O impacto ambiental causado pelo descarte indevido de embalagens a base de polímeros sintéticos, desperta interesse na necessidade de desenvolvimento de embalagens a partir de materiais biodegradáveis. O objetivo deste trabalho foi desenvolver e caracterizar bandejas a base de espuma de amido de mandioca para embalagem de alimentos, obtidas pelo processo de expansão térmica. Foram testadas diferentes proporções de amido e água, como agente expansor, na formulação das espumas de amido. Como plastificantes, foram utilizados o glicerol, sorbitol e poli (álcool vinílico) (PVA), nas concentrações de 2,5; 5,0 e 7,5% m/m sob a massa de amido. A fim de melhorar as características de hidrofilia das espumas, foi aplicado revestimento a base de poli (ácido lático) (PLA) na superfície das bandejas. Testou-se a aplicação das bandejas como embalagem no armazenamento de morangos. A caracterização das embalagens foi realizada através de análises de espessura e densidade, MEV, capacidade de absorção de água (CAA), solubilidade, isotermas de sorção, resistência à tração, ângulo de contato e degradação enzimática. A proporção amido:água que produziu espumas com a melhor combinação entre as características de densidade, capacidade de absorção de água e aspecto visual foi 47,5:52,5. O aditivo que resultou em espumas de amido com a maior possibilidade de aplicação como embalagem para alimentos, por apresentar a menor CAA, ser menos solúvel e com os valores mais baixos de umidade de equilíbrio, foi o PVA em concentração de 5%. A aplicação de revestimento a base de PLA na superfície das bandejas de espuma de amido resultou em diminuição de 86% na CAA das bandejas e ângulo de contato de 84,4° (maior hidrofobia). Os modelos que melhor se ajustam a isoterma de sorção das bandejas sem revestimento são os de GAB e Oswin. Enquanto que para bandejas com revestimento de PLA, o modelo de Oswin apresenta o melhor ajuste. O ensaio de degradação enzimática por amilases microbianas revelou ocorrência da hidrólise do amido nas primeiras 8 horas do teste, indicando característica de biodegradabilidade das bandejas. Os resultados do presente estudo, obtidos através do teste de aplicabilidade das bandejas, comprovam a possibilidade da utilização da embalagem à base de amido de mandioca revestidas com PLA em alimentos com alta atividade de água. / Submitted by Ana Guimarães Pereira (agpereir@ucs.br) on 2015-06-25T13:40:08Z No. of bitstreams: 1 Dissertacao Fernanda Stoffel.pdf: 3201484 bytes, checksum: 203dc77998956c27e8e954d02d1b8c0e (MD5) / Made available in DSpace on 2015-06-25T13:40:08Z (GMT). No. of bitstreams: 1 Dissertacao Fernanda Stoffel.pdf: 3201484 bytes, checksum: 203dc77998956c27e8e954d02d1b8c0e (MD5) / Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul. / The environmental impact caused by improper disposal of packaging materials based on synthetic polymers, arouses interest in development packaging from biodegradable materials. The objective of this study was to develop and characterize trays base of cassava starch foam for food packaging, obtained by the thermal expansion process. Different proportions of starch and water, as blowing agent, in the formulation of the starch foams was tested. As plasticizers were used glycerol, sorbitol, and poly (vinyl alcohol) (PVA) at concentrations of 2.5; 5.0 and 7.5% w/w on starch weight. In order to improve the hydrophilic characteristics of the foams, coating was applied of poly (lactic acid) (PLA) in the surface of the trays. The application of the trays as packaging strawberries in storage was tested. Packaging characterization was performed by thickness and density analysis, SEM, water absorption capacity (WAC), solubility, sorption isotherms, tensile strength, angle of contact and enzymatic degradation. The ratio starch:water who produced foams with the best combination of density, water absorption capacity and visual aspect characteristics was 47.5:52.5. The starch additive resulted in foams with the highest possibility of application as packaging for food for presenting the lowest WAC be less soluble and lowest values equilibrium moisture, was PVA concentration of 5%. The use of PLA-based coating on the surface of the starch foam trays resulted in a decrease of 86% in WAC trays and contact angle of 84.4 ° (higher hydrophobicity). The models that best fit the sorption isotherm of uncoated trays are the GAB and Oswin models. While for trays with PLA coating, Oswin model presents the best fit. The assay of enzymatic degradation by microbial amylases revealed the occurrence of the hydrolysis of starch in the first 8 hours of the test, indicating the biodegradability characteristics of the trays. The results of this study, obtained through the applicability of the test trays prove the possibility of the use of tapioca starch-based packing covered with PLA foods with high water activity.

