Thesis (Ph. D.)--University of Wisconsin--Madison, 1980. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

The phaseolin (phas) promoter drives the copious production of transcripts encoding the protein phaseolin during seed embryogenesis but is silent in vegetative tissues when a nucleosome is positioned over its three phased TATA boxes. Transition from the inactive state in transgenic Arabidopsis leaves was accomplished by ectopic expression of the transcription factor PvALF (Phaseolus vulgaris ABI3-like factor), and application of abscisic acid (ABA). PvALF belongs to a family of seed-specific transcriptional activators that includes the maize viviparious1 (VP1) and the Arabidopsis abscisic acid-insensitive3 (ABI3) proteins. The major goal of the study is to gain insight to the regulation of seed-specific gene expression in three different aspects. First, since ABI3 (homolog of PvALF) is involved in ABA-mediated expression of several seed-specific protein genes in Arabidopsis, understanding its transcriptional regulation will provide insight to the mechanism by which PvALF expression is controlled. To achieve this, ABI3 promoter deletion analysis using either $-glucuronidase (gus) or green fluorescent protein (gfp) reporter gene fusions have identified various regulatory regions within the ABI3 promoter including two upstream activating sequences and a minimal seed specific expression region. In addition, a 405 bp 5' UTR was shown to play a negative role in ABI3 expression, possibly through post-transcriptional mechanisms. Second, placement of PvALF expression under control of an estradiol-inducible promoter permitted chronological ChIP analysis of changes in histone modifications, notably increased acetylation of H3-K9, as phas chromatin is remodeled (potentiated). A different array of changes (trimethylation of H3-K4) is associated with ABA-mediated activation. In contrast, H3-K14 acetylation decreased upon phas potentiation and increased on activation. Whereas decreases in histone H3 and H4 levels were detected during PvALF-mediated remodeling, slight increases occurred following ABA-mediated activation, suggesting the restoration of histone-phas interactions or the redeposition of histones in the phas chromatin. The observed histone modifications thus provide insight to the factors involved in euchromatinization and activation of a plant gene. Finally, ectopically expressed ABI5 and PvALF renders the activation of phas ABA-independent, suggesting ABI5 acts downstream of ABA during phas activation.

O feijão constitui-se em um dos alimentos básicos da população brasileira e proporciona vários benefícios à saúde. No entanto, possui fatores antinutricionais que impedem parte do aproveitamento de alguns de seus nutrientes, destacando-se dentre esses fatores os polifenóis. Estes podem complexar com proteínas, diminuindo a sua digestibilidade e seu aproveitamento pelo organismo. Assim, esta pesquisa teve por objetivo avaliar o potencial antioxidante de feijão comum (Phaseolus vulgaris, L.) e, após a complexação de suas proteínas e polifenóis, avaliar os efeitos dos tratamentos dados e dessa ligação na digestibilidade protéica. Foi utilizado duas cultivares de feijão comum de duas diferentes cores, preto (Xamego) e marrom (BRS9435-Cometa), e uma linhagem de coloração branca (G2358). Os grãos foram analisados na forma crua e cozidos em autoclave, macerados ou não. Foram realizadas análises de: teor de taninos, fenólicos totais, atividade antioxidante, eletroforese, digestibilidade protéica in vitro, e ácidos fenólicos e flavonóides por CLAE. Nas análises dos grãos íntegros, a cultivar BRS9435-Cometa crua apresentou a maior atividade antioxidante e, apesar da maceração e da cocção terem reduzido teor de taninos, compostos fenólicos e atividade antioxidante da maior parte dos feijões, a maceração aumentou significativamente a quantidade de fenólicos totais em feijão preto. Houve alteração no perfil eletroforético da proteína dos grãos após a complexação com os polifenóis. As bandas com maior peso molecular, presentes nas faseolinas puras, apresentaram menor intensidade quando houve a complexação com polifenóis. A digestibilidade das faseolinas de feijões crus variou de 95,67 a 97,66% e a de feijões cozidos de 90,44 a 97,02%. Na digestibilidade de faseolinas puras a maceração seguida de cocção somente exerceu influência significativa em grãos brancos, reduzindo a digestibilidade da faseolina. De modo geral a complexação faseolinapolifenóis aumentou o percentual de digestibilidade. O efeito esperado da complexação