O presente trabalho visou a avaliação do potencial antifúngico de extratos brutos das diferentes partes do vegetal (folha, caule e raiz) de espécies de Piperaceae, os quais foram submetidos ao ensaio de bioautografia frente aos fungos Cladosporium cladosporioides e Cladosporium sphaerospermum. Após a triagem biológica, os extratos brutos das espécies Piper mollicomum, Piper marginatum, Peperomia alata e Peperomia glabella foram selecionados para etapas posteriores de fracionamento e purificação, utilizando-se técnicas cromatográficas. O estudo fitoquímico biomonitorado das espécies selecionadas identificou como metabólitos ativos dois cromenos e uma di-hidrochalcona; duas flavanonas e três fenilpropanóides, sendo um de estrutura inédita, denominado marginatumol; dois policetídeos inéditos, alatanona A e alatanona B; quatro acilresorcinóis e quatro secolignanas, incluindo a peperomina G inédita. Todos os metabólitos isolados apresentaram potencial atividade antifúngica, sendo os policetídeos alatanona A e alatanona B os mais ativos, com atividade superior às substâncias controle (nistatina e miconazol). Assim, os dados obtidos neste estudo permitiram descrever diferentes classes de produtos naturais a partir de uma coleção de extratos de espécies de Piperaceae, com potencial para estudos de relações estrutura-atividade. / The present study was addressed to evaluate the antifungal activity of crude extracts from Piperaceae species in which different parts of the plant (leaves, stems and roots) were assessed by means of bioautography against Cladosporium cladosporioides and Cladosporium sphaerospermum. After antifungal assay, the extratcs from Piper mollicomum, Piper marginatum, Peperomia alata and Peperomia glabella displayed higher growth inhibitory potency and thus were selected for further fractionation and purification using chromatographic techniques. Such dereplication using the bioautographic assay yielded two chromenes and one dihydrochalcone; two flavanones and three phenylpropanoids, including the novel compound marginatumol; two new polyketides, alatanone A and alatanone B; four acylresorcinols and four secolignans, including the novel peperomin G, respectively. The bioautography indicated potential antifungal activity for all isolated metabolites and compounds alatanone A and alatanone B were found the most active in comparison to positive controls (nystatin and miconazole). The bioprospecting approach was proven to be very effective when the isolation of bioactive compounds from a collection of crude extracts is concerned, although not necessarily unraveling the major biosynthetic pathways in each plant species.

Made available in DSpace on 2015-05-14T12:59:56Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 8420094 bytes, checksum: e38d9fce0fc67703d2f3e878474b7c32 (MD5) Previous issue date: 2013-02-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The Piper L. genus is the largest one among the Piperaceae. Containing at least 1000 species, they are located mainly on the neotropical region of Earth and it draws our attention due to its medical, economical, chemical and cultural properties, such as anti-inflamatory, antibacterial and fungicide actions. Piper genus species have been becoming research sources for specific classes of secondary metabolites which play an important role concerning biological activities, including alkaloids, amides, chalcones, chromenes, flavonoids, lignans, terpenes, cyclopentanediones, steroids, porphyrin derivatives and benzoic acid derivatives. Piper caldense C. DC., which is commonly known as Pimenta D arda , is used in Paraíba as sedative, snake venom antidote, anti-toothache agent, and as active in located pain treatments through the use of compresses. This research reports the phytochemical study of P. caldense C. DC. leaves and fruits. Through usual chromatographic methods and spectroscopic techniques such as IR and one and two-dimensional ¹H and ³CNMR techniques and comparison with literature data it was possible to isolate and identify an mixture of the β-sitosterol and sitgmasterol steroids, besides porphyrin substances: 15-hydroxy-A-pophyrinlactone and a phaeophytin. Those fractions were obtained from crude ethanolic extract of P. caldense C. DC. leaves. Three benzoic acid derivatives were isolated from the hexane extract of P. caldense C. DC. fruits. From its leaves and fruits it was possible to obtain seven compunds commonly found it many vegetal species, but for the first time stated from P. caldense C. DC. The hexane and dichloromethane fruit extracts and isolated substances were submitted to