O impacto ambiental causado pelo descarte indevido de embalagens a base de polímeros sintéticos, desperta interesse na necessidade de desenvolvimento de embalagens a partir de materiais biodegradáveis. O objetivo deste trabalho foi desenvolver e caracterizar bandejas a base de espuma de amido de mandioca para embalagem de alimentos, obtidas pelo processo de expansão térmica. Foram testadas diferentes proporções de amido e água, como agente expansor, na formulação das espumas de amido. Como plastificantes, foram utilizados o glicerol, sorbitol e poli (álcool vinílico) (PVA), nas concentrações de 2,5; 5,0 e 7,5% m/m sob a massa de amido. A fim de melhorar as características de hidrofilia das espumas, foi aplicado revestimento a base de poli (ácido lático) (PLA) na superfície das bandejas. Testou-se a aplicação das bandejas como embalagem no armazenamento de morangos. A caracterização das embalagens foi realizada através de análises de espessura e densidade, MEV, capacidade de absorção de água (CAA), solubilidade, isotermas de sorção, resistência à tração, ângulo de contato e degradação enzimática. A proporção amido:água que produziu espumas com a melhor combinação entre as características de densidade, capacidade de absorção de água e aspecto visual foi 47,5:52,5. O aditivo que resultou em espumas de amido com a maior possibilidade de aplicação como embalagem para alimentos, por apresentar a menor CAA, ser menos solúvel e com os valores mais baixos de umidade de equilíbrio, foi o PVA em concentração de 5%. A aplicação de revestimento a base de PLA na superfície das bandejas de espuma de amido resultou em diminuição de 86% na CAA das bandejas e ângulo de contato de 84,4° (maior hidrofobia). Os modelos que melhor se ajustam a isoterma de sorção das bandejas sem revestimento são os de GAB e Oswin. Enquanto que para bandejas com revestimento de PLA, o modelo de Oswin apresenta o melhor ajuste. O ensaio de degradação enzimática por amilases microbianas revelou ocorrência da hidrólise do amido nas primeiras 8 horas do teste, indicando característica de biodegradabilidade das bandejas. Os resultados do presente estudo, obtidos através do teste de aplicabilidade das bandejas, comprovam a possibilidade da utilização da embalagem à base de amido de mandioca revestidas com PLA em alimentos com alta atividade de água. / Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul. / The environmental impact caused by improper disposal of packaging materials based on synthetic polymers, arouses interest in development packaging from biodegradable materials. The objective of this study was to develop and characterize trays base of cassava starch foam for food packaging, obtained by the thermal expansion process. Different proportions of starch and water, as blowing agent, in the formulation of the starch foams was tested. As plasticizers were used glycerol, sorbitol, and poly (vinyl alcohol) (PVA) at concentrations of 2.5; 5.0 and 7.5% w/w on starch weight. In order to improve the hydrophilic characteristics of the foams, coating was applied of poly (lactic acid) (PLA) in the surface of the trays. The application of the trays as packaging strawberries in storage was tested. Packaging characterization was performed by thickness and density analysis, SEM, water absorption capacity (WAC), solubility, sorption isotherms, tensile strength, angle of contact and enzymatic degradation. The ratio starch:water who produced foams with the best combination of density, water absorption capacity and visual aspect characteristics was 47.5:52.5. The starch additive resulted in foams with the highest possibility of application as packaging for food for presenting the lowest WAC be less soluble and lowest values equilibrium moisture, was PVA concentration of 5%. The use of PLA-based coating on the surface of the starch foam trays resulted in a decrease of 86% in WAC trays and contact angle of 84.4 ° (higher hydrophobicity). The models that best fit the sorption isotherm of uncoated trays are the GAB and Oswin models. While for trays with PLA coating, Oswin model presents the best fit. The assay of enzymatic degradation by microbial amylases revealed the occurrence of the hydrolysis of starch in the first 8 hours of the test, indicating the biodegradability characteristics of the trays. The results of this study, obtained through the applicability of the test trays prove the possibility of the use of tapioca starch-based packing covered with PLA foods with high water activity.

