M.Sc. (Botany) / Please refer to full text to view abstract

Andrographis paniculata (AP) is a traditional herbal medicine which is widely used for the treatment of many diseases in Asia. Recently, various biological activities of AP extract or andrographolide (AND), such as immunostimulatory activity, anti-inflammatoryeffect, cytokine induction or deduction, a potential cancer therapeutic agent and T cell activation suppression, have been reported. However the potential ofAP extract or AND to stimulate the specific or adaptive immune response using microbial vaccines has not been determined. In this project, AND has been purifiedfrom AP and its identity confirmed by the melting point, colour test, TLC, UV absorption spectrum, ESI-MS and H-NMR. An aqueous and two ethanolic extracts ofAndrographis paniculata and AND, an active principle of Andrographis paniculata, were investigated for their antimicrobial activity against nine bacterial species in vitro using the disc diffusion method. It was discovered that neither the aqueous extract nor AND were bacteriostatic or bactericidal against S. typhimurium, E. coli, S. sonnei, S. aureus, P. aeruginosa, S. pneumoniae, S. pyogenes, L. pneumophila or B. pertussisbut the two ethanolic extracts of AP were bacteriostatic against L. pneumophila and B. pertussis. It was also observed that the ethanol extract of AP and AND stimulatedboth antibody and cell-mediated immunity (CMI) responses to a killed S. typhimurium vaccine. Mice were vaccinated with either one dose or two doses of killed S. typhimurium vaccine. They were fed two different quantities of an ethanol extract of AP or AND for 14 days in mice immunised with one dose of the vaccine, and for 28 days in mice immunised with two doses of vaccine, respectively. Both the extract and AND significantly increased the IgG antibody titres against S. typhimurium, with theincrease in antibody titres being statistically significant in the two dose vaccine group. Although not statistically significant, there was also a substantial increase in the IgG antibody titres in the one dose vaccine group. Splenocyte cultures from mice fromboth the immunisation groups treated with the extract or AND stimulated with the S. typhimurium lysate showed a significant increase in the production of IFN-γ in both14 and 28 day AP extract or AND treatment groups. The increase indicates the induction of a cell-mediated immune response. To confirm the immunomodulatory potential of AP extract and determine the immunomodulatory potential of AND, experiments were conducted using mouse salmonellosis as a model system. Mice were vaccinated with two doses of killed S. typhimurium vaccine by intraperitoneal(i/p) route and orally dosed with AP extract at 25 mg/kg bodyweight or AND at 4 mg/kg bodyweight for total 28 days, followed by oral challenge with virulent S. typhimurium. Both AP extract and AND substantially increased the survival rate by 50% after mice were challenged with 10 fold of a sublethal dose (1.5x10 6 cfu) of virulent S. typhimurium. They also promoted clearance of S. typhimurium from challenged mice by days 8 or 12 post-challenge with 1.5x10 cfu virulent S. typhimurium respectively. Sera IgG, IgA antibody titres against S. typhimurium and IFN-γ or IL-2 were detected after the mice were challenged by the oral route with a sublethal dose (1.5x10 cfu) of virulent S. typhimurium for 12 days. It was thus concluded that both Andrographis paniculata and AND not only elicited both humoral and cell-mediated immune responses in the mouse model, but also increasedthe protective efficacy against salmonellosis on mice vaccinated with inactivated S. typhimurium. Therefore the ability of Andrographis paniculata and AND to promoteacquired immunity, particularly in inducing CMI may be important in protection against intracellular pathogen infection.