microbiological assay and antifungal activites were detected concerning Candida genus yeast. This test is being done for the first time in the fruits of the studied species. / O gênero Piper L. é o maior da família Piperaceae, compreendendo pelo menos 1000 espécies, que se encontram distribuídas especialmente na região neotropical do globo terrestre e se destaca por suas propriedades medicinais, econômicas, químicas e culturais (anti-inflamatória, antibacteriana, anestésia e fungicida, dentre outros). Espécies do gênero Piper têm-se tornado fontes de pesquisa de classes específicas de metabólitos secundários com marcantes atividades biológicas, incluindo alcaloides, amidas, chalconas, cromenos, flavonoides, lignanas, terpenos e ciclopentanodionas, esteroides, derivados porfirínicos e derivados do ácido benzoico. Piper caldense C. DC., conhecida popularmente como pimenta d arda , é utilizada na Paraíba como sedativa, antídoto para picadas de cobras, para dores de dente, bem como na forma de compressa no local afetado para alívio da dor. Este trabalho reporta o estudo fitoquímico das folhas e frutos de P.caldense C. DC. Utilizando-se métodos cromatográficos usuais e técnicas espectroscópicas de IV e RMN de 1H e 13C uni e bidimensionais e a comparação dos dados com a literatura foi possível isolar e identificar do extrato etanólico bruto das folhas de Piper caldense C.DC., uma mistura de esteroides, o β-sitosterol e o estigmasterol, e duas substâncias porfirínicas, a 15-hidroxi-porfirionolactona A e a feofitina a, e, do extrato hexânico dos frutos, três derivados do ácido benzoico, compreendendo ao total sete substâncias, descritas pela primeira vez em Piper caldense C. DC. Os extratos hexânico e diclorometânico dos frutos e suas substâncias isoladas foram submetidos a ensaios microbiológicos, apresentando atividade antifúngica frente a leveduras do gênero Candida. Este ensaio está sendo feito pela primeira vez nos frutos da espécie estudada.

O presente trabalho visou a avaliação do potencial antifúngico de extratos brutos das diferentes partes do vegetal (folha, caule e raiz) de espécies de Piperaceae, os quais foram submetidos ao ensaio de bioautografia frente aos fungos Cladosporium cladosporioides e Cladosporium sphaerospermum. Após a triagem biológica, os extratos brutos das espécies Piper mollicomum, Piper marginatum, Peperomia alata e Peperomia glabella foram selecionados para etapas posteriores de fracionamento e purificação, utilizando-se técnicas cromatográficas. O estudo fitoquímico biomonitorado das espécies selecionadas identificou como metabólitos ativos dois cromenos e uma di-hidrochalcona; duas flavanonas e três fenilpropanóides, sendo um de estrutura inédita, denominado marginatumol; dois policetídeos inéditos, alatanona A e alatanona B; quatro acilresorcinóis e quatro secolignanas, incluindo a peperomina G inédita. Todos os metabólitos isolados apresentaram potencial atividade antifúngica, sendo os policetídeos alatanona A e alatanona B os mais ativos, com atividade superior às substâncias controle (nistatina e miconazol). Assim, os dados obtidos neste estudo permitiram descrever diferentes classes de produtos naturais a partir de uma coleção de extratos de espécies de Piperaceae, com potencial para estudos de relações estrutura-atividade. / The present study was addressed to evaluate the antifungal activity of crude extracts from Piperaceae species in which different parts of the plant (leaves, stems and roots) were assessed by means of bioautography against Cladosporium cladosporioides and Cladosporium sphaerospermum. After antifungal assay, the extratcs from Piper mollicomum, Piper marginatum, Peperomia alata and Peperomia glabella displayed higher growth inhibitory potency and thus were selected for further fractionation and purification using chromatographic techniques. Such dereplication using the bioautographic assay yielded two chromenes and one dihydrochalcone; two flavanones and three phenylpropanoids, including the novel compound marginatumol; two new polyketides, alatanone A and alatanone B; four acylresorcinols and four secolignans, including the novel peperomin G, respectively. The bioautography indicated potential antifungal activity for all isolated metabolites and compounds alatanone A and alatanone B were found the most active in comparison to positive controls (nystatin and miconazole). The bioprospecting approach was proven to be very effective when the isolation of bioactive compounds from a collection of crude extracts is concerned, although not necessarily unraveling the major biosynthetic pathways in each plant species.

