Le raisin (Vitis vinifera L.) est un fruit charnu non climactérique d’importance économique majeure. De nombreux paramètres, comme la taille, le métabolisme des acides organiques et des sucres, l’équilibre hydrique et la synthèse de métabolites secondaires, affectent la « qualité » de la baie. Les signaux affectant la reprogrammation du métabolisme cellulaire et l’expression des gènes codant des protéines clefs du développement et du mûrissement incluent les hormones et l’accumulation des sucres, présents à des taux variables au cours du développement de la baie. La régulation transcriptionnelle est l’élément essentiel qui contrôle ces modifications majeures. A ce jour, dans le contexte de la régulation du développement du raisin, très peu d’acteurs moléculaires ont pu être clairement caractérisés. Les travaux réalisés dans le cadre de cette thèse ont donc eu pour objectif de mieux comprendre cette régulation par l’identification et la caractérisation de facteurs de transcription régulés transcriptionnellement par le saccharose et exprimés au cours du développement de la baie de raisin. Parmi les cinq facteurs de transcription identifiés, trois n’ont pu être étudiés que de manière préliminaire et deux, VvCEB1 et VvABF2, ont fait l’objet d’une caractérisation plus approfondie. VvCEB1 (Vitis vinifera Cell Elongation bHLH) est exprimé préférentiellement dans la baie de raisin au cours de la phase de maturation. Une approche par transgénèse chez la Vigne et la Tomate montre que la surexpression de ce facteur de transcription affecte à la fois le développement des embryons de Vigne et des fruits de Tomate, en modifiant notamment le nombre et la taille des cellules qui les constituent. Une analyse transcriptionnelle réalisée à partir de ce matériel biologique confirme le rôle de VvCEB1 dans la stimulation de l’expansion cellulaire et suggère l’implication de l’auxine. VvABF2 est régulé transcriptionnellement par l’ABA, une hormone clef impliquée dans les processus de maturation des fruits non climactériques. Les transcrits VvABF2 s’accumulent majoritairement pendant l’étape de maturation. Une approche transcriptomique sur génome complet de Vigne et réalisée à partir de suspensions cellulaires embryogènes surexprimant VvABF2, traitées ou non par l’ABA souligne la stimulation de l’expression des gènes codant les enzymes liées à la maturation du raisin. En effet, des gènes des voies de biosynthèse des composés phénoliques et du ramollissement sont affectés. Enfin, l’analyse phénotypique de plants de Tomate surexprimant VvABF2 renforce l’hypothèse liant ce facteur de transcription au ramollissement : à partir du stade "turning" un ramollissement accéléré du fruit est observé.Prise dans son ensemble cette thèse contribue à une meilleure compréhension des mécanismes de signalisation moléculaires impliqués dans le développement d’un fruit charnu non climactérique. / Grapevine (Vitis vinifera L.) is the most economically important fruit crop in the world. The quality of this non climacteric fruit depends on numerous parameters such as size, organic acids and sugar metabolisms, water balance, and secondary metabolism. During fruit development and ripening, hormones and sugars can act as signals affecting cellular metabolism and expression of key regulatory genes. Up to now, little is known about the gene networks involved in these processes and therefore very few of these regulatory genes have been characterized.The aim of this PhD work was to get a better insight in the mechanisms involved in grape berry development through the identification and characterization of sugar-dependent transcription factors (TFs) that were expressed during fruit development. We identified five TFs, and fully characterized two of them: VvCEB1 and VvABF2.VvCEB1 (Vitis vinifera Cell Elongation bHLH) is preferentially expressed in the berry at the onset of ripening and during the ripening stage. Overexpression of VvCEB1 in grapevine and tomato affected both grape embryo development and plant organ size by modifying their cell number and cell size. A transcriptional analysis performed on these transgenic lines confirmed that VvCEB1 stimulates cell expansion and suggests the involvement of the phytohormone auxin.VvABF2 is transcriptionally regulated by abscisic acid (ABA), a phytohormone known to play a crucial role in non-climacteric fruit ripening. VvABF2 transcripts accumulated more abundantly during the ripening stage. A whole-genome based transcriptomic approach performed on VvABF2-overexpressing grape cells treated or not with ABA emphasizes the up-regulation of genes linked to fruit ripening processes. Indeed, genes encoding enzymes involved in the biosynthesis of phenolic compounds and in softening were affected. Finally, phenotypic analysis of VvABF2-overexpressing tomato fruits reinforces the link between this transcription factor and fruit softening. Indeed, from the “turning” stage, an accelerated fruit softening was observed in the transgenic fruits.Taken together, this PhD work contributed to a better understanding of the molecular mechanisms involved in the ripening of non climacteric fleshy fruit.