Atualmente o consumo de xarope de maltose vem crescendo devido ao seu uso em cervejarias e está substituindo progressivamente os adjuntos amiláceos. O xarope de maltose é, tradicionalmente, produzido por meio da hidrólise ácida e/ou enzimática de amido ou flocos de milho. Este trabalho teve como objetivo analisar a possibilidade de obtenção de maltose a partir de outras matérias-primas amiláceas como a mandioca e a puba (produto derivado da fermentação da mandioca) pela ação da a-amilase bacteriana e da a-amilase fúngica sem que fosse necessária a extração do amido. Amostras de mandioca e puba com 10, 20 e 30% de sólidos foram incubadas com a-amilase bacteriana termoestável durante 10, 20 e 30 minutos a 80 0 C, adicionando-se em seguida, µ-amilase fúngica e incubando-se as amostras durante 48 horas a 55 0 C. O grau de sacarificação, expresso em dextrose equivalente (DE), foi determinado pelo método DNS em vários intervalos de tempo. Glicose e maltose foram determinadas por HPLC após 48 horas de sacarificação. Os resultados mostraram que o tempo de ação da a-amilase bacteriana não causou diferenças significativas no grau de hidrólise entre as amostras, mas a concentração de sólidos influiu significativamente no grau da liquefação das amostras. O comportamento das curvas do grau de sacarificação foi semelhante para todos os tratamentos tanto da mandioca quanto da puba. O conteúdo de maltose nas amostras variou entre 30-60% e a glicose entre 0-10% caracterizando um xarope com alto teor de maltose. A eficiência de hidrólise ficou abaixo do esperado. Entretanto, esse fato pode ser explicado pela utilização da mandioca sem extração prévia do amido e pelas dificuldades na extração dos sólidos por centrifugação. Pode-se afirmar que tanto a mandioca quanto à puba podem ser utilizadas como matéria prima em substituição ao milho na obtenção de xarope de maltose através da hidrólise enzimática. A puba, porém, é de mais fácil manuseio sendo que o tratamento com 20% de sólidos, exposto durante 10 minutos a a-amilase bacteriana proporcionou maior rendimento, atingindo 4,2 kg de maltose e 0,3 kg de glicose por 100 kg de mandioca fresca além de proporcionar menor quantidade de resíduo sólidos de 13,7 kg. / Nowadays the consumption of maltose syrups is increasing due to its utilization in breweries where it replaces starch adjuncts. Traditionally maltose syrup has been produced from cornstarch or pellets using acid and/or enzymatic hydrolysis. The objective of this work was to evaluate the possibility of obtaining maltose from starch sources other than maize, such as cassava roots or puba, a fermented cassava product, without extraction of the starch, using bacterial a-amylase and fungal a-amylase for starch hydrolysis. Cassava and puba samples with 10, 20 and 30% dry matter were incubated with termostable bacterial a-amylase for 10, 20 and 30 minutes at 80 0 C, followed by the addition of fungal a-amylase. The samples were incubated for 48 hours at 55 0 C. The degree of saccharification, expressed as dextrose equivalent (DE), was determined by the DNS method. The glucose and maltose contents of the hydrolysate were determined after 48 hours by HPLC. The results showed that the time of action of a-amylase did not influence the degree of saccharification of the samples but the solids concentration significantly affected the hydrolysis degree. The saccharification degree curves were similar for both cassava and puba. The maltose content of the samples varied between 30-60% and the glucose content between 0-10%, which characterized them as "High maltose syrups". The hydrolysis efficiency was lower than expected. However, this fact could be explained by the use of cassava without extraction of the starch and by the difficulty of extracting the solids by centrifugation. It was concluded that for replacement of corn starch, both cassava roots and puba could be used as raw materials for maltose syrup production by enzymatic hydrolysis. However the puba was easier to handle than cassava. The puba treatment consisting of 20% of solids and 10 minutes exposure to a-amylase gave the highest yield reaching 4.2 kg of maltose and 0.3 kg of glucose per 100 kg of raw cassava, in addition to a lower solids residue of 13.7 kg.