Evolution of Piperales – matK gene and trnK intron sequence data reveal lineage specific resolution contrast. Piperales are one of the largest basal angiosperm orders with a nearly worldwide distribution. The order includes three species rich genera, Piper (ca. 1,000 species), Peperomia (ca. 1,500-1,700 species), and Aristolochia s. l. (ca. 500 species). Sequences of the matK gene and the non-coding trnK group II intron are analysed for a dense set of 105 taxa representing all families (except Hydnoraceae) and all generic segregates (except Euglypha within Aristolochiaceae) of Piperales. A large number of highly informative indels are found in the Piperales trnK/matK dataset. Within a narrow region approximately 500 nt downstream in the matK coding region (CDS), a length variable simple sequence repeat (SSR) expansion segment occurs, in which insertions and deletions have led to short frame-shifts. These are corrected shortly afterwards, resulting in a maximum of 6 amino acids being affected. Furthermore, additional non-functional matK copies were found in Zippelia begoniifolia, which can easily be discriminated from the functional open reading frame (ORF). The trnK/matK sequence data fully resolve relationships within Peperomia, whereas they are not effective within Piper. The resolution contrast is correlated with the rate heterogenity between those lineages. Parsimony, Bayesian and likelihood analyses result in virtually the same topology, and converge on the monophyly of Piperaceae and Saururaceae. Lactoris gains high support as sister to Aristolochiaceae subf. Aristolochioideae, but the different tree inference methods yield conflicting results with respect to the relationships of subfam. Asaroideae. In Piperaceae, a clade formed by the monotypic genus Zippelia and the small genus Manekia (=Sarcorhachis) is sister to the two large genera Piper and Peperomia. Systematics of pipevines – Combining morphological and fast-evolving molecular characters to investigate the relationships within subfamily Aristolochioideae (Aristolochiaceae) A combined phylogenetic analysis of the Aristolochioideae was conducted based on 72 morphological characters and molecular datasets (matK gene, trnK intron, trnL intron, trnL-trnF spacer). The analysis sampled 33 species as the ingroup, including two species of Thottea and 30 species of Aristolochia and the monotypic genus Euglypha, which represent all the infrageneric taxa formally described; Saruma henryi and Asarum caudatum were used as the outgroup. The results corroborate a sister-group relationship between Thottea and Aristolochia, and the paraphyly of Aristolochia with respect to Euglypha that consequently should be included into Aristolochia. Two of the three subgenera within Aristolochia (Isotrema and Pararistolochia) are shown to be monophyletic, whereas the signal obtained from the different datasets about the relationships within subg. Aristolochia is low and conflicting, resulting in collapsed or unsupported branches. The relationship between the New World and the Old World species of subgenus Aristolochia is conflictive because morphological data support these two groups as monophyletic, whereas molecular data show the monophyletic Old World species of Aristolochia nested within the New World species. A sister group relationship is proposed between A. lindneri and pentandrous species, which suggests that a group of five species from central and southern South America (including A. lindneri) could be monophyletic and sister to Aristolochia subsection Pentandrae, a monophyletic taxon consisting of about 35 species from southern USA, Mesoamerica, and the West Indies. Colonisation, phylogeography and evolution of endemism in Mediterranean Aristolochia (Aristolochiaceae). This study provides evidence for a multiple colonisation of the western Old World from Asian ancestors within Aristolochia section Diplolobus (subsection Aristolochia and Podanthemum). Within subsection Podanthemum it is assumed, that the colonisation of the African continent happened at least two times independently. In contrast, for subsection Aristolochia, a rapid morphological radiation in the Near East (or close to this area) with subsequent star like colonisation of the different current distribution areas, which is not paralleled on the molecular level, appears to be more likely. Phylogenetic tree reconstruction is unsupported for these clades, but most clades are highly supported as monophyletic. Interestingly the Mediterranean and temperate Eurasian species, which are morphologically distinct (A. pistolochia, A. clematitis) are not clustering within the main clades, but are independent lineages. Analogue, A. rigida a species from Somalia is well-supported sister to the subsection Aristolochia. Within subsection Podanthemum the colonisation event from an Asian ancestor is clearly traceable, whereas in subsection Aristolochia the path is not traceable, since the ancestors are extinct or not present in the connecting areas. Within the Mediterranean, Near East and Caucasian species of subsection Aristolochia two morphologically and biogeographically well supported groups can be identified: the Near East/Caucasian species and the West Mediterranean species. The previous groupings for the latter, based on morphological characters, could be substantiated only partly by our results. This study provides the first phylogeny of all West Mediterranean species. In addition an independent complex is established including some micro endemic species. The phylogenetic results are discussed with respect to biogeography, and morphology, to give a first insight into the radiation and colonisation of the genus Aristolochia in the Mediterranean region. Universal primers for a large cryptically simple cpDNA microsatellite region in Aristolochia. We provide a new and valuable marker to study species relationships and population genetics in order to trace evolutionary, ecological, and conservational aspects in the genus Aristolochia. Universal primers for amplification and subsequent sequencing of a chloroplast microsatellite locus inside the trnK intron are presented. Utility of the primers has been tested in 32 species representing all clades of Aristolochia, including population studies within the A. pallida complex, A. clusii and A. rotunda. The microsatellite region is characterized as a (AnTm)k repeat of 22–438 bp containing a combination of different repeats arranged as ‘cryptically simple’. Trapped! Pollination of Aristolochia pallida Willd. in the Mediterranean A first study of the pollination biology of a Mediterranean Aristolochia species in its natural habitat is presented. 