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Item Morfo-anatomia e fitoquímica de Cymbopogon densiflorus (Steud.) Stapf e Cymbopogon nardus (L.) Rendle (Poaceae: Panicoideae)(Universidade Federal de Goiás, 2007-09-28) BARBOSA, Lília Cristina de Souza; PAULA, José Realino de; http://lattes.cnpq.br/3191837532986128; REZENDE, Maria Helena; http://lattes.cnpq.br/5093753722360659The genus Cymbopogon Sprengel belong to the Poaceae family and it has 40 species distributed in Tropical and Subtropical Africa, Asia and Australia, although some species went introduced in America. Many species of this genus are cultivated for the extraction of essential oil, from their leaves, with large medicinal, food and industrial importance. The species in focus, Cymbopogon densiflorus (Steud.) Stapf and C. nardus (L.) Rendle are originated from Africa and Asia, respectively. This research had as objective, to broaden the knowledge about the species C. densiflorus and C. nardus, by the morphological and anatomy studies from leaves and culms, phytochemical analysis and essential oil analysis from the leaves. Anatomical studies have been of relevant importance to the pharmacognosy researches, mainly for the identification of many vegetal raw materials. Several times, these raw materials are known by the same popular name or then, they are commercialized with contaminated agents or with other parts of the specie. Through of anatomical analysis, it was checked commons characters, such as leaf lamina and sheath amphistomata, stomatas with guard cells dumbbell and subsidiary cells dome-shape, rares in adaxial surface and abundant in abaxial surface, predominated in intercostal zones. The adaxial and abaxial surfaces had long cells and short cells: cork and dumb-bell and cross-shaped silica cells, these last it is placed in costal zones; macro-hairs and micro-hairs abundant in abaxial surface. In the leaf lamina, bulliforms cells are presents in adaxial surface, they were alternated with fibers in the costal zones and the mesophyll is homogeneous with chlorenchyma radiated to the bundle sheaths and arm cells with walls invaginated that they determined the intervenal distance by one or three cells, characterized Kranz anatomy. Bundle sheaths collateral, of 1st, 2nd and 3rd orders with single vascular bundle sheaths. The cap region is constituted by sclerenchyma and the epidermis has silica cells. However, both species had different anatomical features, as the form of midrib, in the leaf laminas; C. densiflorus showed colourless parenchyma cells in the mesophyll of leaf sheaths, that they do not exist in C. nardus. In the culms, numbers of metaxylem vessels in the each side of protoxylem vessels in vascular bundles: 1, in C. densiflorus, 2 and 3, in C. nardus; and the presence of sclerenchyma cylinder and fistula in C. nardus, absent characters in C. densiflorus. Moreover, in C. densiflorus, while C. nardus showed these characters. The preliminary phytochemistry analysis C. densiflorus and C. nardus leaves evidencied flavonoids, saponins, coumarins and traces of cardioactive glycosides. In the essential oil analysis, C. densiflorus leaves showed trans-p-mentha-1(7),8-dien-2-ol, trans-p-mentha-2-8-dien-1-ol, cis-carveol and cis-p-mentha-2-8-dien-1-ol as majority constituents; while C. nardus leaves had geraniol, citronellol and citronellal. The anatomical characters observed can be important to the taxonomic determinations of species studied, in the genus. Through the results found, it verifies the phytotherapics potential of both species. Future researches in isolation and purify of the secondary metabolites, pharmacologics and toxicologics analysis of extracts and of the essential oil, it will be important to assure the therapeutic efficiency of these.Item Clonagem e expressão do gene da tiorredoxina 1 de Paracoccidioides brasiliensis em Pichia pastoris(Universidade Federal de Goiás, 2010-08-27) CINTRA, Lorena Cardoso; FARIA, Fabrícia Paula de; http://lattes.cnpq.br/3739169267521003; JESUÍNO, Rosália Santos Amorim; http://lattes.cnpq.br/5113656623817587The termodimorphic fungus Paracoccidioides brasiliensis is the etiological agent of paracoccidioidomycosis, a human systemic mycosis of high prevalence in Latin America. P. brasiliensis is exposed to oxidative stress (OS) caused by reactive oxygen species (ROS) produced by the defense cells of the human host. When the invasion by pathogens occurs, the host defense system generates ROS to fight the invader. Inside the human host, P. brasiliensis is phagocytosed by macrophages, facing an extremely hostile