Bioquímica e Biologia Molecular
URI permanente desta comunidadehttps://locus.ufv.br/handle/123456789/11837
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Resultados da Pesquisa
Item Distyly and variation in floral traits in natural populations of Psychotria ipecacuanha (Brot.) Stokes (Rubiaceae)(Brazilian Journal of Botany, 2005-04) Oliveira, Luiz Orlando de; Vieira, Milene F.; Rossi, Ana Aparecida B.Psychotria ipecacuanha is a perennial, medicinal herb that grows in clusters in the understory of humid, shady areas of the Atlantic Rain Forest of southeastern Brazil. The present study characterized the variation in floral traits among 35 clusters from three natural populations of this plant species. Field observations showed that the clusters are isomorphic, that is, a given cluster will either set long-styled or short-styled flowers. Stigmas and anthers are reciprocally placed in each morph, a dimorphism characteristic of distyly. The populations are isoplethic, that is, a given population exhibits an equilibrium 1:1 ratio of floral morphs. Morphometric analyses revealed that anther length, stigma length, corolla diameter, and pollen grain diameter were consistently greater in short-styled flowers, regardless of the population investigated. Significant differences for floral traits in the short-styled morph were found among populations. Floral traits in the long-styled morph also showed some significant differences among populations, but not for stigma height and corolla length. Controlled pollinations carried out in natural populations showed that fruit production was higher after inter-morph pollination. Nevertheless, observations of pollen tube growth in style, and also fruit production after spontaneous self-pollination and intra-morph pollination, indicated partial intramorph compatibility in this plant species.Item Atividade da amilase em rã-touro durante a fase pós-metamórfica(Pesquisa Agropecuária Brasileira, 2005-10) Oliveira, Maria Goreti de Almeida; Lima, William Cardoso; Euclydes, Ricardo Frederico; Braga, Luís Gustavo TavaresO objetivo deste trabalho foi avaliar a atividade da amilase no quimo da rã-touro, Rana catesbeiana Shaw 1802 (3,6 a 200 g). Oitenta e sete animais foram distribuídos em baias-testes com temperatura e fotoperíodo controlados. As rãs selecionadas na fase pós-metamórfica receberam ração comercial extrusada ad libitum. Durante 87 dias de experimento, foram efetuadas 29 coletas em intervalos de 1 a 8 dias. As coletas do conteúdo intestinal foram feitas mediante a insensibilização das rãs em gelo e água e isolamento posterior do intestino delgado das mesmas. Para os testes da atividade da amilase, foram utilizados kits enzimáticos comerciais. A atividade da amilase foi observada a partir do terceiro dia e apresentou aumento até alcançar estabilidade, quando os animais pesavam 4,35 g. Do sétimo dia até o final do experimento, ocorreu a manutenção da estabilidade da atividade (137,44 UI) e atividade específica da amilase (14.688 UI mg-1).Item Desempenho e atividades enzimáticas em girinos de rã-touro(Pesquisa Agropecuária Brasileira, 2008-11) Oliveira, Maria Goreti Almeida; Moura, Guilherme de Souza; Lanna, Eduardo Teixeira Arruda; Garcia, Silvana Lages Ribeiro; Seixas Filho, José Teixeira de; Silva, Lilia Nogueira daO objetivo deste trabalho foi avaliar o desempenho e as atividades das enzimas digestivas de girinos de rã-touro, alimentados com ração comercial durante 60 dias, com um girino por litro, em caixas com 30 L de água. Os parâmetros medidos quinzenalmente foram: sobrevivência, comprimento, peso final, consumo de ração, e a atividade de amilase, lipase e tripsina. O maior crescimento dos girinos foi do 15º ao 45º dia - 20,61 mm. Durante os primeiros 15 dias, os girinos apresentaram a maior taxa de crescimento específico, 16,73% por dia. Do 45º ao 60º dia, os girinos apresentaram o maior ganho de peso, 3,98±0,73 g, o maior consumo de ração, 14,40±1,14 g, e a melhor conversão alimentar, 3,67:1,0±0,18. Houve aumento significativo da atividade enzimática depois do 30º dia. Observaram-se relações entre as atividades específicas das enzimas e os distintos estágios de desenvolvimento dos girinos.Item Reproductive studies in ipecac (Psychotria ipecacuanha (Brot.) stockes; Rubiaceae): pollen development and morphology(Brazilian Archives of Biology and Technology, 2008-09) Oliveira, Luiz Orlando de; Souza, Margarete Magalhães; Martins, Ernane Ronie; Pereira, Telma Nair SantanaThe aim of this work was to carry out the reproductive studies on Brazilian accessions of ipecac, Psychotria ipecacuanha. It presented heterostyly, with brevistylous and longistylous flowers. The pollen development was observed from the sections of the anthers embedded in resin. Anther development was normal as usually observed in dicotyledones, displaying four layers: outer epidermis, endothecium, middle layer and inner