Orientador: Eliana Regina Forni Martins / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-25T23:43:47Z (GMT). No. of bitstreams: 1 Braga_KlenyaRosaRocha_D.pdf: 3638781 bytes, checksum: f3a0b3548e4388cdabb276fb9662c32a (MD5) Previous issue date: 2014 / Resumo: A família Asteraceae (Compositae) possui cerca de 23.000 espécies e 1.600 gêneros. Sua distribuição é cosmopolita, com representantes em todos os continentes. Sua maior diversidade está na América do Sul. A família está bem representada no Brasil (cerca de 300 gêneros e 2.000 espécies), ocorrendo principalmente em formações abertas, como cerrados e campos. Cassini, em 1817 foi o primeiro a propor um sistema de classificação para a família, que foi alterado por vários autores. Atualmente a família está dividida 12 subfamílias e 43 tribos. Dentre os parâmetros que podem ser obtidos a partir de estudos citogenéticos, o número cromossômico é o mais amplamente utilizado. Estima-se que cerca de 61,6% dos gêneros de Asteraceae possuem pelo menos uma contagem de números cromossômicos, sendo relatada grande variação (2n= 4 a ca. 432). O número básico x=9 é o mais frequente. Apesar do número considerável de trabalhos referentes à contagem cromossômica e análise de cariótipos de espécies de Asteraceae, este número ainda é muito pequeno quando comparado com a riqueza de espécies da família. No que diz respeito aos estudos cromossômicos utilizando bandamento cromossômico e hibridização de DNA in situ, os trabalhos são mais escassos. Diante da riqueza de espécies, da diversidade cariotípica da família e da carência de dados citogenéticos para subsidiar a reavaliação das delimitações de seus grupos taxonômicos, este trabalhou objetivou caracterizar o cariótipo de diversas espécies nativas de Asteraceae proveniente da Reserva Biológica Municipal (RBM) da Serra do Japi uma das últimas grandes áreas de floresta contínua do Estado de São Paulo, que apresenta uma grande diversidade de espécies de Asteraceae. Foram feitos a determinação do número cromossômico, o detalhamento de caracteres morfológicos dos cromossomos (tamanho e forma) e a aplicação de técnicas de bandamento CMA/DAPI e técnica de FISH. Foi determinado o número cromossômico de 35 espécies, as quais estão distribuídas em 14 tribos e 26 gêneros, com variação de 2n=16 a 72. As tribos mais bem representadas foram Eupatorieae (9spp.), Vernonieae (6spp.) e Astereae (5spp.). Seis novas contagens foram determinadas e quatro divergiram de relatos anteriores: Bidens subalternans (2n=72), Chromolaena odorata (2n=30), Praxelis kleineoides (2n=30) e Vernonia diffusa (2n=34). Os cariótipos podem ser considerados moderadamente simétricos, com o índice TF% variando de 29 em Praxelis kleineoides (2n=30) a 44 into G. intermedia (2n=20). Dezesseis espécies foram submetidas à técnica de bandamento CMA/DAPI. Houve variação de 2 a 12 bandas CMA+/DAPI-. A técnica de FISH revelou diferenças cariotípicas em cinco espécies estudadas: Conyza sumatrensis var. leotheca, Bidens pilosa, Mikania sp, Vernonia diffusa e V. remotiflora, que apresentaram 2 a 6 sítios de DNAr 45S e 2 a 3 sítios de DNAr 5S. Nossos dados de bandamento CMA/DAPI e FISH são inéditos para todas as espécies estudadas / Abstract: The Asteraceae family (Compositae) comprises around 23.000 species and 1.600 genera. Its distribution is comospolitan, with representatives in all continents. The highest diversity is found in South America. The family is well represented in Brazil (with 300 genera and 2.000 species), mainly occurring in the opened formations, like savannas and fields. Cassini, in 1817 was the first to propose a system of classification for the family, which was later changed by several authors. Nowadays the family is divided in 12 subfamilies and 43 tribes. Among the parameters that can be obtained from cytogenetic studies, the number of chromosomes is the most commonly used. Its estimated that about 61,6% of Asteraceae genera have at least a count chromosome numbers, being reported wide variation (2n= 4 a ca. 432). The basic number x=9 is the most frequent. In spite of considerable number of works on the chromosomal count and karyotpe analysis from species of Asteraceae, this number is still very small when compared with species richness within the family. In respect of chromosomal studies used in the chromosome banding and in situ DNA hybridization, the works are more scarce. Before wealth of species, of karyotype diversity of family and of lack of data cytogenetics to subsidize the revaluation of delimitations of its taxonomic groups, this work aims to characterize the karyotype of several native species of Asteraceae from the Municipal Biological Reserve (MBR) of Japi, one of the last large areas of continuous forest of São Paulo, that has a great diversity of species of Asteraceae. We determined the chromosomal number, detailing the morphological characters of chromosomes (size and shape), and applied techniques of CMA/DAPI banding and FISH. We determined the chromosome number of 35 species, which are distributed in 14 tribes and 26 genera, ranging from 2n = 16-72. The most well represented tribes were Eupatorieae (9spp.), Vernonieae (6spp.) and Astereae (5spp.). Six new counts were determined and four differed from previous scores: Bidens subalternans (2n=72), Chromolaena odorata (2n=30), Praxelis kleineoides (2n=30) and Vernonia diffusa (2n=34). The karyotpes can be considered moderately symmetric, with index TF% ranging from 29 in Praxelis kleineoides (2n=30) to 44 in G. intermedia (2n=20). Sixteen species were submitted to technique of banding CMA/DAPI. There were variations of 2 to 12 CMA+/DAPI- bands. The technique of FISH revealed varied karyotypes in five species: Conyza sumatrensis var. leotheca, B. pilosa, Mikania sp, Vernonia diffusa and V. remotiflora showed 2 to 6 sites of 45S rDNA and 2 to 3 sites of 5S rDNA. Our data of banding CMA/DAPI and FISH are original for all studied species / Doutorado / Biologia Vegetal / Doutora em Biologia Vegetal

