Estabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae)

Detalhes bibliográficos
Autor(a) principal: Moraes, Brenda Amaral de
Data de Publicação: 2015
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UERJ
Texto Completo: http://www.bdtd.uerj.br/handle/1/7959
Resumo: Cleome spinosa is a herbaceous species used in folk medicine, mainly in the Northeast of Brazil. Cultures of adventitious roots were established with root segments (0.5 to 1.0 cm) obtained from two sources of explants: in vitro propagated plants and plants obtained from in vitro germination process. The explants were inoculated in MS liquid medium supplemented with or without the auxin NAA, IAA and IBA in different concentrations (0, 0.5, 1.0, 1.5, 3.0, 5.0 mg.L-1). Cultures were maintained in a growth room under agitation (100 rpm) and photoperiod of 16h light or in the dark, with subcultures every 45 days. Explants from in vitro propagated plants were also cultured in medium containing BAP in combination with auxins, in the presence of sorbitol (alone or in combination with sucrose), in MS medium with a reduction in the total concentration of mineral salts (MS1/2 and MS1/4) and on solidified medium. The results showed that hormonal supplementation was essential for roots multiplication, once cultures maintained in MS0 medium only developed shoots. The medium supplementation with NAA was not efficient for root production and resulted in callus induction. The presence of IAA and IBA was efficient to root multiplication. A significant root proliferation capacity was observed in cultures initiated from explants originating from plants obtained by in vitro germination process. On the other hand, cultures initiated with explants originating from in vitro propagated plants, regardless the different manipulations performed in the culture medium, showed low root multiplication capacity. The highest biomass production was achieved by cultures initiated with explants from in vitro germination plants cultured in medium supplemented with 3.0 mg.L-1 IBA and kept in the dark. Cultures with the highest biomass accumulation were subcultured at each 45 days or 60 days interval to fresh medium with the same composition during three - four subcultures. Cultures maintained at 45 days interval reached the highest biomass accumulation during the second subculture, while cultures maintained at 60 days interval, despite the maintenance of root multiplication along the subcultures, reached the highest production during the first 60 days in culture. Samples from in vivo and in vitro material were extracted with increasingly polar solvents (hexane, dichloromethane, ethyl acetate, methanol) and the extracts were submitted to chromatographic analysis. The analysis by thin layer indicated the presence of terpenes in hexane and dichloromethane extracts obtained with field plants and in vitro materials, and the presence of phenolic componds in ethyl acetate extracts obtained with field plants. Analysis by liquid chromatography coupled to mass spectrometry showed the in vitro production of metabolites not found in field plants, especially flavonoids. The gas chromatography coupled to mass spectrometry indicated the presence of steroids in the extracts obtained with hexane from field plants and root cultures. The results demonstrated the feasibility in the production of in vitro root cultures of C. spinosa and the potential of these biotechnological materials to phytochemical studies, mainly considering the presence under in vitro cultures of metabolites not produced by field plants
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spelling Gurgel, Claudia Simõeshttp://lattes.cnpq.br/5590442186208185Albarello, Normahttp://lattes.cnpq.br/4767211177616413Duarte, Aline Castellarhttp://lattes.cnpq.br/4275030752135552Oliveira, Elisabeth Atalla Mansur dehttp://lattes.cnpq.br/7809321078544787http://lattes.cnpq.br/5039555262972543Moraes, Brenda Amaral de2021-01-05T18:24:36Z2015-05-192015-02-26MORAES, Brenda Amaral de. Estabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae). 2015. 124 f. Dissertação (Mestrado em Conservação e Utilização da Biodiversidade) - Universidade do Estado do Rio de Janeiro, Rio de Janeiro, 2015.http://www.bdtd.uerj.br/handle/1/7959Cleome spinosa is a herbaceous species used in folk medicine, mainly in the Northeast of Brazil. Cultures of adventitious roots were established with root segments (0.5 to 1.0 cm) obtained from two sources of explants: in vitro propagated plants and plants obtained from in vitro germination process. The explants were inoculated in MS liquid medium supplemented with or without the auxin NAA, IAA and IBA in different concentrations (0, 0.5, 1.0, 1.5, 3.0, 5.0 mg.L-1). Cultures were maintained in a growth room under agitation (100 rpm) and photoperiod of 16h light or in the dark, with subcultures every 45 days. Explants from in vitro propagated plants were