A mandioca (Manihot esculenta Crantz) é uma das principais culturas do mundo, havendo grande variabilidade genética. As variedades são classificadas com base na palatabilidade e toxicidade das raízes, em mansas ou doces e bravas ou amargas. Apesar da importância, o potencial da mandioca é pouco explorado, não sendo conhecidos, em nível molecular, os elementos determinantes para as suas características. Assim, pretendeu-se identificar, empregando a 2D-PAGE, proteínas que possam estar associadas com as diferenças físico-químicas das raízes tuberosas de variedades de mesa (IAC 576-70 e IAC 06-01), indústria (Cigana Preta, IAC 12 e IAC 90) e de uso misto (Vassourinha Paulista). Após extração de proteínas e separação por 2D-PAGE, as imagens dos géis foram analisadas no programa Delta2D (DECODON), sendo realizada análise estatística utilizando-se ANOVA (p<0,01), Heat Map e Análises de Componentes Principais (ACP) e de Agrupamentos. Os 146 spots de interesse foram removidos dos géis e suas proteínas digeridas e sequenciadas por espectrometria de massas. Algumas proteínas refletiram as características fenotípicas das variedades em estudo, especialmente entre as de mesa e indústria. Pela ACP, foram explicados 54,54% da variabilidade entre as amostras. A primeira componente separou as variedades exclusivamente de mesa de todas as demais, enquanto a segunda separou a IAC 90 de todas as outras, sendo esta caracterizada por um perfil proteico diferente das demais amostras de uso industrial. A IAC 576-70 e a IAC 12 apresentaram alta correlação positiva, assim como, a Vassourinha e a Cigana. A Análise de Agrupamentos corroborou as informações da ACP, revelando que o proteoma das raízes tuberosas refletiu diferenças fenotípicas entre as variedades. / Cassava (Manihot esculenta Crantz) is a main crop with large genetic variability. The varieties are classified according palatability and toxicity of the roots as sweet or bitter cassavas. Despite its importance, little is known about the molecular basis of phenotypic characteristics. Therefore, this study aimed to identify proteins associated to the differences between the sweet (\'IAC 576-70\' e \'IAC 06-01\'), bitter (\'Cigana Preta\', \'IAC 12\' e \'IAC 90\') and the mixed-use (\'Vassourinha Paulista\') varieties by 2D-PAGE. After the protein extraction and separation by 2D-PAGE, the gel images were analyzed through the software Delta 2D (DECODON), and the statistical analysis were performed with ANOVA (p<0,01), Heat Map, Principal Component Analysis (PCA) and Cluster Analysis. The 146 significant spots were removed from the gels, digested and sequenced by mass spectrometry. Some proteins were related to the physico-chemical characteristics of the varieties, especially between the sweet and the bitter. Variability of the samples was explained at the level of 54,54% by the PCA. The first component separated the sweet varieties from all others while the second one separated the \'IAC 90\' from all others. This variety was characterized by a different protein profile among the bitter cassavas. The \'IAC 576-70\' and the \'IAC 12\' were positively correlated, as well as, \'Vassourinha\' and the \'Cigana\'. Cluster Analysis agreed the PCA information, revealing that the proteomes of the tuberous roots reflected phenotypic differences among the varieties.

Pubagem é o processo de fermentação natural de raízes de mandioca para produção de puba, um alimento tradicional nas regiões Norte e Nordeste do Brasil. Além da fermentação lática, uma ação combinada da pectina-metil-esterase endógena e enzimas microbianas despolimerizantes causa o amolecimento das raízes, que é importante para sua completa desintegração. No entanto, o reconhecimento do ponto final de fermentação e as condições que propiciam puba de boa qualidade não são completamente esclarecidos. O objetivo do trabalho foi estudar a influência da temperatura de fermentação e da adição de enzima como auxiliar de pubagem na textura das raízes e rendimento de puba, procurando associar estes parâmetros para detectar o ponto final desta etapa do processo. Cerca de 1Kg de raízes descascadas e 2 litros de água foram colocados em recipientes plásticos. Diferentes concentrações (0,1 e 2mL) de pectinase comercial de Aspergillus aculeatus/Kg de raiz foram adicionadas em tratamentos separados. Amostras foram incubadas a 25, 30 e 35ºC, por períodos de 48, 72 e 96 horas. Foram efetuadas análises de acompanhamento do processo fermentativo (pH, textura e rendimento) e da composição da farinha de puba obtida. O maior rendimento, de 77% após 72 horas, foi observado no tratamento a 25ºC com 2mL de enzima por Kg de raiz, quando a textura que era de 18,62lbf/g na matériaprima passou para 3,92lbf/g na raiz pubada. Outros rendimentos mais próximos desse