A obtenção de novas cultivares de uvas de mesa e a compreensão da regulação da expressão gênica, associada à formação do fruto de uma cultivar de uva sem semente, constituem pré-requisitos essenciais para o desenvolvimento de ferramentas aplicadas ao melhoramento genético para essas espécies. Com este objetivo, dois trabalhos foram conduzidos a partir do estudo de transcritos derivados de frutos da cultivar (cv) Sultanina de videira. No primeiro trabalho, três bibliotecas subtraídas foram construídas a partir de RNA total de frutos em diferentes estádios de formação, utilizando-se uma metodologia modificada da análise representacional diferencial (RDA), denominada análise representacional de transcritos em grupos (BRAT). Um total de 2.500 fragmentos derivados de transcritos (TDFs) foram identificados e clonados a partir de estádios específicos do desenvolvimento do fruto. Após sequenciamento e análise in silico, 1.554 (62,16%) dos transcritos foram validados de acordo com a qualidade da sequência. A montagem das sequências derivadas de genes expressos (ESTs) resultou em 726 singletos e 69 clusters, com aproximadamente 76% de redundância. Entre os TDFs identificados, onze genes candidatos e dois genes-referência foram selecionados e submetidos a uma análise aprofundada dos seus perfis de expressão temporal por RT-qPCR. A expressão de sete genes foi concordante com os estádios específicos do desenvolvimento dos frutos. Entre os candidatos selecionados os genes a seguir listados são possivelmente os mais promissores quando se considera o desenvolvimento do fruto da cv. Sultanina: (i) VvUBP1 (proteína de ligação a oligouridilatos), (ii) VvRIP1 (proteínas induzidas pelo amadurecimento), (iii) VvP450 (citocromo P450), (iv) VvDOF1 (proteína do tipo Dof); (v) VvERF1 (fator de transcrição responsivo ao etileno), e (vi) VvGID1L1 (receptor de ácido giberélico). Num segundo momento, foi realizado um estudo mais detalhado dos genes Dof de videira. Tais genes correspondem a um grupo de fatores de transcrição específicos de plantas, os quais estão envolvidos na regulação de diferentes funções, tais como, resposta ao estresse, hormônios e luz, sinalização de fitocromo, germinação de sementes e expressão gênica tecido-específica. A família de genes Dof de videira compreende 26 membros, sendo que as sequências de aminoácidos de todos os domínios Dof alinham perfeitamente, incluindo resíduos de cisteína que são críticos para a ligação de zinco e outros resíduos conservados em fatores de transcrição Dof de A. thaliana, O. sativa e outras plantas. Baseado na análise de domínios Dof, sugere-se que estes domínios sejam possivelmente funcionais em videira. A localização física dos genes Dof nos cromossomos de videira foi realizada. Além disso, foi conduzida uma análise filogenética comparando o grupo de genes Dof em videira com os seus homólogos em outras duas eudicotiledôneas completamente sequenciadas, A. thaliana e Populus, permitindo identificar claramente clusters de genes parálogos e ortólogos. Por fim, os perfis de expressão de todos os 26 genes Dof foram estudados por RT-qPCR em nove órgãos de videira. / Table grapes production and kwnolegment about the gene expression regulation associated with the formation of seedless grape cultivar, constitute essential prerequisites for the development of tools applied to genetic improvement of this plant species. To this end, two studies were conducted using the analysis of transcripts derived from fruits of the Sultanina cultivar (cv.) grapevine. In the first study, three subtracted libraries were constructed from total RNA from fruit different developmental stages using a modified methodology of Representational Difference Analysis (RDA), named Bulk Representational Analysis of Transcripts (BRAT). A total of 2,500 transcripts-derived fragments (TDFs) were identified and cloned from specific stages of fruit development. After sequencing and analysis in silico, 1,554 (62.16%) transcripts were validated in accordance with the sequence quality. The assembly of sequences derived from expressed genes (ESTs) resulted in 726 singletons and 69 clusters, with overall EST redundancy of approximately 76%. Among the TDFs identified eleven candidate genes and two reference genes were selected and subjected to a thorough analysis of their temporal expression profiles by RT-qPCR. Among them, seven genes proved to be in agreement with the stage-specific expression. Among candidates selected from those differentially expressed, the genes listed below were considered the most promising when considering the fruit development in grapevine cv. Sultanina: (i) VvUBP1(oligouridylate binding protein), (ii)VvRIP1 (ripening induced protein), (iii) VvP450 (cytochrome P450), (iv) VvDOF1 (Dof-like protein), (v) VvERF1 (ethylene-responsive transcription factor), and (vi) VvGID1L1 (gibberellins receptor). The second study detailed the grapevine Dof gene family. Dof proteins are involved in the regulation of different functions such as plant response to stress, hormones and light phytochrome signaling, seed germination and tissue-specific gene expression. The Dof gene grapevine family includes 26 members. The amino acid sequences deduced from Dof domains matched perfectly including cysteine residues critical for zinc binding and other residues known to be conserved in Dof transcription factors from A. thaliana, O. sativa and other plants. Based on analysis of Dof domains, it is suggested that all grapevine Dof domains are possibly functional. The physical location of Dof genes in grapevine chromosome was determined. A phylogenetic study comparing grapevine Dof genes with their counterparts in two other eudicots completely sequenced, A. thaliana and Populus, allowed us to identify clear clusters of paralogous and orthologs genes. Finally, the expression profiles of all 26 Dof genes was studied by RT-qPCR in nine vegetative and reproductive grapevine organs.

A obtenção de novas cultivares de uvas de mesa e a compreensão da regulação da expressão gênica, associada à formação do fruto de uma cultivar de uva sem semente, constituem pré-requisitos essenciais para o desenvolvimento de ferramentas aplicadas ao melhoramento genético para essas espécies. Com este objetivo, dois trabalhos foram conduzidos a partir do estudo de transcritos derivados de frutos da cultivar (cv) Sultanina de videira. No primeiro trabalho, três bibliotecas subtraídas foram construídas a partir de RNA total de frutos em diferentes estádios de formação, utilizando-se uma metodologia modificada da análise representacional diferencial (RDA), denominada análise representacional de transcritos em grupos (BRAT). Um total de 2.500 fragmentos derivados de transcritos (TDFs) foram identificados e clonados a partir de estádios específicos do desenvolvimento do fruto. Após sequenciamento e análise in silico, 1.554 (62,16%) dos transcritos foram validados de acordo com a qualidade da sequência. A montagem das sequências derivadas de genes expressos (ESTs) resultou em 726 singletos e 69 clusters, com aproximadamente 76% de redundância. Entre os TDFs identificados, onze genes candidatos e dois genes-referência foram selecionados e submetidos a uma análise aprofundada dos seus perfis de expressão temporal por RT-qPCR. A expressão de sete genes foi concordante com os estádios específicos do desenvolvimento dos frutos. Entre os candidatos selecionados os genes a seguir listados são possivelmente os mais promissores quando se considera o desenvolvimento do fruto da cv. Sultanina: (i) VvUBP1 (proteína de ligação a oligouridilatos), (ii) VvRIP1 (proteínas induzidas pelo amadurecimento), (iii) VvP450 (citocromo P450), (iv) VvDOF1 (proteína do tipo Dof); (v) VvERF1 (fator de transcrição responsivo ao etileno), e (vi) VvGID1L1 (receptor de ácido giberélico). Num segundo momento, foi realizado um estudo mais detalhado dos genes Dof de videira. Tais genes correspondem a um grupo de fatores de transcrição específicos de plantas, os quais estão envolvidos na regulação de diferentes funções, tais como, resposta ao estresse, hormônios e luz, sinalização de fitocromo, germinação de sementes e expressão gênica tecido-específica. A família de genes Dof de videira compreende 26 membros, sendo que as sequências de aminoácidos de todos os domínios Dof alinham perfeitamente, incluindo resíduos de cisteína que são críticos para a ligação de zinco e outros resíduos conservados em fatores de transcrição Dof de A. thaliana, O. sativa e outras plantas. Baseado na análise de domínios Dof, sugere-se que estes domínios sejam possivelmente funcionais em videira. A localização física dos genes Dof nos cromossomos de videira foi realizada. Além disso, foi conduzida uma análise filogenética comparando o grupo de genes Dof em videira com os seus homólogos em outras duas eudicotiledôneas completamente sequenciadas, A. thaliana e Populus, permitindo identificar claramente clusters de genes parálogos e ortólogos. Por fim, os perfis de expressão de todos os 26 genes Dof foram estudados por RT-qPCR em nove órgãos de videira. / Table grapes production and kwnolegment about the gene expression regulation associated with the formation of seedless grape cultivar, constitute essential prerequisites for the