Mono e diacilgliceróis são produtos empregados na indústria alimentícia, farmacêutica, cosmética e química como emulsificantes e melhoradores de viscosidade de produtos alimentícios, cosméticos e farmacêuticos. No entanto, a forma mais usual de obtê-los é por síntese química, o que acaba rendendo produtos finais caros e com atributos de qualidade, rendimento e de aplicabilidade tecnológica inferiores aos esperados. A busca por formas de obtenção mais racionais, eficientes e com melhor padrão de qualidade destes produtos foi o objetivo principal do trabalho, por meio de hidrólise parcial enzimática, que necessita de condições de reação mais brandas. Foram avaliadas a hidrólise enzimática descontína, empregando como substrato a trioleína técnica, e a hidrólise enzimática descontínua-alimentada, usando como substrato o óleo de girassol médio oléico. Foi utilizada, em ambos processsos, a lipase imobilizada sn-1,3 específica Lipozyme RM IM (de Rhizomucor miehei). A caracterização dos padrões e dos substrados, bem como o acompanhamento da formação dos produtos da hidrólise enzimática foram feitos por determinação da porcentagem de hidrólise, cromatografia em camada delgada (TLC), dos perfis das curvas de fusão e cristalização por calorimetria diferencial de varredura (DSC), cromatografia gasosa (CG) e cromatografia de exclusão de tamanho de alto desempenho (HPSEC). Os parâmetros de hidrólise descontínua foram o tempo de reação, a temperatura e a concentração inicial de substrato. Os parâmetros de hidrólise descontínua-alimentada foram tempo de enchimento e intervalo de alimentação de substrato. Para as respostas analíticas de porcentagem de hidrólise e de composição de frações lipídicas foi aplicado um modelo de regressão múltipla com base em metodologia de superfície de resposta. Os resultados experimentais observados nas reações de hidrólise enzimática descontínua de trioleína técnica mostraram de 24,7 a 34,2% de mono e diacilgliceróis (para 5% de óleo na emulsão) e de 21,4 a 33,6% de mono e diacilgliceróis (para 20% de óleo na emulsão). Os resultados experimentais observados nas reações de hidrólise enzimática descontínua-alimentada de óleo de girassol médio oléico (para 15% de óleo na emulsão), mostraram de 7,9 a 31,8% de mono e diacilgliceróis. Os modelos de superfície de resposta foram considerados significativos e preditivos. As hidrólises obtidas no formato descontínuo e descontínuo-alimentado permitiram efetivamente a obtenção de frações de mono/ diacilgliceróis com vários graus de eficiência de conversão e com corretas identificação e quantificação das frações de lipídios procuradas. As correlações feitas entre porcentagem de hidrólise e entalpias de cristalização e fusão, corroboradas com os resultados qualitativos e/ou quantitativos diretos obtidos na cromatografia de camada delgada (TLC) e de HPSEC, demonstraram que estes atributos podem positivamente indicar a ocorrência efetiva de reação de hidrólise, além de auferir uma escala de desempenho de reação alinhada com o previsto na literatura, à medida que são aumentadas a temperatura, o tempo de hidrólise e a porcentagem inicial de substrato oleoso, sob regime descontínuo, e que puderam ser melhoradas, de forma inovadora, sob parâmetros de tempo total de alimentação e de intervalo de alimentação, sob regime descontínuo-alimentado. A hidrólise parcial enzimática de triacilgliceróis utilizando lipase imobilizada sn-1,3 específica pode ser considerada uma alternativa às vias químicas para a produção de misturas de mono e diacilgliceróis para utilização como aditivos químicos. / Mono and diacylglycerols are products used in the food, pharmaceutical, cosmetic and chemical industries as emulsifiers and viscosity improvers for food products, cosmetics and pharmaceuticals. However, the most usual forms of obtaining them are by chemical synthesis, which ends up yielding expensive final products with attributes of quality, yield and technological applicability lower than expected. The search for more rational, efficient and better quality standards of these products was the aim of the work, through partial enzymatic hydrolysis, which requires milder reaction conditions. Discontinuous enzymatic hydrolysis was evaluated using technical triolein as substrate and discontinuous-fed enzymatic hydrolysis using as the substrate the mid oleic sunflower oil. In both processes, immobilized lipase sn-1,3 specific Lipozyme RM IM (from Rhizomucor miehei) was used. The characterization of the patterns and substrates, as well as the monitoring of the formation of the products from the enzymatic hydrolysis were made by determining the percentage of hydrolysis, thin layer chromatography (TLC), profiles of the melting and crystallization curves by differential scanning calorimetry ( DSC), gas chromatography (GC) and high performance size exclusion chromatography (HPSEC). The parameters of discontinuous hydrolysis were the reaction time, the temperature and the initial substrate concentration. The parameters of discontinuous-fed hydrolysis were filling time and substrate feed interval. For the analytical responses of hydrolysis percentage and composition of lipid fractions a multiple regression model was applied based on response surface methodology. The experimental results observed in the reactions of discontinuous enzymatic hydrolysis of technical triolein indicated amounts of mono- and diacylglycerols from 24.7 to 34.2% (for 5% of oil in the emulsion) and from 21.4 to 33.6% for mono and diacylglycerols with 20% oil in the emulsion. The experimental results observed in the reactions of discontinuous-fed enzymatic hydrolysis of mid oleic sunflower oil (for 15% oil in the emulsion), showed from 7.9 to 31.8% of mono and diacylglycerols. Response surface models were considered significant and viii predictive. The hydrolysis obtained in the discontinuous and discontinuous-fed form allowed to obtain fractions of mono / diacylglycerols with various degrees of conversion efficiency and with correct identification and quantification of the lipid fractions sought. The correlations between the percentage of hydrolysis and enthalpies of crystallization and fusion, corroborated with the qualitative and / or quantitative direct results obtained in thin layer chromatography (TLC) and HPSEC, showed that these attributes can positively indicate the effective occurrence of reaction of Hydrolysis, in addition to achieving a reaction performance scale in line with the literature, as the temperature rate, the hydrolysis time and the initial percentage of oily substrate are increased under a discontinuous regime and can be improved, in a innovative form, under parameters of total filling time and feeding interval, under a fed-batch regime. The partial enzymatic hydrolysis of triacylglycerols using specific sn-1,3-specific immobilized lipase may be considered an alternative to the chemical pathways for the production of mono- and diacylglycerol blends for use as chemical additives.