valor, 75,7 e 74,7%, foram obtidos respectivamente na fermentação natural a 35ºC e na amostra tratada com 2mL de enzima incubada a 30ºC/48 horas. Os valores de textura foram de 2,50lbf/g para o primeiro tratamento e 2,89lbf/g para o segundo. A relação entre textura e rendimento foi significativa para a temperatura de 30ºC e todos os períodos de 72 horas. A associação entre os dados de textura e rendimento permite concluir que quando a textura alcança valores da ordem de 4lbf/g os rendimentos de puba são maiores, indicando o final do processo. Os rendimentos mais baixos ocorreram para os valores de textura próximos a 2,30lbf/g de amostra, geralmente obtidos em fermentações mais longas e com maior concentração de enzima. / "Retting" is a natural fermentation of cassava roots for the production of puba, a traditional food in the North and Northeast of Brazil. Besides the lactic fermentation, the combined action of the endogenous pectin-methyl-esterase and depolymeryzyng enzymes, causes a softening of the roots which is important for its complete disintegration. However, the recognition of the end point of fermentation and the conditions that lead to good quality puba are not completely understood. The objective of this work was to study the influence of the temperature and enzyme concentration on the yield and texture, aiming at associating these parameters to detect the end point of fermentation. The physicochemical properties of the puba flour were also determined. About 1Kg of peeled roots and 2 liters of water were placed in plastic containers. One or two milliliters of commercial pectinase from Aspergillus aculeatus were added / Kg root, in separate treatments. In the control treatment no enzyme was added. Samples were incubated at 25, 30 and 35ºC and portions removed after 48, 72 and 96 hours for the determination of pH, texture, yield and dry matter. Fibers, starch, total soluble sugars and amylose were determined in the flour of the dried puba. The highest yield of 77% was obtained after 72 hours for the treatment at 25ºC with 2mL of enzyme/kg of root, when the texture values decreased from 18.62lbf/g (raw material) to 3.92lbf/g (retted root). A high yield of 75.7% was also observed in same period, for the natural fermentation at 35ºC, when texture values of 2.50lbf/g were reached. The relation between texture and yield was significant for a temperature of 30ºC and a period of 72 hours. The association between the texture and yield data suggests that when the texture reaches values of the order of 4lbf/g, the yields are higher, indicating the end of the process. The yields were lower when texture values were close to 2.30lbf/g, usually achieved for long fermentations and higher enzyme concentrations.

Made available in DSpace on 2014-07-29T15:22:52Z (GMT). No. of bitstreams: 1 Dissertacao Janaina Pereira.pdf: 781786 bytes, checksum: b3c5556878ceb315e8986893961a68f8 (MD5) Previous issue date: 2010-07-14 / The cassava bagasse, subproduct of the processing of the cassava starch, has been used mainly as animal feed. The main characteristic is to present high moisture content when in natura, starch content and dietary fiber. This study aimed to characterize the cassava starch physical and chemically, cassava bagasse in natura and cassava bagasse dehydrated and the biscuits formulated with different proportions of cassava bagasse dehydrated, with an sensorial biscuits analysis. Made, previous, the dehydration of cassava meal, then has been elaborated some biscuits formulas with concentrations of cassava from 0% (BP), 2% (BF2), 4% (BF4) 6% (BF6) and 8% (BF8), instead of cassava starch. The biscuits were analyzed for their preference, acceptability, physical and chemical characteristics and microbial quality. Data were analyzed by analysis of variance, Tukey test at 5% level of significance. The cassava starch presented content (90.65 g 100 g-1), carbohydrate (89,34 g 100 g-1) and zinc (0.88 g 100 g-1) and, lower ash content ( 0.13 g 100 g-1), acidity (1.01 mL of 0.1 NaOH 100 g-1), acid factor (0.28 mL of hydrochloric acid), phosphorus (45.00 g 100 g-1) , potassium (786.67 g 100 g-1) and calcium (200 g 100 g-1). The in natura cassava meal showed high levels of moisture (85.60 g 100 g-1). The cassava bagasse dehydrated showed high starch content (75,04 g 100 g-1), dietary fiber (60.27 g 100 g-1), phosphorus (64.00 g 100 g-1), calcium (500.00 g 100 g-1), magnesium (200 g 100 g-1) and manganese (4.00 g 100 g-1) and, reduced lipid content (4.49 g 100 g-1) and ash (1.98 g 100 g-1). The dietary fiber content and color of the biscuits increased with the addition of cassava bagasse dehydrated on the formulation, and the dietary fiber content ranged from 5.47 g 100 g-1 to 11.40 g 100 g-1. The specific volume of the biscuits