183 flowers of Aristolochia pallida were investigated, which in total contained 73 arthropods, dominated by two groups of Diptera, Sciaridae (37%) and Phoridae (19%). However, only Phoridae are regarded as potential pollinators, since pollen has been found exclusively on the body surfaces of these insects. All Phoridae belong to the genus Megaselia and are recognised as four undescribed species. The measurements of flower and insect dimensions suggest that size is an important constrain for successful pollination: 1) the insects must have a definitive size for being able to enter the flower and 2) must be able to get in touch with the pollen. Only very few insect groups found in Aristolochia pallida fulfil these size requirements. However, size alone is not a sufficient constrain as too many fly species of the same size might be trapped but not function as pollinators. Instead, specific attraction is required as otherwise pollen is lost. Since all trapped Phoridae are males, a chemical attraction (pheromones) is proposed as an additional constrain. Since A. pallida flowers are protogynous, the record of Megaselia loaded with pollen found in a flower during its female stage proves that this insect must have been visited at least one different flower during its male stage before. Further on, this observation provides strong evidence that the flowers are cross-pollinated. All these factors indicate a highly specialised pollination of Aristolochia pallida by Megaselia species.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-01-26T13:52:37Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_MetodoIdentificacaoGenes.pdf: 963765 bytes, checksum: 9cb1e27661c2b5e611915be85e11786d (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-01-27T13:31:03Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_MetodoIdentificacaoGenes.pdf: 963765 bytes, checksum: 9cb1e27661c2b5e611915be85e11786d (MD5) / Made available in DSpace on 2017-01-27T13:31:03Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_MetodoIdentificacaoGenes.pdf: 963765 bytes, checksum: 9cb1e27661c2b5e611915be85e11786d (MD5) Previous issue date: 2015-09-28 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A pimenta do reino (Piper nigrum) é uma planta de alto interesse biotecnológico, alvo de pesquisas tanto para a exploração do metabolismo quanto para o melhoramento relacionado a problemas fitopatológicos, além do entendimento da evolução das angiospermas basais, grupo ancestral ao qual ela pertence. O avanço nos métodos de sequenciamento de nova geração proporcionou o acesso ao patrimônio genético de plantas não modelo possibilitando a abertura de novas perspectivas biotecnológicas. A identificação de genes não homólogos restritos a certas espécies, denominados genes taxonomicamente restritos (GTRs), é um alvo biotecnológico prioritário, especialmente em espécies e grupos divergentes e ancestrais. Este trabalho tem por objetivo estabelecer um método de identificação de GTRs a partir de dados de RNA-seq e de validar a abordagem num conjunto de dados de pimenta do reino. O método consiste na filtragem de transcritos em várias etapas, de forma que os transcritos anotados e os falsos positivos são retirados, e os dados restantes sem informações moleculares são classificados como potenciais GTRs. A aplicação da abordagem em dados de transcriptoma de pimenta do reino (35.631 transcritos) resultou em 22.661 transcritos anotados por similaridade. Os transcritos não anotados nessa primeira análise foram processados na ferramenta TRAPID, obtendo 12.895 transcritos não anotados. A avaliação dos transcritos para detecção de falsos positivos resultou em 245 transcritos verdadeiros, que foram analisados quando a presença de RNA não codificante, sendo encontrados 204 transcritos sem identificação. Ao final da aplicação do método restaram 71 transcritos não anotados com regiões codificantes de proteínas, assinalados como potenciais GTRs. A caracterização desses potenciais GTRs em pimenta do reino pode fornecer novas informações sobre o mecanismo molecular dessa espécie e talvez elucidar vias para o estabelecimento de cultivares tolerantes a doenças. / Black pepper (Piper nigrum) is a biotechnologically interesting plant, research target for both metabolism exploration and improvement related to phytopathological problems, in addition to understanding the evolution of basal angiosperms, ancestral group to which it belongs. With the technological revolution, the next generation sequencing offered access to genetic heritage of non model plants enabling the opening of new biotechnological perspective. The identification of non homologous genes restricted to certain species, called taxonomically restricted genes (TRGs), is a primary biotechnological target, especially in species and groups that are divergent and ancestral. This study aims to establish a method for TRGs identification from RNA-seq data and to validate the approach a dataset for black pepper. The method consists in filtering the transcripts in several stages, so that the annotated transcripts and false positives are removed, and the remaining data without molecular information are classified as potential TRGs. The application of this approach to a black pepper transcriptome dataset (35,631 transcripts) resulted in 22,661 transcripts annotated by similarity. The transcripts that were not annotated in this first analysis were processed in the TRAPID tool, resulting in 12,895 transcripts not annotated. The evaluation of transcripts for false positive detection resulted in 245 true transcripts that were analyzed for the presence of non-coding RNA, resulting in 204 unidentified transcripts. At the end of the method application 71 non annotated transcripts remained with coding regions of protein, indicating potential TRGs. The characterization of these potential TRGs in black pepper can provide new information about the molecular mechanism of this specie and perhaps elucidate pathways for the establishment of cultivars tolerant to disease.