Made available in DSpace on 2017-05-12T14:46:45Z (GMT). No. of bitstreams: 1 Bruna.pdf: 2218151 bytes, checksum: 72c1c9063129258699329dd11860a24c (MD5) Previous issue date: 2013-02-21 / There are currently several studies aiming to convert plant biomass, which offers low cost and high generation in a sustainable and alternative energy source, thus adding economic value to the material. Lignocellulosic fibers, after passing through specific pretreatment of hydrolysis, originate fermentable sugars that can be biotransformed on second-generation ethanol. The breaking of this complex results in two main fractions, the cellulosic and the hemicellulosic fractions. The cellulosic one is still a challenge due to the cost, since it requires a pretreatment with high concentrations of acids and high thermal energy, and an enzymatic treatment too. The hemicellulosic fraction is more easily removable and it is composed primarily of xylose, but this sugar is not easily fermentable by traditional yeasts. The Pichia stipitis yeast has been studied due to its ability to use pentoses, appearing to be promising yeast for further industrial application. For this reason, this work aimed to evaluate fermentation parameters such as pH, temperature and agitation for ethanol production by Pichia stipitis ATCC 58376 in a medium consisting of the hemicellulosic hydrolyzate of sunflower meal, a by-product of sunflower oil extraction. In addition, the effects of inoculum adaptation were evaluated through the cultivation of cells in the hydrolyzed one and the reuse of the cells recovered at the end of the fermentation process. Sunflower meal was subjected to acid hydrolysis with sulfuric acid 6% (m/v), 120 atm in 20 min, resulting in a hemicellulosic hydrolyzate with 8.06 g.l-1 of glucose, 49.93 g.L-1 of xylose, 8.67 g.L-1 of arabinose, 3.55 g.L-1 of acetic acid, 0.031 g.L-1 of hydroxymethylfurfural, 0.033 g.L-1 of furfural, and 0.89 g.L-1 of phenols. The hydrolyzed one was detoxified by pH changes and adsorption with activated carbon to reduce or eliminate compounds formed during the hydrolysis, which are toxic to microbial metabolism. After this treatment, there was a reduction of 58% of acetic acid, and furfural, hydroxymethylfurfural and phenols were not detected in the hydrolyzate. Fermentations were conducted according to a factorial 33- DCCR, with 8 major tests, 6 axial and 3 central, totalizing 17 tests in different conditions of pH, temperature and agitation. Data analysis was performed using the STATISTICA 8.0 program. The tests showed no significant difference between them until the period of 72 hours, since in the test 14 (pH 5.0, 30ºC and 234 rpm) and in the 15, 16 and 17 central points (pH 5.0, 30° C and 150 rpm) it was verified the largest concentrations of ethanol, 13.31 g.L-1 and 13.28 g.L-1 respectively after 84 hours, regardless of the condition of the inoculum preparation. Efficiency and productivity were the same for the last five tests (13, 14, 15, 16 and 17): 0.45 g. g-1 and 0.185 g.l-1 .h-1after 84 hours of fermentation. The optimization of the process led to a final production of ethanol of 13.92 g.l-1, 4.38% higher than test 13, yield equivalent to the theoretical, 0, 51g.g-1, and productivity of 0.165 g.L-1.h-1 after 84 hours of fermentation. / Atualmente há vários estudos com objetivo de converter a biomassa vegetal, a qual apresenta baixo custo e alta geração, em uma fonte alternativa e sustentável de energia, agregando valor econômico à matéria. As fibras lignocelulósicas, após passarem por pré-tratamento específico de hidrólise, originam açúcares fermentescíveis que podem ser biotransformados em etanol de segunda geração. A quebra desse complexo resulta em duas frações principais: a celulósica e a hemicelulósica. A fração celulósica é ainda um desafio devido ao custo, já que esta requer um pré-tratamento com altas concentrações de ácidos e grande energia térmica e ainda um tratamento enzimático. A fração hemicelulósica é mais facilmente extraível e constituída principalmente por xilose, porém este açúcar não é facilmente fermentável por leveduras tradicionais. A levedura Pichia stipitis está sendo muito estudada devido a sua capacidade de utilizar pentoses, mostrando ser muito promissora para aplicação industrial. Por essa razão, este trabalho teve como objetivo avaliar parâmetros fermentativos como pH, temperatura e agitação para a produção de etanol por Pichia stipitis ATCC 58376, em meio constituído de hidrolisado hemicelulósico de farelo de girassol, um sub-produto da extração do óleo de girassol. Além disso, foram avaliados os efeitos da adaptação do inóculo pelo cultivo das células no hidrolisado e o reaproveitamento das células separadas ao final de repetições do processo fermentativo. O farelo de girassol foi submetido à hidrolise ácida com ácido sulfúrico 6% (m/v), 120 atm, por 20 min., resultando num hidrolisado hemicelulósico com 8,06 g.L-1 de glicose, 49,93 g.L-1 de xilose, 8,67 g.L-1 de arabinose, 3,55 g.L-1 de ácido acético, 0,031 g.L-1 de hidroximetilfurfural, 0,033 g.L-1 de furfural e 0,89 g.L-1 de fenóis. O hidrolisado foi destoxificado por alterações de pH e adsorção com carvão ativado, a fim de reduzir e/ou eliminar compostos formados durante a hidrólise, que são tóxicos ao metabolismo microbiano. Após esse tratamento, houve redução de 58% de ácido acético e o hidroximetilfurfural, furfural e fenóis não foram detectados no hidrolisado. As fermentações foram conduzidas segundo um esquema fatorial 33 - DCCR, com 8 ensaios principais, 6 axiais e 3 centrais, totalizando 17 ensaios, em diferentes condições de pH, temperatura e agitação. A análise dos dados foi realizada empregando-se o programa STATISTICA 8.0. Os ensaios apresentaram diferença significativa entre si até o período de 72 horas, sendo que no ensaio 14 (pH 5,0, 30ºC e 234 rpm) e nos ensaios 15, 16 e 17 nos pontos centrais (pH 5,0, 30 ºC e 150 rpm) foram verificadas as maiores concentrações de etanol, 13,31 g.L-1 e 13,28 g.L-1, respectivamente, em 84 horas e independentemente da condição de preparo do inóculo. O rendimento e a produtividade foram iguais para os cinco últimos ensaios (13, 14, 15,16 e 17), 0,45 g.g-1 e 0,185 g.L-1.h-1 em 84 horas de fermentação. A otimização do processo levou a uma produção final de etanol de 13,92 g.L-1, 4,38% superior ao ensaio 13, rendimento equivalente ao teórico: 0,51g.g-1 e produtividade de 0,165 g.L-1.h-1 em 84 horas de fermentação.