Las células endoteliales (CE) son responsables de la síntesis y liberación de sustancias vasoactivas ante un estímulo neuronal, hormonal o físico. Procesos patológicos, factores ambientales o el envejecimiento del individuo, dañan las CE y alteran su función. Se denomina disfunción endotelial (DE) a la capacidad disminuída de las CE de inducir vasorrelajación ante el estímulo agonista colinérgico. Los AINEs son fármacos antiinflamatorios, analgésicos y antipiréticos que bloquean la COX. Experimentos preliminares han mostrado que el clonixinato de lisina (Clx), un AINE antranílico, induce DE aguda en anillos de aorta in vitro, por un mecanismo no dilucidado. El extracto de Vitis vinifera (EVv) es rico en flavonoides, compuestos con acción antioxidante. Se intenta determinar si el EVv es capaz de prevenir in vitro la DE inducida por Clx. Se montaron anillos aórticos de ratas en baños termorregulados para la determinación isométrica de fuerza, asignándose al azar en tres grupos: control, grupo 1 (G1) y grupo 2 (G2). El G1 se incubó con 100 µM Clx y el G2 con 5,35 ng/mL EVv junto con 100 µM Clx. Determinamos y comparamos la DE en los anillos aórticos precontraídos con fenilefrina respecto del control. El G1 mostró una notable DE la que fue significativamente (p< 0,05) prevenida por la incubación con EVv. Los resultados obtenidos muestran que el EVv puede prevenir la DE inducida por Clx in vitro. Si bien, se requieren otros experimentos para explicar el mecanismo por el cual Clx induce DE y de prevención del EVv, este fitofármaco se perfila como una alternativa de protección vascular para los pacientes que utilizan crónicamente el Clx.

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Submitted by Gabriela Lopes (gmachadolopesufpel@gmail.com) on 2017-03-23T16:17:49Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação final (1).pdf: 1606741 bytes, checksum: 01f63a4e690a8f3143c6fd2940d71195 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-03-28T20:11:25Z (GMT) No. of bitstreams: 2 Dissertação final (1).pdf: 1606741 bytes, checksum: 01f63a4e690a8f3143c6fd2940d71195 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-03-28T20:11:25Z (GMT). No. of bitstreams: 2 Dissertação final (1).pdf: 1606741 bytes, checksum: 01f63a4e690a8f3143c6fd2940d71195 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-02-22 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / A Região da Campanha Gaúcha vêm se destacando na produção de uvas Vitis vinifera L. para a elaboração de vinhos finos. Estendendo-se ao longo da fronteira com o Uruguai, tem como principais referências os municípios de Candiota, Bagé, Dom Pedrito e Santana do Livramento. Entre as atividades de grande importância no manejo cultural da vitivinicultura podemos destacar a poda seca. Normalmente realizada no período de repouso que abrange os meses de julho a setembro, a poda seca exige uma grande demanda de mão de obra. Portanto é necessário coletar todas as informações possíveis sobre o ciclo fenológico da cultura facilitando o manejo e escalonando a mão de obra disponível. O trabalho teve como objetivo avaliar o comportamento fenológico e a qualidade pós colheita das cultivares Merlot, Cabernet Sauvignon e Tannat sobre diferentes épocas de poda seca. O experimento foi conduzido em um vinhedo comercial localizado no município de Candiota, Rio Grande do Sul na safra 2015/2016. O delineamento experimental foi inteiramente casualizado constando de quatro tratamentos e três repetições, onde os tratamentos foram as diferentes épocas de poda seca realizadas em maio, junho, julho e agosto, compondo os tratamentos 1, 2, 3 e 4 respectivamente. As avaliações fenológicas observadas ao longo do ciclo da cultura foram: brotação, floração, frutificação e início da maturação. Após a colheita as variáveis avaliadas no mosto foram os sólidos solúveis totais, açúcares, acidez titulável, pH e densidade. Para o cálculo de produção foram avaliados o número total de cachos por tratamento, número de cachos por planta, massa total dos cachos por tratamento e massa média dos cachos por planta. Os dados foram submetidos à análise de variância (Anova) e comparação de médias pelo teste de Tukey a 5%. A videira „Tannat‟ podada em agosto teve um comportamento fenológico mais tardio em relação aos demais tratamentos. Após a brotação, as plantas podadas em junho desenvolveram-se mais rápidas que as demais. As diferentes épocas de poda seca não interferiram na produção. Quando a poda seca for realizada no mês de agosto a colheita da „Tannat‟ pode ser mais tardia. Para „Cabernet Sauvignon‟ a poda antecipada do mês de maio permanece mais tempo em dormência, brotando ao mesmo tempo que as plantas podadas em julho. A poda realizada em junho antecipa a brotação podendo provocar prejuízos ocasionados pelas geadas primaveris. A poda realizada no mês de agosto obteve maior rendimento por planta quando comparada a época tradicional no mês de julho. A poda seca em diferentes épocas não influenciou nas características analíticas do mosto. Na videira „Merlot‟, a poda seca realizada no mês de junho antecipou a brotação e quando realizada no mês de agosto obteve número total e massa total de cachos superior à realizada no mês de junho. As diferentes