In this study two medicinal plant species, namely Artemisia afra Jacq. ex. Willd and Elytropappus rhinocerotis Less. (L.f) (= Dicerotamnus rhinocerotis Koekemoer), both belonging to the family Asteraceae, have been investigated and different compounds isolated and characterised. Both species are important plants used in traditional medicine in general in Africa and particularly in South Africa. A. afra, commonly called "African wormwood" is one of 400 species belonging to the genus Artemisia and it is the only one indigenous to Africa. E. rhinocerotis is one of eight Elytropappus species which are all restricted to the Cape floristic region. The aim of this study was to investigate the phytochemistry of these species. In total fifteen compounds were isolated and characterised. From the E. rhinocerotis extract, four known compounds, labdanolic acid, methyl labdanolate, 6, 7- dimethoxycoumarin and a sesquiterpene viridiflorol were isolated. These compounds were not previously reported from E. rhinocerotis. Two different chemotypes of A. afra were studied and eleven compounds were isolated. These compounds include sesquiterpenes such as taurin, artesin, maritimin, artemin, norsantolinifolide, santolinifolide A, and reynosin, a flavonoid, 5- hydroxy-7,4-dimethoxyflavone, a coumarin called scopoletin or 7-hydroxy-6- methoxycoumarin and other aromatic compounds such as p-hydroxyacetophenone, and 2,4-dihydroxy-6-methoxyacetophenone. Except for taurin, scopoletin and 5-hydroxy-7, 4- dimethoxyflavone , none of these other compounds has been reported previously from A. afra. This study has shown that A. afra contains a large number of sesquiterpenoids, mostly from the eudesmane-type. Structural elucidation of different compounds was performed using mainly NMR spectroscopy. Other methods used for identification include LC-MS and infrared spectroscopy. The major compound, labdanolic acid, is known to selectively inhibit cyclooxygenase-2, an enzyme associated with inflammation. The presence of labdanolic acid in the plant may account for its traditional use as an anti-inflammatory. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

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O processo de transformação microbiana de metabólitos secundários é uma técnica emergente no Brasil e que apresenta enorme potencial para obtenção de substâncias biotransformadas com novas e diferentes moléculas, as quais podem apresentar melhor desempenho de suas atividades biológicas. O objetivo do presente projeto foi efetuar transformações microbianas de metabólitos secundários oriundos da espécie vegetal Viguiera robusta Gardn. (família Asteraceae), com ênfase nas lactonas sesquiterpênicas (LSTs), para posterior avaliação de sua citotoxicidade frente a linhagens celulares. As etapas de isolamento e purificação de LSTs foram realizadas através de métodos cromatográficos e a elucidação estrutural através de métodos espectroscópicos. A avaliação da atividade citotóxica dos metabólitos biotransformados foi proposta em virtude de estudos iniciais indicarem potencial citotóxico da LST budleína A frente às linhagens SK-BR-3 (adenocarcinoma de mama) e células leucêmicas JURKAT. / The microbial transformation of secondary metabolites is an emerging technique in Brazil with high potential to obtain biotransformed compounds with new and different molecules which can show enhanced biological activities. Herein, microbial transformations of secondary metabolites from Viguiera robusta Gardn. (family Asteraceae), with emphasis in sesquiterpene lactones (STLs), were proposed with the aim to evaluate their cytotoxic activity against tumor cells lines. Isolation and purification of STLs were carried out through chromatographic methods and structural elucidation by spectroscopic methods. The evaluation of cytotoxic activity of the biotransformed metabolites was proposed due to the fact that previous studies indicated potential cytotoxic activity of the STLs budlein A against SK-BR-3 and JURKAT cells lines.