development of tools applied to genetic improvement of this plant species. To this end, two studies were conducted using the analysis of transcripts derived from fruits of the Sultanina cultivar (cv.) grapevine. In the first study, three subtracted libraries were constructed from total RNA from fruit different developmental stages using a modified methodology of Representational Difference Analysis (RDA), named Bulk Representational Analysis of Transcripts (BRAT). A total of 2,500 transcripts-derived fragments (TDFs) were identified and cloned from specific stages of fruit development. After sequencing and analysis in silico, 1,554 (62.16%) transcripts were validated in accordance with the sequence quality. The assembly of sequences derived from expressed genes (ESTs) resulted in 726 singletons and 69 clusters, with overall EST redundancy of approximately 76%. Among the TDFs identified eleven candidate genes and two reference genes were selected and subjected to a thorough analysis of their temporal expression profiles by RT-qPCR. Among them, seven genes proved to be in agreement with the stage-specific expression. Among candidates selected from those differentially expressed, the genes listed below were considered the most promising when considering the fruit development in grapevine cv. Sultanina: (i) VvUBP1(oligouridylate binding protein), (ii)VvRIP1 (ripening induced protein), (iii) VvP450 (cytochrome P450), (iv) VvDOF1 (Dof-like protein), (v) VvERF1 (ethylene-responsive transcription factor), and (vi) VvGID1L1 (gibberellins receptor). The second study detailed the grapevine Dof gene family. Dof proteins are involved in the regulation of different functions such as plant response to stress, hormones and light phytochrome signaling, seed germination and tissue-specific gene expression. The Dof gene grapevine family includes 26 members. The amino acid sequences deduced from Dof domains matched perfectly including cysteine residues critical for zinc binding and other residues known to be conserved in Dof transcription factors from A. thaliana, O. sativa and other plants. Based on analysis of Dof domains, it is suggested that all grapevine Dof domains are possibly functional. The physical location of Dof genes in grapevine chromosome was determined. A phylogenetic study comparing grapevine Dof genes with their counterparts in two other eudicots completely sequenced, A. thaliana and Populus, allowed us to identify clear clusters of paralogous and orthologs genes. Finally, the expression profiles of all 26 Dof genes was studied by RT-qPCR in nine vegetative and reproductive grapevine organs.

Doctor of Philosophy / Department of Horticulture, Forestry, and Recreation Resources / Khatamian Houchang / This project investigated the physiological and biochemical responses of three grapevine genotypes to a deficit irrigation strategy termed partial rootzone drying (PRD). The principle objectives of the project were to (1) establish if the response to PRD is a unique vine physiological response; (2) to investigate the effect of PRD on berry maturation and composition; (3) effect of PRD on vine water-use efficiency; (4) investigate the hormonal signal involved in the PRD mechanism. In addition, the project investigated the adaptation of these three grapevine genotypes to Kansas climatic conditions. The irrigation project was conducted in both a greenhouse at Kansas State University, Manhattan, KS, and field conditions in an experimental vineyard at the John C. Pair Horticultural Research Center, Haysville, Kansas. The greenhouse study was replicated four times (6 months each run) and the field study ran from the 2005/2006 season through the 2006/2007 season. Application of PRD to vines resulted in a unique physiological response distinct from other established deficit irrigation procedures such as regulated deficit irrigation (RDI); however, the overall physiological responses of the vines were defined by the environmental conditions of the vineyard where the experiment occurred. Reduced water availability, low to medium vigor vines, restricted root development, and high evaporative demand were all factors in the vines response to PRD. Under these conditions, PRD did maintain the yield at deficit irrigation rates compared to fully irrigated vines, and was responsible for physiological changes in the vine that could have long-term implications for yield stability. The application of PRD resulted in an improvement in plant water status and maintenance of leaf function as measured by photosynthesis, stomatal conductance and leaf water potential throughout the season.