épocas de poda seca não diferenciaram os resultados das variáveis físico-químicas da „Merlot‟. / The Campanha Gaucha region has been highlighted due to production of Vitis vinifera L. grape to make fine wines. This region extends along the border of Brazil and Uruguay whose main cities are Candiota, Bagé, Dom Pedrito and Santana do Livramento. Between the activities of utmost importance in management of viticulture activity, the dry pruning can be emphasized. In general, the dry pruning is performed in rest period, i. e. between the months of July and September, and requires a high labor demand. Thus, it is necessary to collect all the obtainable information on the phenological cycle of this culture to make easier the management and scaling of the available labor. The work aims to evaluate the phenological behavior and the quality of postharvest of the cultivars Merlot, Cabernet Sauvignon and Tannat, in different periods of dry pruning. The experiments were carried out in a commercial vineyard located in the city of Candiota, Rio Grande do Sul, during the harvest 2015/2016. The experimental design was a completely randomized design consisting of four treatments and three replications, where the treatments were the different dry pruning times performed in May, June, July and August, composing treatments 1, 2, 3 and 4 respectively. The phenological evaluations observed during the crop cycle were: sprouting, flowering, fruiting and beginning of maturation. After the harvest the variables evaluated in the wort were the total soluble solids, sugars, titratable acidity, pH and density. The total number of bunches per treatment, number of bunches per plant, total mass of bunches per treatment and average mass of bunches per plant were evaluated for the production calculation. Data were submitted to analysis of variance (Anova) and comparison of means by the Tukey test at 5%. The 'Tannat' vine pruned in August had a later phenological behavior in relation to the other treatments. After sprouting, the plants pruned in June developed faster than the others. The different seasons of dry pruning did not interfere with production. When dry pruning is performed in August, the 'Tannat' harvest may be later. For 'Cabernet Sauvignon' the early pruning of the month of May remains longer in dormancy, sprouting at the same time as the plants pruned in July. The pruning done in June anticipates the sprouting and can cause damages caused by the spring frosts. The pruning done in August obtained higher yield per plant when compared to the traditional season in July. Dry pruning at different times did not influence the analytical characteristics of the must. In the 'Merlot' grapevine, the dry pruning carried out in the month of June anticipated the sprouting and when it was carried out in the month of August it obtained a total number and a total mass of bunches higher than the one realized in the month of June.The different seasons of dry pruning did not differentiate the results of the physical-chemical variables of 'Merlot'.

Both the physiological and biochemical control of budburst in the grapevine, Vitis Vinifera L. were investigated. It was found that the accuracy of a predictive model for grapevine budburst based on ambient temperature was limited under the experimental conditions. There was a significant correlation of 4.7 ± 0.3 days between the days of maximal xylem exudation and budburst over the 3 years of investigation. The co-relationships between daily xylem exudate volume and a range of environmental parameters were considered. It was found that soil temperature was highly correlated against daily xylem exudation. Ambient temperature and soil moisture were significantly correlated with xylem exudation, however the coefficients of correlation were much lower than that of soil temperature. Rainfall showed only a very limited correlation with daily xylem exudate flow. Seasonal variations in the pH and the carbohydrate and inorganic nutrient concentrations of xylem exudate were investigated. Exudate carbohydrate concentrations fell from 660 µM before the day of maximal xylem exudation to zero levels within 4 weeks. Xylem exudate pH was found to consistently fall to a minimum at the time of maximal exudate flow. Exudate concentrations of the metallic cofactors Ca, K, Mg, Mn and Zn varied directly with daily exudate flow, suggesting some sort of flow-dependent mobilisation of these nutrients. A growth promontory oligosaccharide fraction was prepared by partial acid hydrolysis of grapevine primary cell wall material. This fraction significantly increased control growth of the Lemna minor L. bioassay over a limited ‘window’ of bioactivity. A growth inhibitory oligosaccharide fraction, similar in activity to abscisic acid was isolated from grapevine xylem exudate prior to budburst. The exudate concentration or efficacy of this substance declined after budburst such that there was no apparent growth inhibition. A model is proposed for grapevine budburst whereby an oligosaccharide growth inhibitor is gradually removed from the xylematic stream under the effects of soil temperature, allowing the surge of metabolic activity and vegetative growth that constitute budburst.