Mono e diacilgliceróis são produtos empregados na indústria alimentícia, farmacêutica, cosmética e química como emulsificantes e melhoradores de viscosidade de produtos alimentícios, cosméticos e farmacêuticos. No entanto, a forma mais usual de obtê-los é por síntese química, o que acaba rendendo produtos finais caros e com atributos de qualidade, rendimento e de aplicabilidade tecnológica inferiores aos esperados. A busca por formas de obtenção mais racionais, eficientes e com melhor padrão de qualidade destes produtos foi o objetivo principal do trabalho, por meio de hidrólise parcial enzimática, que necessita de condições de reação mais brandas. Foram avaliadas a hidrólise enzimática descontína, empregando como substrato a trioleína técnica, e a hidrólise enzimática descontínua-alimentada, usando como substrato o óleo de girassol médio oléico. Foi utilizada, em ambos processsos, a lipase imobilizada sn-1,3 específica Lipozyme RM IM (de Rhizomucor miehei). A caracterização dos padrões e dos substrados, bem como o acompanhamento da formação dos produtos da hidrólise enzimática foram feitos por determinação da porcentagem de hidrólise, cromatografia em camada delgada (TLC), dos perfis das curvas de fusão e cristalização por calorimetria diferencial de varredura (DSC), cromatografia gasosa (CG) e cromatografia de exclusão de tamanho de alto desempenho (HPSEC). Os parâmetros de hidrólise descontínua foram o tempo de reação, a temperatura e a concentração inicial de substrato. Os parâmetros de hidrólise descontínua-alimentada foram tempo de enchimento e intervalo de alimentação de substrato. Para as respostas analíticas de porcentagem de hidrólise e de composição de frações lipídicas foi aplicado um modelo de regressão múltipla com base em metodologia de superfície de resposta. Os resultados experimentais observados nas reações de hidrólise enzimática descontínua de trioleína técnica mostraram de 24,7 a 34,2% de mono e diacilgliceróis (para 5% de óleo na emulsão) e de 21,4 a 33,6% de mono e diacilgliceróis (para 20% de óleo na emulsão). Os resultados experimentais observados nas reações de hidrólise enzimática descontínua-alimentada de óleo de girassol médio oléico (para 15% de óleo na emulsão), mostraram de 7,9 a 31,8% de mono e diacilgliceróis. Os modelos de superfície de resposta foram considerados significativos e preditivos. As hidrólises obtidas no formato descontínuo e descontínuo-alimentado permitiram efetivamente a obtenção de frações de mono/ diacilgliceróis com vários graus de eficiência de conversão e com corretas identificação e quantificação das frações de lipídios procuradas. As correlações feitas entre porcentagem de hidrólise e entalpias de cristalização e fusão, corroboradas com os resultados qualitativos e/ou quantitativos diretos obtidos na cromatografia de camada delgada (TLC) e de HPSEC, demonstraram que estes atributos podem positivamente indicar a ocorrência efetiva de reação de hidrólise, além de auferir uma escala de desempenho de reação alinhada com o previsto na literatura, à medida que são aumentadas a temperatura, o tempo de hidrólise e a porcentagem inicial de substrato oleoso, sob regime descontínuo, e que puderam ser melhoradas, de forma inovadora, sob parâmetros de tempo total de alimentação e de intervalo de alimentação, sob regime descontínuo-alimentado. A hidrólise parcial enzimática de triacilgliceróis utilizando lipase imobilizada sn-1,3 específica pode ser considerada uma alternativa às vias químicas para a produção de misturas de mono e diacilgliceróis para utilização como aditivos químicos. / Mono and diacylglycerols are products used in the food, pharmaceutical, cosmetic and chemical industries as emulsifiers and viscosity improvers for food products, cosmetics and pharmaceuticals. However, the most usual forms of obtaining them are by chemical synthesis, which ends up yielding expensive final products with attributes of quality, yield and technological applicability lower than expected. The search for more rational, efficient and better quality standards of these products was the aim of the work, through partial enzymatic hydrolysis, which requires milder reaction conditions. Discontinuous enzymatic hydrolysis was evaluated using technical triolein as substrate and discontinuous-fed enzymatic hydrolysis using as the substrate the mid oleic sunflower oil. In both processes, immobilized lipase sn-1,3 specific Lipozyme RM IM (from Rhizomucor miehei) was used. The characterization of the patterns and substrates, as well as the monitoring of the formation of the products from the enzymatic hydrolysis were made by determining the percentage of hydrolysis, thin layer chromatography (TLC), profiles of the melting and crystallization curves by differential scanning calorimetry ( DSC), gas chromatography (GC) and high performance size exclusion chromatography (HPSEC). The parameters of discontinuous hydrolysis were the reaction time, the temperature and the initial substrate concentration. The parameters of discontinuous-fed hydrolysis were filling time and substrate feed interval. For the analytical responses of hydrolysis percentage and composition of lipid fractions a multiple regression model was applied based on response surface methodology. The experimental results observed in the reactions of discontinuous enzymatic hydrolysis of technical triolein indicated amounts of mono- and diacylglycerols from 24.7 to 34.2% (for 5% of oil in the emulsion) and from 21.4 to 33.6% for mono and diacylglycerols with 20% oil in the emulsion. The experimental results observed in the reactions of discontinuous-fed