environment due to nitric oxide and hydrogen peroxide. The Trx1 is an intracellular redox protein, which participates in the maintenance of cell redox homeostasis, both in terms of OS as reducer. It is ubiquitous and is characterized by typical CXXC active site, responsible for oxidation, reduction, or isomerization of proteins disulfide bonds. In a previous work, it was isolated, characterized and cloned into expression vector pGEX-4T-3 cDNA coding for TRX1 of P. brasiliensis (accession number AY376435). The recombinant protein (recPbTRX1) was produced and partially purified and the yeast cells of P. brasiliensis showed increased expression of the gene coding for PbTRX1 in response to OS. This study aimed the heterologous expression of cDNA of a thioredoxin of the fungus P. brasiliensis in Pichia pastoris, in order to obtain it in larger amounts for their subsequent biochemical characterization and application in biotechnological processes. The P. brasiliensis thioredoxin 1 (trx1) cDNA was obtained via PCR using the plasmid pGEX-Trx1 as template and cloned into expression vector pHIL-D2 and pPIC9 (for intracellular and extracellular expression). The insertion of the interested gene in the correct orientation was verified by sequencing and the homology was observed with Trx1 P. brasiliensis. These vectors were used to transform the P. pastoris yeast strain SMD1168 with his4- genotype. The presence of the cassette s expression was confirmed in the yeast s genome. No transformants able to secrete the protein from the building with the vector pPIC9 were detected and the intracellular production was carried from the pHIL-D2 vector.Item Identificação de isolados de Trichoderma spp. utilizando marcadores do tipo RAPD e DNA Barcode(Universidade Federal de Goiás, 2010-03-03) SANTOS, Patrícia Ribeiro dos; COELHO, Alexandre Siqueira Guedes; http://lattes.cnpq.br/0840926305216925; ULHOA, Cirano Jose; http://lattes.cnpq.br/8368469162867277Biological control is a natural process that regulates the number of individuals existing in a population, by the action of another individual called "natural enemies or biological control agent (parasitoids, predators and pathogens), causing biotic mortality. Fungi that act as antagonists of phytopathogenic fungi have been used to control the disease, and 90% of these applications have been made using strains of fungi of the genus Trichoderma. The genus Trichoderma (Ascomycetes) Hypocreales, belonging to the class and brings Hifomicetos species that are among the soil fungi most commonly found in nature. There are several problems related to the nomenclature of individuals of this gender. Molecular tools such as markers and DNA Barcode has been used in identification of Trichoderma but also another fungi. The objective of this work was to identify isolates of Trichoderma spp. using only molecular tools. Analysis using markers make possible the observation of the high degree of genetic variability between these individuals, but proved problematic when the sample size was increased. The identification of individuals using DNA Barcode was possible only for a few individuals who show the need for the high quality sequences obtained by sequencing. The phylogenetic analysis was extremely difficult because is time consuming and required much time to perform the analysis. However by Neighbor-joining analysis was observed that when using databases such as BLAST error rate in identifying these species is high due to deposition of sequences of isolates incorrectly identified.Item Morfologia, Anatomia foliar e Fitoquímica de espécies de Eriope Humb. &Bonpl. ex Benth. (Lamiaceae) ocorrentes em Goiás(Universidade Federal de Goiás, 2008-06-26) SCHLIEWE, Marcos Augusto; PAULA, José Realino de; http://lattes.cnpq.br/3191837532986128; REZENDE, Maria Helena; http://lattes.cnpq.br/5093753722360659Eriope with about 30 species is a genus of the family Lamiaceae, subfamily Nepetoideae, tribe Ocimae and subtribe Hyptidinae, out of this 20 species are restricted to the campos rupestres of the Brazil. In Goiás, the Chapada dos Veadeiros, Serra de Caldas, Serra dos Cristais, Serra Dourada and Serra dos Pireneus are typical rupestres environments. Beyond these places, in environments of cerrado, in the cities of Colinas do Sul, Goiânia, Mineiros, Planaltina de Goiás, Uruaçu and Vianópolis were located specimens of Eriope. The aim of this work is to supply new morphologic, anatomical and phytochemistry data to contribute with the taxonomy of Eriope. Eight taxa were collected: Eriope complicata Mart. ex Benth., Eriope crassipes