decreased with the increasing of increment of cassava bagasse dehydrated in the formulation. There was no significant difference between the biscuits, as the preference. The biscuits had good acceptability for taste and appearance and microbiological quality according to the standards required by legislation. It is concluded that cassava biscuits made with cassava meal is a dried product with good nutritional potential and good acceptability. / O farelo de mandioca, subproduto do processamento da fécula de mandioca, vem sendo usado, principalmente, na alimentação animal. Entre suas principais características destaca-se o elevado teor de umidade, quando in natura, o teor de amido e o conteúdo de fibras alimentares. Este trabalho objetivou caracterizar físico e quimicamente a fécula de mandioca, o farelo de mandioca in natura e desidratado e os biscoitos de polvilho, formulados com diferentes proporções de farelo de mandioca desidratado, bem como analisá-los sensorialmente. Realizou-se, previamente, a desidratação do farelo de mandioca, em seguida elaborou-se formulações de biscoitos com concentrações deste de 0% (BP), 2% (BF2), 4% (BF4), 6% (BF6) e 8% (BF8), em substituição à fécula de mandioca. Os biscoitos foram analisados quanto à preferência, aceitabilidade, características físicas e químicas e qualidade microbiológica. Os dados foram analisados por meio da análise de variância, teste de Tukey, em nível de 5% de significância. A fécula de mandioca apresentou teor de amido de 90,65 g 100 g-1, carboidrato 89,34 g 100 g-1, zinco 0,88 g 100 g-1, teor de cinzas de 0,13 g 100 g-1, acidez de 1,01 mL de NaOH 0,1 100 g-1, fator ácido 0,28 mL de ácido clorídrico, fósforo 45,00 g 100 g-1, potássio 786,67 g 100 g-1 e cálcio 200 g 100 g-1. O farelo de mandioca in natura apresentou elevado teor de umidade (85,60 g 100 g-1). O farelo de mandioca desidratado apresentou elevado teor de amido (75,04 g 100 g-1), fibra alimentar (60,28 g 100 g-1), fósforo (64,00 g 100 g-1), cálcio (500,00 g 100 g-1), magnésio (200 g 100 g-1) e manganês (4,00 g 100 g-1) e reduzido teor de lipídio (4,49 g 100 g-1) e cinzas (1,98 g 100 g-1). O teor de fibra alimentar e a cor dos biscoitos aumentaram, significativamente, com a adição do farelo de mandioca desidratado nas formulações, sendo que o teor de fibra alimentar variou de 5,47 g 100 g-1 a 11,40 g 100 g-1. O volume específico dos biscoitos diminuiu com o aumento de incremento de farelo de mandioca desidratado na formulação. Não houve diferença significativa entre os biscoitos, quanto à preferência. Os biscoitos apresentaram boa aceitabilidade para aparência e sabor e qualidade microbiológica, estando de acordo com os padrões exigidos pela legislação. Conclui-se que biscoitos de polvilho elaborados com farelo de mandioca desidratado, constituem um produto com bom potencial nutricional e de boa aceitabilidade.

The response of cassava (Manihot esculenta Crantz) to water stress was studied in two distinct sites (Tucson, Arizona and Santander de Quilichao, Colombia). A third experiment was conducted in Palmira, Colombia to evaluate the relationship between photosynthesis, relative humidity and yield of cassava cultivars HCol 1684. Plants stressed after 2 months of growth in Tucson reduced growth by reducing leaf formation, expansion and leaf area. Reductions in LAI due to stress imposed to MCol 1684 in Quilichao were attained by reducing leaf expansion rates and leaf formation in plants stressed after 3 months of age. Plants stressed later increased leaf fall and did not reduce leaf formation. Stress reduced dry matter production in all cultivars, especially when given to young plants. Stress also altered the patterns of dry matter partitioning in 3-month-old plants of MCol 1684, but not in those stressed at 6 months. Transpiration and diffusive conductances of MCol 1684 were reduced after 40 days of stress. These parameters were correlated to photosynthesis and leaf temperatures for non-stressed plants, and additionally with relative humidity in the plants stressed after 3 months of growth. Interestingly, leaf temperatures were lower in stressed plants of MCol 1684, which suggests that stressed cassava plants can avoid excessive heating caused by stomatal closure simply changing leaf orientation and increasing reflectance. Leaf water potential was slightly reduced by stress in Tucson except for MVen 218. Stressed plants of MCol 1684 in Quilichao showed lower values of (L) than those of non-stressed plants after 30-40 days of treatment, suggesting an adaptation of stressed plants acquired during the stress period. The effects of air humidity on stomatal functioning of MCol 1684 seems to be strong as suggested by the dependence of transpiration, conductances and photosynthesis on relative humidity.