Dinitro-ortho-cresol (500, 1000, and 2000 ppm) and dormant oil (2.5, 5.0, and 10%) were applied to Thompson Seedless and Perlette grapevines alone and in combination to enhance budbreak and fruitfulness. Applications were made immediately after pruning. Use of these materials during the winter, immediately after pruning, had no effect on either fruitfulness or budbreak in Perlette or Thompson Seedless. Thompson Seedless cuttings collected in August were exposed to six temperatures (7, 13, 18, 24, 29 and 35 C) for five time periods (2, 4, 6, 8 and 12 weeks). After temperature treatment the cuttings were planted in the greenhouse at 24 (+OR-) 1 C. Cuttings exposed to 24, 29 and 35 C broke sooner than those exposed to 7, 13, 18 C at all time periods. The percentage of the bud openings of cuttings at 29 C for a period of 2 and 4 weeks was 95% and 100% respectively. The optimum temperature for budburst was 29 C. Gibberellic acid, Thiourea, dinitro-ortho-sec-butyl-phenol (DINOSEB), potassium nitrate and Endothal were used at two concentrations alone and in combination, to break rest of Thompson Seedless buds. Thiourea at 2% and DINOSEB at 1000 ppm alone were the only treatments which gave a higher percentage of bud opening after 20 forcing days at 25 (+OR-) 1 C temperature. Similar results were obtained from cuttings taken in both winter and summer. Three times after harvest, Perlette and Thompson Seedless were defoliated using the senesce enhancer Endothal. Defoliation times (4, 8, and 12 weeks) after harvest were used. In half of the treatments, regrowth was controlled with Endothal. Gibberellic acid (1000 ppm) Thiourea (2%) and DINOSEB (2000 ppm) were applied at the time of defoliation. The four- and twelve-week defoliation periods with new vine growth controlled for improving budbreak and fruitfulness were best. Growth regulators did not improve the defoliation treatments. The best treatments hastened budbreak by 10 days, more than doubled vine fruitfulness, and increased sugar content in berries resulting in a 10-day earlier harvest than in the control. Results were similar in both Thompson Seedless and Perlette. Thompson Seedless and Cardinal vines grown under Arizona desert conditions were defoliated 4 weeks after harvest with 2000 ppm Endothal. Two weeks later they were pruned and treated with 1000 ppm gibberellic acid, 2% Thiourea, and 1000 ppm DINOSEB. Cardinal and Thompson Seedless vines produced a second commercial crop in December of the same year. Thiourea (2%) and 1000 ppm DINOSEB did not have a significant effect; however, 1000 ppm Gibberellic acid reduced the number of clusters per vine.

Memoria para optar al título profesional de Ingeniera Agrónoma Mención Fruticultura / Con el objetivo de describir el comportamiento de una serie de variables que influyen en el rendimiento, en función del vigor de las plantas y apoyada en una planilla electrónica, se realizó un estudio durante la temporada 2010/2011 en un parronal de la variedad Flame Seedless, ubicado en la localidad de La Arena, Región de Atacama. Se trabajó con una planilla electrónica que integra los factores vegetativos y productivos que afectan el rendimiento de las plantas y permitiendo realizar la estimación de cosecha expresada en cajas exportables de 8,2 kg, diferenciando los datos de acuerdo al vigor de las plantas que compone el cuartel.

