Caracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabica

Detalhes bibliográficos
Autor(a) principal: Figueiredo, S??rgio Araujo
Data de Publicação: 2011
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UCB
Texto Completo: https://bdtd.ucb.br:8443/jspui/handle/123456789/50
Resumo: ??-galactosidases are a class of glycosyl-hydrolases that act on the plant cell primary walls, hydrolyzing ??-D-galactose at the nonreducing ends of ??-D-galactosides present in several biological molecules. Initially a characterization of the monosaccharides present in the primary wall of the pericarp and endosperm of Coffea arabica fruits at different ripening stages was performed, identifying the polysaccharides present in these regions, along with the putative carbohydrate target for the ??-galactosidase. In parallel, a molecular and a biochemical characterization of ??-galactosidase was performed. A partial characterization of ??-galactosidase genomic DNA structure, along with a transcription analysis and an in vitro and in situ biochemical activity were performed, identifying peaks of expression in the early stages of growth and in fully ripe fruit. Finally, in order to evaluate the ??-galactosidase effects on coffee fruit ripening, C. arabica calli were transformed by biolistic using RNA interference approach, in order to obtain genetically modified coffee plants with a silenced ??-galactosidase expression. Three transgenic calli growing on selective medium containing ammonium glufosinate were obtained, two of which contained the ??-galactosidase gene fragment. These calli are under embryogenic regeneration and the resulting seedlings will be further analyzed in order to confirm the presence of the transgenes and to assess of the effects of ??-galactosidase gene silencing on coffee fruit ripening.
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spelling Arag??o, Francisco Jos?? LimaCPF:30860997120aragao@cenargen.embrapa.brCPF:57970823679http://lattes.cnpq.br/3564213039969253Figueiredo, S??rgio Araujo2016-06-24T04:00:22Z2011-07-122011-06-03FIGUEIREDO, S??rgio Araujo. Caracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabica. 2011. 244 f. Tese (Doutorado em Ci??ncias Gen??micas, Gen??tica Molecular e de Popula????es, Biotecnologia Molecular) - Universidade Cat??lica de Bras??lia, Bras??lia, 2011.https://bdtd.ucb.br:8443/jspui/handle/123456789/50??-galactosidases are a class of glycosyl-hydrolases that act on the plant cell primary walls, hydrolyzing ??-D-galactose at the nonreducing ends of ??-D-galactosides present in several biological molecules. Initially a characterization of the monosaccharides present in the primary wall of the pericarp and endosperm of Coffea arabica fruits at different ripening stages was performed, identifying the polysaccharides present in these regions, along with the putative carbohydrate target for the ??-galactosidase. In parallel, a molecular and a biochemical characterization of ??-galactosidase was performed. A partial characterization of ??-galactosidase genomic DNA structure, along with a transcription analysis and an in vitro and in situ biochemical activity were performed, identifying peaks of expression in the early stages of growth and in fully ripe fruit. Finally, in order to evaluate the ??-galactosidase effects on coffee fruit ripening, C. arabica calli were transformed by biolistic using RNA interference approach, in order to obtain genetically modified coffee plants with a silenced ??-galactosidase expression. Three transgenic calli growing on selective medium containing ammonium glufosinate were obtained, two of which contained the ??-galactosidase gene fragment. These calli are under embryogenic regeneration and the resulting seedlings will be further analyzed in order to confirm the presence of the transgenes and to assess of the effects of ??-galactosidase gene silencing on coffee fruit ripening.As ??-galactosidases sao uma classe de glicosil-hidrolases que atuam na parede primaria das celulas vegetais, hidrolisando residuos ??-D-galactosis de extremidades nao redutoras de ??-D galactosideos presentes em diversas moleculas biologicas. Inicialmente foi feita uma caracterizacao dos monossacarideos presentes na parede primaria do pericarpo e do endosperma de frutos de Coffea arabica em distintas fases de maturacao, identificando os polissacarideos presentes nesta regiao celular, juntamente com os provaveis carboidratos-alvo para as ??-galactosidases. Em paralelo, foi realizada uma caracterizacao molecular e bioquimica das ??-galactosidases. Foi realizada uma caracteriza????o parcial da estrutura do seu DNA genomico, juntamente com uma analise do nivel de transcricao e da atividade bioquimica in vitro e in situ foram realizadas, identificando picos de expressao nas fases iniciais de crescimento e nos frutos completamente maduros. Por fim, visando avaliar os efeitos das ??-galactosidases na maturacao de frutos de cafe, calos embriogenicos de C. arabica foram transformados por biobalistica, utilizando a tecnica do RNA interferente, com a finalidade de obtencao de plantas geneticamente modificadas de cafeeiro para o silenciamento da expressao do gene das ??