enzymatic hydrolysis of mid oleic sunflower oil (for 15% oil in the emulsion), showed from 7.9 to 31.8% of mono and diacylglycerols. Response surface models were considered significant and viii predictive. The hydrolysis obtained in the discontinuous and discontinuous-fed form allowed to obtain fractions of mono / diacylglycerols with various degrees of conversion efficiency and with correct identification and quantification of the lipid fractions sought. The correlations between the percentage of hydrolysis and enthalpies of crystallization and fusion, corroborated with the qualitative and / or quantitative direct results obtained in thin layer chromatography (TLC) and HPSEC, showed that these attributes can positively indicate the effective occurrence of reaction of Hydrolysis, in addition to achieving a reaction performance scale in line with the literature, as the temperature rate, the hydrolysis time and the initial percentage of oily substrate are increased under a discontinuous regime and can be improved, in a innovative form, under parameters of total filling time and feeding interval, under a fed-batch regime. The partial enzymatic hydrolysis of triacylglycerols using specific sn-1,3-specific immobilized lipase may be considered an alternative to the chemical pathways for the production of mono- and diacylglycerol blends for use as chemical additives.

Sunflower is grown worldwide for oil production. In Australia, sunflower is under intense competition from canola as an oil crop. Confectionery sunflower aimed for the overseas snack food market is a potential opportunity for Australian farmers due to its ability to provide off-season supply to the major market located mainly in the northern hemisphere. To serve this aim, new confectionery sunflower hybrids adapted to Australian production system with drought tolerance, large achenes, high self-fertility and rust resistance are of paramount importance. The objectives of this project were to understand the genetic inheritance and genetic linkage relationships of the above traits and also to identify linked DNA markers with the potential for marker-assisted selection (MAS). The inheritance of achene-length was studied in a 12 x 12 diallel and a 14 x 12 North Carolina design II (NC II) mating design. Achene-length was evaluated under three environments for each mating design. Genetic correlation among all five achene-size related traits (achene-length, -width, - thickness and also kernel-length and -width) and 100-achene weight were moderate to high. Narrow-and broad-sense heritability for achene-length was moderate to high and similar for each design. The general combining ability (GCA) and maternal effects were found to be the main genetic factors controlling achene-length and each accounted for around 45% of the total genotypic variance. The inheritance of self-fertility (SF) was assessed with the same 12 x 12 diallel mating design evaluated in two environments. The 12 parents also displayed various levels of SF in addition to a wide range of achene-length. Negative genetic correlations between SF and all five achene-size related traits and 100-achene weight were observed. Modelling achene-length as a covariate revealed that certain pre-pollination floret characteristics represented by achene-length at maturity strongly affects SF in sunflower. When the effect of achene-length was removed statistically as a covariate, the specific combining ability (SCA) effect increased dramatically and became the major factor (other than pre-pollination floret characteristics) for SF determination and accounted for 65% of the total variance. Long achene genotypes tended to have low SF and a positive SCA effect was observed more frequently in F1 hybrids that involved small achene lines. The broad sense heritability was moderate (0.663 ± 0.017), while the estimate of narrow-sense heritability was nearly zero. A segregating F6:7 recombinant inbred line population with 239 progeny was developed from a cross between HaR4, a low delta (Δ), high transpiration efficiency (TE), sunflower line with rust resistance but relatively low SF, and SA52 with all the characteristics in the opposite direction. The population was evaluated in two environments for three TE-related traits including Δ, specific leaf weight (SLW) and chlorophyll content (CC) and four agronomic traits i.e. days to flowering (DTF), plant height (PlantH), SF and rust resistance (RustR). A linkage map 1074.6 cM in length with 138 SSR and 37 ISSR markers consisting of 19 linkage groups was constructed. Spatially adjusted BLUPs were used in QTL analysis for individual and combined environments. Altogether, 10 QTL were identified for Δ, SLW and CC at individual environments, and 7 were detected by a combined analysis. Sixteen QTL were identified for 4 agronomic traits and 14 of them were confirmed by a combined analysis. Significant linkage among all three TE related traits was discovered on linkage group 4 (LG4), where alleles from one parent had the same direction of influence on TE which suggested either coupling phase linkage or pleiotropy. QTL for SF and RustR were closely linked on LG13. An F2 population with 502 progeny developed from a cross between Jumbo, a large achene confectionery sunflower line, and RIL25, an oilseed small achene line, was evaluated for five achene-size related traits in a single field trial. Using a subset of 178 progeny a framework genetic linkage map, 899.7 cM in length, containing 128 SSR markers and 23 ISSR markers was constructed with 18 linkage groups. Based on the results of a simulated QTL analysis, a modified selective genotyping strategy was employed for QTL analysis with 178 F2 individuals (128 randomly selected, 25 shortest achene F2’s and 25 longest achene F2’s). Altogether 18 achene-size related QTL with small effects were identified. Achene-size related genes characteristically expressed additive gene action although several alleles were partially recessive towards the large achene parent. The 18 identified achene-size related QTL were validated in the same diallel mating experiment mentioned above. Significant marker-trait associations were detected for 15 QTL. In addition to 12 significant alleles detected from the F2 population, 12 additional alleles associated with achene-size traits were revealed after the analysis with the diallel. These markers are ready to be used in MAS for confectionery sunflower breeding. Great steps have been made in obtaining essential knowledge of inheritance and linkage of target traits for breeding confectionery sunflower adapted to Australian production environments. The identified markers can be used in MAS and further enhance the breeding process.