Los hongos de la familia Botryosphaeriaceae causan daños en la madera de la vid en regiones vitícolas de todo el mundo. Para caracterizar las distintas enfermedades de madera de la vid en planta adulta y para conocer el impacto de estos patógenos en Cataluña, se llevó a cabo un muestreo en viñedos afectados de diferentes denominaciones de origen catalanas. Los resultados confirmaron la existencia en Cataluña de las tres enfermedades de la vid más comunes en planta adulta (yesca, eutipiosis y BDA [black dead arm]), y evidenciaron asimismo la dificultad para diagnosticarlas, debido a la múltiple y frecuente concurrencia de éstas sobre una misma planta. Las especies fúngicas aisladas con mayor frecuencia fueron Phaeomoniella chlamydospora, Fomitiporia mediterranea, Eutypa lata y distintas especies de Botryosphaeriaceae. Dentro de esta familia se identificaron seis especies, siendo por orden de abundancia Diplodia seriata, Neofusicoccum parvum, Dothiorella viticola, Botryosphaeria dothidea, N. luteum y N. vitifusiforme. La controversia taxonómica que presenta la familia Botryosphaeriaceae, debido a las similitudes morfológicas entre taxones, se resolvió con el estudio combinado de la morfología de los conidios de los anamorfos y la información molecular de distintas regiones del ADN. Mediante la técnica de la PCR-cooperativa y el empleo posterior de una sonda de hibridación específica, se desarrolló una herramienta molecular de gran sensibilidad para la detección rápida y rutinaria de las especies de Botryosphaeriaceae. La sonda se diseñó sobre la región nuclear del ITS (espaciadores internos de transcripción) del ARNr, e identificó de forma específica a un grupo de siete especies de Botryosphaeriaceae (las seis aisladas de vides catalanas más Diplodia mutila). Profundizando en la caracterización de la familia, se estudió la capacidad de cinco especies (B. dothidea, Di. seriata, Do. viticola, N. luteum y N. parvum) para producir micotoxinas. Todos los aislados estudiados produjeron compuestos hidrofílicos fitotóxicos, aunque N. luteum y N. parvum destacaron por la fitotoxicidad que también mostraron sus compuestos lipofílicos. Esto corroboró los resultados de las pruebas de patogenicidad, en las que N. luteum y N. parvum mostraron una gran virulencia. Dothiorella viticola, N. vitifusiforme y Di. seriata mostraron una virulencia débil o nula, mientras que B. dothidea mostró un carácter patogénico intermedio. Sobre la susceptibilidad del hospedador a los distintos patógenos, se observó que el portainjerto Ru140 fue el menos susceptible y el 41B el más susceptible, mientras que las seis variedades de uva ensayadas (Cabernet-Sauvignon, Garnacha, Macabeo, Parellada, Tempranillo y Xarel·lo) no mostraron diferencias claras de susceptibilidad. Se constató que algunas especies vegetales que comparten su hábitat con la vid, como almendros, encinas, jaras, melocotoneros y nogales, son susceptibles a algunas especies de Botryosphaeriaceae, y que por tanto podrían actuar como hospedadores alternativos de los patógenos. De los factores abióticos que podrían afectar al desarrollo de las enfermedades se estudió el estrés hídrico. Los resultados obtenidos mostraron que tanto el estrés hídrico como el patógeno N. parvum alteraron parámetros fisiológicos y de crecimiento de la vid, aunque no se observó un efecto combinado de ambos sobre la planta. Además, la longitud de las necrosis vasculares causadas por N. parvum no aumentó significativamente con el estrés hídrico, confirmando que N. parvum es un patógeno primario y no un patógeno de debilidad. / Species in the fungal family Botryosphaeriaceae are known for causing grapevine trunk diseases in grape-producer regions worldwide. Declining vineyards were surveyed in Catalonia (NE Spain) to characterize those diseases on adult plants and to identify their causing agents. Most comon grapevine trunk diseases (esca, Eutypa dieback and black dead arm disease) were confirmed to occur in Catalonia. However, concurrence of symptoms in the same plant, belonging to at least two different diseases, often difficults disease diagnosis. Most frequently isolated fungal pathogens were Phaeomoniella chlamydospora, Fomitiporia mediterranea, Eutypa lata and several species of Botryosphaeriaceae. Six botryosphaeriaceous species were identified, in decreasing abundance order: Diplodia seriata, Neofusicoccum parvum, Dothiorella viticola, Botryosphaeria dothidea, N. luteum and N. vitifusiforme. The controversy in the taxonomy of Botryosphaeriaceae due to confusing morphological similarities among the species was resolved by combining the study of conidial morphology and multigene nucleotide