Submitted by Cleide Dantas (cleidedantas@ufpa.br) on 2014-12-10T00:11:12Z No. of bitstreams: 2 license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Dissertacao_AvaliacaoPotencialMicrobicida.pdf: 2937381 bytes, checksum: 7a4f1498597c6eb85a59a088c3577ee3 (MD5) / Approved for entry into archive by Ana Rosa Silva (arosa@ufpa.br) on 2014-12-11T14:47:37Z (GMT) No. of bitstreams: 2 license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Dissertacao_AvaliacaoPotencialMicrobicida.pdf: 2937381 bytes, checksum: 7a4f1498597c6eb85a59a088c3577ee3 (MD5) / Made available in DSpace on 2014-12-11T14:47:37Z (GMT). No. of bitstreams: 2 license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Dissertacao_AvaliacaoPotencialMicrobicida.pdf: 2937381 bytes, checksum: 7a4f1498597c6eb85a59a088c3577ee3 (MD5) Previous issue date: 2014 / Há necessidade da pesquisa para a descoberta de novas drogas antimicrobianas contra estes patógenos multirresistentes que veem crescendo, progressivamente, durante os últimos anos. As potencialidades de uso das plantas medicinais contra estes microrganismos se encontram em estado de evolução inicial, seja como recurso terapêutico, seja como fundamento para o surgimento de novos fármacos. Também vale atentar para o fato de que a biodiversidade amazônica constitui fator relevante no estado para a pesquisa destas novas terapêuticas baseadas nos recursos vegetais disponíveis. Tratando-se de uma prática que estimula a participação direta da população no processo saúde-doença, a realização da avaliação da espécie Ayapana triplinervis V., que é uma planta de prevalência amazônica e de uso tradicional, mostra-se essencial, principalmente para preconizar a realização de prospecções fitoquímicas. Além de possibilitar a caracterização de seus componentes e de frações do material vegetal, assim como determinar avaliações farmacológicas destes produtos, tais como atividade antimicrobiana, a fim de determinar a eficácia e garantir a segurança ao uso de plantas medicinais, e de seus derivados e produtos fitoterápicos.

<p>The genus <i>Arnica</i> (Asteraceae) was investigated for phylogenetic relationships and sesquiterpene lactone (STL) content with the aims to trace the evolutionary history of the genus and to investigate possible congruencies between DNA sequence data, secondary chemistry, and biological activity. </p><p>Complex evolutionary patterns in <i>Arnica</i> are evident from phylogenetic analyses of chloroplast regions (the <i>rpl16</i> and <i>rps16</i> introns and the <i>psbA–trnH</i>, <i>ycf4–cemA</i>, and <i>trnT–L</i> spacers), nuclear ribosomal regions (the internal and external transcribed spacers) and the nuclear low-copy DNA region coding for the second largest subunit of RNA polymerase II (<i>RPB2</i>) between exons 17 and 23. Polymorphism was detected in nuclear ribosomal and low-copy regions<i>,</i> likely caused by polyploidy and agamospermy. Lineage sorting and/or hybridization is a possible explanation for incongruencies between topologies of the different DNA regions. None of the five subgenera in <i>Arnica</i> constitute a monophyletic group according to any of our analyses. </p><p>Sesquiterpene lactone profiles were compared to nuclear ribosomal DNA data using phylogenetic inference and principal component analysis for 33 accessions of 16 species. Clusters supported by both STL chemistry and ribosomal DNA sequence data consist of multiple accessions of the same species (e.g.<i> A montana </i>and<i> A. longifolia</i>), indicating that these species are well defined both genetically and chemically, based on our sampling. Support for subspecies classification of <i>A. chamissonis</i> and <i>A. parryi</i> was found in chemical data. For the first time STLs are reported from subtribe Madiinae, sister to Arniciinae.</p><p>Anti-inflammatory properties, as measured by inhibition of human neutrophil elastase release from neutrophils and inhibition of the binding of transcription factor NF-κB to DNA, were investigated for extracts of 12 <i>Arnica</i> species. <i>Arnica montana</i>, <i>A. chamissonis</i> and <i>A. longifolia</i> accessions show high inhibitory effects in both bioassays. Generally, species with a more diverse STL chemistry also possess the strongest inhibitory activity in the bioassays.</p>

