Le développement de la baie de raisin est un processus dynamique présentant une courbe de croissance sigmoïde avec deux phases de croissance séparée par une phase de latence. Il se caractérise par une biosynthèse coordonnée de métabolites primaires et secondaires. À la fin de la phase de latence, un phénomène appelé véraison se produit, au cours duquel le fruit commence à prendre de la couleur et le processus de maturation est initié. Les anthocyanes sont responsables de la coloration des baies et la régulation de leur biosynthèse été largement étudiée. Cependant, peu d'études ont porté sur la caractérisation métabolique et génétique de variétés de baies de couleurs fortement contrastées, dans un même fond génétique. En combinant la technologie RNAseq et des analyses métabolomiques, nous avons effectué une caractérisation comparée de deux variétés de raisin de table : Chimenti Globe (CG, rose pâle) et Red Globe (RG, rouge foncé). L'originalité de ce modèle est que CG a été généré à partir d'un événement de mutation spontanée de RG, dans un vignoble de production, permettant ainsi l’étude de la régulation de la biosynthèse des anthocyanes dans un même fond génétique. L'analyse du contenu métabolique des baies a démontré l'importance des stades de développement, de la véraison à la maturation, dans les deux variétés de raisin. En particulier, des différences marquées dans les concentrations de certains métabolites de la voie des phénylpropanoïdes (shikimate, UDP-glucose, phénylalanine) et du tréhalose-6-phosphate ont été mises en évidence en post-véraison. De plus, les différences entre les variétés étaient dues à des changements dans les métabolites liés à la biosynthèse du saccharose et de l'anthocyanine. Les baies de CG ne contenaient que des anthocyanines dihydroxylées (péonidine et cyanidine), et aucune quantité détectable d’anthocyanes trihydroxylées (malvidine, delphinidine et pétunidine), qui sont abondamment présentes dans les pellicules des baies de RG. Ceci explique le phénotype de couleur rose pâle des baies de CG. Une analyse transcriptomique globale par RNAseq montre que 109 gènes sont exprimés de façon différentielle chez CG, en comparaison avec RG, y compris de nombreux gènes liés au métabolisme des flavonoïdes. Notamment, 11 gènes codant pour des 3'5'-hydroxylases flavonoïde, une enzyme-clé pour la biosynthèse des anthocyanes trihydroxylées, sont réprimés dans CG. A partir de cette analyse, un gène candidat pour la régulation de la voie de biosynthèse des anthocyanes a été sélectionné : le gène Cytb5, qui code pour un cytochrome b5, non caractérisé à ce jour chez la Vigne. La surexpression de Cytb5 par transgénèse dans des vignes hybrides V. berlandieri x V. rupestris cv. 110R suggère fortement un rôle clé pour ce gène dans la régulation de la biosynthèse des anthocyanes chez la vigne : les embryons obtenus présentaient une forte coloration rouge, indiquant la présence d’anthocyanes en quantités importantes dans les tissus végétatifs. De plus les embryons transgéniques ont un développement accéléré par rapport aux embryons sauvages. Ces travaux ont permis de mieux comprendre la régulation de l’accumulation des anthocyanes responsables de la coloration des baies de raisin, et plus largement, la régulation du métabolisme des flavonoïdes. / El desarrollo de la uva es un proceso dinámico caracterizado por una curva de crecimiento doble sigmoidea, separada por una fase lag, en donde ocurre una biosíntesis coordinada de metabolitos primarios y secundarios. Al final de la fase lag, ocurre un fenómeno llamado pinta correspondiente al comienzo de la coloración de la baya e iniciándose también el proceso de maduración. Las antocianinas son las responsables de la coloración de la piel de las bayas y su regulación ha sido ampliamente estudiada. Sin embargo, pocos estudios se han enfocado en la caracterización genética y metabólica, utilizando variedades contrastantes de color de piel. Utilizando análisis metabólico y la tecnología de RNA-seq, se realizó una nueva caracterización comparativa de dos uvas de mesa, Chimenti Globe (CG) y Red Globe (RG), que poseen un color de piel de la baya contrastante: CG tiene un color rojizo claro y RG posee un color morado. La originalidad de este modelo es que CG fue generada en un evento espontáneo de campo desde una rama de una planta RG. Por lo tanto, el background genético responsable del cambio de color es el mismo. El análisis del contenido metabólico de las pieles de las bayas reveló la importancia de las etapas de desarrollo, pinta y maduración, en ambas variedades en estudio. En particular, la diferencia en la concentración de metabolitos de la ruta fenilpropanoide, tales como shikimato y fenilalanina y otras moléculas como UDP-glucosa y trehalosa-6-fosfato, entre otros. Asimismo, las diferencias entre las variedades estuvieron dadas por cambios relacionados con la biosíntesis de sacarosa y antocianinas. CG solo contenía antocianinas dihidroxiladas, cianidina y peonidina, y no las del tipo trihidroxiladas, malvidina, delfinidina y petunidina, lo cual fue consistente con el fenotipo del color de piel observado. A partir del análisis transcriptómico, generamos un heatmap con 109 genes expresado diferencialmente en CG en comparación con RG, siendo muchos de estos asociados a la ruta biosíntesis de flavonoides. Además, observamos que 11 copias del gen flavonoide 3'5'-hidroxilasa, que codifica una enzima clave para la biosíntesis de antocianinas trihidroxiladas, no estaban inducidas en CG. A partir de este análisis, se seleccionó un gen candidato para contribuir en el estudio de la ruta de biosíntesis de antocianinas: citocromo b5 (Cytb5) que codifica una proteína clave donadora de electrones no caracterizada en vides. La sobreexpresión de Cytb5 en embriones V. berlandieri x V. rupestris cv. 110R sugirió fuertemente la participación en la ruta, ya que los embriones transgénicos exhibieron un color rojizo e incluso, un desarrollo acelerado en comparación con el control. Con estos resultados, hemos sido capaces de proporcionar información sobre la regulación de las antocianinas en vides responsables de la coloración de la piel de las bayas, abriendo nuevos terrenos en la búsqueda de reguladores moleculares de la vía flavonoide.