Thesis (Ph. D.)--Miami University, Dept. of Botany, 2005. / Title from second page of PDF document. Includes bibliographical references.

Made available in DSpace on 2014-06-11T19:26:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-10-13Bitstream added on 2014-06-13T18:30:23Z : No. of bitstreams: 1 ferreira_mi_me_botfca.pdf: 413587 bytes, checksum: 89293821d623c0c862c4d0148e01479d (MD5) / Piper aduncum é conhecida popularmente como pimenta-de-macaco, aperta-ruão, jaborandi do mato, pimenta-do-fruto-ganchoso, falso-jaborandi, jaguarandi, matico e pimenta-longa. Seu uso na medicina popular é amplamente relatado para doenças ginecológicas, desordens intestinais, diurético, carminativo, digestivo, para males do fígado, no combate a erisipela e tratamento de úlceras crônicas. A espécie é rica em óleo essencial, e este apresenta atividade inseticida, larvicida, antimicrobiana, antifúngica, antitumoral e apresenta baixa toxicidade. Porém na área agronômica os estudos com a espécie são bastante escassos. Modificações nos níveis de luminosidade a que uma espécie está submetida podem acarretar diferentes respostas em suas características fisiológicas, bioquímicas, anatômicas e de crescimento e, consequentemente, podem alterar a sua composição química e atividade biológica. Desse modo, o presente estudo tem como objetivo avaliar o desempenho ecofisiológico e a produção de biomassa, além da produtividade e qualidade do óleo essencial de folhas e raízes de Piper aduncum sob diferentes condições de intensidade luminosa, contribuindo para o conhecimento e conservação das plantas com potencial medicinal e para o desenvolvimento de futuras pesquisas. O delineamento experimental utilizado foi o inteiramente casualizado, com três tratamentos e sete repetições, sendo que cada parcela constou de cinco plantas. Os tratamentos consistiram em três níveis de luminosidade: 0% (pleno sol), 50% e 70% de sombreamento, tendo sido avaliados aos 177 dias. As plantas apresentaram melhor desempenho ecofisiológico sob sombreamento de 50% e 70%, considerando-se as maiores taxas de concentração interna de CO2 e condutância estomática, porém a assimilação... / Piper aduncum is popularly known as the “pimenta-de-macaco, apertaruão, jaborandi do mato, pimenta-do-fruto-ganchoso, falso-jaborandi, jaguarandi, matico e pimenta-longa”. Its use in folk medicine is widely reported to gynecological diseases, intestinal disorders, diuretic, carminative, digestive, for ailments of the liver, in fighting to erysipelas and treatment of chronic ulcers. The species is rich in essential oil, this activity presents insecticide, larvicide, antimicrobial, antifungal, antitumor and has low toxicity. But agronomy studies with the species are scarce. Changes in light levels to which a species is subjected may cause different responses in their physiological, biochemical, anatomical and growth and hence may alter the chemical composition and biological activity. Thus, this study aims to evaluate the ecophysiological and biomass production, as well as productivity and quality of essential oil of leaves and roots of Piper aduncum under different light intensity, contributing to the knowledge and conservation of plants with potential medicine and the development of future research. The experimental design was completely randomized design with three treatments and seven replicates, each plot consisted of five plants. The treatments 4 consisted of three levels of lightness: 0% (full sun), 50% and 70%, having been assessed at 177 days. The results allowed to say that the plants showed better ecophysiological performance under 50% and 70% of shade, considering the higher rates of internal CO2 concentration and stomatal conductance, but showed no statistical difference in CO2 assimilation, and showed lowest transpiration in full sun. In relation to biometric parameters, the plants showed higher dry mass of stem, height and leaf area to 70%, and the total dry mass and dry mass of leaves were not... (Complete abstract click electronic access below)