The genus Arnica (Asteraceae) was investigated for phylogenetic relationships and sesquiterpene lactone (STL) content with the aims to trace the evolutionary history of the genus and to investigate possible congruencies between DNA sequence data, secondary chemistry, and biological activity. Complex evolutionary patterns in Arnica are evident from phylogenetic analyses of chloroplast regions (the rpl16 and rps16 introns and the psbA–trnH, ycf4–cemA, and trnT–L spacers), nuclear ribosomal regions (the internal and external transcribed spacers) and the nuclear low-copy DNA region coding for the second largest subunit of RNA polymerase II (RPB2) between exons 17 and 23. Polymorphism was detected in nuclear ribosomal and low-copy regions, likely caused by polyploidy and agamospermy. Lineage sorting and/or hybridization is a possible explanation for incongruencies between topologies of the different DNA regions. None of the five subgenera in Arnica constitute a monophyletic group according to any of our analyses. Sesquiterpene lactone profiles were compared to nuclear ribosomal DNA data using phylogenetic inference and principal component analysis for 33 accessions of 16 species. Clusters supported by both STL chemistry and ribosomal DNA sequence data consist of multiple accessions of the same species (e.g. A montana and A. longifolia), indicating that these species are well defined both genetically and chemically, based on our sampling. Support for subspecies classification of A. chamissonis and A. parryi was found in chemical data. For the first time STLs are reported from subtribe Madiinae, sister to Arniciinae. Anti-inflammatory properties, as measured by inhibition of human neutrophil elastase release from neutrophils and inhibition of the binding of transcription factor NF-κB to DNA, were investigated for extracts of 12 Arnica species. Arnica montana, A. chamissonis and A. longifolia accessions show high inhibitory effects in both bioassays. Generally, species with a more diverse STL chemistry also possess the strongest inhibitory activity in the bioassays.

Las proteasas aspárticas típicas de plantas (o tipo pepsina) han sido escasamente estudiadas en comparación con sus contrapartes de origen no vegetal. Una excepción a esta afirmación, lo constituyen las cardosinas (Cynara cardunculus) usadas en la manufactura de quesos y la fitepsina (Hordeum vulgare), que al día de hoy continúa siendo una de las pocas proteasas de esta clase caracterizada estructuralmente y por tanto, el modelo utilizado en esta área. Una de las principales razones por las cuales las PAs de plantas no han sido objeto de mayor cantidad de estudios es su escasa abundancia natural. Ésto resulta especialmente cierto en el caso de sus precursores, los cuales contienen el llamativo dominio PSI (inserto específico de plantas), único entre las PAs. En un intento por identificar nuevas fuentes vegetales de PAs típicas, extractos de flores de varias especies pertenecientes a la familia Asteraceae que crecen en Argentina fueron probados en ensayos de coagulación de leche. Una de las especies que mostró esta capacidad coagulante sugiriendo por tanto la presencia de PAs, fue Cirsium vulgare -comúnmente conocido como cardo negro y considerada una maleza en los campos de nuestra región. Fundamentalmente debido a su amplia disponibilidad y a la ausencia de estudios reportados sobre sus PAs, se decidió emplear esta especie de la familia Asteraceae como material vegetal. En este contexto, el presente trabajo intenta generar un aporte al conocimiento de las PAs de origen vegetal mediante el estudio de procirsina, una proenzima de C. vulgare perteneciente a la familia A1, y del dominio PSI contenido en su secuencia.

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El género Stomatanthes pertenece a la familia Asteraceae, tribu Eupatorieae. Fue descripto en el año 1970 por King y Robinson como una segregación de Eupatorium sensu lato, el cual contiene cerca de 1000 especies. Originalmente fue definido para acomodar una especie de Eupatorium (Eupatorium africanum) de África, y posteriormente se agregaron 12 especies sudamericanas y dos más fueron reconocidas para África. Stomatanthes comprende 17 especies que se distribuyen en dos áreas disyuntas: Sureste de América del Sur (Brasil, Paraguay, Uruguay, Bolivia, Argentina) y África (centro y este). La mayor parte de sus especies (13) son sudamericanas. Como resultado de la revisión sistemática, todas las especies de Stomatanthes, fueron por primera vez correctamente descriptas e ilustradas, aportándose mapas de distribución, datos ecológicos, fenológicos y clave para la identificación de las distintas especies del género. Se describe una nueva especie: S. reticulatus. Se define un nombre nuevo para S. pernambucensis (S. bracteatus nov. nom). Se seleccionan 5 lectotipos y 1 neotipo. Se reconocen como sinónimos por primera vez los siguientes nombres: Kuhnia glabrata. var.α, Kuhnia glabrata var.β, Eupatorium oblongifolium f. odoratissima, E. polycephalum var. pinnatifidum. Se resuelven problemas nomenclaturales complejos en la especie Stomatanthes polycephalus, y se enmienda su descripción original. Se excluyen de Stomatanthes por primera vez: Eupatorium bracteatum var. reticulatum (=Bejaranoa balansae) y Eupatorium oblongifolium var. paraguariensis.