A atividade antioxidante (AOX) de vinhos é decorrente, dentre outros fatores, da presença de substâncias fenólicas, como as antocianinas que, além de contribuírem para a formação da cor, devido a sua capacidade redutora, são capazes de neutralizar moléculas de radicais-livres. No presente trabalho, 92 amostras de vinhos tintos da safra 2008/2009, provenientes de diversas regiões do país, agrupadas de acordo com a sua similaridade (UF e fisiografia), foram analisados quanto à cor, através de parâmetros de luz, saturação e tonalidade; à concentração de antocianinas totais (por espectrofotometria diferencial e por CLAE); à caracterização dos pigmentos (por CLAE); à concentração de polifenóis totais (IPT) (pelo método de Folin Ciocalteau’s); e à atividade antioxidante (por DPPH+), sendo que as duas últimas características foram avaliadas sob a forma de evolução temporal. Os resultados mostram que, em relação ao perfil evolutivo, pelo menos um dos grupos difere (p<0,05) nos tempos. A região Vale do Rio São Francisco (VSF) exibiu, tanto os mais elevados valores de IPT em todos os tempos de análise [(tzero=4.516; t25=3.516; t35=2.475; t45=2.027; t55=1.598) mg.L-1 GAE], quanto a mais alta AOX, nos quatro primeiros dos cinco momentos avaliados. A região Oeste do Paraná (OPR) apresentou os mais baixos resultados para ambos os parâmetros considerados em todos os tempos medidos, sendo que os valores de IPT corresponderam a 47, 52, 54, 58 e 63% e os valores de AOX a 49, 55, 57, 61 e 79% dos valores encontrados para VSF. No decorrer das 55 semanas avaliadas, as diferenças entre os grupos tende a diminuir. Em relação aos pigmentos, 12 padrões comerciais permitiram comparar os vinhos no que diz respeito as presença e a quantificação destes compostos. À exceção de dois, todos os vinhos analisados por CLAE confirmaram a malvidina-3-monoglicosídeo (Mv-3G), como o composto antociânico predominante. O Vale dos Vinhedos (Vvi) apresentou os maiores conteúdos, tanto do pigmento majoritário da uva (Mv-3G – 38,66 μ.L-1), quanto da cianidina-3-monoglicosídeo (Cn- 3G – 0,914 μ.L-1), composto antociânico mais relatado em alimentos. Correlações fortes e muito fortes foram observadas entre IPT e AOX e fortes entre antocianinas totais (expressas em Mv-3G por espectrofotometria diferencial) e Mv-3G (quantificada por CLAE). ANOVA mostrou que, em CLAE, não houve diferenças (p<0,1) entre as regiões, indicando que variações internas nos grupos, sobressaíram-se às evidenciadas pelas regiões, não sendo possível caracterizá-las conforme o tipo nem a concentração de antocianinas. / The antioxidant capacity (AOX) of wines is due, among other factors the phenolic substances, such as anthocyanins that, beyond to color contribution, present scavenging properties, having effect on radical oxygen species (ROS). In this study, 92 samples of 2008/2009 vintage Brazilian red wines, from dissimilar production areas, were grouped according to similarities (Federal State and physiographic attributes) and analyzed in terms of color, through light, saturation and hue parameters; in terms of total anthocyanin concentration (by pH-differential spectrophotometric methods and high performance liquid chromatography (HPLC)); in terms of pigment characterization (HPLC); in terms of total polyphenols concentration (IPT) (Folin Ciocalteau’s method); and in terms of its scavenging properties (DPPH+), being the last two attributes analyzed over time evolution profile. The results showed that for the evolution profile, at least one of the groups differed (p<0,05) in the evaluated times. The “Vale do Rio São Francisco” (VSF) region exhibited not only the most elevated values for IPT in all analyzed sampling [(tzero=4,516; t25=3,516; t35=2,475; t45=2,027; t55=1,598) mg.L-1 GAE], as the most AOX in the first four from the five evaluated moments. The “Oeste do Paraná” (OPR) presented the lowest results for both IPT and AOX parameters, in all measuring times, and the IPT values corresponded to 47, 52, 54, 58 e 63%, while the AOX values to 49, 55, 57, 61 e 79% from values found for VSF. Over the 55 weeks, the differences among groups tended to diminish. Considering the pigments, 12 commercial standards allowed to compare wines based on presence and concentration of these compounds. Except for two samples, all the HPLC-analyzed wines confirmed the malvidin-3-monoglucoside, as the anthocyanin most abounding in wines. The VVi exhibited the highest levels, even for the grape major pigment (Mv- 3G – 38.66 μ.L-1), as for cianidin-3-monoglucoside (Cn-3G – 0.914 μ.L-1), the most reported anthocyanin in food. Strong and very strong correlations were found among IPT and AOX, while total anthocyanins (in Mv-3G base, by pH-differential analysis) and Mv-3G concentration (by HPLC) exhibited strong linear associations. ANOVA showed that, in terms of chromatographic analysis, no differences (p<0,1) among regions were found, indicating that inside-groups variations were high enough to not allow the characterization according to qualitative and quantitative anthocyanin profile.