Memoria para optar al título profesional de Ingeniero Agrónomo / Considerando la relevancia de la uva de mesa en el norte del país y las restricciones asociadas a los suelos con bajo contenido de materia orgánica, se hace necesario el uso de enmiendas orgánicas para mejorar las condiciones físicas del suelo, con el propósito de aumentar o mantener los rendimientos en el tiempo. En el presente trabajo se evaluó la acción sinérgica del compost con un cultivo de cobertura, sobre las propiedades físicas del suelo y algunas variables de desarrollo de la vid. El estudio se realizó en Vicuña, Región de Coquimbo, sobre un suelo Typic Haplocambid cultivado con un parrón de uva de mesa variedad Queen Rose. Considerando unidades experimentales de 7 plantas en la hilera en un diseño en bloques al azar, en noviembre de 2014 se realizaron las aplicaciones de compost (5,7 ton ha-1) y en mayo de 2015 se realizó la siembra de haba (500.000 plantas ha- 1). Se generaron así los tratamientos testigo (T0), suelo acondicionado con compost (T1) y suelo acondicionado con compost más cultivo de cobertura (T2). Al cabo de seis meses (septiembre de 2015) se evaluó por horizonte genético la densidad aparente, distribución de tamaño de poros, resistencia mecánica, estabilidad de los micro agregados y conductividad hidráulica, además del rendimiento y crecimiento de raíces. Los manejos generaron una tendencia a incrementar el agua aprovechable del suelo, alcanzando en el rango de 0-60 cm de profundidad, niveles de 61,8; 70,3 y 67,4 mm en los tratamientos T0, T1 y T2, respectivamente. Junto con esto, se mejoró la porosidad gruesa en superficie, especialmente con compost, aunque sin diferencias estadísticas significativas. Por otra parte, la mayor continuidad porosa se generó en el tratamiento de compost con haba (T2), el cual logró los mayores niveles de conductividad hidráulica, la que alcanzó niveles de 2,3; 3,5 y 6,9 cm h-1 en T0, T1 y T2, respectivamente. En cuanto a la resistencia mecánica, en general los valores se encontraron por debajo del valor crítico de 80 N. Finalmente, hubo un aumento en el peso promedio de racimo en T1 de un 5,9% con respecto al testigo, aunque sin lograr diferencias estadísticas significativas, por lo que es necesario continuar estudiando las alternativas de cultivos de cobertura para potenciar el efecto del compost sobre las propiedades físicas del suelo.

Muscadinia rotundifolia est une espèce de la famille des Vitaceae. C’est un sous-genre du genre Vitis, le deuxième sous-genre étant celui des Euvitis qui comprend l’espèce cultivée Vitis vinifera (2n=38). M. rotundifolia (2n=40) est une source de résistance aux maladies très importante pour l’amélioration de la vigne. Son génome commence seulement à être décrit avec deux cartes génétiques récemment publiées. Ma thèse a consisté à utiliser des ressources génomiques chez M. rotundifolia cv Regale (banque BAC, collection de séquence d’extrémités de BAC ou BES et séquences de BACs) pour caractériser le génome de cette espèce en comparaison avec celui de V. vinifera. Les résultats obtenus ne montrent pas de différence importante entre les génomes des deux espèces en termes de composition du génome en bases (GC%), en séquences codantes ou en éléments répétés. De même, à une échelle globale, la famille de gènes NBS-LRR semble être similaire en termes de nombre et de balance entre les sous-familles. A une échelle plus fine cependant (carte physique et séquences de BAC), des remaniements relativement importants sont observés dans des régions portant cette famille de gènes, aboutissant parfois à des contenus différents en gènes, de région normalement homologues : duplication différentielles de gènes, présence/absence de gènes. / Muscadinia Rotundifolia is a species of the Vitaceae family. It is a sub-genus of the Vitis genus along with the Euvitis sub-genus, which the cultivated species Vitis vinifera belongs to. M. rotundifolia (2n=40) is a very important source of resistance to diseases in grapevine breeding programs. Its genome is only starting to be described with the recent publication of two genetic maps. The present study aimed at using M. rotundifolia cv Regale genomic resources (BAC library, BAC end sequences or BES, BAC sequences) in order to characterize the genome of this species in comparison with the genome of V. vinifera. The results showed that there is no striking difference between the two species in term of base composition (GC %), repeats frequency and gene space. The NBS LRR gene family also seems to be globally quite similar between the two species in terms of numbers and balance between subfamilies. At a finer scale (physical map and BAC sequence), frequent rearrangements are observed in genomic regions carrying the NBS-LRR gene family sometimes clearly associated with a different gene content between the two species in homologous regions: differential gene duplication, presence/absence of genes.

