria Fernandeshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787017A5Tavares, Mara Garciahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4798782P4Dias, Luiz Antonio dos Santoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763137P6Waldschmidt, Ana Mariahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4784763Z72015-03-26T12:10:34Z2008-11-062015-03-26T12:10:34Z2008-07-28LOPES, Denilce Meneses. Molecular analysis in Melipona rufiventris and Melipona mondury (Hymenoptera: Apidae). 2008. 96 f. Tese (Doutorado em Análises quantitativas e moleculares do Genoma; Biologia das células e dos tecidos) - Universidade Federal de Viçosa, Viçosa, 2008.http://locus.ufv.br/handle/123456789/231Melipona mondury and M. rufiventris are two species of indigenous stingless bees which are very similar morphologically. Studies carried out with those species and different kinds of molecular markers revealed low genetic variability. Besides, populations of some regions revealed patterns which suggest that more than one species are receiving the same scientific name. Today, however, several methodologies make it possible to differentiate species and evaluate the genetic structure and the genic flow, besides supplying the basis for the understanding of other important aspects such as the genome evolution of different groups and organisms. So, in this work, it was tried to optimize tools that could help populational and evolutionary studies of the two mentioned species, which could also be used with other bee species. In order to achieve this, several methodologies were used: (1) the drawing of primers microsatellites for Melipona mondury and M. rufiventris and the evaluation of their amplification in other stingless bee species; (2) comparison between the polymorphism found with the specific primers and that of the heterologous primers; (3) analysis of the cytochrome B sequence and (4) standardization of the flow cytometry protocol, in order to determine the DNA content of bees and its use on M. rufiventris, M. mondury and Scaptotrigona xantrotricha. Using the ISSR technique, it was possible to achieve the drawing of 10 primers microsatellites for M. rufiventris and 11 for M. mondury, which presented the following genetic diversity values: H=0,47 and H=0,38, respectively. Those values were higher than those observed in amplifications in which it was used primers drawn for M. bicolor (H=0,16 and H=0,18 for M. mondury and M. rufiventris, respectively). In the analysis of cytochrome B sequences, eleven haplotypes were found which allowed the separation of three different groups, showing that could there are more than one species in these regions. Finally, the standardized methodology for the determination of the bee DNA content furnished good quality histograms, with variation coefficient values of 2,87-4,14%. It demonstrates the usefulness of this methodology in studies involving bees. The DNA 1C content values found for females of M. rufiventris, M. mondury and S. xantotricha were C = 0.93 pg, C = 0,95 pg and C = 0,44 pg, respectively, which reveals that the size of the genome of the two species of Melipona was about twice the size of the one belonging to S. xantotricha. This variation may show differences in the genomic structure of the species analyzed. However, further studies should be carried out to investigate the genome of other species of Melipona and other genders of stingless bees, in order to verify the processes involved in the expansion and contraction of their genomes.Melipona mondury e M. rufiventris são duas espécies de abelhas indígenas sem ferrão morfologicamente bastante semelhantes. Estudos realizados com essas espécies utilizando diferentes tipos de marcadores moleculares mostraram baixa variabilidade genética. Além disso, populações de algumas regiões possuem características que sugerem que mais de uma espécie está recebendo o mesmo nome científico. Neste trabalho procurou-se otimizar ferramentas que pudessem ser utilizadas em estudos populacionais e evolutivos das duas espécies supracitadas, bem como de outras espécies de abelhas. Para isto, foram (1) desenhados primers microssatélites para Melipona mondury e M. rufiventris os quais foram também avaliados em outras espécies de meliponíneos, (2) foram comparados os niveis de polimorfismo encontrado utilizando primers específicos e primers heterólogos, (3) o gene do citocromo B foi parcialmente amplificado e a seqüência do mesmo foi comparada em populações das duas espécies e (4) padronizado um protocolo de citometria de fluxo, para determinação do conteúdo de DNA de abelhas e aplicação do