Sunflower is grown worldwide for oil production. In Australia, sunflower is under intense competition from canola as an oil crop. Confectionery sunflower aimed for the overseas snack food market is a potential opportunity for Australian farmers due to its ability to provide off-season supply to the major market located mainly in the northern hemisphere. To serve this aim, new confectionery sunflower hybrids adapted to Australian production system with drought tolerance, large achenes, high self-fertility and rust resistance are of paramount importance. The objectives of this project were to understand the genetic inheritance and genetic linkage relationships of the above traits and also to identify linked DNA markers with the potential for marker-assisted selection (MAS). The inheritance of achene-length was studied in a 12 x 12 diallel and a 14 x 12 North Carolina design II (NC II) mating design. Achene-length was evaluated under three environments for each mating design. Genetic correlation among all five achene-size related traits (achene-length, -width, - thickness and also kernel-length and -width) and 100-achene weight were moderate to high. Narrow-and broad-sense heritability for achene-length was moderate to high and similar for each design. The general combining ability (GCA) and maternal effects were found to be the main genetic factors controlling achene-length and each accounted for around 45% of the total genotypic variance. The inheritance of self-fertility (SF) was assessed with the same 12 x 12 diallel mating design evaluated in two environments. The 12 parents also displayed various levels of SF in addition to a wide range of achene-length. Negative genetic correlations between SF and all five achene-size related traits and 100-achene weight were observed. Modelling achene-length as a covariate revealed that certain pre-pollination floret characteristics represented by achene-length at maturity strongly affects SF in sunflower. When the effect of achene-length was removed statistically as a covariate, the specific combining ability (SCA) effect increased dramatically and became the major factor (other than pre-pollination floret characteristics) for SF determination and accounted for 65% of the total variance. Long achene genotypes tended to have low SF and a positive SCA effect was observed more frequently in F1 hybrids that involved small achene lines. The broad sense heritability was moderate (0.663 ± 0.017), while the estimate of narrow-sense heritability was nearly zero. A segregating F6:7 recombinant inbred line population with 239 progeny was developed from a cross between HaR4, a low delta (Δ), high transpiration efficiency (TE), sunflower line with rust resistance but relatively low SF, and SA52 with all the characteristics in the opposite direction. The population was evaluated in two environments for three TE-related traits including Δ, specific leaf weight (SLW) and chlorophyll content (CC) and four agronomic traits i.e. days to flowering (DTF), plant height (PlantH), SF and rust resistance (RustR). A linkage map 1074.6 cM in length with 138 SSR and 37 ISSR markers consisting of 19 linkage groups was constructed. Spatially adjusted BLUPs were used in QTL analysis for individual and combined environments. Altogether, 10 QTL were identified for Δ, SLW and CC at individual environments, and 7 were detected by a combined analysis. Sixteen QTL were identified for 4 agronomic traits and 14 of them were confirmed by a combined analysis. Significant linkage among all three TE related traits was discovered on linkage group 4 (LG4), where alleles from one parent had the same direction of influence on TE which suggested either coupling phase linkage or pleiotropy. QTL for SF and RustR were closely linked on LG13. An F2 population with 502 progeny developed from a cross between Jumbo, a large achene confectionery sunflower line, and RIL25, an oilseed small achene line, was evaluated for five achene-size related traits in a single field trial. Using a subset of 178 progeny a framework genetic linkage map, 899.7 cM in length, containing 128 SSR markers and 23 ISSR markers was constructed with 18 linkage groups. Based on the results of a simulated QTL analysis, a modified selective genotyping strategy was employed for QTL analysis with 178 F2 individuals (128 randomly selected, 25 shortest achene F2’s and 25 longest achene F2’s). Altogether 18 achene-size related QTL with small effects were identified. Achene-size related genes characteristically expressed additive gene action although several alleles were partially recessive towards the large achene parent. The 18 identified achene-size related QTL were validated in the same diallel mating experiment mentioned above. Significant marker-trait associations were detected for 15 QTL. In addition to 12 significant alleles detected from the F2 population, 12 additional alleles associated with achene-size traits were revealed after the analysis with the diallel. These markers are ready to be used in MAS for confectionery sunflower breeding. Great steps have been made in obtaining essential knowledge of inheritance and linkage of target traits for breeding confectionery sunflower adapted to Australian production environments. The identified markers can be used in MAS and further enhance the breeding process.

