Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-10-04T11:49:58Z No. of bitstreams: 2 Tese - Darlene Ana de Paula Vieira - 2015.pdf: 5053649 bytes, checksum: eac1ec3280ae5a3470b6e2b602bcc3eb (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-10-04T11:50:10Z (GMT) No. of bitstreams: 2 Tese - Darlene Ana de Paula Vieira - 2015.pdf: 5053649 bytes, checksum: eac1ec3280ae5a3470b6e2b602bcc3eb (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-10-04T11:50:10Z (GMT). No. of bitstreams: 2 Tese - Darlene Ana de Paula Vieira - 2015.pdf: 5053649 bytes, checksum: eac1ec3280ae5a3470b6e2b602bcc3eb (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-11-16 / The tomato cultivars (Solanum lycopersicum L.) intended for processing, must provide a number of features, and one of them is that the fruit has adequate physical and chemical quality of the preparation of derivatives. The aim of this study was to determine the biometric characteristics, physico-chemical, the fruit mechanical and textural and anatomical of the fruit epidermis, the lycopene content and β-carotene, the color instrumental parameters L*, a* and b* and ripe whole fruit for digital image (RGB), and the physical and chemical characteristics the fruit (pH, TA, TSS and TSS/TA). Classify cultivars by neural networks using the physical, physico-chemical and RGB values for digital images. Employing entirely randomized design for all analyzes. The results were submitted to analysis of variance, the physico-chemical (pH, total acidity, TSS, TSS/TA) by Anova One Way, and the pigments and color by Anova Factorial. Means were compared using the Tukey test at 5% significance. Pearson correlation was established between all the answers, and the mathematical transformed were still applied in the following answers: L*, a* and b* of instrumental analysis and image. Among the cultivars in 2013, they had better set of other mechanical properties were AP533, IT761, HY37 and H9992. A mixture of hexane, acetone and ethanol has proved to be more efficient in the extraction of lycopene and β-carotene that only acetone. Regarding the instrumental color of the fruit did not differ, however, the image for color evaluation methods obtained values of how light or dark, and coordinates of chromaticity a* and b* greater than the instrumental method with the Hunterlab closer the chromaticity that the values of the Cielab. From an industrial point of view, wish to tomato cultivars for processing with higher TSS values TA and lycopene and a* and lower pH and L*. Whereas the TSS is critical to industrial yield, stood out the IT761 cultivars BRSena, AP543 and H9992. In 2014 five cultivars were analyzed, and the fruit pericarp thickness of greater show greater firmness of the skin, and less moisture high pectin content. more firm fruits in the standing position have a higher modulus of elasticity of the skin. Among the cultivars studied, those who had better set of mechanical properties were the TC2736 and CVR2909, which also had lower longitudinal diameter, peduncle scar, fresh and volume. The characteristics observed in the pith of tomato fruit clearly showed the disruption of the cell wall during the fourth stage of maturation of the cells related to the loss of fruit firmness. The method for color evaluation image using the Cielab scale obtained Hue angle values and a*/b* very close to those found instrumentally by colorimeter, indicating that the color analyzes digital image using these two parameters may be a practical means and low cost, to inspect the fruit color. / As cultivares de tomate (Solanum lycopersicum L.) destinadas à industrialização, devem apresentar uma série de características, e uma delas é que o fruto tenha qualidade físico-químicas adequada à preparação dos produtos derivados. O objetivo deste trabalho foi determinar as características biométricas, físico-químicas, mecânico-texturais do fruto e anatômicas da epiderme do fruto, os teores de licopeno e β-caroteno, os parâmetros instrumentais de cor L*, a* e b* e do fruto inteiro maduro por imagem digital (RGB), além das características físico-químicas (pH, AT, SST e SST/AT) do fruto. Classificar as cultivares por redes neurais, utilizando as características físicas, físico-químicas