Submitted by Viviane Lima da Cunha (viviane@biblioteca.ufpb.br) on 2016-03-29T14:36:11Z No. of bitstreams: 1 arquivototal.pdf: 5620328 bytes, checksum: cc12fb802b3be6b2f1e66e8519ace0cf (MD5) / Made available in DSpace on 2016-03-29T14:36:11Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 5620328 bytes, checksum: cc12fb802b3be6b2f1e66e8519ace0cf (MD5) Previous issue date: 2015-02-26 / The species Peperomia obtusifolia (L.) A. Dietr, which belongs to the Piperaceae family, is a well-known plant, distributed from Mexico to South America. Previous phytochemical studies demonstrated the presence of compounds from several classes, such as: lignans, flavonoids, amides, phenolic compounds and chromenes, which demonstrated interesting biological activities. This work describes the metabolomic analysis of Peperomia obtusifolia, which study in real time, the metabolism of this species in many kinds of scenarios. However, to accomplish this study, firstly it was developed a standard protocol, capable of preparing the samples in a reproducible way, enabling the following metabolomic studies to generate reliable results. Later, it was conducted the metabolomic study of Peperomia obtusifolia and its varieties, by fingerprinting analysis, in order to characterize the metabolomic profiles of each one of them. Furthermore, it was also evaluated the behavior of the species against the stress induced by the herbivory of Monoplatus sp. In general, the analyzes were conducted by NMR of 1H (which the data was interpreted by PCA), mass spectrometry, HPLC, GC and its hyphenated techniques. As a result, it was found that, despite the fact that the Peperomia obtusifolia varieties belong to the same species, they had different metabolomes. Regarding the study of the biotic stress against the plant, changes were detected on the metabolomic profile of the attacked plants, indicating a response against the herbivore attack. As a major result of this research, it can be highlighted the fact that the same changes were also detected on the metabolome of the intact plants closed to the attacked ones, indicating a sort of interplant communication by volatile compounds, capable to induce the neighborhood yet not attacked, to be prepared for an imminent attack, giving them a better chance of survival. / A espécie Peperomia obtusifolia (L.) A. Dietr, pertencente à família Piperaceae, é uma planta amplamente conhecida, cuja ocorrência vai desde o México até a América do Sul. Estudos fitoquímicos anteriores revelaram a presença de compostos de diversas classes, como: lignanas, flavonoides, amidas, compostos fenólicos e cromenos, os quais demonstraram atividade biológica bastante promissora. Este trabalho aborda a análise metabolômica de Peperomia obtusifolia, que visa estudar em tempo real o metabolismo desta espécie em diversos tipos de cenários. Entretanto, para que este estudo fosse realizado, primeiramente foi desenvolvido um protocolo padrão, capaz de preparar amostras de maneira reprodutível, possibilitando assim uma análise metabolômica que gerasse resultados fidedignos. Posteriormente foi realizado o estudo metabolômico de Peperomia obtusifolia e suas variedades, por análise de fingerprinting, que visa caracterizar os perfis metabólicos de cada organismo. Além disso, também foi avaliado o metaboloma da espécie frente ao estresse biótico provocado pela herbivoria do besouro Monoplatus sp. As análises das amostras foram realizadas por meio de RMN de 1H (cujos dados foram interpretados por PCA), espectrometria de massas, CLAE, CG e técnicas hifenadas. Como resultado, descobriu-se que apesar das variedades de Peperomia obtusifolia pertencerem à mesma espécie, elas possuíam metabolomas diferentes. Já em relação ao estudo de estresse biótico aplicado à planta, foram detectadas nas plantas predadas, alterações nos perfis metabolômicos, configurando-se uma resposta contra o ataque provocado. Considera-se um resultado de maior importância, o fato de que plantas intactas que estavam próximas às predadas, responderam ao ataque de maneira semelhante, indicando assim, uma comunicação inter-plantas por meio de compostos voláteis, capaz de induzir a vizinhança ainda não atacada a se preparar para reagir ao ataque, possibilitando assim, maior chance de sobrevivência.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-01-26T14:15:37Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_ProducaoAvaliacaoAtividade.pdf: 1531568 bytes, checksum: fa545ee0497fdfbb6c535111d161a8bc (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-01-27T13:40:46Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_ProducaoAvaliacaoAtividade.pdf: 1531568 bytes, checksum: fa545ee0497fdfbb6c535111d161a8bc (MD5) / Made available in DSpace on 2017-01-27T13:40:46Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_ProducaoAvaliacaoAtividade.pdf: 1531568 bytes, checksum: fa545ee0497fdfbb6c535111d161a8bc (MD5) Previous issue date: 2015-08-06 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Muitas espécies do gênero Piper apresentam ampla distribuição na Amazônia e diversas aplicações biológicas devido a grande diversidade estrutural de seus metabólitos secundários. A espécie Piper divaricatum, é endêmica da Amazônia e produz em seu óleo essencial elevadas concentrações de metileugenol (50 – 90%), um fenilpropanóide