also cultured in medium containing BAP in combination with auxins, in the presence of sorbitol (alone or in combination with sucrose), in MS medium with a reduction in the total concentration of mineral salts (MS1/2 and MS1/4) and on solidified medium. The results showed that hormonal supplementation was essential for roots multiplication, once cultures maintained in MS0 medium only developed shoots. The medium supplementation with NAA was not efficient for root production and resulted in callus induction. The presence of IAA and IBA was efficient to root multiplication. A significant root proliferation capacity was observed in cultures initiated from explants originating from plants obtained by in vitro germination process. On the other hand, cultures initiated with explants originating from in vitro propagated plants, regardless the different manipulations performed in the culture medium, showed low root multiplication capacity. The highest biomass production was achieved by cultures initiated with explants from in vitro germination plants cultured in medium supplemented with 3.0 mg.L-1 IBA and kept in the dark. Cultures with the highest biomass accumulation were subcultured at each 45 days or 60 days interval to fresh medium with the same composition during three - four subcultures. Cultures maintained at 45 days interval reached the highest biomass accumulation during the second subculture, while cultures maintained at 60 days interval, despite the maintenance of root multiplication along the subcultures, reached the highest production during the first 60 days in culture. Samples from in vivo and in vitro material were extracted with increasingly polar solvents (hexane, dichloromethane, ethyl acetate, methanol) and the extracts were submitted to chromatographic analysis. The analysis by thin layer indicated the presence of terpenes in hexane and dichloromethane extracts obtained with field plants and in vitro materials, and the presence of phenolic componds in ethyl acetate extracts obtained with field plants. Analysis by liquid chromatography coupled to mass spectrometry showed the in vitro production of metabolites not found in field plants, especially flavonoids. The gas chromatography coupled to mass spectrometry indicated the presence of steroids in the extracts obtained with hexane from field plants and root cultures. The results demonstrated the feasibility in the production of in vitro root cultures of C. spinosa and the potential of these biotechnological materials to phytochemical studies, mainly considering the presence under in vitro cultures of metabolites not produced by field plantsCleome spinosa é uma espécie herbácea de uso na medicina popular, especialmente no Nordeste do Brasil. A cultura in vitro de raízes adventícias da espécie foi iniciada com segmentos radiculares (0,5 e 1,0 cm) obtidos a partir de duas fontes de explantes: plantas propagadas in vitro e plantas oriundas do processo de germinação in vitro. Os explantes foram inoculados em meio MS líquido suplementado ou não com as auxinas ANA, AIA e AIB em diferentes concentrações (0,5; 1,0; 1,5; 3,0; 5,0 mg.L-1). As culturas foram mantidas em sala de crescimento, sob agitação (100 rpm) e sob fotoperíodo de 16h ou no escuro, com subculturas a cada 45 dias. Explantes oriundos de plantas propagadas in vitro também foram cultivados em meio contendo a citocinina BAP em associação com auxinas, na presença de sorbitol (isoladamente ou em associação com sacarose), em meio MS contendo redução na concentração total de sais minerais (MS1/2 e MS1/4) e em meio sólido. Os resultados mostraram que a adição de auxinas ao meio de cultura foi essencial à multiplicação das raízes, uma vez que em meio MS0 ocorreu significativo desenvolvimento de brotos. A suplementação com ANA não foi eficiente para a produção de raízes e acarretou no calejamento dos explantes, enquanto que a presença de AIA e AIB resultaram na multiplicação das raízes. Ainda assim, independentemente das manipulações realizadas no meio de cultivo, a capacidade de multiplicação mostrou-se reduzida. Uma expressiva multiplicação de raízes foi observada em culturas iniciadas a partir de explantes oriundos de plantas obtidas por germinação in vitro. A maior produção de biomassa foi alcançada em culturas iniciadas com segmentos radiculares de plantas obtidas por germinação in vitro cultivadas em meio contendo com 3,0 mg.L-1 de AIB e mantidas no escuro. Culturas estabelecidas nas melhores condições para acúmulo de biomassa foram acompanhadas por três subculturas, sendo avaliados períodos de cultura de 45 dias e de 60 dias. Culturas mantidas a intervalos de 45 dias apresentaram maior produção de raízes durante a segunda subcultura, enquanto que para o intervalo de 60 dias, embora tenha sido observada a capacidade de multiplicação das raízes, a maior produção de biomassa ocorreu nos primeiros 60 dias de cultivo. A partir de