Memoria para optar al Título Profesional de Ingeniera Agrónoma / Chile es el principal exportador de uva de mesa en el mundo, y a nivel nacional este frutal representa la mayor superficie plantada. Frecuentemente las raíces de vid deben desarrollarse en suelos con características físicas desfavorables, provocando una merma en el rendimiento. Una forma de abordar este problema es implementar el uso de portainjertos, sin embargo, no se dispone de suficientes antecedentes específicos que indiquen cómo éstos responden a distintas condiciones del suelo. Uno de los factores a considerar es la humedad del suelo, que, dependiendo de su nivel, puede inducir en las raíces respuestas asociadas al crecimiento y desarrollo o también a la producción de sustancias como el etileno. En consecuencia, se requiere más información sobre el comportamiento de los portainjertos frente a distintos niveles de riego. Los objetivos planteados en esta investigación fueron, determinar la producción de etileno en raíces de tres portainjertos de vid en tres condiciones de humedad del suelo, y evaluar el efecto de la cantidad de agua sobre el desarrollo de brotes y el desarrollo radical de estos portainjertos. El estudio se llevó a cabo en macetas de 20 L, con una mezcla de arena gruesa, perlita y suelo franco arenoso. Los tratamientos correspondieron a la combinación de portainjerto (“Harmony”, “Paulsen 1103” y “Ramsey”) y humedad del suelo (50%, 100% y 150 % de la “capacidad de maceta” de Harmony, efectuando riegos mediante goteros con descargas de 2, 4 y 6 L h-1 respectivamente). Se utilizó un diseño experimental completamente aleatorizado con estructura factorial de tratamiento 3x3 y seis repeticiones. Se evaluó el etileno producido por las raíces, el crecimiento semanal de brotes y la masa final de raíces. Los resultados no mostraron diferencias en el etileno emitido por raíces de “Harmony”, “Paulsen 1103” y “Ramsey” en ninguna fecha de evaluación, independiente del nivel de riego. La longitud final de brotes solo presentó diferencias entre los niveles de riego, siendo el efectuado a 150% de la “capacidad de maceta” el que mostró mayor crecimiento, independiente del portainjerto utilizado. A partir de la tercera semana se evidenciaron diferencias en el largo de brotes entre tratamientos. “Ramsey” regado a 50% de la “capacidad de maceta” se destacó por presentar los menores valores, permaneciendo así hasta el final de la temporada, mientras que “Paulsen 1103” y “Ramsey” regados a 150% de la “capacidad de maceta” mostraron los mayores crecimientos de brotes desde la sexta semana de evaluación. Los sistemas radicales de los tres portainjertos no presentaron diferencias en masa fresca y seca, independiente del agua suministrada.

Bibliography: p. 227-253. / xvii, 254 p. : ill. (chiefly col.), photos (col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / This project sought to examine the spatial and temporal distribution, as well as the composition, of the flavan-3-ols, proanthocyanidins and flavonols in winegrapes throughout berry development. In addition, the author sought to explore how modifying bunch exposure to light would impact upon flavonoid biosynthesis in the grape berry by comparing the flavonoid composition of shaded and exposed grapes. The expression of key genes in the flavonoid biosynthetic pathway was also investigated to understand how the pathway is regulated during berry development. / Thesis (Ph.D.)--University of Adelaide, School of Agriculture and Wine, Discipline of Wine and Horticulture, 2004

Reproduction de : Thèse doctorat : Biologie et physiologie végétales : Reims : 2007. / Titre provenant de l'écran titre. Bibliogr. f. 123-154.