e valores de RGB por imagens digitais. Empregando-se delineamento inteiramente casualizado para todas as análises. Os resultados foram submetidos à análise de variância, os físico-químicos (pH, acidez total, SST, SST/AT) por Anova One Way, e os de pigmentos e de cor por Anova Fatorial. As médias foram comparadas através do teste Tukey a 5% de significância. Correlação de Pearson foi estabelecida entre todas as respostas obtidas, e ainda foram aplicadas as transformadas matemáticas nas seguintes respostas: L*, a* e b* da análise instrumental e de imagem. Entre as cultivares estudadas em 2013, as que apresentaram melhor conjunto de propriedades mecânicas foram AP533, IT761, HY37 e H9992. A mistura de hexano, acetona e etanol mostrou ser mais eficiente na extração de licopeno e β-caroteno que somente a acetona. Em relação à cor instrumental do fruto não se diferenciaram, no entanto, os métodos de avaliação de cor por imagem obtiveram valores de quão claro e escuro e coordenadas de cromacidade a* e b* maiores que o método instrumental, sendo o Hunterlab mais aproximado do colorimétrico, que os valores do Cielab. Do ponto de vista da indústria, desejam-se cultivares de tomate para processamento com maiores valores de STT, AT, licopeno e a* e menores de pH e L*. Considerando que o teor de SST é fundamental para o rendimento industrial, destacaram-se as cultivares IT761, BRSena, AP543 e H9992. Em 2014 foram analisadas cinco cultivares, sendo que os frutos com espessura do pericarpo maior apresentam maior firmeza da pele, menor umidade e maior teor de pectina. Frutos mais firmes na posição em pé possuem módulo de elasticidade da pele maior. Entre as cultivares estudadas, as que apresentaram melhor conjunto de propriedades mecânicas foram a TC2736 e a CVR2909, que também apresentaram menores diâmetro longitudinal, cicatriz do pedúnculo, massa fresca e volume. As características observadas no mesocarpo do fruto de tomate mostraram claramente a desestruturação da parede celular durante o quarto estádio de maturação das células, relacionadas com a perda de firmeza dos frutos. O método de avaliação de cor por imagem utilizando a escala Cielab obteve valores de ângulo Hue e relação a*/b* bem próximos aos encontrados instrumentalmente pelo colorímetro, indicando que as análises de cor por imagem digital usando este dois parâmetros poderão ser um meio prático e de baixo custo, para inspecionar a cor dos frutos.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2017-06-07T11:55:12Z No. of bitstreams: 2 Dissertação - Yuri de Oliveira Castro - 2017.pdf: 2073976 bytes, checksum: 2173380278c4268f1f75502613d21754 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-06-07T11:55:32Z (GMT) No. of bitstreams: 2 Dissertação - Yuri de Oliveira Castro - 2017.pdf: 2073976 bytes, checksum: 2173380278c4268f1f75502613d21754 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-06-07T11:55:32Z (GMT). No. of bitstreams: 2 Dissertação - Yuri de Oliveira Castro - 2017.pdf: 2073976 bytes, checksum: 2173380278c4268f1f75502613d21754 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-02-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Encourage the adventitious roots can bring increments to the culture of tomatoes for industrial processing (TI). The objective of this study was to determine the effect of positions transplanting and ages of adventitious roots in changes in plant development in TI productivity. Three experiments were carried out arranged in factorial scheme (3 x 3 + 2): three positions of transplanting (collect level, at the level of the cotyledon and at the level of the first leaf fully expanded) in three ages of change (30, 60 and 90 days after sowing-DAS) and a free additional position (at the level of the last leaf fully expanded) for the ages of 60 and 90 DAS. The changes of the 30 did not present last leaf fully expanded and did not receive the last position of transplanting. In experiment 1 (one) the contribution of the zones or regions Annals (collect the cotyledon; the cotyledon to the first leaf; the first sheet to the last leaf fully expanded) to the number of roots adventitious issued. The Count took place every two days after transplanting (DAT) and was held until the 