com propriedade antioxidante e fungicida. Devido às suas potenciais aplicações, o objetivo deste estudo foi o estabelecimento do cultivo in vitro e a comparação da biossintese dos metabólitos secundários e propriedades antioxidantes com o cultivo in vivo. Para o estabelecimento do cultivo in vitro foram utilizados ápices caulinares cultivados em meio Murashige e Skoog , acrescido do regulador de crescimento BAP 0,5 mg.mL-1. Para o cultivo in vivo, foram propagadas microestacas em casa de vegetação no substrato vermiculita e adição de solução nutritiva de Murashige e Skoog. Os compostos voláteis identificados nas folhas das plântulas cultivadas in vivo foram metileugenol, β-elemeno e E-β-ocimeno, os quais não diferiram do cultivo in vivo, com exceção dos 90 dias. O cultivo in vitro das raízes não se mostrou eficiente para produção de fenilpropanóides e apresentou um perfil bastante diferenciado em relação ao cultivo in vivo de terpenos. De uma maneira geral, para as plantas cultivadas in vitro não houve diferença estatística significativa no teor de compostos fenólicos e na atividade antioxidante das folhas. No entanto, a atividade antioxidante das raízes foi bastante expressiva. Os resultados obtidos reforçam a hipótese de que plantas regeneradas in vitro podem sintetizar metabólitos semelhantes a planta-mãe e manter suas propriedades biológicas. / Many species of the genus Piper are widely distributed in the Amazon and various biological applications because of large structural diversity of its secondary metabolites. The species Piper divaricatum, is endemic in the Amazon and produces in its essential oil high concentrations of methyleugenol (50-90%), an phenylpropanoid with antioxidant and fungicidal properties. Because of its potential applications, the objective of this study was to establish the in vitro cultivation and comparing the biosynthesis of secondary metabolites and antioxidant properties with the in vivo cultivation. For establish the in vitro culture were used shoot apexes on Murashige e Skoog medium with addition of regulator BAP 0.5 mg.mL-1. For in vivo cultivation, micropiles were propagated in the greenhouse in vermiculite and adding nutritious Murashige e Skoog solution. The volatile compounds identified in the leaves of seedlings grown in vivo were methyleugenol, β-elemene and E-β-ocimene, which did not differ from in vivo cultivation, with the exception of 90 days. The in vitro culture of roots was not efficient to produce phenylpropanoids and presented a very different profile compared to the in vivo cultivation of terpenes. In general, for plants in vitro cultivated there was no statistically significant difference in the phenolics compounds content and antioxidant activity in the leaves. However, the antioxidant activity of roots was significant. The results support the hypothesis that in vitro regenerated plants can synthesize metabolites similar the matrix plant and maintain their biological properties.

Submitted by Nathalya Silva (nathyjf033@gmail.com) on 2017-05-30T15:35:08Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AvaliacaoAtividadeAntinociceptiva.pdf: 1956189 bytes, checksum: 7517747ee93d0e00de15afa1deb6685f (MD5) / Approved for entry into archive by Edisangela Bastos (edisangela@ufpa.br) on 2017-06-06T21:41:17Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AvaliacaoAtividadeAntinociceptiva.pdf: 1956189 bytes, checksum: 7517747ee93d0e00de15afa1deb6685f (MD5) / Made available in DSpace on 2017-06-06T21:41:17Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AvaliacaoAtividadeAntinociceptiva.pdf: 1956189 bytes, checksum: 7517747ee93d0e00de15afa1deb6685f (MD5) Previous issue date: 2016-10-13 / A Peperomia pellucida (Piperaceae) é uma planta herbácea comumente encontrada nos continentes americano e asiático. Na Amazônia a espécie é conhecida pelo nome de erva-de-jabuti. Esta planta é utilizada, na medicina popular, para tratar uma vasta gama de sintomas e doenças, tais como conjuntivite, dor de cabeça, asma, úlcera gástrica, inflamação. Este estudo visou analisar a atividade antinociceptiva e anti-inflamatória dessa substância, assim como elucidar seu mecanismo de ação. Para todos os ensaios foram utilizados camundongos albinos suíços, machos (25 - 40 g), que foram inicialmente tratados com a pellucidina A nas doses de 0,5; 1 e 5 mg/kg (i.p.), submetidos a avaliação locomotora pelo ensaio de campo aberto e a modelos animais de dor aguda, como os testes de contorção abdominal induzida por ácido acético, testes da formalina e o ensaio da placa-quente. Em seguida, foi realizado o teste de contorção abdominal induzida por ácido acético para a elucidação do mecanismo de ação, no