tapetum. The pollen was bicellular and filled with starch at the microspore stage. Pollen morphology was studied using SEM, which showed pollen polymorphism within and between the two floral morphs. Five types of pollen with reticulate or perforate exine were identified. The characteristics showed that the sexual process was as important as the vegetative propagation for the reproduction of this species.Item Identification and in silico analysis of the Citrus HSP70 molecular chaperone gene family(Genetics and Molecular Biology, 2007) Fietto, Luciano G.; Costa, Maximiller D. L.; Cruz, Cosme D.; Souza, Alessandra A.; Machado, Marcos A.; Fontes, Elizabeth P. B.The completion of the genome sequencing of the Arabidopsis thaliana model system provided a powerful molecular tool for comparative analysis of gene families present in the genome of economically relevant plant species. In this investigation, we used the sequences of the Arabidopsis Hsp70 gene family to identify and annotate the Citrus Hsp70 genes represented in the CitEST database. Based on sequence comparison analysis, we identified 18 clusters that were further divided into 5 subgroups encoding four mitochondrial mtHsp70s, three plastid csHsp70s, one ER luminal Hsp70 BiP, two HSP110/SSE-related proteins and eight cytosolic Hsp/Hsc70s. We also analyzed the expression profile by digital Northern of each Hsp70 transcript in different organs and in response to stress conditions. The EST database revealed a distinct population distribution of Hsp70 ESTs among isoforms and across the organs surveyed. The Hsp70-5 isoform was highly expressed in seeds, whereas BiP, mitochondrial and plastid HSp70 mRNAs displayed a similar expression profile in the organs analyzed, and were predominantly represented in flowers. Distinct Hsp70 mRNAs were also differentially expressed during Xylella infection and Citrus tristeza viral infection as well as during water deficit. This in silico study sets the groundwork for future investigations to fully characterize functionally the Citrus Hsp70 family and underscores the relevance of Hsp70s in response to abiotic and biotic stresses in Citrus.Item Qualidade nutricional e escore químico de aminoácidos de diferentes fontes protéicas(Food Science and Technology, 2006-01) Pires, Christiano Vieira; Oliveira, Maria Goreti de Almeida; Costa, Neuza Maria Brunoro; Rosa, José CésarAs proteínas são moléculas essenciais para aos organismos animais, devendo, portanto, estar presentes na alimentação em quantidades adequadas. Além do aspecto quantitativo deve-se levar em conta o aspecto qualitativo, isto é, seu valor nutricional, que dependerá de sua composição, digestibilidade, biodisponibilidade de aminoácidos essenciais, ausência de toxicidade e de fatores antinutricionais. O objetivo deste trabalho foi avaliar a digestibilidade in vivo, o escore químico de aminoácidos (EQ) e o escore químico de aminoácido corrigido pela digestibilidade protéica (PDCAAS) das seguintes fontes de proteína: carne de rã sem osso, carne de rã com osso, carne de rã mecanicamente separada (CMS), carne bovina, ovo em pó, caseína, trigo, milho, soja convencional, soja isenta de inibidor de tripsina Kunitz e de lipoxigenases (soja KTI-LOX-), proteína texturizada de soja (PTS) e feijão. As proteínas de origem animal apresentaram maiores valores de digestibilidade que as de origem vegetal. Carne de rã sem osso apresentou a proteína com maior digestibilidade protéica de todas as proteínas estudadas, não diferindo, entretanto, da caseína, CMS, carne bovina e rã com osso. Das proteínas de origem animal, a do ovo em pó foi aquela que apresentou menor digestibilidade protéica. Nenhuma das proteínas de origem animal apresentou aminoácidos essenciais limitantes quando comparadas com o padrão da FAO/WHO. Feijão, soja convencional, soja KTI-LOX- e PTS, tiveram os aminoácidos sulfurados (metionina+cisteína) como limitantes. Enquanto que para trigo e milho, o aminoácido mais limitante foi a lisina. Soja KTI-LOX- e PTS apresentaram valores de PDCAAS superiores aos da soja convencional, mostrando uma possível elevação na qualidade protéica da soja melhorada geneticamente e da soja processada.Item Purification and characterization of an a-galactosidase from Aspergillus fumigatus(Brazilian Archives of Biology and Technology, 2005-03) Rezende, Sebastião Tavares de; Guimarães, Valéria Monteze; Rodrigues, Marília de Castro; Felix, Carlos RobertoAspergillus fumigatus secreted invertase (b-fructofuranosidase) and a-galactosidase enzymatic activities able to hydrolyzing raffinose oligosaccharides (RO). a-Galactosidase was induced by galactose, melibiose and raffinose, but galactose was the most efficient inducer. It was purified by gel filtration and two ion exchange chromatographies and showed Mw of 54.7 kDa. The purified enzyme showed maximal activity against p-nitrophenyl-a-D-galactopyranoside (pNPGal) at