20º DAT. The experiment was designed entirely with four replicates and plots sub-plot. In experiment 2 (two) to evaluate the architecture of the root system of seedlings of thee through images of the roots of plants cultivated in rizontrons (phenotyping platform of high performance) and analyzed by software WinRHIZO PRO 2013. The experiment was designed in complete randomized blocks (DBC) with five repetitions. In experiment 3 (three) measured periodically the leaf area and dry mass of aerial part of plants, equations of partition and fotoassimilados allocation. The experiment was designed in DBC with six repetitions. The area between the collect and the cotyledon presented the highest number of adventitious roots in all ages of change, with quadratic responses to 60 and 90 of seedlings and seedlings for 30 DAS. The issue of roots beyond the 20º day may be related to better hormonal and nutritional condition of 30 seedlings of compared with the seedlings of 60 and 90 DAS. The depth of the root system is linked to a greater leaf area, although the contribution of photosynthesis for the adventitious roots become less relevant than other factors such as nutritional status of changes. The leaf area and dry mass of plant organs were influenced by age of change. Seedlings of 30 and 60 DAS the present greater leaf area, dry mass accumulation and distribution of roots when compared with 90 DAS the seedlings. Productivity did not differ between the ages of 30 and 60 DAS and was higher if compared to 90 DAS seedlings. / Incentivar o enraizamento adventício pode trazer incrementos à cultura do tomate para processamento industrial (TI). Objetivou-se neste estudo determinar o efeito de posições de transplantio e idades de muda no enraizamento adventício, no desenvolvimento vegetal e na produtividade de TI. Foram realizados três experimentos: experimento 1 (um): em estufa com mudas transplantadas em tubetes para produção de mudas florestais; experimento 2 (dois): em casa de vegetação com mudas transplantadas em rizotron; experimento 3 (três): em campo aberto. No experimento 1 (um) avaliou-se a contribuição das zonas ou regiões caulinares (do coleto ao cotilédone; do cotilédone à primeira folha; da primeira folha à última folha totalmente expandida) para o número de raízes adventícias emitidas. A contagem ocorreu a cada dois dias após o transplantio (DAT) e foram realizadas ate o 20º DAT. O experimento foi delineado inteiramente ao acaso com quatro repetições. No experimento 2 (dois) avaliou-se a arquitetura do sistema radicular de mudas de TI através de imagens das raízes de plantas cultivadas em rizontrons (plataforma de fenotipagem de alto desempenho) e analisadas pelo software WinRHIZO PRO 2013. O experimento foi delineado em blocos completos casualizados (DBC) com cinco repetições em esquema fatorial adicional 3 x 3 + 2, no qual, mudas de 30, 60 e 90 DAS foram transplantadas em 3 níveis (coleto, cotilédone e primeira folha) e as idades de 60 e 90 DAS receberam um nível adicional de transplantio cada (última folha). No experimento 3 (três) mediu-se periodicamente a área foliar e a massa seca da parte aérea das plantas, obtendo-se equações de partição e alocação de fotoassimilados. O experimento foi delineado em DBC com seis repetições em esquema fatorial adicional 3 x 3 + 2 (idem ao experimento 2). A zona entre o coleto e o cotilédone apresentou o maior número de raízes adventícias em todas as idades de muda, com respostas quadráticas para mudas de 60 e 90 DAS e linear para mudas de 30 DAS. A emissão de raízes além do 20º dia pode estar relacionada à melhor condição hormonal e nutricional das mudas de 30 DAS comparada com as mudas de 60 e 90 DAS. A profundidade do sistema radicular está ligada a uma maior área foliar, apesar de a contribuição da fotossíntese para o enraizamento adventício ser menos relevante que outros fatores como o estado nutricional da muda. A área foliar e a massa seca dos órgãos vegetais foram influenciadas pela idade da muda. Mudas de 30 e 60 DAS apresentam maior área foliar, acúmulo de massa seca e distribuição de raízes quando comparadas com mudas de 90 DAS. A produtividade não diferiu entre as idades de 30 e 60 DAS e foi maior se comparada às mudas de 90 DAS.