As raízes de Pfaffia paniculata têm sido popularmente utilizadas com vários propósitos, assim como adjuvante na terapia contra o câncer. Recentemente demonstramos que as raízes pulverizadas de Pfaffia paniculata causam uma atenuação do crescimento do tumor de Ehrlich em sua forma ascítica. Os objetivos do presente estudo foram caracterizar os efeitos do extrato etanólico, resíduos aquoso ou butanólico de Pfaffia paniculata sobre o desenvolvimento do tumor de Ehrlich, assim como investigar seus possíveis mecanismos. Em um primeiro experimento, foi demonstrado que camundongos machos Swiss, portadores do tumor de Ehrlich ascítico, tratados com resíduo aquoso ou butanólico, apresentaram uma maior sobrevida em relação aos animais controle. Nos experimentos com o tumor na forma sólida ou ascítica, os animais receberam, diariamente, o resíduo butanólico por 16 dias (experimento com o tumor sólido) ou 15 dias (experimento com o tumor ascítico). No 8º dia de tratamento, foram inoculados com 2,5 x 106 células tumorais, no coxim plantar esquerdo, ou com 5,0 x 106 células tumorais, Intraperitonealmente. O tumor ascítico foi avaliado, 7 dias após a inoculação do tumor, pela quantificação do volume de fluído ascítico, concentração de células tumorais e número total de células tumorais. O tumor sólido foi avaliado pela mensuração diária das patas e pela morfometria da área de necrose em meio à massa tumoral em cortes corados por HE, e proliferação celular, por imunoistoquímica, 8 dias após a inoculação do tumor. Nesses experimentos, nenhum destes parâmetros avaliados apresentaram alterações nos camundongos tratados com este resíduo. A fim de se avaliar a toxicidade do resíduo butanólico, os animais foram tratados com este resíduo por 14 dias. Os animais tratados com este resíduo apresentaram uma discreta diminuição de ganho de peso. Não houve evidências de toxicidade hepática e renal, pela mensuração das atividades de enzimas indicativas de necrose e pelo exame histopatológico de figado e rim, onde não se observou necrose. Em outro experimento, alguns parâmetros da atividade macrofágica, 24 após a inoculação do tumor, foram avaliados, por citometria de fluxo. Os animais receberam o resíduo butanólico por 7 dias, e foram inoculados com 5 x 106 células tumorais, intraperitonealmente. O burst oxidativo ativado por Staphylococcus aureus foi menor em animais tratados com o resíduo, porém não foram observadas diferenças ma fagocitose e burst oxidativo espontâneo ou ativado por PMA. Por fim, estudos in vitro foram realizados. Os efeitos deste resíduo sobre a viabilidade, medido pelo ensaio do MTT, e sobre o ciclo celular, utilizando citometria de fluxo, de células do tumor de Ehrlich foram avaliados. As maiores concentrações do resíduo levaram a uma diminuição da viabilidade e um aumento de morte celular. Assim, o tratamento com este resíduo, in vivo, causou uma atenuação do desenvolvimento tumoral, provavelmente devido a uma diminuição na formação do fluído ascítico ou a morte celular. Os efeitos deste resíduo foram mais pronunciados em cultura celular. / The roots of Pfaffia paniculata have been popularly used for various purposes, as well as an adjuvant for cancer therapy. Recently we have shown that the powdered roots of Pfaffia paniculata promote an attenuation of the Ehrlich tumor growth in its ascitic form. The aims of the present study were to characterize the effects of the ethanolic extract, aqueous or butanolic residues of Pfaffia paniculata on Ehrlich tumor development in mice, as well as investigate its possible mechanisms. In a first experiment, it was shown that aqueous or butanolic residues-treated Swiss mice bearing Ehrlich ascites tumor presented a higher survival time than control animal. In the experiments with the tumor in its solid or ascitic forms, the animals were given, daily, the butanolic residue for 16 days (experiment with solid tumor) or 15 days (experiment with ascitic tumor). On the 8o day of treatment, they were inoculated with 2,5 x 106 tumor cells, on the left footpad, or with 5 x 106 tumor cells, intraperitoneally. The ascitic tumor was evaluated, 7 days after the inoculation of the tumor, by the quantification of the volume of the ascitic fluid, concentration of tumor cells and total number of tumor cells. The solid tumor was evaluated by the daily measurement of the footpads and morphometry of the necrosis within the tumor area on HE stained slices, and cell proliferation by immunohistochemistry, 8 days after the inoculation of the tumor. In these experiments, none of these parameters showed any alterations in mice treated with the residue. In order to evaluate the toxicity of butanolic residue, the animals were treated with this residue (200mg/Kg) for 14 days. The animals treated with the residue showed a discrete decrease on weight gain. There were no evidences of hepatic and renal toxicity, by measurement of enzymes activities of necrosis, and also lack of necrosis by histopathological analysis of liver and kidney. In another experiment, some parameters of macrofagic activity, 24 hours after the inoculation of tumor, were evalueted, by flow citometry. The animals received the butanolic residue for 7 days and then were inoculated with 5 x 106 tumor cells, intraperitoneally. The oxidative burst activated by Staphylococcus aureus was reduced on animals treated with the residue, but no differences on fagocytosis and spontaneous or oxidative burst esimulated by PMA were found. Finally, in vitro studies were performed. The effects of this residue on the cell viability, measured by the MTT assay, and on the cell cycle, by flow cytometry, in Ehrlich tumor cells were evaluated. The highest concentrations of the residue caused a decrease of the cell viability and an increase of death cell. Thus, the treatment with this residue, in vivo, caused an attenuation of the tumor development, likely due to a decrease in the ascitic fluid formation or cell death. The effects of this residue on cell death were more pronounced in cultured cells. These results point to a novel agent for the cancer therapy, but further studies must be performed to elucidate the mechanisms responsible for these effects.