-galactosidases. Foram obtidos tres calos transgenicos crescendo em meio seletivo com glufosinato de amonio, dentre os quais dois continham o fragmento deste gene. Estes calos encontram-se em fase de formacao de embrioes somaticos e as plantulas resultantes desta regeneracao serao analisadas, a posteriori, para confirmacao da presenca dos transgenes e avaliacao dos efeitos do silenciamento do gene das ??-galactosidases sobre a maturacao de frutos de cafe.Made available in DSpace on 2016-06-24T04:00:22Z (GMT). No. of bitstreams: 1 Sergio Araujo Figueiredo.pdf: 5051471 bytes, checksum: 845523d46a114f99e92e4c1f07b80e2a (MD5) Previous issue date: 2011-06-03application/pdfhttps://bdtd.ucb.br:8443/jspui/retrieve/1941/Sergio%20Araujo%20Figueiredo.pdf.jpgporUniversidade Cat??lica de Bras??liaPrograma de P??s-Gradua????o em Ci??ncias Gen??micas e BiotecnologiaUCBBRCi??ncias Gen??micas, Gen??tica Molecular e de Popula????es, Biotecnologia Molecularcaf??polissacar??deosbiotecnologiacoffea arabicaB-galactosidasecell wallpolysaccharidesinterfering RNAbiolisticCNPQ::CIENCIAS BIOLOGICAS::GENETICACaracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabicainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UCBinstname:Universidade Católica de Brasília (UCB)instacron:UCBORIGINALSergio Araujo Figueiredo.pdfapplication/pdf5051471https://200.214.135.178:8443/jspui/bitstream/123456789/50/1/Sergio+Araujo+Figueiredo.pdf845523d46a114f99e92e4c1f07b80e2aMD51THUMBNAILSergio Araujo Figueiredo.pdf.jpgSergio Araujo Figueiredo.pdf.jpgimage/jpeg5416https://200.214.135.178:8443/jspui/bitstream/123456789/50/2/Sergio+Araujo+Figueiredo.pdf.jpgf493a942110142846983641f6e221c80MD52123456789/502018-11-08 15:07:39.037Biblioteca Digital de Teses e Dissertaçõeshttps://bdtd.ucb.br:8443/jspui/
dc.title.por.fl_str_mv Caracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabica
title Caracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabica
spellingShingle Caracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabica
Figueiredo, S??rgio Araujo
caf??
polissacar??deos
biotecnologia
coffea arabica
B-galactosidase
cell wall
polysaccharides
interfering RNA
biolistic
CNPQ::CIENCIAS BIOLOGICAS::GENETICA
title_short Caracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabica
title_full Caracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabica
title_fullStr Caracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabica
title_full_unstemmed Caracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabica
title_sort Caracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabica
author Figueiredo, S??rgio Araujo
author_facet Figueiredo, S??rgio Araujo
author_role author
dc.contributor.advisor1.fl_str_mv Arag??o, Francisco Jos?? Lima
dc.contributor.advisor1ID.fl_str_mv CPF:30860997120
dc.contributor.advisor1Lattes.fl_str_mv aragao@cenargen.embrapa.br
dc.contributor.authorID.fl_str_mv CPF:57970823679
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/3564213039969253
dc.contributor.author.fl_str_mv Figueiredo, S??rgio Araujo
contributor_str_mv Arag??o, Francisco Jos?? Lima
dc.subject.por.fl_str_mv caf??
polissacar??deos
biotecnologia
topic caf??
polissacar??deos
biotecnologia
coffea arabica
B-galactosidase
cell wall
polysaccharides
interfering RNA
biolistic
CNPQ::CIENCIAS BIOLOGICAS::GENETICA
dc.subject.eng.fl_str_mv coffea arabica
B-galactosidase
cell wall
polysaccharides
interfering RNA
biolistic
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::GENETICA
dc.description.abstract.eng.fl_txt_mv ??-galactosidases are a class of glycosyl-hydrolases that act on the plant cell primary walls, hydrolyzing ??-D-galactose at the nonreducing ends of ??-D-galactosides present in several biological molecules. Initially a characterization of the monosaccharides present in the primary wall of the pericarp and endosperm of Coffea arabica fruits at different ripening stages was performed, identifying the polysaccharides present in these regions, along with the putative carbohydrate target for the ??-galactosidase. In parallel, a molecular and a biochemical characterization of ??-galactosidase was performed. A partial characterization of ??-galactosidase genomic DNA structure, along with a transcription analysis and an in vitro and in situ biochemical activity were performed, identifying peaks of expression in the early stages of growth and in fully ripe fruit. Finally, in order to evaluate the ??-galactosidase effects on coffee fruit ripening, C. arabica calli were transformed by biolistic using RNA interference approach, in order to obtain genetically modified coffee plants with a silenced ??-galactosidase expression. Three transgenic calli growing on selective medium containing ammonium glufosinate were obtained, two of which contained the ??-galactosidase gene fragment. These calli are under embryogenic regeneration and the resulting seedlings will be further analyzed in order to confirm the presence of the transgenes and to assess of the effects of ??-galactosidase gene silencing on coffee fruit ripening.