mesmo em M. rufiventris, M. mondury e Scaptotrigona xantotricha. Utilizando-se a técnica de ISSR foram desenhados 10 primers microssatélites para M. rufiventris e 11 para M. mondury e a diversidade genética nessas espécies foi estimada utilizando esses primers e os valores encontrados foram H = 0,47 e H = 0,38, respectivamente. Estes valores são superiores àqueles observados em amplificações nas quais se utilizaram primers desenhados para M. bicolor (H = 0,16 e H = 0,18 para M. mondury e M. rufiventris, respectivamente). Na análise de seqüências do citocromo B encontraram-se onze haplótipos que permitiram a separação de dois grupos distintos, evidenciando a existência de mais de uma espécie nas localidades estudadas. Por fim, a metodologia padronizada para determinação do conteúdo de DNA de abelhas forneceu histogramas, com valores de coeficiente de variação de 2,87 a 4,14%, o que demonstra a utilidade da mesma em estudos envolvendo abelhas. Os valores de conteúdo de DNA 1C encontrados para fêmeas de M. rufiventris, M. mondury e S. xantotricha foram C = 0.93 pg, C = 0,95 pg e C = 0,44 pg, respectivamente, o que demonstra que o tamanho do genoma das duas espécies de Melipona foi cerca de duas vezes maior que o de S. xantotricha. Esta variação pode refletir diferenças na estrutura genômica das espécies analisadas. Entretanto, estudos adicionais deverão ser realizados no sentido de estudar o genoma de outras espécies de Melipona, bem como de outros gêneros de abelhas sem ferrão, a fim de verificar os processos envolvidos na expansão e/ou contração de seus genomas.Universidade Federal de Viçosaapplication/pdfporUniversidade Federal de ViçosaDoutorado em Biologia Celular e EstruturalUFVBRAnálises quantitativas e moleculares do Genoma; Biologia das células e dos tecidosHymenopteraMicrossatélitesCytbCitometria de fluxoHymenopteraMicrosatellitesCytbFlow cytometryCNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERALAnálises moleculares em Melipona rufiventris e Melipona mondury (Hymenoptera: Apidae)Molecular analysis in Melipona rufiventris and Melipona mondury (Hymenoptera: Apidae)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf584301https://locus.ufv.br//bitstream/123456789/231/1/texto%20completo.pdfce14d873d81b8a9a70e990fc1d11c7fcMD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain155743https://locus.ufv.br//bitstream/123456789/231/2/texto%20completo.pdf.txt1c9c334fdd62f4c661ff9f2dfc7e2999MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3593https://locus.ufv.br//bitstream/123456789/231/3/texto%20completo.pdf.jpg44ad11b6852dfada719da9dcaa99670fMD53123456789/2312016-04-06 23:00:16.174oai:locus.ufv.br:123456789/231Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-07T02:00:16LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Análises moleculares em Melipona rufiventris e Melipona mondury (Hymenoptera: Apidae)
dc.title.alternative.eng.fl_str_mv Molecular analysis in Melipona rufiventris and Melipona mondury (Hymenoptera: Apidae)
title Análises moleculares em Melipona rufiventris e Melipona mondury (Hymenoptera: Apidae)
spellingShingle Análises moleculares em Melipona rufiventris e Melipona mondury (Hymenoptera: Apidae)
Lopes, Denilce Meneses
Hymenoptera
Microssatélites
Cytb
Citometria de fluxo
Hymenoptera
Microsatellites
Cytb
Flow cytometry
CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
title_short Análises moleculares em Melipona rufiventris e Melipona mondury (Hymenoptera: Apidae)
title_full Análises moleculares em Melipona rufiventris e Melipona mondury (Hymenoptera: Apidae)
title_fullStr Análises moleculares em Melipona rufiventris e Melipona mondury (Hymenoptera: Apidae)
title_full_unstemmed Análises moleculares em Melipona rufiventris e Melipona mondury (Hymenoptera: Apidae)
title_sort Análises moleculares em Melipona rufiventris e Melipona mondury (Hymenoptera: Apidae)
author Lopes, Denilce Meneses
author_facet Lopes, Denilce Meneses
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4707735U1
dc.contributor.author.fl_str_mv Lopes, Denilce Meneses
dc.contributor.advisor-co1.fl_str_mv Campos, Lúcio Antonio de Oliveira
dc.contributor.advisor-co1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783908P9
dc.contributor.advisor-co2.fl_str_mv Salomão, Tânia Maria Fernandes
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787017A5
dc.contributor.advisor1.fl_str_mv Tavares, Mara Garcia
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4798782P4
dc.contributor.referee1.fl_str_mv Dias, Luiz Antonio dos Santos
dc.contributor.referee1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763137P6