A atividade antioxidante (AOX) de vinhos é decorrente, dentre outros fatores, da presença de substâncias fenólicas, como as antocianinas que, além de contribuírem para a formação da cor, devido a sua capacidade redutora, são capazes de neutralizar moléculas de radicais-livres. No presente trabalho, 92 amostras de vinhos tintos da safra 2008/2009, provenientes de diversas regiões do país, agrupadas de acordo com a sua similaridade (UF e fisiografia), foram analisados quanto à cor, através de parâmetros de luz, saturação e tonalidade; à concentração de antocianinas totais (por espectrofotometria diferencial e por CLAE); à caracterização dos pigmentos (por CLAE); à concentração de polifenóis totais (IPT) (pelo método de Folin Ciocalteau’s); e à atividade antioxidante (por DPPH+), sendo que as duas últimas características foram avaliadas sob a forma de evolução temporal. Os resultados mostram que, em relação ao perfil evolutivo, pelo menos um dos grupos difere (p<0,05) nos tempos. A região Vale do Rio São Francisco (VSF) exibiu, tanto os mais elevados valores de IPT em todos os tempos de análise [(tzero=4.516; t25=3.516; t35=2.475; t45=2.027; t55=1.598) mg.L-1 GAE], quanto a mais alta AOX, nos quatro primeiros dos cinco momentos avaliados. A região Oeste do Paraná (OPR) apresentou os mais baixos resultados para ambos os parâmetros considerados em todos os tempos medidos, sendo que os valores de IPT corresponderam a 47, 52, 54, 58 e 63% e os valores de AOX a 49, 55, 57, 61 e 79% dos valores encontrados para VSF. No decorrer das 55 semanas avaliadas, as diferenças entre os grupos tende a diminuir. Em relação aos pigmentos, 12 padrões comerciais permitiram comparar os vinhos no que diz respeito as presença e a quantificação destes compostos. À exceção de dois, todos os vinhos analisados por CLAE confirmaram a malvidina-3-monoglicosídeo (Mv-3G), como o composto antociânico predominante. O Vale dos Vinhedos (Vvi) apresentou os maiores conteúdos, tanto do pigmento majoritário da uva (Mv-3G – 38,66 μ.L-1), quanto da cianidina-3-monoglicosídeo (Cn- 3G – 0,914 μ.L-1), composto antociânico mais relatado em alimentos. Correlações fortes e muito fortes foram observadas entre IPT e AOX e fortes entre antocianinas totais (expressas em Mv-3G por espectrofotometria diferencial) e Mv-3G (quantificada por CLAE). ANOVA mostrou que, em CLAE, não houve diferenças (p<0,1) entre as regiões, indicando que variações internas nos grupos, sobressaíram-se às evidenciadas pelas regiões, não sendo possível caracterizá-las conforme o tipo nem a concentração de antocianinas. / The antioxidant capacity (AOX) of wines is due, among other factors the phenolic substances, such as anthocyanins that, beyond to color contribution, present scavenging properties, having effect on radical oxygen species (ROS). In this study, 92 samples of 2008/2009 vintage Brazilian red wines, from dissimilar production areas, were grouped according to similarities (Federal State and physiographic attributes) and analyzed in terms of color, through light, saturation and hue parameters; in terms of total anthocyanin concentration (by pH-differential spectrophotometric methods and high performance liquid chromatography (HPLC)); in terms of pigment characterization (HPLC); in terms of total polyphenols concentration (IPT) (Folin Ciocalteau’s method); and in terms of its scavenging properties (DPPH+), being the last two attributes analyzed over time evolution profile. The results showed that for the evolution profile, at least one of the groups differed (p<0,05) in the evaluated times. The “Vale do Rio São Francisco” (VSF) region exhibited not only the most elevated values for IPT in all analyzed sampling [(tzero=4,516; t25=3,516; t35=2,475; t45=2,027; t55=1,598) mg.L-1 GAE], as the most AOX in the first four from the five evaluated moments. The “Oeste do Paraná” (OPR) presented the lowest results for both IPT and AOX parameters, in all measuring times, and the IPT values corresponded to 47, 52, 54, 58 e 63%, while the AOX values to 49, 55, 57, 61 e 79% from values found for VSF. Over the 55 weeks, the differences among groups tended to diminish. Considering the pigments, 12 commercial standards allowed to compare wines based on presence and concentration of these compounds. Except for two samples, all the HPLC-analyzed wines confirmed the malvidin-3-monoglucoside, as the anthocyanin most abounding in wines. The VVi exhibited the highest levels, even for the grape major pigment (Mv- 3G – 38.66 μ.L-1), as for cianidin-3-monoglucoside (Cn-3G – 0.914 μ.L-1), the most reported anthocyanin in food. Strong and very strong correlations were found among IPT and AOX, while total anthocyanins (in Mv-3G base, by pH-differential analysis) and Mv-3G concentration (by HPLC) exhibited strong linear associations. ANOVA showed that, in terms of chromatographic analysis, no differences (p<0,1) among regions were found, indicating that inside-groups variations were high enough to not allow the characterization according to qualitative and quantitative anthocyanin profile.