As raízes de Pfaffia paniculata têm sido popularmente utilizadas com vários propósitos, assim como adjuvante na terapia contra o câncer. Recentemente demonstramos que as raízes pulverizadas de Pfaffia paniculata causam uma atenuação do crescimento do tumor de Ehrlich em sua forma ascítica. Os objetivos do presente estudo foram caracterizar os efeitos do extrato etanólico, resíduos aquoso ou butanólico de Pfaffia paniculata sobre o desenvolvimento do tumor de Ehrlich, assim como investigar seus possíveis mecanismos. Em um primeiro experimento, foi demonstrado que camundongos machos Swiss, portadores do tumor de Ehrlich ascítico, tratados com resíduo aquoso ou butanólico, apresentaram uma maior sobrevida em relação aos animais controle. Nos experimentos com o tumor na forma sólida ou ascítica, os animais receberam, diariamente, o resíduo butanólico por 16 dias (experimento com o tumor sólido) ou 15 dias (experimento com o tumor ascítico). No 8º dia de tratamento, foram inoculados com 2,5 x 106 células tumorais, no coxim plantar esquerdo, ou com 5,0 x 106 células tumorais, Intraperitonealmente. O tumor ascítico foi avaliado, 7 dias após a inoculação do tumor, pela quantificação do volume de fluído ascítico, concentração de células tumorais e número total de células tumorais. O tumor sólido foi avaliado pela mensuração diária das patas e pela morfometria da área de necrose em meio à massa tumoral em cortes corados por HE, e proliferação celular, por imunoistoquímica, 8 dias após a inoculação do tumor. Nesses experimentos, nenhum destes parâmetros avaliados apresentaram alterações nos camundongos tratados com este resíduo. A fim de se avaliar a toxicidade do resíduo butanólico, os animais foram tratados com este resíduo por 14 dias. Os animais tratados com este resíduo apresentaram uma discreta diminuição de ganho de peso. Não houve evidências de toxicidade hepática e renal, pela mensuração das atividades de enzimas indicativas de necrose e pelo exame histopatológico de figado e rim, onde não se observou necrose. Em outro experimento, alguns parâmetros da atividade macrofágica, 24 após a inoculação do tumor, foram avaliados, por citometria de fluxo. Os animais receberam o resíduo butanólico por 7 dias, e foram inoculados com 5 x 106 células tumorais, intraperitonealmente. O burst oxidativo ativado por Staphylococcus aureus foi menor em animais tratados com o resíduo, porém não foram observadas diferenças ma fagocitose e burst oxidativo espontâneo ou ativado por PMA. Por fim, estudos in vitro foram realizados. Os efeitos deste resíduo sobre a viabilidade, medido pelo ensaio do MTT, e sobre o ciclo celular, utilizando citometria de fluxo, de células do tumor de Ehrlich foram avaliados. As maiores concentrações do resíduo levaram a uma diminuição da viabilidade e um aumento de morte celular. Assim, o tratamento com este resíduo, in vivo, causou uma atenuação do desenvolvimento tumoral, provavelmente devido a uma diminuição na formação do fluído ascítico ou a morte celular. Os efeitos deste resíduo foram mais pronunciados em cultura celular. / The roots of Pfaffia paniculata have been popularly used for various purposes, as well as an adjuvant for cancer therapy. Recently we have shown that the powdered roots of Pfaffia paniculata promote an attenuation of the Ehrlich tumor growth in its ascitic form. The aims of the present study were to characterize the effects of the ethanolic extract, aqueous or butanolic residues of Pfaffia paniculata on Ehrlich tumor development in mice, as well as investigate its possible mechanisms. In a first experiment, it was shown that aqueous or butanolic residues-treated Swiss mice bearing Ehrlich ascites tumor presented a higher survival time than control animal. In the experiments with the tumor in its solid or ascitic forms, the animals were given, daily, the butanolic residue for 16 days (experiment with solid tumor) or 15 days (experiment with ascitic tumor). On the 8o day of treatment, they were inoculated with 2,5 x 106 tumor cells, on the left footpad, or with 5 x 106 tumor cells, intraperitoneally. The ascitic tumor was evaluated, 7 days after the inoculation of the tumor, by the quantification of the volume of the ascitic fluid, concentration of tumor cells and total number of tumor cells. The solid tumor was evaluated by the daily measurement of the footpads and morphometry of the necrosis within the tumor area on HE stained slices, and cell proliferation by immunohistochemistry, 8 days after the inoculation of the tumor. In these experiments, none of these parameters showed any alterations in mice treated with the residue. In order to evaluate