materiais in vivo e in vitro foram realizadas extrações com solventes de polaridade crescente (hexano, diclorometano, acetato de etila e metanol) e os extratos foram submetidos a avaliações cromatográficas. As análises por cromatografia em camada delgada mostraram a presença de terpenos nos extratos obtidos com hexano e diclorometano, tanto em material obtido a campo como naqueles produzidos in vitro e de compostos fenólicos nos extratos em acetato de etila obtidos a partir de material de campo. Pelas análises por cromatografia líquida associada à espectrometria de massas foi possível observar a presença de flavonoides nas culturas in vitro, não detectados no material de campo. As análises por cromatografia de fase gasosa associada à espectrometria de massas apontaram a presença de esteroides nos extratos em hexano de raízes coletadas a campo e nas culturas in vitro de raízes. Os resultados obtidos mostraram a viabilidade da produção de culturas de raízes in vitro para a espécie C. spinosa e o potencial deste material para a produção de substâncias bioativas, algumas não encontradas em material coletado a campoSubmitted by Boris Flegr (boris@uerj.br) on 2021-01-05T18:24:36Z No. of bitstreams: 1 Brenda_Dissertacao_PPGBV_completa.pdf: 4916062 bytes, checksum: db57360fa8a6cfbec891a4afea77a483 (MD5)Made available in DSpace on 2021-01-05T18:24:36Z (GMT). No. of bitstreams: 1 Brenda_Dissertacao_PPGBV_completa.pdf: 4916062 bytes, checksum: db57360fa8a6cfbec891a4afea77a483 (MD5) Previous issue date: 2015-02-26Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade do Estado do Rio de JaneiroPrograma de Pós-Graduação em Biologia VegetalUERJBRCentro Biomédico::Instituto de Biologia Roberto Alcantara GomesAuxinPhenolic compoundsChromatographyCrude extractTerpenesAuxinasCompostos fenólicosCromatografiaExtrato brutoTerpenos.Fisiologia vegetalTecidos vegetais - Cultura e meios de culturaFitoquímicosPlantas medicinaisCNPQ::CIENCIAS BIOLOGICAS::BOTANICAEstabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae)Establishment of in vitro root cultures and evaluation of secondary metabolites production of Cleome spinosa Jacq. (Cleomaceae)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UERJinstname:Universidade do Estado do Rio de Janeiro (UERJ)instacron:UERJORIGINALBrenda_Dissertacao_PPGBV_completa.pdfapplication/pdf4916062http://www.bdtd.uerj.br/bitstream/1/7959/1/Brenda_Dissertacao_PPGBV_completa.pdfdb57360fa8a6cfbec891a4afea77a483MD511/79592024-02-26 15:49:12.096oai:www.bdtd.uerj.br:1/7959Biblioteca Digital de Teses e Dissertaçõeshttp://www.bdtd.uerj.br/PUBhttps://www.bdtd.uerj.br:8443/oai/requestbdtd.suporte@uerj.bropendoar:29032024-02-26T18:49:12Biblioteca Digital de Teses e Dissertações da UERJ - Universidade do Estado do Rio de Janeiro (UERJ)false
dc.title.por.fl_str_mv Estabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae)
dc.title.alternative.eng.fl_str_mv Establishment of in vitro root cultures and evaluation of secondary metabolites production of Cleome spinosa Jacq. (Cleomaceae)
title Estabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae)
spellingShingle Estabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae)
Moraes, Brenda Amaral de
Auxin
Phenolic compounds
Chromatography
Crude extract
Terpenes
Auxinas
Compostos fenólicos
Cromatografia
Extrato bruto
Terpenos.
Fisiologia vegetal
Tecidos vegetais - Cultura e meios de cultura
Fitoquímicos
Plantas medicinais
CNPQ::CIENCIAS BIOLOGICAS::BOTANICA
title_short Estabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae)
title_full Estabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae)
title_fullStr Estabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae)
title_full_unstemmed Estabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae)
title_sort Estabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae)
author Moraes, Brenda Amaral de
author_facet Moraes, Brenda Amaral de
author_role author
dc.contributor.advisor1.fl_str_mv Gurgel, Claudia Simões
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/5590442186208185
dc.contributor.advisor-co1.fl_str_mv Albarello, Norma
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/4767211177616413
dc.contributor.referee1.fl_str_mv Duarte, Aline Castellar
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/4275030752135552
dc.contributor.referee2.fl_str_mv Oliveira, Elisabeth Atalla Mansur de
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/7809321078544787
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/5039555262972543
dc.contributor.author.fl_str_mv Moraes, Brenda Amaral de
contributor_str_mv Gurgel, Claudia Simões
Albarello, Norma
Duarte, Aline Castellar
Oliveira, Elisabeth Atalla Mansur de
dc.subject.eng.fl_str_mv Auxin
Phenolic compounds
Chromatography
Crude extract
Terpenes
topic Auxin
Phenolic compounds
Chromatography
Crude extract
Terpenes
Auxinas
Compostos fenólicos
Cromatografia
Extrato bruto
Terpenos.