faseolina-frações de polifenóis em reduzir a digestibilidade da faseolina foi muito discreto e muitas vezes inexistente, e o de aumento da digestibilidade pela cocção com ou sem maceração também foi limitado. Para os extratos brutos dos feijões, os resultados de DPPH variaram de 2,96 a 14,04 mg de TEAC.g-1 de extrato, e os de ABTS entre 13,70 e 29,51 mg de TEAC.g-1 de extrato. Em ambos os métodos de avaliação da atividade antioxidante, os resultados foram maiores para a maioria dos extratos das cultivares marrom e preta. Quanto às análises realizadas por CLAE, para ácidos fenólicos contatou-se que todos os extratos avaliados possuem os ácidos gálico, clorogênico, vanílico e sinápico, e que os dois primeiros foram os encontrados em maiores teores; praticamente todos os flavonóides foram detectados em quase todas as amostras, com exceção do Caempferol, que somente foi detectado nas amostras maceradas e cozidas. Para extratos de feijões brancos, os flavonóides encontrados em maiores concentrações foram a catequina e o caempferol-3-rutinosídeo; extratos de feijões marrons apresentaram predominantemente a catequina e o caempferol-3-glicosídeo; e em extratos de feijões pretos, predominantemente a catequina, caempferol-3-glicosídeo e quercetina-3-glicosídeo. De maneira geral, o efeito do tratamento térmico aumentou a atividade antioxidante, bem como a concentração dos compostos fenólicos avaliados nos extratos de feijões. / Common bean is one of the staples of the brazilian population and provides numerous health benefits. Nevertheless, it has antinutritional factors that prevent the use of some part of their nutrients, especially polyphenols. These can complex with proteins, decreasing their digestibility and utilization by the body. Thus, this research aimed to evaluate the antioxidant potential of common bean (Phaseolus vulgaris L.) and, after complexation of proteins and polyphenols, assess the effects of treatments given and of this complexation on protein digestibility. Was used two common bean cultivars in two different colors, black (Xamego) and brown (BRS9435-Comet), and a line of white color (G2358). The grains were analyzed as raw and cooked in an autoclave, soaked or not. The analyses that were performed are: content of tannins, total phenolics, antioxidant activity, electrophoresis, in vitro protein digestibility, and phenolic acids and flavonoids by HPLC. In the analyses of whole grains, raw BRS9435- Comet had the highest antioxidant activity and, despite soaking and cooking have shown low levels of tannins, phenolic compounds and antioxidant activity of most beans, soaking significantly increased the amount of total phenolic in black beans. There were changes in the electrophoretic profile of protein from grains by complexation with polyphenols. The bands with higher molecular weight, present in pure phaseolins, had a lower intensity when there was complexation with polyphenols. The digestibility of phaseolins from raw beans ranged from 95.67 to 97.66% and from cooked beans from 90.44 to 97.02%. Digestibility of pure phaseolins from soaked beans only exerted significant influence on white grains, reducing the digestibility of phaseolin. In general, the complexation of phaseolin and polyphenols increased percentage of digestibility. The expected effect of complexation of fractions of polyphenols and phaseolins in reduce the digestibility of phaseolins was very low and often non-existent, and the increase in the digestibility by cooking with or without soaking was also limited. For crude extracts of the beans, the results of DPPH ranged from 2.96 to 14.04 mg TEAC.g-1 extract, and of ABTS between 13.70 and 29.51 mg TEAC.g-1 extract. In both methods of assessing antioxidant activity, the results were higher for most of the extracts of brown and black cultivars. For analyses performed by HPLC it was found for phenolic acids that all extracts evaluated have the gallic acid, chlorogenic, vanillic and sinapic, and the first two were found at higher levels. Practically all flavonoids were detected in almost all samples, except for kaempferol, wich was only detected in soaked and cooked samples. For extracts from white beans, flavonoids found in highest concentrations were catechin and kaempferol-3-rutinoside; brown beans extracts showed mostly catechin and kaempferol-3- glucoside, and in extracts of black beans, mainly catechin, kaempferol-3-glucoside and quercetin-3-glucoside. In general, the effect of heat treatment increased the antioxidant activity and concentration of phenolic compounds evaluated in extracts of beans.