Benth., Eriope crassipes Benth. subsp. crassipes var. acutifolia Benth., Eriope crassipes Benth. subsp. crassipes var. macrophylla Benth., Eriope crassipes Benth. subsp. cristalinae Harley, Eriope foetida A. St. Hil. ex Benth., Eriope velutina Epling and one taxon that probaly it s a new species endemic of Cristalina-GO region. The subspecies Eriope crassipes Benth. subsp. cristalinae Harley presented morphologic and anatomical characteristics differential among group, in this way the suggestion is the manutetion specific status Eriope cristalinae (Harley) C. T. Rizzini and in synonim Eriope crassipes Benth. subsp. cristalinae Harley. Identification keys were elaborate using morphologic and anatomical data for the taxa in study. The species presented anthraquinone flavonic and saponin heterosides; triterpenes and fenolic compound. In the essential oils 58 compounds were identificated, α-copaen, Aromadendrene and copaen-4α-ol were identificated in all the taxa. Key words: morpho-anatomical,Item Efeitos tóxicos e genotóxicos do herbicida Roundup Transorb® em Guppy (Poecilia reticulata) submetido a tratamento agudo(Universidade Federal de Goiás, 2011-01-31) SOUZA FILHO, José de; SABÓIA-MORAIS, Simone Maria Teixeira de; http://lattes.cnpq.br/6723881044959716The effects of toxic, mutagenic and genotoxic on target organs and the fish genome has been the subject of many studies, especially those that seek to establish these bodies and responses of genes to environmental stimuli. Histopathological studies, mutagenic, genotoxic and Poecilia reticulata (Guppy) were motivated by the scarcity of data in the literature concerning the effects of the formulation of the herbicide Roundup Transorb®. With everything, we sought to ascertain the effects produced by this formulation that is widely used in the Midwest of the country. This study aimed to evaluate the toxicity and mutagenic and genotoxic herbicide R. Transorb® in liver, gill and erythrocytes of the guppy, calculating and applying the LC50 test micronucleus (MN) and Comet assay (AC). The toxicity bioassays were performed to calculate the LC50;12-96h, to evaluate the effects of sublethal concentrations of herbicide subjected to acute treatment. We used 36 adult fish, weighing on average 0.496 g ± 0.28 g to calculate the LC50 and 75 adult fish also carry the MN and AC. During exposure to the herbicide was observed fish behavior at all concentrations and controlled chemical and physical changes of water. The erythrocytes were obtained by centrifugation of the gills being added to precipitate fetal calf serum, and then dripped on the slides to make the smear and electrophoresis. For the analysis of plates, 75.000 cells were counted and stipulate the frequency of occurrence of Nuclear Morphological Changes (AMNs) and AC were analyzed for 100 cells per specimen. The LC50; 12, 24, 48, 72 e 96h of R. Transorb® were 11:24, 8:55, 6.5, 6.10 and 5.65μl/L, respectively, indicating that this species is very sensitive to the herbicide studied in relation to other tropical species analyzed. These factors may be related to different formulations of herbicides and their respective surfactants. It was found that the mortality rate of fish poisoned by the herbicide increased as exposure concentrations increased. The values of LC50,12-96h show a downward trend over time, suggesting that during periods of reduced exposure will require greater amounts of herbicide to cause mortality of 50% of the population of animals. Animals exposed to the herbicide showed behavioral changes varied, such as aggressiveness, irritability, loss of the escape reflex, darkening of the body surface and banging against the wall of the aquarium. In addition, R. Transorb® hitológicas induced liver changes appreciably impairing the normal functioning of this organ. In MN and EC was possible to detect significant genotoxic and mutagenic effects in erythrocytes and gill cells, occurring gradual increase in the number of cells with AMNs and ADN damage under higher concentrations,indicating concentration-dependent effect. This herbicide has acute toxicity to guppy, promoting behavioral changes, possibly acting in a manner damaging to the liver but also in the nervous system and gill. These results also suggest that the formulation of this herbicide has genotoxic and mutagenic in erythrocytes and gill cells detected by the MN and AC. Therefore, it confirms the high risk of herbicide to the environment, its contamination may present a strong threat to populations of fish and other organisms both vertebrates and invertebrates and human health.