Made available in DSpace on 2017-07-10T17:40:52Z (GMT). No. of bitstreams: 1 Sidiane_Coltro_Roncato.pdf: 1950234 bytes, checksum: ab588be5ce6968a641dc1b750a09907e (MD5) Previous issue date: 2015-08-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The objective of this work was to study the effect of Crambe abyssinica leaf extract on Meloidogyne incognita and Meloidogyne javanica control in tomato plants, and the possible effect of allyl isothiocyanate component in this interaction. Five experiments were conducted. The first experiment was carried out with both Meloidogyne species in vitro: dried leaves of crambe (200 mg L-1) were added in the solvents: water for infusion and grinding, acetone, water + ethanol, methyl alcohol, hexane and chloroform. After the evaporation rotative, the aqueous extracts were analyzed in vitro for nematicide activity (hatching, mortality and motility of second stage juveniles (J2)). After the extracts with increased activity were tested at doses of 200, 300, 400 and 500 mg L-1 of crambe dry leaves. To quantify the allyl isothiocyanate in the extracts was used high performance liquid chromatography (HPLC). The second and third experiment were with M. incognita and hydroalcoholic extract to 250 mg L-1 was used, and each experiment was repeated twice, being the second time with the two best times of the first. The first repetition of the second experiment was conducted in factorial design (3x4+1) in a greenhouse, with three extract application routes: leaf; soil; leaf+soil, in four periods: before inoculation; on inoculation; after inoculation; and weekly up to 45 days and an additional treatment (inoculated and untreated control). In the second repetition, the factorial design was (3x2+1), three routes of application as mentioned above, and two periods (after inoculation and weekly). The third experiment includes the same treatments, however, to the application routes is added the immersion of the tomato roots. The fourth and fifth experiments were with M. javanica, using the same methodology described for the experiments with M. incognita. The results derived from the extracts of hydroalcoholic solution, methanol and trituration were more effective in the action against nematodes, however, the hydroalcoholic one caused higher mortality, with 93% and 64% for M. incognita and M. javanica, respectively. The greatest effect was on the dose of 250 mg L-1, and only the extracts obtained by methanol solution and the hydroalcoholic presented allyl isothiocyanate. When the hydroalcoholic extract was tested in a greenhouse in tomato plants, to M. incognita the applications via soil and weekly did not differ statistically from the applications soil+leaf for the analyzed variables. When the immersion of the tomato plants rooting system was added, weekly applications routes via root+soil and root+soil+leaf stood out, and nematode population in the soil was reduced by 75.55% and 71.95%, respectively, however, the second experiment confirmed the control only to the application via soil+root+leaf. For M. javanica, applications of the extract via soil+leaf weekly caused reduction of total galls on 46.15%. When added the immersion of the root system, the weekly application via root+soil+leaf reduced the egg masses and total galls on 60.95% and 27.95%, respectively. Therefore, crambe extract has potential to make up the integrated management of M. javanica and M. incognita on tomato plants / O objetivo deste trabalho foi estudar o efeito do extrato de folhas de Crambe abyssinica no controle de Meloidogyne incognita e Meloidogyne javanica em tomateiro e, possível atuação do componente alil isotiocianato nesta interação. Foram conduzidos cinco experimentos. O primeiro experimento foi realizado com ambas as espécies de Meloidogyne in vitro: folhas secas de crambe (200 mg L-1) foram adicionadas nos solventes: água para infusão e trituração, acetona, água+álcool etílico, álcool metílico, hexano e clorofórmio. Após a evaporação rotativa, os extratos aquosos foram analisados in vitro para atividade nematicida (eclosão, mortalidade e motilidade de juvenis de segundo estádio (J2)). Após, os extratos com maior atividade foram testados nas doses 200, 300, 400 e 500 mg L-1 de folhas secas de crambe. Para quantificar o alil isotiocianato nos extratos foi utilizada cromatografia líquida de alta eficiência (HPLC). O segundo e terceiro experimento foram com M. incognita e extrato hidroalcoólico à 250 mg L-1 foi utilizado e, cada experimento foi repetido duas vezes, sendo a segunda vez com as duas melhores épocas da primeira. A primeira repetição do segundo experimento foi conduzido em esquema fatorial (3x4+1) em casa de vegetação, com três vias de aplicação