dc.description.abstract.por.fl_txt_mv As ??-galactosidases sao uma classe de glicosil-hidrolases que atuam na parede primaria das celulas vegetais, hidrolisando residuos ??-D-galactosis de extremidades nao redutoras de ??-D galactosideos presentes em diversas moleculas biologicas. Inicialmente foi feita uma caracterizacao dos monossacarideos presentes na parede primaria do pericarpo e do endosperma de frutos de Coffea arabica em distintas fases de maturacao, identificando os polissacarideos presentes nesta regiao celular, juntamente com os provaveis carboidratos-alvo para as ??-galactosidases. Em paralelo, foi realizada uma caracterizacao molecular e bioquimica das ??-galactosidases. Foi realizada uma caracteriza????o parcial da estrutura do seu DNA genomico, juntamente com uma analise do nivel de transcricao e da atividade bioquimica in vitro e in situ foram realizadas, identificando picos de expressao nas fases iniciais de crescimento e nos frutos completamente maduros. Por fim, visando avaliar os efeitos das ??-galactosidases na maturacao de frutos de cafe, calos embriogenicos de C. arabica foram transformados por biobalistica, utilizando a tecnica do RNA interferente, com a finalidade de obtencao de plantas geneticamente modificadas de cafeeiro para o silenciamento da expressao do gene das ??-galactosidases. Foram obtidos tres calos transgenicos crescendo em meio seletivo com glufosinato de amonio, dentre os quais dois continham o fragmento deste gene. Estes calos encontram-se em fase de formacao de embrioes somaticos e as plantulas resultantes desta regeneracao serao analisadas, a posteriori, para confirmacao da presenca dos transgenes e avaliacao dos efeitos do silenciamento do gene das ??-galactosidases sobre a maturacao de frutos de cafe.
description ??-galactosidases are a class of glycosyl-hydrolases that act on the plant cell primary walls, hydrolyzing ??-D-galactose at the nonreducing ends of ??-D-galactosides present in several biological molecules. Initially a characterization of the monosaccharides present in the primary wall of the pericarp and endosperm of Coffea arabica fruits at different ripening stages was performed, identifying the polysaccharides present in these regions, along with the putative carbohydrate target for the ??-galactosidase. In parallel, a molecular and a biochemical characterization of ??-galactosidase was performed. A partial characterization of ??-galactosidase genomic DNA structure, along with a transcription analysis and an in vitro and in situ biochemical activity were performed, identifying peaks of expression in the early stages of growth and in fully ripe fruit. Finally, in order to evaluate the ??-galactosidase effects on coffee fruit ripening, C. arabica calli were transformed by biolistic using RNA interference approach, in order to obtain genetically modified coffee plants with a silenced ??-galactosidase expression. Three transgenic calli growing on selective medium containing ammonium glufosinate were obtained, two of which contained the ??-galactosidase gene fragment. These calli are under embryogenic regeneration and the resulting seedlings will be further analyzed in order to confirm the presence of the transgenes and to assess of the effects of ??-galactosidase gene silencing on coffee fruit ripening.
publishDate 2011
dc.date.available.fl_str_mv 2011-07-12
dc.date.issued.fl_str_mv 2011-06-03
dc.date.accessioned.fl_str_mv 2016-06-24T04:00:22Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
status_str publishedVersion
format doctoralThesis
dc.identifier.citation.fl_str_mv FIGUEIREDO, S??rgio Araujo. Caracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabica. 2011. 244 f. Tese (Doutorado em Ci??ncias Gen??micas, Gen??tica Molecular e de Popula????es, Biotecnologia Molecular) - Universidade Cat??lica de Bras??lia, Bras??lia, 2011.
dc.identifier.uri.fl_str_mv https://bdtd.ucb.br:8443/jspui/handle/123456789/50
identifier_str_mv FIGUEIREDO, S??rgio Araujo. Caracteriza????o bioqu??mica e molecular da ??-Galactosidade durante a matura????o de frutos de coffea arabica. 2011. 244 f. Tese (Doutorado em Ci??ncias Gen??micas, Gen??tica Molecular e de Popula????es, Biotecnologia Molecular) - Universidade Cat??lica de Bras??lia, Bras??lia, 2011.
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dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Cat??lica de Bras??lia
dc.publisher.program.fl_str_mv Programa de P??s-Gradua????o em Ci??ncias Gen??micas e Biotecnologia
dc.publisher.initials.fl_str_mv UCB
dc.publisher.country.fl_str_mv BR
dc.publisher.department.fl_str_mv Ci??ncias Gen??micas, Gen??tica Molecular e de Popula????es, Biotecnologia Molecular
publisher.none.fl_str_mv Universidade Cat??lica de Bras??lia
dc.source.none.fl_str_mv reponame:Biblioteca Digital de Teses e Dissertações da UCB
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