the toxicity of butanolic residue, the animals were treated with this residue (200mg/Kg) for 14 days. The animals treated with the residue showed a discrete decrease on weight gain. There were no evidences of hepatic and renal toxicity, by measurement of enzymes activities of necrosis, and also lack of necrosis by histopathological analysis of liver and kidney. In another experiment, some parameters of macrofagic activity, 24 hours after the inoculation of tumor, were evalueted, by flow citometry. The animals received the butanolic residue for 7 days and then were inoculated with 5 x 106 tumor cells, intraperitoneally. The oxidative burst activated by Staphylococcus aureus was reduced on animals treated with the residue, but no differences on fagocytosis and spontaneous or oxidative burst esimulated by PMA were found. Finally, in vitro studies were performed. The effects of this residue on the cell viability, measured by the MTT assay, and on the cell cycle, by flow cytometry, in Ehrlich tumor cells were evaluated. The highest concentrations of the residue caused a decrease of the cell viability and an increase of death cell. Thus, the treatment with this residue, in vivo, caused an attenuation of the tumor development, likely due to a decrease in the ascitic fluid formation or cell death. The effects of this residue on cell death were more pronounced in cultured cells. These results point to a novel agent for the cancer therapy, but further studies must be performed to elucidate the mechanisms responsible for these effects.

Made available in DSpace on 2014-06-11T19:32:39Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-12-17Bitstream added on 2014-06-13T19:03:09Z : No. of bitstreams: 1 vilela_gf_dr_botfca.pdf: 1103988 bytes, checksum: accc5fdd3c2e93121b5bc3f01c2a9a80 (MD5) / Este estudo realizou a caracterização agronômica e análise do princípio ativo de cinco acessos da espécie medicinal Hebanthe eriantha, um recurso extrativista com a variabilidade genética ameaçada. A espécie Hebanthe eriantha, anteriormente denominada Pfaffia paniculata e depois Hebanthe paniculata, a partir de 2008 recebeu a atual denominação. Este estudo é uma parceria entre o Laboratório de Plantas Medicinais da UNESP-Botucatu, o Pólo Vale do Paraíba em Pindamonhangaba da Agência Paulista de Tecnologia dos Agronegócios (APTA) e as Divisões de Agrotecnologia e Química do Centro Pluridisciplinar de Pesquisas Químicas Biológicas e Agrícolas (CPQBA) da UNICAMP. Inicialmente, foi instalada uma coleção de trinta e nove acessos da espécie coletados na região do Vale do Paraíba no CPQBA e no Pólo Vale do Paraíba. Foram realizados testes de propagação por estaquia desses acessos e a partir dos resultados foi instalado um ensaio experimental a campo em blocos casualizados com cinco acessos e sete repetições. Foram avaliados o peso fresco/seco da parte aérea, o peso fresco/seco da raiz, o comprimento da maior haste, a área foliar, o índice de espuma e o teor de ácido fáfico das amostras. O acesso 1800 apresentou um peso seco de raiz, peso seco da parte aérea e área foliar superior aos demais acessos. O teor de ácido fáfico não diferiu entre os acessos e o acesso 1801 apresentou a maior média (0,64% m/m) seguido do acesso 1800 com uma média de (0,63% m/m). Existe uma correlação positiva entre área foliar e peso seco da parte aérea e peso seco de raízes entre os cinco acessos. / This survey realized a agronomic caracterization and active compounds analysis of five Hebanthe eriantha medicinal plant accessions, a extractivist resource on risk of genetic variability detriment. Hebanthe eriantha, before named Pfaffia paniculata and Hebanthe paniculata, in 2008 receive actual denomination. This study is a association between UNESPBotucatu Medicinal Plants Laboratory, Paraiba Valley Center (APTA) and CPQBAUNICAMP Agrotecnology Division. In first, it was established a collection of thirty nine accessions of the species collected on Paraiba Valley. Shoot development using cuttings was studied and a field experiment using randomized complete blocks with five accession and seven replications was used. The stems and leaves wet/dry weight, roots wet/dry weight, biggest stem length, foliar area, lather index and pfaffic acid content samples was avaliated. The accession 1800 had a higher root dry weight, stems and leaves dry weight and foliar area than others accessions. Pfaffic acid content was similar of anothers accessions and 1801 accession had highest media (0,64% m/m) and 1800 accession media (0,63% m/m). There are a positive correlation between accessions foliar area and stems and leaves dry weight with roots dry weight.