pH 4.5-5.5 and 55 °C, and retained about 80% of the original activity after incubation for 90 minutes at 50ºC. The KM for pNPGal was 0.3 mM. Melibiose was hydrolyzed by the enzyme but raffinose was very poor substrate.Item Extracellular nucleotide metabolism in Leishmania: influence of adenosine in the establishment of infection(Microbes and Infection, 2008-07) Fietto, Juliana Lopes Rangel; Marques-da-Silva, Eduardo de Almeida; Oliveira, Jamile Camargos de; Figueiredo, Amanda Braga; Lima Júnior, Djalma de Souza; Carneiro, Cláudia Martins; Afonso, Luís Carlos CroccoLeishmaniasis is a parasitic disease with a variety of clinical forms, which are related to the Leishmania species involved. In the murine model, Leishmania amazonensis causes chronic non-healing lesions in Leishmania braziliensis- or Leishmania major-resistant mouse strains. In this study, we investigated the involvement of the pathway of extracellular nucleotide hydrolysis, with special focus on the role of extracellular adenosine, in the establishment of Leishmania infection. Our results show that the more virulent parasite—L. amazonensis—hydrolyzes higher amounts of ATP, ADP and AMP than the two other species, probably due to the higher expression of membrane NTPDase. Corroborating the idea that increased production of adenosine is important to lesion development and establishment of tissue parasitism, we observed that increased 5′-nucleotidase activity in L. braziliensis or addition of adenosine at the moment of infection with this parasite resulted in an increase in lesion size and parasitism as well as a delay in lesion healing. Furthermore, inhibition of adenosine receptor A2B led to decreased lesion size and parasitism. Thus, our results suggest that the conversion of ATP, a molecule with pro-inflammatory activity, into adenosine, which possesses immunomodulatory properties, may contribute to the establishment of infection by Leishmania.Item Protein digestibility, protease activity, and post-embryonic development of the velvetbean caterpillar (Anticarsia gemmatalis) exposed to the trypsin-inhibitor benzamidine(Pesticide Biochemistry and Physiology, 2006-09) Pilon, Anderson M.; Guedes, Raul Narciso C.; Oliveira, Maria Goreti A.Protein digestibility, proteolytic activity, and post-embryonic development of Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae) were assessed in larvae reared on artificial diet containing 0.00, 0.25, 0.50, and 0.75% (w/w) of the synthetic trypsin inhibitor benzamidine. Diet consumption was affected by the inhibitor when the insects were exposed to 0.50% benzamidine showing a 4-day delay and a 70%-higher peak of consumption. Larva weight gain was also affected by benzamidine and again the results of 0.50% benzamidine were unexpected due to the worst performance of the insects at this inhibitor concentration and not at 0.75% benzamidine. These patterns of consumption and weight gain were however consistent with the results of protein digestibility, which affects larvae mortality and adult emergence. The insect proteolytic activity was also affected by benzamidine, particularly at 0.50%. These results indicate that the insects are able to circumvent the potentially harmful effects of the inhibitor since at the highest concentration the negative impact is mitigated.Item Expression of genes encoding cytosolic and endoplasmic reticulum HSP90 proteins in the aquatic fungus Blastocladiella emersonii(Gene, 2008-03-31) Fietto, Luciano G.; Pugliese, Luciana; Georg, Raphaela C.; Gomes, Suely L.HSP90 proteins are important molecular chaperones involved in multiple cellular processes. This work reports the characterization of cDNAs encoding two distinct HSP90 proteins (named HSP90A and HSP90B) from the chytridiomycete Blastocladiella emersonii. Deduced amino acid sequences of HSP90A and HSP90B exhibit signatures of the cytosolic and endoplasmic reticulum (ER) HSP90 proteins, respectively. A genomic clone encoding HSP90A was also characterized indicating the presence of a single intron of 184 bp interrupting the coding region, located near the amino-terminus of the protein. Expression of both HSP90A and HSP90B genes increases significantly during heat shock at 38 °C, with highest induction ratios observed in cells stressed during germination of the fungus. Changes in the amount of HSP90A transcript were also evaluated during B. emersonii life cycle at physiological temperature (27 °C), and its levels were found to increase both during germination and sporulation of the fungus. HSP90A protein levels were analyzed during B. emersonii life cycle and significant changes were observed only during sporulation. Furthermore, during heat stress a large increase in the amount of HSP90A protein was observed. Induction of HSP90A and HSP90B genes during heat stress indicates the importance of both genes in the response to high temperature in B. emersonii.