O feijão constitui-se em um dos alimentos básicos da população brasileira e proporciona vários benefícios à saúde. No entanto, possui fatores antinutricionais que impedem parte do aproveitamento de alguns de seus nutrientes, destacando-se dentre esses fatores os polifenóis. Estes podem complexar com proteínas, diminuindo a sua digestibilidade e seu aproveitamento pelo organismo. Assim, esta pesquisa teve por objetivo avaliar o potencial antioxidante de feijão comum (Phaseolus vulgaris, L.) e, após a complexação de suas proteínas e polifenóis, avaliar os efeitos dos tratamentos dados e dessa ligação na digestibilidade protéica. Foi utilizado duas cultivares de feijão comum de duas diferentes cores, preto (Xamego) e marrom (BRS9435-Cometa), e uma linhagem de coloração branca (G2358). Os grãos foram analisados na forma crua e cozidos em autoclave, macerados ou não. Foram realizadas análises de: teor de taninos, fenólicos totais, atividade antioxidante, eletroforese, digestibilidade protéica in vitro, e ácidos fenólicos e flavonóides por CLAE. Nas análises dos grãos íntegros, a cultivar BRS9435-Cometa crua apresentou a maior atividade antioxidante e, apesar da maceração e da cocção terem reduzido teor de taninos, compostos fenólicos e atividade antioxidante da maior parte dos feijões, a maceração aumentou significativamente a quantidade de fenólicos totais em feijão preto. Houve alteração no perfil eletroforético da proteína dos grãos após a complexação com os polifenóis. As bandas com maior peso molecular, presentes nas faseolinas puras, apresentaram menor intensidade quando houve a complexação com polifenóis. A digestibilidade das faseolinas de feijões crus variou de 95,67 a 97,66% e a de feijões cozidos de 90,44 a 97,02%. Na digestibilidade de faseolinas puras a maceração seguida de cocção somente exerceu influência significativa em grãos brancos, reduzindo a digestibilidade da faseolina. De modo geral a complexação faseolinapolifenóis aumentou o percentual de digestibilidade. O efeito esperado da complexação faseolina-frações de polifenóis em reduzir a digestibilidade da faseolina foi muito discreto e muitas vezes inexistente, e o de aumento da digestibilidade pela cocção com ou sem maceração também foi limitado. Para os extratos brutos dos feijões, os resultados de DPPH variaram de 2,96 a 14,04 mg de TEAC.g-1 de extrato, e os de ABTS entre 13,70 e 29,51 mg de TEAC.g-1 de extrato. Em ambos os métodos de avaliação da atividade antioxidante, os resultados foram maiores para a maioria dos extratos das cultivares marrom e preta. Quanto às análises realizadas por CLAE, para ácidos fenólicos contatou-se que todos os extratos avaliados possuem os ácidos gálico, clorogênico, vanílico e sinápico, e que os dois primeiros foram os encontrados em maiores teores; praticamente todos os flavonóides foram detectados em quase todas as amostras, com exceção do Caempferol, que somente foi detectado nas amostras maceradas e cozidas. Para extratos de feijões brancos, os flavonóides encontrados em maiores concentrações foram a catequina e o caempferol-3-rutinosídeo; extratos de feijões marrons apresentaram predominantemente a catequina e o caempferol-3-glicosídeo; e em extratos de feijões pretos, predominantemente a catequina, caempferol-3-glicosídeo e quercetina-3-glicosídeo. De maneira geral, o efeito do tratamento térmico aumentou a atividade antioxidante, bem como a concentração dos compostos fenólicos avaliados nos extratos de feijões. / Common bean is one of the staples of the brazilian population and provides numerous health benefits. Nevertheless, it has antinutritional factors that prevent the use of some part of their nutrients, especially polyphenols. These can complex with proteins, decreasing their digestibility and utilization by the body. Thus, this research aimed to evaluate the antioxidant potential of common bean (Phaseolus vulgaris L.) and, after complexation of proteins and polyphenols, assess the effects of treatments given and of this complexation on protein digestibility. Was used two common bean cultivars in two different colors, black (Xamego) and brown (BRS9435-Comet), and a line of white color (G2358). The grains were analyzed as raw and cooked in an autoclave, soaked or not. The analyses that were performed are: content of tannins, total phenolics, antioxidant activity, electrophoresis, in vitro protein digestibility, and phenolic acids and flavonoids by HPLC. In the analyses of whole grains, raw BRS9435- Comet had the highest antioxidant activity and, despite soaking and cooking have shown low levels of tannins, phenolic compounds and antioxidant activity of most beans, soaking significantly increased the amount of total phenolic in black beans. There were changes in the electrophoretic profile of protein from grains by complexation with polyphenols. The bands with higher molecular weight, present in pure phaseolins, had a lower intensity when there was complexation with polyphenols. The digestibility of phaseolins from raw beans ranged from 95.67 to 97.66% and from cooked beans from 90.44 to 97.02%. Digestibility of pure phaseolins from soaked beans only exerted significant influence on white grains, reducing the digestibility of phaseolin. In general, the complexation of phaseolin and polyphenols increased percentage of digestibility. The expected effect of complexation of fractions of polyphenols and phaseolins in reduce the digestibility of phaseolins was very low and often non-existent, and the increase in the digestibility by cooking with or without soaking was also limited. For crude extracts of the beans, the results of DPPH ranged from 2.96 to 14.04 mg TEAC.g-1 extract, and of ABTS between 13.70 and 29.51 mg TEAC.g-1 extract. In both methods of assessing antioxidant activity, the results were higher for most of the extracts of brown and black cultivars. For analyses performed by HPLC it was found for phenolic acids that all extracts evaluated have the gallic acid, chlorogenic, vanillic and sinapic, and the first two were found at higher levels. Practically all flavonoids were detected in almost all samples, except for kaempferol, wich was only detected in soaked and cooked samples. For extracts from white beans, flavonoids found in highest concentrations were catechin and kaempferol-3-rutinoside; brown beans extracts showed mostly catechin and kaempferol-3- glucoside, and in extracts of black beans, mainly catechin, kaempferol-3-glucoside and quercetin-3-glucoside. In general, the effect of heat treatment increased the antioxidant activity and concentration of phenolic compounds evaluated in extracts of beans.