Increasing fuel prices, stricter government policies, and technological developments made it possible to seek for renewable alternatives, called biofuels, to petroleum fuel. Biodiesel, a biofuel that is produced from chemically mixing animal fat, vegetable oils, or recycled restaurant grease with alcohol and catalyst, is gaining popularity in recent years as a substitute for petroleum diesel. Ninety percent (90%) of U.S. biodiesel industry makes use of soybean oil as its feedstock. However, soybean oil alone cannot meet such a huge demand on biofuel production. Hence, it is important to identify and get more information about other feedstocks, specifically on its effects on the performance and exhaust emissions of diesel engines. The purpose of this study is to investigate the performance and emissions of two diesel engines operating on different biodiesel fuels (i.e. canola oil, sunflower oil, safflower oil, peanut oil, and chicken fat) and compare them to the performance and emissions when the engine is operated on soybean oil-based biodiesel and petroleum-based diesel. Results indicated that an engine operating on biodiesel generates a little less power and torque at any given speed than one running on diesel. Such power and torque loss were attributed to the biodiesel's lower energy content. The lower heating value (energy content) of biodiesel can be reflected in the specific fuel consumption, i.e., to generate the same power, more biodiesel is needed. The reduction in torque and power of less than 10% indicates that in some cases biodiesel has better combustion than diesel. Unfortunately, the high efficiency of combustion may give rise to increased combustion temperature which may lead to higher exhaust emissions. The gradual decrease in the total hydrocarbon and CO2 emissions, as blends were increased from B20 to B100, was also found to be an indication of better combustion using biodiesel fuels than petroleum diesel. However, NOx emissions were higher, predominantly at low speeds for most biodiesel and blends and therefore may require some additives or engine modifications/or adjustments to equalize the NOx emissions of diesel. Other emissions particularly SO2 were lower than standards require.

社交媒體在最近的社會運動中扮演了非常重要的角色，例如阿拉伯 之春、西班牙的Indignants 抗議運動，以及在北美的佔領華爾街運動， 皆凸顯出數位社交媒體的特殊地位與它對促進社會運動的貢獻。而 此篇碩士論文主要探討社交媒體在台灣的「太陽花運動」中所擔任 的角色。文中主張了社交媒體與「新媒體」已經是這個運動的延伸 與成功的重要因素。除了台灣年輕人常用的Facebook、Line 等社交 媒體之外，另一個使用社交媒體的重要因子是抗議者對台灣傳統媒 體的不信任。一般公民也可以透過「新媒體」成為抗議活動的一部 分，發揮和主流媒體相同的影響力。這些「新媒體」會為台灣更清 晰的民主鋪路，並同時提高年輕公民更多的政治意識。本論文的研 究包含來自Facebook 與PTT 還有「新媒體」的資料，此外，此論文 亦透過與太陽花學運參與者的採訪，評估他們在運動時使用社交媒 體的狀況。 / Social media plays a crucial role in recent social movements. The Arab Spring, the Indignants protest in Spain and the Occupy Wall Street movement in North America highlighted the special role of digital social media networks and its contribution to the facilitation of protest movements. This master thesis examines the role of the social media in the “Sunflower Student Movement” in Taiwan. It argues that social media and the “new media” have been a crucial factor to the prolongation and success of this movement. Besides the high penetration rate of social media, such as Facebook and LINE, among young Taiwanese, another important factor for protestors to use social media was the mistrust in the traditional Taiwanese media. Through the “new media” ordinary citizens could become a part of the protest and exert the same influence as the mainstream media. The “new media” may pave the way for a more transparent democracy in Taiwan and raise more political awareness among the young citizens. The research of this thesis includes information collected from Facebook and PTT as well as the “new media”. Additionally, interviews among participants of the Sunflower Student Movement were conducted to assess their use of social media during the movement.