A atividade antioxidante (AOX) de vinhos é decorrente, dentre outros fatores, da presença de substâncias fenólicas, como as antocianinas que, além de contribuírem para a formação da cor, devido a sua capacidade redutora, são capazes de neutralizar moléculas de radicais-livres. No presente trabalho, 92 amostras de vinhos tintos da safra 2008/2009, provenientes de diversas regiões do país, agrupadas de acordo com a sua similaridade (UF e fisiografia), foram analisados quanto à cor, através de parâmetros de luz, saturação e tonalidade; à concentração de antocianinas totais (por espectrofotometria diferencial e por CLAE); à caracterização dos pigmentos (por CLAE); à concentração de polifenóis totais (IPT) (pelo método de Folin Ciocalteau’s); e à atividade antioxidante (por DPPH+), sendo que as duas últimas características foram avaliadas sob a forma de evolução temporal. Os resultados mostram que, em relação ao perfil evolutivo, pelo menos um dos grupos difere (p<0,05) nos tempos. A região Vale do Rio São Francisco (VSF) exibiu, tanto os mais elevados valores de IPT em todos os tempos de análise [(tzero=4.516; t25=3.516; t35=2.475; t45=2.027; t55=1.598) mg.L-1 GAE], quanto a mais alta AOX, nos quatro primeiros dos cinco momentos avaliados. A região Oeste do Paraná (OPR) apresentou os mais baixos resultados para ambos os parâmetros considerados em todos os tempos medidos, sendo que os valores de IPT corresponderam a 47, 52, 54, 58 e 63% e os valores de AOX a 49, 55, 57, 61 e 79% dos valores encontrados para VSF. No decorrer das 55 semanas avaliadas, as diferenças entre os grupos tende a diminuir. Em relação aos pigmentos, 12 padrões comerciais permitiram comparar os vinhos no que diz respeito as presença e a quantificação destes compostos. À exceção de dois, todos os vinhos analisados por CLAE confirmaram a malvidina-3-monoglicosídeo (Mv-3G), como o composto antociânico predominante. O Vale dos Vinhedos (Vvi) apresentou os maiores conteúdos, tanto do pigmento majoritário da uva (Mv-3G – 38,66 μ.L-1), quanto da cianidina-3-monoglicosídeo (Cn- 3G – 0,914 μ.L-1), composto antociânico mais relatado em alimentos. Correlações fortes e muito fortes foram observadas entre IPT e AOX e fortes entre antocianinas totais (expressas em Mv-3G por espectrofotometria diferencial) e Mv-3G (quantificada por CLAE). ANOVA mostrou que, em CLAE, não houve diferenças (p<0,1) entre as regiões, indicando que variações internas nos grupos, sobressaíram-se às evidenciadas pelas regiões, não sendo possível caracterizá-las conforme o tipo nem a concentração de antocianinas. / The antioxidant capacity (AOX) of wines is due, among other factors the phenolic substances, such as anthocyanins that, beyond to color contribution, present scavenging properties, having effect on radical oxygen species (ROS). In this study, 92 samples of 2008/2009 vintage Brazilian red wines, from dissimilar production areas, were grouped according to similarities (Federal State and physiographic attributes) and analyzed in terms of color, through light, saturation and hue parameters; in terms of total anthocyanin concentration (by pH-differential spectrophotometric methods and high performance liquid chromatography (HPLC)); in terms of pigment characterization (HPLC); in terms of total polyphenols concentration (IPT) (Folin Ciocalteau’s method); and in terms of its scavenging properties (DPPH+), being the last two attributes analyzed over time evolution profile. The results showed that for the evolution profile, at least one of the groups differed (p<0,05) in the evaluated times. The “Vale do Rio São Francisco” (VSF) region exhibited not only the most elevated values for IPT in all analyzed sampling [(tzero=4,516; t25=3,516; t35=2,475; t45=2,027; t55=1,598) mg.L-1 GAE], as the most AOX in the first four from the five evaluated moments. The “Oeste do Paraná” (OPR) presented the lowest results for both IPT and AOX parameters, in all measuring times, and the IPT values corresponded to 47, 52, 54, 58 e 63%, while the AOX values to 49, 55, 57, 61 e 79% from values found for VSF. Over the 55 weeks, the differences among groups tended to diminish. Considering the pigments, 12 commercial standards allowed to compare wines based on presence and concentration of these compounds. Except for two samples, all the HPLC-analyzed wines confirmed the malvidin-3-monoglucoside, as the anthocyanin most abounding in wines. The VVi exhibited the highest levels, even for the grape major pigment (Mv- 3G – 38.66 μ.L-1), as for cianidin-3-monoglucoside (Cn-3G – 0.914 μ.L-1), the most reported anthocyanin in food. Strong and very strong correlations were found among IPT and AOX, while total anthocyanins (in Mv-3G base, by pH-differential analysis) and Mv-3G concentration (by HPLC) exhibited strong linear associations. ANOVA showed that, in terms of chromatographic analysis, no differences (p<0,1) among regions were found, indicating that inside-groups variations were high enough to not allow the characterization according to qualitative and quantitative anthocyanin profile.