Student Number : 9610284H - MSc dissertation - School of Molecular and Cell Biology - Faculty of Science / Cassava Mosaic Disease is the most devastating disease affecting cassava (Manihot esculenta Crantz) crops worldwide. This disease is associated with eight species of geminiviruses, all belonging to the genus Begomovirus of the family Geminiviridae. In South Africa, in particular, CMD is caused by South African cassava mosaic virus (SACMV). Currently, there are no adequate methods for control of this disease as mechanisms within virus-host interactions are poorly understood. This brings about the need for development of virus-disease control strategies. This study was therefore conducted to identify the host’s response to an invading virus. The model plant, Arabidopsis was chosen as it is a well-characterized plant system, with expression databases readily available as its entire genome has been sequenced. This study was conducted, firstly, to phenotypically determine if Arabidopsis was resistant or susceptible to SACMV infection, and secondly, to identify the host’s response to pathogen infection on a molecular level through gene expression studies utilizing microarrays. Results from the symptomatology study revealed that Arabidopsis plants were fully symptomatic 28 days post-inoculation, displaying characteristic disease symptoms such as stunting, yellowing, and leaf deformation. This indicated that Arabidopsis was susceptible to SACMV infection. Microarray analyses revealed 86 differentially expressed genes, of which 48 showed up-regulation and 38 down-regulation. Relative quantification real-time PCR was performed on selected genes to confirm these results. Many up-regulated genes were shown to be primarily involved in a general stress response induced by the host, whereas those genes that were downregulated seemed to be involved in more specific responses to viral invasion, probably a consequence of suppression of host genes by SACMV to enhance its own replication. The majority of genes identified fell under the predominant functional categories involved in metabolism, transcription, and transport. To our knowledge, this is the first study in which a DNA geminivirus has been used in a host-pathogen interaction utilizing microarrays.

ABSTRACT Cassava is currently the second most important source of carbohydrates on the African continent. In the last two decades, cassava crops have been severely affected by outbreaks of cassava mosaic disease (CMD). South African cassava mosaic virus (SACMV) has been associated with CMD outbreaks in the Mpumalanga province. Advances in post-transcriptional gene silencing (PTGS) technology have provided promising new strategies for the engineering of virus resistance in plants. Inverted repeat (IR) constructs are currently the most potent inducers of PTGS, however, these constructs are inherently unstable. The purpose of this study was to develop IR constructs with an improved stability for the efficient induction of PTGS in plants. Two mismatched inverted repeat constructs, one targeting the SACMV BC1 open reading frame, the other targeting the Maize streak virus (MSV) AC1 open reading frame, were successfully created. Sodium bisulfite was used to deaminate cytosine residues on the sense arm of the constructs. The resulting number of GT mismatches was seemingly sufficient to stabilize the linear conformation of the IR constructs, as they were efficiently propagated by E.coli DH5!, and subsequently behaved like linear DNA molecules. Furthermore, it was found that the number of mismatches on the BC1 construct (17.5%) was ideal, as the subsequent stability of the predicted RNA hairpin was not affected. Due to the higher number of mismatches on the AC1 construct (23.5%), it was found that the loop region of the RNA hairpin was marginally destabilized. Despite this, long stretches of stable dsRNA were still produced from the AC1 IR construct, and is likely to induce PTGS. Interestingly, it was observed that the mismatched IR constructs, although still replicated in E.coli, were marginally destabilized in Agrobacterium. Therefore, it was deduced that the stability of a mismatched IR construct may be influenced by the particular intracellular environment of an organism. Due to the recalcitrance of cassava to transformation, a model plant system, Nicotiana benthamiana, was used to screen constructs for toxicity, stability, and efficiency of PTGS induction. Agrobacteriummediated transformation and regeneration of N. benthamiana was optimized, and 86% transformation efficiency was achieved when using leaf disk explants. It was found that the addition of an ethylene scrubber, potassium permanganate, substantially increased the rate of regeneration by reducing the frequency of hyperhydritic plants. Transgene iv integration was confirmed by PCR amplification of the hptII gene in the T-DNA region. Transgene expression was confirmed by screening for GUS and GFP reporter genes. No toxic responses to the transgene have been observed thus far. Studies are currently underway to confirm the stability of the mismatched IR constructs in N. benthamiana. PAGE Northern blotting is being done, as the detection of siRNAs derived from the transgene will confirm that constructs are functional. In addition, infectivity assays are underway to determine the efficacy of BC1 knockdown by a stably integrated construct. Due to the enhanced stability of mismatched IR constructs, they may be an appealing alternative to currently available intron-spliced, or exact matched hairpin systems.