Sunflower is grown worldwide for oil production. In Australia, sunflower is under intense competition from canola as an oil crop. Confectionery sunflower aimed for the overseas snack food market is a potential opportunity for Australian farmers due to its ability to provide off-season supply to the major market located mainly in the northern hemisphere. To serve this aim, new confectionery sunflower hybrids adapted to Australian production system with drought tolerance, large achenes, high self-fertility and rust resistance are of paramount importance. The objectives of this project were to understand the genetic inheritance and genetic linkage relationships of the above traits and also to identify linked DNA markers with the potential for marker-assisted selection (MAS). The inheritance of achene-length was studied in a 12 x 12 diallel and a 14 x 12 North Carolina design II (NC II) mating design. Achene-length was evaluated under three environments for each mating design. Genetic correlation among all five achene-size related traits (achene-length, -width, - thickness and also kernel-length and -width) and 100-achene weight were moderate to high. Narrow-and broad-sense heritability for achene-length was moderate to high and similar for each design. The general combining ability (GCA) and maternal effects were found to be the main genetic factors controlling achene-length and each accounted for around 45% of the total genotypic variance. The inheritance of self-fertility (SF) was assessed with the same 12 x 12 diallel mating design evaluated in two environments. The 12 parents also displayed various levels of SF in addition to a wide range of achene-length. Negative genetic correlations between SF and all five achene-size related traits and 100-achene weight were observed. Modelling achene-length as a covariate revealed that certain pre-pollination floret characteristics represented by achene-length at maturity strongly affects SF in sunflower. When the effect of achene-length was removed statistically as a covariate, the specific combining ability (SCA) effect increased dramatically and became the major factor (other than pre-pollination floret characteristics) for SF determination and accounted for 65% of the total variance. Long achene genotypes tended to have low SF and a positive SCA effect was observed more frequently in F1 hybrids that involved small achene lines. The broad sense heritability was moderate (0.663 ± 0.017), while the estimate of narrow-sense heritability was nearly zero. A segregating F6:7 recombinant inbred line population with 239 progeny was developed from a cross between HaR4, a low delta (Δ), high transpiration efficiency (TE), sunflower line with rust resistance but relatively low SF, and SA52 with all the characteristics in the opposite direction. The population was evaluated in two environments for three TE-related traits including Δ, specific leaf weight (SLW) and chlorophyll content (CC) and four agronomic traits i.e. days to flowering (DTF), plant height (PlantH), SF and rust resistance (RustR). A linkage map 1074.6 cM in length with 138 SSR and 37 ISSR markers consisting of 19 linkage groups was constructed. Spatially adjusted BLUPs were used in QTL analysis for individual and combined environments. Altogether, 10 QTL were identified for Δ, SLW and CC at individual environments, and 7 were detected by a combined analysis. Sixteen QTL were identified for 4 agronomic traits and 14 of them were confirmed by a combined analysis. Significant linkage among all three TE related traits was discovered on linkage group 4 (LG4), where alleles from one parent had the same direction of influence on TE which suggested either coupling phase linkage or pleiotropy. QTL for SF and RustR were closely linked on LG13. An F2 population with 502 progeny developed from a cross between Jumbo, a large achene confectionery sunflower line, and RIL25, an oilseed small achene line, was evaluated for five achene-size related traits in a single field trial. Using a subset of 178 progeny a framework genetic linkage map, 899.7 cM in length, containing 128 SSR markers and 23 ISSR markers was constructed with 18 linkage groups. Based on the results of a simulated QTL analysis, a modified selective genotyping strategy was employed for QTL analysis with 178 F2 individuals (128 randomly selected, 25 shortest achene F2’s and 25 longest achene F2’s). Altogether 18 achene-size related QTL with small effects were identified. Achene-size related genes characteristically expressed additive gene action although several alleles were partially recessive towards the large achene parent. The 18 identified achene-size related QTL were validated in the same diallel mating experiment mentioned above. Significant marker-trait associations were detected for 15 QTL. In addition to 12 significant alleles detected from the F2 population, 12 additional alleles associated with achene-size traits were revealed after the analysis with the diallel. These markers are ready to be used in MAS for confectionery sunflower breeding. Great steps have been made in obtaining essential knowledge of inheritance and linkage of target traits for breeding confectionery sunflower adapted to Australian production environments. The identified markers can be used in MAS and further enhance the breeding process.