Orientadores: Maria Lígia Rodrigues Macedo, Maria das Graças Machado Freire / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-21T18:00:01Z (GMT). No. of bitstreams: 1 Martins_CarlosHenriqueZanini_M.pdf: 3256360 bytes, checksum: 92dcd99d5853476244d7668b4ceb774b (MD5) Previous issue date: 2007 / Resumo: O resumo poderá ser visualizado no texto completo da tese digital / Abstract: The abstract is available with the full electronic document / Mestrado / Bioquimica / Mestre em Biologia Funcional e Molecular

Submitted by LUCAS DE PAULA RAMOS null (lucas93paula@hotmail.com.br) on 2017-01-20T14:09:20Z No. of bitstreams: 1 Dissertação (correções biblioteca).pdf: 4172563 bytes, checksum: 3cfab803e9b5a294021bdd02082d4128 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2017-01-24T17:04:02Z (GMT) No. of bitstreams: 1 ramos_lp_me_sjc.pdf: 4172563 bytes, checksum: 3cfab803e9b5a294021bdd02082d4128 (MD5) / Made available in DSpace on 2017-01-24T17:04:02Z (GMT). No. of bitstreams: 1 ramos_lp_me_sjc.pdf: 4172563 bytes, checksum: 3cfab803e9b5a294021bdd02082d4128 (MD5) Previous issue date: 2016-12-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O objetivo do trabalho foi investigar se os extratos de Pfaffia paniculata K. e Juglans regia L. possuem ação antifúngica, antibacteriana e toxicidade celular, com testes in vitro. Para os testes antifúngicos foram utilizadas cepas ATCC de Candida spp., e para os testes antibacterianos foram utilizadas cepas ATCC de Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans e Pseudomonas aeruginosa. Para a atividade antimicrobiana primeiramente foram determinados os valores da Concentração Inibitória Mínima (CIM) e da Concentração Microbicida Mínima (CMM) dos extratos pelo método de microdiluição em caldo, segundo Clinical and Laboratory Standards Institute (CLSI). Os micro-organismos que apresentaram CMM foram selecionados para os testes em biofilme, no qual foi preparado em fundo de placa com 96 poços, por 48 h. Após os biofilmes foram tratados por 5 min. utilizando as concentrações de 200, 100 e 50 mg dos extratos. Para mensuração da biomassa foi utilizado o teste de Cristal violeta (CV), e para avaliar a atividade metabólica foi utilizado o teste de MTT. A citotoxicidade foi avaliada sobre fibroblastos gengivais humanos (FMM-1) utilizando os mesmos parâmetros de tratamento utilizados para os testes em biofilmes. Foram avaliadas a viabilidade celular pelos testes de MTT, vermelho neutro e cristal violeta. Os dados obtiveram distribuição normal e foram analisados por ANOVA e teste de Tukey, com significância de 5% (p<0.05%). O extrato de P. paniculata demostrou ação antifúngica em biofilmes, com reduções médias de 29,4 e 42,7% nos testes de CV e MTT. Já a ação antibacteriana foi restrita a S. mutans e P. aeruginosa com reduções médias de 15,7 e 28,6% nos respectivos testes. O extrato de J. regia também demostrou ação antifúngica com redução média de 22,2% na biomassa e 31,4% na atividade metabólica. A ação antimicrobiana ficou restrita a P. aeruginosa com reduções médias de 17,7 e 15,6%, indicados pelos testes de CV e MTT. Quanto a citotoxicidade, a média entre os três testes realizados, indicou que