Submitted by Gabriela Lopes (gmachadolopesufpel@gmail.com) on 2016-10-04T17:45:24Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese.Maria Inês Machado.pdf: 1755709 bytes, checksum: e4832900f39993c29a1573433a9d0c93 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2016-10-11T20:26:22Z (GMT) No. of bitstreams: 2 Tese.Maria Inês Machado.pdf: 1755709 bytes, checksum: e4832900f39993c29a1573433a9d0c93 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-10-11T20:26:22Z (GMT). No. of bitstreams: 2 Tese.Maria Inês Machado.pdf: 1755709 bytes, checksum: e4832900f39993c29a1573433a9d0c93 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2014-07-16 / Sem bolsa / O mercado brasileiro para produtos de frutas de clima temperado é bastante promissor, porém existem desuniformidades na qualidade de frutas, falhas no processamento, que favorecem o desperdício tanto no campo quanto na indústria, gerando a necessidade na correção de lotes defeituosos. Assim como outros frutos, o pêssego apresenta compostos bioativos, mas que ainda pouco têm se estudado, principalmente a estabilidade destes compostos frente ao processamento. Assim, o objetivo deste estudo foi avaliar o impacto do processamento de pêssego sobre o conteúdo de compostos bioativos em pêssego em calda e pessegada e avaliar a estabilidade destes compostos durante o período de estocagem destes produtos. Para isto foram utilizadas três cultivares de pêssegos (Prunus pérsica): Santa Áurea, Esmeralda e Maciel, frutos provenientes de cinco produtores da região de Pelotas/RS, e cultivados sob sistema de rastreabilidade da fruta, sendo avaliados durante duas safras consecutivas. Os pêssegos foram processados na forma de doce em calda e de pessegada, estocados e posteriormente avaliados. Foram realizadas as análises de pH, acidez titulável (AT), sólidos solúveis (SS), relação SS/AT, total de carotenoides, carotenoides individuais, total de compostos fenólicos, compostos fenólicos individuais, vitamina C e total de antocianina s. A cultivar Esmeralda destacou-se entre as demais estudadas demonstrando relação SS/AT:20; representando excelente qualidade para processamento, seguido da cultivar Santa Áurea e Maciel. Ocorreram perdas significativas de compostos bioativos durante a elaboração e estocagem de pêssego em calda, sendo a vitamina C (40% perdas) e os carotenoides (77%perdas) os componentes mais instáveis. Após elaboração de pessegada não foram verificadas perdas significativas de compostos fenólicos sendo intensificadas durante o período de estocagem, atingindo valores acima de 57% de perdas. A escolha de cultivares para elaboração de pêssego em calda e doce em massa apresentou valorização das características sensoriais, físico-químicas e bioativas. / The Brazilian market for products of temperate climate fruits is very promising, but there is no uniformity on fruit quality, processing failures that favor waste in the field and in the industry, generating correction of defective lots. As other fruits, peach presents bioactive compounds, but so far still not studied, mainly the stability of these compounds against processing. The objective of this study was to evaluate the impact of peach processing on the content of bioactive compounds in peach compotes and in sweet peach mass, and evaluate the stability of these compounds during the storage of these products. For this, three cultivars of peach (Prunus persica L.) were used: Santa Áurea, Esmeralda and Maciel, fruits were obtained from five producers of Pelotas / RS, and grown under fruit traceability system, being evaluated during two consecutive harvests. The peaches were processed as sweet peach mass and peach compote, stored and later evaluated. Analyzes of pH, titratable acidity (TA), soluble solids (SS), SS / TA, total carotenoids, individual carotenoids, total phenolic compounds, individual phenolic compounds, vitamin C and total anthocyanin were analysed. The Esmeralda cultivar stood out among the others showing SS / TA ratio: 20; representing excellent quality for processing, followed by cv. Santa Aurea and Maciel. Significant losses of bioactive compounds during the preparation and storage of peaches compotes were observed, with vitamin C (40% loss) and carotenoids (77% loss) the most unstable components. After sweet peach mass processing, no significant losses of phenolic compounds was observed, but the losses were intensified during storage period, where values above 57% losses were observed. The choice of cultivars for preparation of peaches compotes and sweet peach mass, enhanced the sensory, physicochemical and bioactive characteristics.

The diploma thesis deals with the preparation of extract and concentrate of anthocyanin dyes from aronia moldings (Aronia melanocarpa). It also determines chemical properties important for human´s health, which are important also for further possible use of extract/concentrate as natural red food coloring. A series of extractions of dried and powdered aronia moldings was carried out in the experimental part of the thesis, aiming for the optimization the conditions of the extraction. Extracts were characterized by content of total anthocyans, content of total fenolic substances and total antioxidant activity. The best laboratory conditions for extraction were determined as follows: amount of moldings 12,5 g, composition of extraction agent ethanol:water 1:1, volume of extraction mixture 60 ml and time of the extraction 11 hours. Prepared extracts were concentrated on vacuum evaporator. It has been found that the temperature of the water bath in the vacuum evaporator can not exceed 40 °C. At higher temperatures there is significant degradation of anthocyanins. Furthemore, the content of individual anthocyanic dyes in extract and concentrate, from aronia moldings, was quantified by HPLC. The major anthocyanin, cyanidine-3-galactoside, was contained in the concentrate at 923,6 ± 8,5 mg·l-1, the other specified anthocyanin, minor cyanidine-3-glucoside, was present at a concentration of 53,5 ± 0,3 mg·l-1.