qual os animais foram tratados na dose padrão de 5 mg/kg (i.p.) e para análise anti-inflamatória do composto foi utilizado o modelo de granuloma induzido por pelotas de algodão, no qual os animais foram tratados na dose de 10 mg/kg (i.p.). O composto, não demonstrou alterar a deambulação dos animais em nenhuma das doses administradas. No ensaio de contorção a Pellucidina A conseguiu inibir em 43% o número de contorções abdominais na dose de 1 mg/kg, e 65% na dose 5 mg/kg. No teste da formalina, observou-se efeito antinociceptivo apenas dose de 5 mg/kg, com redução de 68% no tempo de lambida da pata do animal na fase inflamatória. Os animais tratados com a Pellucidina A e submetidos ao ensaio da placa-quente não demonstram possuir qualquer alteração em seu tempo de latência sobre a placa, apresentando uma resposta similar aos animais tratados apenas com a solução veículo. Para a elucidação do mecanismo de ação a Pellucidina A foi administrada na dose padrão de 5 mg/kg (i.p.) e associada com a Indometacina (5 mg/kg i.p.), o NS-398 (10 mg/kg i.p.), a cipro-heptadina (0,5 mg/kg i.p.), a naloxona (1mg/kg i.p.) e o L-NAME (5 mg/kg i.p). A Pellucidina A manifestou uma ação sinérgica quando associada com cipro-heptadina e L-NAME, apresentando uma diminuição no padrão de contorções abdominais em 97% quando associado com a cipro-heptadina e 96% com L-NAME. Na análise da ação da Pellucidina A (10 mg/kg i.p.) no teste do granuloma induzido por pelotas de algodão, a Pellucidina A apresentou atividade anti-inflamatória, reduzido em 24% a formação de granuloma nos camundongos tratados. Os resultados obtidos corroboram com a hipótese de que a Pellucidina A apresenta atividade analgésica, possuindo ação periférica, capaz de interferir no processo inflamatório, atuando como um possível agonista glicocorticóide. / The Peperomia pellucida (Piperaceae) is an herbaceous plant commonly found in the American and Asian continents. In the Amazon the species is known by the name of erva-de-jabuti. This plant is used in folk medicine to treat a wide range of symptoms and diseases such as conjunctivitis, headache, asthma, gastric ulcer, inflammation and arthritis. Pellucidin A is an isolated compound of the species Peperomia pellucida and this study aimed to analyze the antinociceptive and anti-inflammatory activity of this compound, as well as to elucidate its mechanism of action. For the assays, male albino mice (25-40 g) were used, which were initially treated with pellucidin A at the doses of 0.5; 1 and 5 mg / kg (i.p.) and subjected to locomotor evaluation by the open field test and animal models of acute pain, such as acetic acid-induced abdominal writhing tests, formalin tests and the hot plate test. The acetic acid-induced abdominal contortion test was realized to elucidate the mechanism of action in which the animals were treated at the standard dose of 5 mg/kg (i.p.) and for anti-inflammatory analysis of the compound was used model of granuloma induced by pellets of cotton, in which the animals were treated in the dose of 10 mg/kg (i.p.). The compound did not show capacity to change the ambulation of the animals at any of the administered doses. In the contortion test, pellucidin A was able to inhibit the number of abdominal writhings in 43% at the dose of 1 mg/kg, and 65% at the dose of 5 mg/kg. In the formalin test, an antinociceptive effect was observed at the dose of 5 mg/kg, with a 68% reduction in the lymph time of the animal's paw in the inflammatory phase, showing a similar response to Indomethacin used at a dose of 10 mg/kg as positive control for this phase. Animals treated with pellucidin A and subjected to the hot plate assay did not show any change in their latency time on the plate, showing a similar response to the animals treated just with the vehicle solution. For the elucidation of the action mechanism, the pellucidin A was administered at the standard dose of 5 mg/kg (i.p.) and associated with Indomethacin (5 mg/kg i.p.), NS-398 (10 mg/kg i.p.) cyproeptadine (0.5 mg/kg i.p.), naloxone (1 mg/kg i.p.) and L-NAME (5 mg/kg i.p.). The pellucidin A has shown a synergistic action when associated with cyproeptadine and L-NAME, with a decrease in the pattern of abdominal writhing by 97% when associated with cyproheptadine and 96% with L-NAME. In the analysis of the action of pellucidin A (10 mg/kg i.p.) in the granuloma test induced by cotton pellets, pellucidin A presented anti-inflammatory activity, reduced granuloma formation in 24% in the treated mice. The results confirm the hypothesis that pellucin A presents analgesic activity capable of interfering in the inflammatory process, acting as a possible glucocorticoid agonist.