após exposição ao extrato de P. paniculata 58,8% das células continuaram viáveis e para J. regia a viabilidade foi de 65,1%. Conclui-se queo extrato de P. paniculata demostrou ação antifúngica sobre todas as cepas de Candida spp. testadas e demostrou ação antibacteriana para P. aeruginosa e S. mutans. As concentrações de 200, 100 e 50 mg do extrato demostraram ser citotóxicas conforme nova diretriz de toxicidade. J. regia demostrou ação antifúngica sobre todas as cepas de Candida spp. testadas e demostrou ação antibacteriana sobre P. aeruginosa. Apenas a concentração de 200 mg do extrato se mostrou tóxica a FMM-1. / The aim of this study was to investigate whether extracts of Pfaffia paniculata K. and Juglans regia L. have antifungal, antibacterial and cellular toxicity, with in vitro tests. ATCC strains of Candida spp. Were used for antifungal tests, and ATCC strains of Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans and Pseudomonas aeruginosa were used for the antibacterial tests. For the antimicrobial activity, the values of the Minimum Inhibitory Concentration (MIC) and the Minimal Microbicidal Concentration (CMM) of the extracts were determined by the microdilution method in broth, according to Clinical and Laboratory Standards Institute (CLSI). The microorganisms that presented CMM were selected for the biofilm tests, in which it was prepared on a 96-well plate bottom for 48 h. After the biofilms were treated for 5 min. Using the concentrations of 200, 100 and 50 mg of the extracts. To measure the biomass, the Violet Crystal test (CV) was used, and the MTT test was used to evaluate the metabolic activity. Cytotoxicity was assessed on human gingival fibroblasts (FMM-1) using the same treatment parameters used for biofilm tests. Cell viability was evaluated by the MTT, neutral red and violet crystal tests. The data obtained normal distribution and were analyzed by ANOVA and Tukey test, with significance of 5%. The extract of P. paniculata showed antifungal action in biofilms, with average reductions of 29.4 and 42.7% in CV and MTT tests; The antibacterial action was restricted to S. mutans and P. aeruginosa with mean reductions of 15.7 and 28.6% in the respective tests. The extract of J. regia also demonstrated antifungal action with an average reduction of 22.2% in biomass and 31.4% in metabolic activity. The antimicrobial action was restricted to P. aeruginosa with mean reductions of 17.7 and 15.6%, indicated by CV and MTT tests. As for cytotoxicity, the mean of the three tests carried out indicated that after exposure to P. paniculata extract 58.8% of the cells remained viable and for viability the viability was 65.1%. In conclusion the extract of P. paniculata showed antifungal action on all strains of Candida spp. Tested and demonstrated antibacterial action for P. aeruginosa and S. mutans. The concentrations of 200, 100 and 50 mg of the extract proved to be cytotoxic according to the new toxicity guideline. J. regia demonstrated antifungal action on all strains of Candida spp. Tested and demonstrated antibacterial action on P. aeruginosa. Only the 200 mg concentration of the extract was shown to be toxic to FMM-1.