dc.contributor.referee2.fl_str_mv Waldschmidt, Ana Maria
dc.contributor.referee2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4784763Z7
contributor_str_mv Campos, Lúcio Antonio de Oliveira
Salomão, Tânia Maria Fernandes
Tavares, Mara Garcia
Dias, Luiz Antonio dos Santos
Waldschmidt, Ana Maria
dc.subject.por.fl_str_mv Hymenoptera
Microssatélites
Cytb
Citometria de fluxo
topic Hymenoptera
Microssatélites
Cytb
Citometria de fluxo
Hymenoptera
Microsatellites
Cytb
Flow cytometry
CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
dc.subject.eng.fl_str_mv Hymenoptera
Microsatellites
Cytb
Flow cytometry
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
description Melipona mondury and M. rufiventris are two species of indigenous stingless bees which are very similar morphologically. Studies carried out with those species and different kinds of molecular markers revealed low genetic variability. Besides, populations of some regions revealed patterns which suggest that more than one species are receiving the same scientific name. Today, however, several methodologies make it possible to differentiate species and evaluate the genetic structure and the genic flow, besides supplying the basis for the understanding of other important aspects such as the genome evolution of different groups and organisms. So, in this work, it was tried to optimize tools that could help populational and evolutionary studies of the two mentioned species, which could also be used with other bee species. In order to achieve this, several methodologies were used: (1) the drawing of primers microsatellites for Melipona mondury and M. rufiventris and the evaluation of their amplification in other stingless bee species; (2) comparison between the polymorphism found with the specific primers and that of the heterologous primers; (3) analysis of the cytochrome B sequence and (4) standardization of the flow cytometry protocol, in order to determine the DNA content of bees and its use on M. rufiventris, M. mondury and Scaptotrigona xantrotricha. Using the ISSR technique, it was possible to achieve the drawing of 10 primers microsatellites for M. rufiventris and 11 for M. mondury, which presented the following genetic diversity values: H=0,47 and H=0,38, respectively. Those values were higher than those observed in amplifications in which it was used primers drawn for M. bicolor (H=0,16 and H=0,18 for M. mondury and M. rufiventris, respectively). In the analysis of cytochrome B sequences, eleven haplotypes were found which allowed the separation of three different groups, showing that could there are more than one species in these regions. Finally, the standardized methodology for the determination of the bee DNA content furnished good quality histograms, with variation coefficient values of 2,87-4,14%. It demonstrates the usefulness of this methodology in studies involving bees. The DNA 1C content values found for females of M. rufiventris, M. mondury and S. xantotricha were C = 0.93 pg, C = 0,95 pg and C = 0,44 pg, respectively, which reveals that the size of the genome of the two species of Melipona was about twice the size of the one belonging to S. xantotricha. This variation may show differences in the genomic structure of the species analyzed. However, further studies should be carried out to investigate the genome of other species of Melipona and other genders of stingless bees, in order to verify the processes involved in the expansion and contraction of their genomes.
publishDate 2008
dc.date.available.fl_str_mv 2008-11-06
2015-03-26T12:10:34Z
dc.date.issued.fl_str_mv 2008-07-28
dc.date.accessioned.fl_str_mv 2015-03-26T12:10:34Z
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dc.identifier.citation.fl_str_mv LOPES, Denilce Meneses. Molecular analysis in Melipona rufiventris and Melipona mondury (Hymenoptera: Apidae). 2008. 96 f. Tese (Doutorado em Análises quantitativas e moleculares do Genoma; Biologia das células e dos tecidos) - Universidade Federal de Viçosa, Viçosa, 2008.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/231
identifier_str_mv LOPES, Denilce Meneses. Molecular analysis in Melipona rufiventris and Melipona mondury (Hymenoptera: Apidae). 2008. 96 f. Tese (Doutorado em Análises quantitativas e moleculares do Genoma; Biologia das células e dos tecidos) - Universidade Federal de Viçosa, Viçosa, 2008.
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dc.publisher.department.fl_str_mv Análises quantitativas e moleculares do Genoma; Biologia das células e dos tecidos
publisher.none.fl_str_mv Universidade Federal de Viçosa
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