Fisiologia vegetal
Tecidos vegetais - Cultura e meios de cultura
Fitoquímicos
Plantas medicinais
CNPQ::CIENCIAS BIOLOGICAS::BOTANICA
dc.subject.por.fl_str_mv Auxinas
Compostos fenólicos
Cromatografia
Extrato bruto
Terpenos.
Fisiologia vegetal
Tecidos vegetais - Cultura e meios de cultura
Fitoquímicos
Plantas medicinais
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::BOTANICA
description Cleome spinosa is a herbaceous species used in folk medicine, mainly in the Northeast of Brazil. Cultures of adventitious roots were established with root segments (0.5 to 1.0 cm) obtained from two sources of explants: in vitro propagated plants and plants obtained from in vitro germination process. The explants were inoculated in MS liquid medium supplemented with or without the auxin NAA, IAA and IBA in different concentrations (0, 0.5, 1.0, 1.5, 3.0, 5.0 mg.L-1). Cultures were maintained in a growth room under agitation (100 rpm) and photoperiod of 16h light or in the dark, with subcultures every 45 days. Explants from in vitro propagated plants were also cultured in medium containing BAP in combination with auxins, in the presence of sorbitol (alone or in combination with sucrose), in MS medium with a reduction in the total concentration of mineral salts (MS1/2 and MS1/4) and on solidified medium. The results showed that hormonal supplementation was essential for roots multiplication, once cultures maintained in MS0 medium only developed shoots. The medium supplementation with NAA was not efficient for root production and resulted in callus induction. The presence of IAA and IBA was efficient to root multiplication. A significant root proliferation capacity was observed in cultures initiated from explants originating from plants obtained by in vitro germination process. On the other hand, cultures initiated with explants originating from in vitro propagated plants, regardless the different manipulations performed in the culture medium, showed low root multiplication capacity. The highest biomass production was achieved by cultures initiated with explants from in vitro germination plants cultured in medium supplemented with 3.0 mg.L-1 IBA and kept in the dark. Cultures with the highest biomass accumulation were subcultured at each 45 days or 60 days interval to fresh medium with the same composition during three - four subcultures. Cultures maintained at 45 days interval reached the highest biomass accumulation during the second subculture, while cultures maintained at 60 days interval, despite the maintenance of root multiplication along the subcultures, reached the highest production during the first 60 days in culture. Samples from in vivo and in vitro material were extracted with increasingly polar solvents (hexane, dichloromethane, ethyl acetate, methanol) and the extracts were submitted to chromatographic analysis. The analysis by thin layer indicated the presence of terpenes in hexane and dichloromethane extracts obtained with field plants and in vitro materials, and the presence of phenolic componds in ethyl acetate extracts obtained with field plants. Analysis by liquid chromatography coupled to mass spectrometry showed the in vitro production of metabolites not found in field plants, especially flavonoids. The gas chromatography coupled to mass spectrometry indicated the presence of steroids in the extracts obtained with hexane from field plants and root cultures. The results demonstrated the feasibility in the production of in vitro root cultures of C. spinosa and the potential of these biotechnological materials to phytochemical studies, mainly considering the presence under in vitro cultures of metabolites not produced by field plants
publishDate 2015
dc.date.available.fl_str_mv 2015-05-19
dc.date.issued.fl_str_mv 2015-02-26
dc.date.accessioned.fl_str_mv 2021-01-05T18:24:36Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv MORAES, Brenda Amaral de. Estabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae). 2015. 124 f. Dissertação (Mestrado em Conservação e Utilização da Biodiversidade) - Universidade do Estado do Rio de Janeiro, Rio de Janeiro, 2015.
dc.identifier.uri.fl_str_mv http://www.bdtd.uerj.br/handle/1/7959
identifier_str_mv MORAES, Brenda Amaral de. Estabelecimento de culturas de raízes in vitro e avaliação da produção de metabólitos secundários de Cleome spinosa Jacq. (Cleomaceae). 2015. 124 f. Dissertação (Mestrado em Conservação e Utilização da Biodiversidade) - Universidade do Estado do Rio de Janeiro, Rio de Janeiro, 2015.
url http://www.bdtd.uerj.br/handle/1/7959
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dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
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dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade do Estado do Rio de Janeiro
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Biologia Vegetal
dc.publisher.initials.fl_str_mv UERJ
dc.publisher.country.fl_str_mv BR
dc.publisher.department.fl_str_mv Centro Biomédico::Instituto de Biologia Roberto Alcantara Gomes
publisher.none.fl_str_mv Universidade do Estado do Rio de Janeiro
dc.source.none.fl_str_mv reponame:Biblioteca Digital de Teses e Dissertações da UERJ
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