Sunflower is grown worldwide for oil production. In Australia, sunflower is under intense competition from canola as an oil crop. Confectionery sunflower aimed for the overseas snack food market is a potential opportunity for Australian farmers due to its ability to provide off-season supply to the major market located mainly in the northern hemisphere. To serve this aim, new confectionery sunflower hybrids adapted to Australian production system with drought tolerance, large achenes, high self-fertility and rust resistance are of paramount importance. The objectives of this project were to understand the genetic inheritance and genetic linkage relationships of the above traits and also to identify linked DNA markers with the potential for marker-assisted selection (MAS). The inheritance of achene-length was studied in a 12 x 12 diallel and a 14 x 12 North Carolina design II (NC II) mating design. Achene-length was evaluated under three environments for each mating design. Genetic correlation among all five achene-size related traits (achene-length, -width, - thickness and also kernel-length and -width) and 100-achene weight were moderate to high. Narrow-and broad-sense heritability for achene-length was moderate to high and similar for each design. The general combining ability (GCA) and maternal effects were found to be the main genetic factors controlling achene-length and each accounted for around 45% of the total genotypic variance. The inheritance of self-fertility (SF) was assessed with the same 12 x 12 diallel mating design evaluated in two environments. The 12 parents also displayed various levels of SF in addition to a wide range of achene-length. Negative genetic correlations between SF and all five achene-size related traits and 100-achene weight were observed. Modelling achene-length as a covariate revealed that certain pre-pollination floret characteristics represented by achene-length at maturity strongly affects SF in sunflower. When the effect of achene-length was removed statistically as a covariate, the specific combining ability (SCA) effect increased dramatically and became the major factor (other than pre-pollination floret characteristics) for SF determination and accounted for 65% of the total variance. Long achene genotypes tended to have low SF and a positive SCA effect was observed more frequently in F1 hybrids that involved small achene lines. The broad sense heritability was moderate (0.663 ± 0.017), while the estimate of narrow-sense heritability was nearly zero. A segregating F6:7 recombinant inbred line population with 239 progeny was developed from a cross between HaR4, a low delta (Δ), high transpiration efficiency (TE), sunflower line with rust resistance but relatively low SF, and SA52 with all the characteristics in the opposite direction. The population was evaluated in two environments for three TE-related traits including Δ, specific leaf weight (SLW) and chlorophyll content (CC) and four agronomic traits i.e. days to flowering (DTF), plant height (PlantH), SF and rust resistance (RustR). A linkage map 1074.6 cM in length with 138 SSR and 37 ISSR markers consisting of 19 linkage groups was constructed. Spatially adjusted BLUPs were used in QTL analysis for individual and combined environments. Altogether, 10 QTL were identified for Δ, SLW and CC at individual environments, and 7 were detected by a combined analysis. Sixteen QTL were identified for 4 agronomic traits and 14 of them were confirmed by a combined analysis. Significant linkage among all three TE related traits was discovered on linkage group 4 (LG4), where alleles from one parent had the same direction of influence on TE which suggested either coupling phase linkage or pleiotropy. QTL for SF and RustR were closely linked on LG13. An F2 population with 502 progeny developed from a cross between Jumbo, a large achene confectionery sunflower line, and RIL25, an oilseed small achene line, was evaluated for five achene-size related traits in a single field trial. Using a subset of 178 progeny a framework genetic linkage map, 899.7 cM in length, containing 128 SSR markers and 23 ISSR markers was constructed with 18 linkage groups. Based on the results of a simulated QTL analysis, a modified selective genotyping strategy was employed for QTL analysis with 178 F2 individuals (128 randomly selected, 25 shortest achene F2’s and 25 longest achene F2’s). Altogether 18 achene-size related QTL with small effects were identified. Achene-size related genes characteristically expressed additive gene action although several alleles were partially recessive towards the large achene parent. The 18 identified achene-size related QTL were validated in the same diallel mating experiment mentioned above. Significant marker-trait associations were detected for 15 QTL. In addition to 12 significant alleles detected from the F2 population, 12 additional alleles associated with achene-size traits were revealed after the analysis with the diallel. These markers are ready to be used in MAS for confectionery sunflower breeding. Great steps have been made in obtaining essential knowledge of inheritance and linkage of target traits for breeding confectionery sunflower adapted to Australian production environments. The identified markers can be used in MAS and further enhance the breeding process.

Made available in DSpace on 2014-06-11T19:30:31Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-08-21Bitstream added on 2014-06-13T18:40:47Z : No. of bitstreams: 1 fabregat_thp_dr_jabo.pdf: 345602 bytes, checksum: 5347d7da2b9921eae2231b13232ccc57 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / O objetivo deste estudo foi avaliar o efeito da utilização de dietas contendo diferentes ingredientes fibrosos (farelo de soja, casca de soja, farelo de girassol e polpa cítrica, este ultimo avaliado em dois níveis de inclusão) sobre o tempo de trânsito gastrointestinal, as digestibilidades da proteína e da energia, o desempenho, a composição corporal, os parâmetros bioquímicos, os índices organométricos e a morfometria intestinal de juvenis de pacu (Piaractus mesopotamicus). Os peixes alimentados com as dietas contendo farelo de soja e farelos de girassol tiveram os melhores resultados de digestibilidade e desempenho, sem apresentar alterações fisiológicas e metabólicas. A inclusão casca de soja na dieta atrasou o tempo de trânsito gastrointestinal, piorou a digestibilidade da proteína e da energia, prejudicou o desempenho, diminuiu as proteínas totais sangüíneas e aumentou a porcentagem de lipídeo muscular. A dieta contendo 45% polpa cítrica acelerou o tempo de trânsito gastrointestinal, piorou o desempenho, provocou alterações na composição corporal e na morfometria intestinal. Cada ingrediente fibroso, em função de sua composição de fibras alimentares e carboidratos complexos, altera o funcionamento do sistema digestório e o aproveitamento de nutrientes, com conseqüências sobre a fisiologia, o metabolismo e o desempenho dos juvenis de pacu / This study aim was to evaluate the effects of diets containing different fiber sources (soybean meal, soybean hulls, sunflower meal and citrus pulp, this last evaluated at two inclusion levels) on gastrointestinal transit time, protein and energy digestibility of protein, performance, body composition, biochemical parameters, organometric index and intestinal morphology of juvenile pacu (Piaractus mesopotamicus). Fish fed diets containing soybean meal and sunflower meal had best digestibility and performance results, with no physiological and metabolic diseases. The inclusion of soybean hulls in the diet delayed gastrointestinal transit time, decreased the digestibility of protein and energy, worsened performance, reduced total blood proteins and increased the percentage of muscle lipid. Diet containing 45% citrus pulp accelerated gastrointestinal transit time, worsened performance, led to changes in body composition and affects the intestinal morphology. Each fibrous ingredient, depending on dietary fiber and complex carbohydrates composition, differently affects the digestive system functioning and nutrient use, with consequences on physiology, metabolism and performance of juvenile pacu