Thesis (MScAgric (Viticulture and Oenology))--University of Stellenbosach, 2010. / ENGLISH ABSTRACT: Cabernet Sauvignon, the most planted red wine cultivar in South Africa, is prone to vigorous growth with low yields. The aim of the study was to describe how Cabernet Sauvignon grapevines react to climate and irrigation within the Swartland region. Such knowledge would assist growers in decisions regarding long term as well as short term cultivation practices. This study was part of a larger project carried out by the Infruitec-Nietvoorbij institute of the Agricultural Research Council at Stellenbosch to determine effects of soil type and climate on yield and wine quality of Cabernet Sauvignon. The larger project was carried out in selected grape growing regions, i.e. Stellenbosch, Swartland, Lower Olifants River and Lower Orange River. Due to the proximity to the Atlantic Ocean, the study area in the Swartland region could be divided into two climatic regions for viticulture. Grapevines near Philadelphia closer to the ocean experienced less water constraints compared to those further inland near Wellington. Variation in stem water potential could also be related to soil water matric potential. Climate tended to have a more pronounced effect on the grapevine response to water constraints further inland than closer to the ocean. Vegetative growth, berry size and yield depended on water constraints experienced by the grapevines. In the warmer climate, severe constraints reduced yield. In the warmer climate, grapes started to ripen earlier than those in the cooler climate. Sugar concentration (mg/mL) was highest where grapevines experienced moderate water constraints. These seemingly balanced grapevines had the highest sugar accumulation, probably due to optimum photosynthesis and carbohydrate utilization. Low water constraints increased vegetative growth which could have been a sink for sugar loading. In addition to sugar loading, degree Balling (°B) increases could also have been due to a concentration effect where water constraints reduced berry volume. Therefore, °B is probably not a representative indicator of grapevine functioning. Anthocyanin biosynthesis, as quantified on a per berry basis, showed that sugar and anthocyanin could be co-regulated, with anthocyanin biosynthesis reaching a plateau when the sugar content per berry reached 200 mg/mL to 220 mg/mL. At véraison, the most intense grape colour occurred where grapevines experienced moderate water constraints, i.e. single drip line at Wellington and no irrigation at Philadelphia. However, at harvest grapes from the cooler climate tended to have more intense colour and higher phenolics, indicating that lower temperatures favoured anthocyanin biosynthesis. These results supported earlier findings that grapevine water status influences berry volume and dynamics of berry ripening. Water constraints tended to increase sensorial wine colour intensity, as well as wine fullness. Moderate water constraints at both localities resulted in the best sensorial wine quality. Yet there were indications that too severe water constraints could be detrimental to wine quality. Irrigation can be used to manipulate grapevine growth in warmer climates, but might be less effective in cooler climates. In warmer climates, moderate water constraints required to achieve balanced grapevine functioning can be obtained with single drip irrigation, but this might not be the case in cooler climates. / AFRIKAANSE OPSOMMING: Cabernet Sauvignon, die mees aangeplante rooiwynkultivar in Suid-Afrika, is geneig tot kragtige groei met lae opbrengste. Die doelwit van hierdie studie was om te beskryf hoe Cabernet Sauvignon-wingerdstokke reageer op klimaat en besproeiing in die Swartland-streek. Hierdie kennis sal kwekers help wanneer hulle besluite moet neem oor langtermyn sowel as korttermyn verbouingspraktyke. Hierdie studie was deel van ‘n groter projek deur die Infruitec-Nietvoorbij Instituut van die Landbounavorsingsraad op Stellenbosch om die effekte van grondtipe en klimaat op die opbrengs en wynkwaliteit van Cabernet Sauvignon te bepaal. Die groter projek is in geselekteerde wingerdverbouingstreke uitgevoer, nl. Stellenbosch, Swartland, Benede Olifantsrivier en Benede Oranjerivier. As gevolg van die nabyheid daarvan aan die Atlantiese Oseaan kon die studiegebied in die Swartland-streek in twee klimaatstreke vir wingerdbou verdeel word. Wingerdstokke by Philadelphia, wat nader aan die oseaan is, het minder waterstremming ervaar as dié verder binnelands naby Wellington. Veranderinge in stamwaterpotensiaal hou moontlik ook verband met die grondwater- matrikspotensiaal. Klimaat het ‘n groter effek op die reaksie van die wingerdstok op waterstremming verder binnelands as nader aan die oseaan. Vegetatiewe groei, korrelgrootte en opbrengs was afhanklik van die waterstremminge wat deur die wingerdstokke ervaar is. In die warmer klimaat het die ernstige stremminge opbrengs verminder. In die warmer klimaat begin druiwe vroeër ryp word as in die koeler klimaat. Suikerkonsentrasie (mg/ml) was die hoogste waar wingerde matige waterstremming ervaar het. Hierdie skynbaar gebalanseerde wingerdstokke het die hoogste suikerakkumulasie vertoon, moontlik as gevolg van optimum fotosintese en koolhidraatverbruik. Lae waterstremming het vegetatiewe groei verhoog, wat ook ‘n vraagpunt vir suikerlading kon wees. Benewens suikerlading kon verhogings in grade Balling (°B) ook moontlik aan ‘n konsentrasie-effek toegeskryf word in terme waarvan waterstremming die korrelvolume verminder het. °B is dus moontlik nie ‘n verteenwoordigende indikator van wingerdstokfunksionering nie. Antosianienbiosintese, soos gekwantifiseer op ‘n per-korrel basis, het getoon dat suiker en antosianien saam gereguleer kon word, en dat antosianienbiosintese ‘n plato bereik het wanneer die suikerinhoud per korrel 200 mg/mL tot 220 mg/mL was. By deurslaan het die mees intense druifkleur voorgekom waar die wingerdstokke matige waterstremming ervaar het, d.w.s. enkel druplyn op Wellington en geen besproeiing op Philadelphia. Teen oes was die druiwe in die koeler klimate egter geneig om meer intense kleur en meer fenole te bevat, wat aandui dat laer temperature antosianienbiosintese bevoordeel. Hierdie resultate ondersteun vroeër bevindings dat die waterstatus van die wingerdstok ‘n invloed op korrelvolume en die dinamika van korrelrypwording het. Waterstremming neig om die sensoriese wynkleurintensiteit te verhoog, asook die volheid van die wyn. Matige waterstremming op beide liggings het aanleiding gegee tot die beste sensoriese wynkwaliteit. Tog was daar aanduidings dat waterstremming wat te straf was, nadelig kon wees vir wynkwaliteit. Besproeiing kan gebruik word om wingerdgroei in warmer klimate te manipuleer, maar is moontlik minder effektief in koeler klimate. In warmer klimate kan die matige waterstremming wat benodig is vir gebalanseerde wingerdstokfunksionering, verkry word deur enkel drupbesproeiing, maar dit is moontlik nie die geval in koeler klimate nie.