data. A sensitive tool for a rapid detection of Botryosphaeriaceae species was developed based on the co-operational PCR technique and the use of a hybridization probe. The probe was designed to target the ITS region (internal transcribed spacers) of the rRNA repeat. It identified specifically the above Botryosphaeriaceae species isolated from grapevine plus Diplodia mutila. Five botryosphaeriaceuos species were tested for mycotoxin production, namely B. dothidea, Di. seriata, Do. viticola, N. luteum and N. parvum. All species produced hydrofilic phytotoxic compounds, and N. luteum and N. parvum were also shown to produce low-weight lipophilic toxins. These results support the high virulence showed by N. luteum and N. parvum in the patogenicity test. Virulence of Dothiorella viticola, N. vitifusiforme and Di. seriata was weak to low, while B. dothidea showed an intermediate virulence. In terms of host susceptibility, rootstock 140Ru was the less susceptible to the botryosphaeriaceous fungi tested while 41B was the most susceptible one. However, no great differences in susceptibility were observed among the six grapevine varieties tested (Cabernet Sauvignon, Grenache, Macabeo, Parellada, Tempranillo and Xarel·lo). Some plant species from the surroundings of vineyards, namely almond, peach and walnut trees, holm oak and rockroses, were shown to be susceptible to some Botryosphaeriaceae species. It is hypothesized that these species could act as alternative hosts for those pathogens. Hydric stress in combination with N. parvum infection was studied as a potential factor that could affect disease development. Physiological and growth parameters of grapevine were modified by both hydric stress and N. parvum main effects, but no combined effects were detected. Vascular necroses were similar in both stressed and non-stressed grapevines, therefore N. parvum is not considered as a weakness pathogen.

Thesis (Ph. D.)--University of Adelaide, Dept. of Horticulture, Viticulture and Oenology, 1999. / Includes bibliographical references (leaves 138-158).

Memoria para optar al Título Profesional de Ingeniero Agrónomo / La presente investigación, de carácter exploratorio, tuvo como principal propósito contribuir al desarrollo de la industria vitivinícola chilena, a partir del análisis de atributos con potencial de mejoramiento genético en Vitis vinifera L. En base a esto, y a los objetivos específicos propuestos, el estudio se subdividió en cuatro niveles, siendo estos: vitivinicultor, enólogo, gerente comercial, y consumidor. En primer lugar, fue necesario identificar los principales problemas asociados al manejo productivo de la vid, y a aquellos presentados en los procesos de vinificación. Para esto, se levantó información por medio de encuestas dirigidas a vitivinicultores y enólogos, con un cuestionario especifico para cada nivel. El segundo objetivo consistió en evaluar la disposición de las viñas por invertir en un plan de mejoramiento genético, para lo cual, se construyó un cuestionario especialmente dirigido a gerentes comerciales. Para finalizar, se caracterizó a los consumidores de vino embotellado, respecto a variables descriptivas de consumo, sociodemográficas, y funcionales en relación al mejoramiento genético.

Tesis para optar al título profesional de Ingeniero Agrónomo y al grado de Magíster en Enología y Vitivinicultura / La unión de distintos factores y propiedades de un alimento determinan la calidad y aceptabilidad del mismo frente al consumidor, siendo el color uno de los factores organolépticos más importantes a la hora de seleccionar y evaluar un alimento. El motivo por el cual el color es un índice de evaluación importante es porque éste sufre procesos de degradaciones después de un tiempo prolongado de almacenamiento y le sigue a menudo un deterioro desagradable del alimento que puede reducir la aceptación del consumidor (Araya,2006).

This thesis deals with the comparison of anthocyanins in blue grape varieties. It has been compared 23 varieties of Vitis vinifera L. and 9 interspecific varieties from the harvest 2015. The grapes come from the wine region Mikulov and from the wine village Lednice. Literary part deals with the chemical structure of anthocyanins and their occurrence in grapes, further accumulation of anthocyanins and influencing factors, physico-chemical factors affecting the stability of anthocyanins, change of anthocyanins during the vinification and extraction of anthocyanins and measurement of color parameters. The practical part is especially focused on compare the content of anthocyanins and their profile in varieties of Vitis vinifera L. and interspecific varieties.