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dc.contributor.authorLópez-Vázquez, Ana Luisa-
dc.date.available2020-03-12-
dc.date.available2020-03-13T14:14:19Z-
dc.date.issued2019-04-30-
dc.identifier.urihttp://repositorioinstitucional.uea.edu.br//handle/riuea/2279-
dc.description.abstractDuroia macrophylla Huber (Rubiaceae) is a species that produces important secondary metabolites, highlighting the indole alkaloids with antitumoral and antituberculosis activities. The objective of this work was to establish seedling cultures, calluses and cell suspensions in vitro of the species, as well as to evaluate the effect of various elicitors on the chemical profile of calluses of D. macrophylla. Seeds from D. macrophylla fruits were used as explants for in vitro germination and seedling growth. The seeds were submitted to different treatments and were tested at concentrations of 0, 2, 5 and 10 mg.L-1 of gibberellic acid (GA3) for the induction of germination in Murashige and Skoog (MS) medium. The germinated seedlings were then used as explants in which different concentrations of plant growth regulators (PGRs) were tested, such as α-naphthalene acetic acid (ANA) and 2,4-dichlorophenoxyacetic acid ( 2,4-D) and the cytokinins N-Benzyl-9- (2-tetrahydropyranyl)-adenine (BAP) and kinetin (KIN) for callus induction. All treatments were exposed to two light conditions: 24 h in the dark and photoperiod of 16/8 h (light / dark), both with a constant temperature of 26 ºC. The best response (70%) for the induction of semi-friable callus (SC) was in the presence of growth regulators BAP and ANA at the concentration of 2 mg.L-1 for both. The 24 h condition in the dark was significant for the formation of white and semi-friable callus for all combinations of PGRs and from these callus cell suspensions were established. Growth curves were defined and the phytochemical analyzes of both cultures were performed by TLC and 1H NMR showing differences in growth and chemical profile of both. From the fractionation of the methanolic extract of the callus was isolated an iridoide-glycoside that is still in the process of elucidation. This is the first report of the presence of iridoids in the species. The effect of different concentrations of the NaCl, KCl, AlCl3, SNP (NO), ABA and different UV exposure times on the chemical profile and iridoid production in the callus was evaluated. The results were evaluated by chemometrics analysis: Principal Component Analysis (PCA)/ Fuzzy and Hierarchical Cluster Analysis (HCA) and fuzziThe elicitors SNP and AlCl3 were shown to have a greater effect on the chemical profile and the production of the iridoid in the calluses, whereas NaCl, KCl and ABA inhibited its production. The results obtained in this work demonstrate the significant potential of the use of elicitation in calluses of the species D. macrophylla and encourage the continuation of the study aiming the induction of the production of the active metabolites of the plant.pt_BR
dc.languageporpt_BR
dc.publisherUniversidade do Estado do Amazonaspt_BR
dc.rightsAcesso Abertopt_BR
dc.rightsAtribuição-NãoComercial-SemDerivados 3.0 Brasil*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/br/*
dc.subjecttecidos vegetaispt_BR
dc.subjectIridoidespt_BR
dc.subjectElicitaçãopt_BR
dc.titleAprimoramento da obtenção de calos e suspensões celulares de Duroia macrophylla Huber (Rubiaceae) e avaliação dos elicitores NaCl, KCl, AlCl3, SNP(NO), ABA e luz UV na produção de metabólitos secundáriospt_BR
dc.typeDissertaçãopt_BR
dc.date.accessioned2020-03-13T14:14:19Z-
dc.contributor.advisor1Nunez, Cecilia Veronica-
dc.contributor.advisor1Latteshttp://lattes.cnpq.br/2046473694108264pt_BR
dc.contributor.referee1Nunez, Cecilia Veronica-
dc.contributor.referee1Latteshttp://lattes.cnpq.br/2046473694108264pt_BR
dc.contributor.referee2Rapôso, Nádia Verçosa de Medeiros-
dc.contributor.referee2Latteshttp://lattes.cnpq.br/5791520737203814pt_BR
dc.contributor.referee3Rocha , Waldireny Caldas-
dc.contributor.referee3Latteshttp://lattes.cnpq.br/0161052060648197pt_BR
dc.creator.Latteshttp://lattes.cnpq.br/8550051372960788pt_BR
dc.description.resumoA espécie Duroia macrophylla Huber (Rubiaceae) é produtora de metabólitos secundários importantes destacando-se os alcaloides indólicos com atividade antitumoral e antituberculose. Este trabalho teve como objetivo estabelecer culturas de plântulas, calos e suspensões in vitro da espécie, assim como avaliar o efeito de diversos elicitores no perfil químico dos calos de D. macrophylla. Foram usados como explantes as sementes dos frutos de D. macrophylla para a germinação e crescimento das plântulas in vitro. As sementes foram submetidas a diferentes tratamentos e foram testadas nas concentrações de 0, 2, 5 e 10 mg.L-1 de ácido giberélico (AG3) para a indução da germinação em meio Murashige e Skoog (MS). Após, foram usadas como explantes as folhas das plântulas germinadas, nas quais foram testadas distintas concentrações de reguladores de crescimento de plantas (PGRs), sendo eles: as auxinas ácido 𝜶-naftaleno acético (ANA) e o ácido 2,4-diclorofenoxiacético (2,4-D) e as citocininas N-Benzil-9-(2- tetrahidropiranil)-adenina (BAP) e a cinetina (KIN) para a indução de calos. Todos os tratamentos foram expostos a duas condições de luz: 24 h no escuro e fotoperiodo de 16/8 h (claro/escuro) e ambas com temperatura constante de 26 ºC. A melhor resposta (70%) para a indução de calos semi-friável (CS) foi na presença dos reguladores de crescimento BAP e ANA na concentração de 2 mg.L- 1 para ambos. A condição de 24 h no escuro foi significativa para a formação de calo branco e semifriável para todas as combinações de PGRs e a partir desses calos foram estabelecidas suspensões celulares. Foram definidas as curvas de crescimento e realizadas as análises fitoquímicas de ambas as culturas por CCDC e RMN de 1H apresentando diferenças no crescimento e no perfil químico de ambas. Do fracionamento do extrato metanólico dos calos foi isolado um iridoide glicosilado que ainda se encontra em processo de elucidação. Este é o primeiro relato da presença de iridoides na espécie. Foi avaliado o efeito de distintas concentrações dos elicitores NaCl, KCl, AlCl3, SNP (NO), ABA e diferentes tempos de exposição a raios UV, no perfil químico e na produção do iridoide nos calos. Os resultados foram avaliados por meio de análises quimiométricas: análise de componentes principais (PCA)/ Fuzzy e análise hierárquica de clusters (HCA). Os elicitores SNP e AlCl3 mostraram ter maior efeito no perfil químico e na produção do iridoide nos calos, enquanto NaCl, KCl e ABA inibiram a produção do mesmo. Os resultados encontrados neste trabalho demonstram o grande potencial do uso da elicitação nos calos da espécie D. macrophylla e estimulam a continuação do estudo visando a indução da produção dos metabólitos ativos da planta.pt_BR
dc.publisher.countryBrasilpt_BR
dc.publisher.programPós-Graduação em Biotecnologia e Recursos Naturaispt_BR
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