Muitas respostas moduladas pela luz durante o desenvolvimento das plantas também são reguladas por hormônios vegetais, levando à hipótese da interação entre ambos os fatores. Uma ferramenta valiosa para testar tal interação seria o uso de mutantes fotomorfogenéticos e hormonais, bem como duplos mutantes combinando ambos. Em tomateiro, embora sejam disponíveis mutantes com alterações na biossíntese de fotorreceptores e/ou na transdução do sinal da luz, bem como mutantes no metabolismo e/ou sensibilidade hormonal, esses estão presentes em cultivares diferentes, o que pode limitar seu uso de modo integrado e a construção de duplos mutantes. No presente trabalho, foram introgredidas em uma única cultivar de tomateiro, Micro-Tom (cv. MT), dezenove mutações afetando a biossíntese ou a resposta a fitocromo, bem como aos hormônios auxina (AUX), citocinina (CK), giberelina (GA), ácido abscísico (ABA), etileno (ET) e brassinoesteróides (BR). Tomando-se vantagem de tal coleção, duas respostas notadamente controladas tanto pela luz quanto por hormônios foram estudadas: alongamento e acúmulo de antocianinas em hipocótilos. Para tal, foram utilizadas as seguintes abordagens: i) tratamentos exógenos de diferentes classes hormonais em mutantes fotomorfogenéticos, ii) observação de hipocótilos de mutantes hormonais crescidos na luz e no escuro, iii) observação de duplos mutantes combinando mutações hormonais e fotomorfogenéticas. Assim, o acúmulo de antocianinas foi promovido pela CK e ABA e inibido pela GA, concordando com a redução no mutante deficiente em ABA (notabilis ou not) e no mutante hipersensível à GA (procera ou pro). Apesar do mutante com baixa sensibilidade à AUX (diageotropica ou dgt) acumular exageradamente antocianinas, a aplicação exógena não evidenciou o papel da AUX, sendo, porém, coerente com a sugestão de que esse mutante possui um balanço AUX/CK voltado para CK. Tanto a aplicação exógena quanto a avaliação nos mutantes epinastic (epi), super produção de ET, e Never ripe (Nr), baixa sensibilidade ao ET, sugerem uma função limitada desse hormônio na biossíntese de antocianinas. Na luz e no escuro, AUX, CK, ABA e ET exógenos resultaram na inibição do alongamento do hipocótilo, sendo coerente com a promoção em dgt (luz), promoção em sit (luz), inibição em epi (luz e escuro). Por outro lado, GA promoveu o alongamento corroborando a promoção em pro. Contrariando o efeito exógeno da CK, brt reduziu o alongamento na luz e no escuro. No escuro, o único mutante que apresentou alongamento do hipocótilo superior a MT foi o mutante deficiente na biossíntese do phy (aurea ou au). A utilização de duplos mutantes combinando phy- e alterações hormonais mostrou uma interação aditiva (au epi, au Nr, au dgt e au sit), sinergística (au pro) e epinástica (au brt) no acúmulo de antocianinas e alongamento do hipocótilo na luz, porém nessa última resposta, au dgt e au sit indicaram uma interação sinergística. Juntos, esses resultados indicam que, embora phy possui vias distintas da AUX, ET, ABA e GA no controle do acúmulo de antocianinas e alongamento do hipocótilo, parece que esse fotorreceptor partilha vias comuns com CK em ambas as respostas. / Many responses regulated by light during plant development are also regulated by plant hormones, suggesting an interaction between these factors. One important approach to test this hypothesis is the use of photomorphogenic and hormonal mutants and double mutant analysis. Mutants with altered photoreceptor biosynthesis, light signal transduction, hormonal metabolism and hormonal sensitivity are available in tomato. However, since they are in different cultivars, this can be a limitation for their use in a comprehensive study, as well as, for the construction of double mutants. In this work we performed the introgression of nineteen mutations in a single cultivar of tomato, Micro- Tom (cv. MT). These mutations affect biosynthesis or response to phytochrome (phy), auxin (AUX), cytokinin (CK), gibberellin (GA), abscisic acid (ABA), ethylene (ET) and brassinosteroid (BR). Using this collection of hormone mutants, we studied two responses which are controlled by light and hormones: elongation and anthocyanin accumulation in hypocotyls. For this purpose, we used three approaches: i) hormonal treatment in the photomorphogenic mutants, ii) measurement of hypocotyl lengths from hormonal mutants grown under light and dark conditions and iii) double mutant (photomorphogenic-hormonal) analysis. Anthocyanin accumulation was promoted by CK and ABA and inhibited by GA. This is in accordance with the reduction of anthocyanin accumulation in the ABA deficient mutant (not) and in the GA hypersensitive mutant (pro). Although the diageotropica (dgt), auxin-insensitive mutant, showed a high anthocyanin accumulation, the addition of auxin did not supported a role for this hormone in anthocyanin accumulation. On the other hand, this could be due to a low auxin-tocytokinin ratio presented by dgt. Data from mutants with altered metabolism and sensitivity of ethylene, epinastic (epi) and Never ripe (Nr) respectively, and from plants treated with this hormone suggest a limited role of ethylene in the anthocyanin biosynthesis. Exogenous AUX, CK, ABA and ET inhibited the hypocotyl elongation. This is coherent with the promotion of hypocotyl elongation in dgt and sit mutants under light conditions and inhibition of hypocotyl elongation in the epi mutant in the light and dark. On the other hand, GA promoted the hypocotyl elongation corroborating the same effect seen in pro. The brt mutant showed a reduced hypocotyl elongation in light and dark conditions, which contradicts the effect of exogenous cytokinin. The phytochromedeficient aurea (au) mutant was the only one to show an enhanced hypocotyl elongation in the dark compared to the wild type (MT). The combination between photomorphogenic and hormonal mutants (double mutants) showed additive (au epi, au Nr, au dgt e au sit), synergistic (au pro) and epistatic (au brt) interactions considering the anthocyanin accumulation and hypocotyl elongation. Synergistic interaction was observed in the elongation hypocotyl of the au dgt and au sit double mutants. These results indicate that phy and CK may share some signaling/metabolic pathways in the control of anthocyanin accumulation and hypocotyl elongation. On the other hand, our data do not support an interaction between phy and the hormones AUX, ET, ABA and GA in the control of hypocotyls elongation or anthocyanin accumulation.