Submitted by MARCOS LEANDRO TEIXEIRA DE OLIVEIRA (marcosteixeira@ufv.br) on 2018-08-13T17:46:27Z No. of bitstreams: 1 texto completo.pdf: 2650973 bytes, checksum: 095b915c3c693f319fe4151f3ac2a950 (MD5) / Made available in DSpace on 2018-08-13T17:46:27Z (GMT). No. of bitstreams: 1 texto completo.pdf: 2650973 bytes, checksum: 095b915c3c693f319fe4151f3ac2a950 (MD5) Previous issue date: 2017-03-31 / Fundação de Amparo à Pesquisa do Estado de MInas Gerais / Polygala paniculata is a medicinal species, whose root extract is used in the treatment of dislocations, trauma and ophidian accidents, therapeutic properties corroborated by science. The medicinal characteristic of plants is usually related to the presence of secretory structures specialized in the synthesis, accumulation and secretion of compounds with therapeutic activity. There are reports of trichomes, cavities and secretory cells of essential oils in P. paniculata. However, little is known about the development, chemical composition and secretory process of these structures. Species with medicinal value have been the target of different biotechnological techniques such as in vitro cultivation, with a view to maximizing the production of bioactive compounds. Species respond in different ways to the manipulation of in vitro conditions, such as light quality, gelling agents and sucrose concentration, which can result in changes in the quality of the plants and, consequently, the secretion produced. Thus, the present study aimed to (1) identify and analyze the development and secretory sites of essential oils; (2) to investigate the subcellular aspects and mechanisms of synthesis, accumulation and release of secretion; (3) Compare the ultrastructural aspects between the identified secretory sites and (4) to evaluate the effects of manipulation of in vitro conditions (light quality, gelling agents and sucrose concentration) on the leaf anatomy of P. paniculata. Polygala paniculata presents three secretory sites of essential oils: trichomes in leaves and stems, cavities in leaves and cortical cells in roots. The development of trichomes and cavities involves the process of programmed cell death. Trichomes, cavities and cortical root cells have the secretion composed of essential oils. The subcellular characteristics of the secretory structures studied are similar and are related to the chemical nature of the secretion. In the in vitro culture, the factors evaluated altered the leaf morphoanatomy of P. paniculata. Fluorescent lamp treatments induced the development of seedlings and the anatomical differentiation similar to that observed in ex situ plants. Treatments with sucrose concentration of 30 g L -1 in agar in 15 gL -1 in Phytagel induced increase in leaf thickness by increasing the cell volume of epidermal cells, lacunar parenchyma and reduction of intercellular spaces. The present study elucidated the structural and viiimechanism of the secretion of essential oils in P. paniculata. The effect of the in vitro conditions on the growth and foliar anatomy of the species can appreciate and subsist to improve in protocols of in vitro cultivation of the species. / Polygala paniculata is a medicinal species, whose root extract is used in the treatment of dislocations, trauma and ophidian accidents, therapeutic properties corroborated by science. The medicinal characteristic of plants is usually related to the presence of secretory structures specialized in the synthesis, accumulation and secretion of compounds with therapeutic activity. There are reports of trichomes, cavities and secretory cells of essential oils in P. paniculata. However, little is known about the development, chemical composition and secretory process of these structures. Species with medicinal value have been the target of different biotechnological techniques such as in vitro cultivation, with a view to maximizing the production of bioactive compounds. Species respond in different ways to the manipulation of in vitro conditions, such as light quality, gelling agents and sucrose concentration, which can result in changes in the quality of the plants and, consequently, the secretion produced. Thus, the present study aimed to (1) identify and analyze the development and secretory sites of essential oils; (2) to investigate the subcellular aspects and mechanisms of synthesis, accumulation and release of secretion; (3) Compare the ultrastructural aspects between the identified secretory sites and (4) to evaluate the effects of manipulation of in vitro conditions (light quality, gelling agents and sucrose concentration) on the leaf anatomy of P. paniculata. Polygala paniculata presents three secretory sites of essential oils: trichomes in leaves and stems, cavities in leaves and cortical cells in roots. The development of trichomes and cavities involves the process of programmed cell death. Trichomes, cavities and cortical root cells have the secretion composed of essential oils. The subcellular characteristics of the secretory structures studied are similar and are related to the chemical nature of the secretion. In the in vitro culture, the factors evaluated altered the leaf morphoanatomy of P. paniculata. Fluorescent lamp treatments induced the development of seedlings and the anatomical differentiation similar to that observed in ex situ plants. Treatments with sucrose concentration of 30 g L -1 in agar in 15 gL -1 in Phytagel induced increase in leaf thickness by increasing the cell volume of epidermal cells, lacunar parenchyma and reduction of intercellular spaces. The present study elucidated the structural and mechanism of the secretion of essential oils in P. paniculata. The effect of the in vitro conditions on the growth and foliar anatomy of the species can appreciate and subsist to improve in protocols of in vitro cultivation of the species.