Foram desenvolvidas embalagens biodegradáveis ativas tipo espuma, obtidas a partir de fécula de mandioca, capazes de aumentar a vida-de-prateleira de frutas minimamente processadas. Para tanto, agentes ativos que evitam a proliferação de fungos, a perda de peso e, também, forneçam uma indicação visual (mudança de cor da embalagem) da qualidade do produto embalado, foram adicionados às embalagens de fonte renovável. As espumas foram obtidas pelos processos de extrusão e termoprensagem de uma massa de fécula, gel (fécula suspensa em água) e aditivos (plastificantes e/ou agentes ativos). As formulações de espumas variaram quanto: ao tipo e concentrações de plastificantes (glicerol e polietilenoglicol PEG 300); às quantidades de sólido (fécula de mandioca em pó); aos diferentes tratamentos da fécula (modificações química por acetilação e físicas irradiação e pré-gelatinização); à procedência da fécula (da Bahia e do Paraná); ao reaproveitamento de espumas descartadas; às diferentes concentrações de agentes ativos (ácido l-ascórbico, cloreto de cálcio, sorbato de potássio e permanganato de potássio) e quanto à adição de corantes naturais (extratos de beterraba e repolho roxo). Foram avaliadas, principalmente, as propriedades mecânicas e de barreira das diferentes formulações de espumas de fécula de mandioca. Os resultados indicaram que quanto maior a concentração de fécula mais resistente é a espuma. As espumas contendo o plastificante PEG 300 se mostraram bastante flexíveis, apresentaram boa resistência à ruptura e absorveram pouca água. A pré-gelatinização da fécula proporcionou melhores propriedades mecânicas e de barreira para as espumas em relação aos outros tratamentos dados à fécula de mandioca. Já a adição de agentes ativos à massa não proporcionou tais melhoras às embalagens. As espumas de fécula de mandioca produzidas pelo processo de extrusão seguido de termoprensagem foram mais resistentes à ruptura do que as produzidas somente por termoprensagem. / Active biodegradable packaging foams type were developed, obtained from cassava starch, to increase the shelf-life of minimally processed fruits. For this, active agents that prevent the proliferation of fungi, weight loss and, still, provide a visual indication (change of packaging color) of the packaged product quality, were added to the renewable packaging. The foams were obtained by extrusion and thermopressing processes of a mixture of starch, gel (starch suspended in water) and additives (plasticizers and/or active agents). The foams formulations varied as: the type and concentrations of plasticizers (glycerol and polyethylene glycol PEG 300); the concentrations of solid (cassava starch powder); the different treatments of starch (chemical, for acetylation, and physical modifications radiation and pre-gelatinization); the starch suppliers (from Bahia and Paraná); the reuse of discarded foams; the different active agents (l-ascorbic acid, calcium chloride, potassium sorbate and potassium permanganate) and the addition of natural dyes (beet and cabbage extracts). It was evaluated, mainly, the mechanical and barrier properties of the different formulations of cassava starch foams. The results indicated that the higher starch concentration more resistant become the foam. The foams containing the plasticizer PEG 300 proved to be very flexible, had good tensile strength and absorbed less water. The pre-gelatinization of starch provided better mechanical and barrier properties to foams in comparison with other treatments of cassava starch. The addition of active agents to the mix did not provide such improvements to packaging. The cassava starch foams produced by extrusion followed by thermopressing were more resistant to breakage than those produced only by thermopressing.