The effects of varying levels of sulphur dioxide (SO₂) on the cold maceration process was investigated with Pinot noir (Vitis vinifera L.) wine. The effects of these varying levels on the wines composition and colour parameters were examined. Cold maceration is a technique whereby grapes are crushed and placed at low temperatures (4 - lO°C) in the presence 50 - 150 mgL⁻¹ SO₂. This process is believed to provide a medium for the extraction of water soluble phenolic compounds, rather than the alcoholic extraction employed in normal fermentations. The extraction of these phenolic compounds was monitored from the juice through to six months of bottle age. The changes were measured using both Spectrophotometric and High Performance Liquid Chromatographic (HPLC) procedures. Cold maceration wines were found to be not significantly different to the control wine in all compositional parameters other than titrateable acidity which was found to be less than the control for all the cold maceration wines. The unsulphured cold maceration wine was not significantly different from the control wine in any of the spectral measurements except natural degree of ionisation, in which it was higher, and total phenolics, in which it was lower. These results indicate that the cold maceration process alone does not alter the extraction of phenolic compounds. The HPLC analysis of the wine confirmed the spectral results indicating that their were no significant differences in the levels of extraction of anthocyanins. The sulphured cold maceration wines were significantly greater than the control in visible colour, colour density, total anthocyanins, natural degree of ionisation, ionised anthocyanins and total phenolics. These results followed similar patterns with wine ageing, at six months these wines were still significantly greater in all the measurements apart from natural degree of ionisation. The results for the sulphured cold maceration wines indicates that SO₂ is acting as a solvent for the extraction of phenolic compounds including anthocyanins. The 50 mgL⁻¹ SO₂ cold maceration wine had similar colour and phenolic content to the 100 mgL⁻¹ SO₂ cold maceration wine at bottling, at six months the 50 mgL⁻¹ SO₂ cold maceration wine still retained a similar colour to the 100 mgL⁻¹ SO₂ cold maceration wine but had vastly reduced anthocyanin content. This indicates that for the grapes utilised in this study the most appropriate level of addition at cold maceration would be 50 mgL⁻¹ of SO₂. With grapes of differing phenolic content the level of addition required will vary.

Two pre-fermentation treatments were investigated in Pinot noir (Vitis vinifera L.) wines. The effects of cold maceration and carbonic maceration on the wines' composition, colour parameters and sensory properties were examined. Cold maceration is a winemaking technique used to increase non-alcoholic extraction in Pinot noir winemaking prior to fermentation. It involves holding crushed grapes with approximately 100-150 mg l⁻¹ SO₂ at low temperatures and is thought to increase the colour, aroma and flavour of the resulting wines. Carbonic maceration uses whole bunches that have undergone anaerobic metabolism to produce characteristically fruity and spicy wines. Pre-fermentation cold maceration produces wines that are higher in titratable acidity and monomeric anthocyanin content, but lower in colour density, hue and polymeric pigments. Reducing the maceration temperature below 10°C has little effect. Carbonic maceration produces wines that are lower in titratable acidity, monomeric anthocyanin content, and colour density but are higher in colour hue and amount of polymeric pigments. Quantitative descriptive analysis was used to define the effects of these pre-fermentation maceration treatments on the sensory characteristics of the resulting wine. Trained panel members found that there were no discernable sensory differences in the compositional parameters despite measurable chemical differences. Investigation into the aroma and flavour characteristics of the wines found that carbonic maceration produces wines that were lower in berry aroma and higher in acetate or ester-type aromas than the control wines. These wines were considered to have specific raspberry, floral, sugar, cherry and chemical aromas. This chemical note was also observed in the flavour of the carbonic maceration wines. The temperature of the cold maceration process has no major effect on the aroma and flavour of the resulting wines. However, the 10°C maceration was higher in woody/tobacco aroma than the 4°C maceration, and the 10°C treatment was also higher in bitter flavour than all the other treatments. Cold maceration wines were found to have specific mixed berry, dried fruit and sweet-oxidised aroma characters, together with a blackberry flavour note.