Coastal dune vegetation plays a key role in dune formation and stabilization through sediment trapping and erosion control. To restore degraded dunes, revegetation of dune building species is critical. Planting density has been found to effect growth of marsh species, with closer plantings alleviating stress through facilitation. As coastal dunes are high stress environments, it is expected that dune species may also exhibit facilitative interactions based on the Stress Gradient Hypothesis. Therefore, planting grasses in clumped configurations may lead to more successful dune revegetation. The objective of this research was to determine how planting density affects the growth of two dominant dune grasses along the US Atlantic coast, Ammophila breviligulata and Uniola paniculata, through field surveys of natural distribution, density, and a manipulation study of planting densities. Natural distribution differed between the two species with A. breviligulata occurring at lower dunes and U. paniculata occurring at higher dunes. Ammophila breviligulata occurred more densely than U. paniculata. Planting density of U. paniculata had an effect on growth parameters (shoot length, stem number, and ramet number) but not survival with dispersed plantings (50 cm apart) having higher growth than clumped plantings. The effect of density planting on growth parameters may impact dune building processes. Sparsely planted U. paniculata may have greater sediment capture compared to densely planted U. paniculata due to greater stem number and biomass resulting in taller, steeper dunes.

O objetivo do presente trabalho foi estudar a biologia e elaborar a tabela de vida de fertilidade de Diaphorina citri (Kuwayama, 1908) (Hemiptera: Liviidae) em duas espécies de Murraya [Murraya paniculata (L.) Jack e Murraya koenigii (L.) Sprengel] e estudar a biologia de Tamarixia radiata Waterston, 1922 (Hymenoptera: Eulophidae) em psilídeos provenientes das duas espécies de Murraya citadas, visando fornecer subsídios ao controle da praga em programas de MIP. Foram realizados experimentos em confinamento (sem chance de escolha) para avaliar o efeito de duas espécies de Murraya sobre a alimentação e fertilidade de D. citri, bem como a biologia da referida espécie de psilídeo nos dois hospedeiros. Também foi avaliado o efeito do hospedeiro no parasitismo sobre D. citri e na biologia de T. radiata. Pode-se concluir que: D. citri preferiu se alimentar em M. koenigii, porém para a oviposição o hospedeiro preferido foi M. paniculata. No entanto, os demais parâmetros observados indicaram que M. paniculata é o hospedeiro mais adequado para o desenvolvimento do psilídeo. Com base na tabela de vida de fertilidade, a taxa liquida de reprodução (R0), de D. citri, foi 3,1 vezes maior quando criado em M. paniculata; os demais parâmetros, exceto o tempo médio de cada geração (T), também favoreceram o desenvolvimento do inseto quando criado em M. paniculata. O desenvolvimento e parasitismo de T. radiata sobre ninfas de D. citri foram afetados quando criadas nas duas espécies, sendo observados melhores resultados quando os psilídeos foram criados em M. koenigii em testes de confinamento. Os resultados obtidos poderão fornecer subsídios ao controle e manejo do psilídeo transmissor do HLB. / The aim of the present research was to evaluate the biology and fertility life table of Diaphorina citri (Kuwayama, 1908) (Hemiptera: Liviidae) comparatives in the two species of Murraya [Murraya paniculata (L.) Jack e Murraya koenigii (L.) Sprengel], and to evaluate the biology of Tamarixia radiata Waterston, 1922 (Hymenoptera: Eulophidae) in the two species studied in confinement, aiming to provide subsidies for pest control in Integrated Pest Management (IPM). Experiments were carried out in confinement (no choice) to evaluate the effect of the two species of Murraya on the feeding and fertility of D. citri, as well as the biology of Asian Citrus Psyllid (ACP) in two hosts. Biology and the effect of host on parasitism of T. radiata on nymphs of D. citri was evaluated. It can be concluded that: D. citri preferred to feed on Murraya koenigii (L.) Sprengel, but for oviposition the preferred host was Murraya paniculata (L.) Jack. However, the other parameters observed indicate that M. paniculata is the most suitable host for the development of the psyllid. According to the fertility life table, the reproduction net rate (R0) of D. citri was 3.1 times higher if reared on M. paniculata; the other parameters, except the mean time of each generation (T), also supported results when it was reared on M. paniculata. The development and parasitism of T. radiata on nymphs of D. citri were affected when reared on different hosts, and better results were observed when T. radiata reared on M. koenigii. The results obtained may provide subsidies for the control and management of the HLB.