do extrato: foliar; solo; foliar+solo, em quatro épocas: antes da inoculação; na inoculação; após a inoculação; e semanalmente até 45 dias e um tratamento adicional (testemunha inoculada e não tratada). Na segunda repetição, o esquema fatorial foi (3x2+1), três vias de aplicação como citado anteriormente, e duas épocas (após a inoculação e semanalmente). O terceiro experimento contempla os mesmos tratamentos, porém, nas vias de aplicação é adicionado a imersão das raízes de tomateiro. O quarto e quinto experimentos foram com M. javanica, com a mesma metodologia descrita para os experimentos com M. incognita. Os resultados dos extratos originados da solução hidroalcoólica, metanólica e por trituração apresentaram-se mais efetivos na ação contra os nematoides, porém, o hidroalcoólico ocasionou maior mortalidade, com 93% e 64% para M. incognita e M. javanica, respectivamente. O maior efeito foi na dose de 250 mg L-1, e somente os extratos obtidos por solução metanólica e o hidroalcoólica apresentaram alil isotiocianato. Quando o extrato hidroalcoólico foi testado em casa de vegetação em plantas de tomateiro, para M. incognita as aplicações via solo e semanalmente não diferiram estatisticamente das aplicações via solo+folha para as variáveis analisadas. Quando foi adicionada a imersão do sistema radicular do tomateiro, aplicações semanais com as vias raiz+solo e raiz+solo+folha se destacaram, e a população do nematoide no solo foi reduzida em 75,55% e 71,95%, respectivamente, porém, o segundo experimento confirmou o controle apenas para a aplicação via raiz+solo+folha. Para M. javanica, aplicações do extrato via solo+folha semanalmente ocasionaram redução de galhas totais em 46,15%. Quando adicionada a imersão do sistema radicular, a aplicação semanal via raiz+solo+folha reduziu massa de ovos e galhas totais em 60,95% e 27,95%, respectivamente. Portanto, o extrato de crambe tem potencial para compor o manejo integrado de M. javanica e M. incognita em tomateiro

Atualmente, o HLB é considerado a principal doença que acomete as plantas cítricas. Diante desse fator, pesquisas por cultivares resistentes a esta doença são necessárias. A transformação genética via Agrobacterium, juntamente com o uso de plantas modelos, tem sido uma alternativa para verificação do funcionamento dos genes em resposta a patógenos, isto porque as plantas modelos possuem como característica ciclo de vida curto e alto poder de regeneração. Assim sendo, objetivou-se com o presente trabalho, a transformação genética via Agrobacterium tumefaciens, de tomate Micro-Tom (Solanum lycopersicum L.) e de laranja doce, com os genes que codificam as proteínas PR-8 e CDR-1, isolados a partir de Citrus simensis. Os cotilédones provenientes de sementes germinadas in vitro de tomate Micro-Tom foram utilizados como fonte de explante para os experimentos de transformação genética com os genes PR-8 e CDR-1. Esses explantes foram subcultivados até o aparecimento de brotos regenerantes e posteriormente plantas transgênicas, as quais foram aclimatizadas e levadas a casa-de-vegetação. A transgenia foi confirmada por PCR e o número de inserções do gene por Southern blot. As plantas foram cultivadas até a obtenção da geração T1. Simultaneamente, foram realizados experimentos de transformação genética em segmentos de epicótilo, provenientes de sementes de laranja ‘Hamlin’ germinadas in vitro, com o gene CDR-1, a fim de se obter plantas transgênicas e sua caracterização. Paralelamente, foi realizada a construção da curva padrão pela análise de qPCR para identificação de Pseudomonas syringae pv. tomato. Foram obtidas treze plantas transgênicas de tomate Micro-Tom com o gene PR-8 e três com o gene CDR-1. As eficiências de transformação foram em torno de 0,38 a 1,98%. Três plantas de tomate Micro-Tom transgênicas com o gene PR-8 foram caracterizadas por Southern blot e o número de inserções variou de 1 a 3. Dezenove plantas transgênicas de laranja ‘Hamlin’ com o gene CDR-1 foram obtidas através dos experimentos de transformação genética. A eficiência de transformação foi de 2,06 a 5,96%. Dessas, apenas uma foi caracterizada por Southern blot apresentando 1 número de cópia do DNA no genoma da planta. / HLB is currently considered the main disease affecting citrus plants. Given this factor, research for cultivars resistant to this disease is needed. Genetic transformation via Agrobacterium with the use of model plants has been an alternative for checking the gene function in response to pathogens, because these model plants have as characteristic a short life cycle and high power of regeneration. Therefore, the aim of this work was to produce transgenic plants, via Agrobacterium tumefaciens, of Micro-Tom tomato (Solanum lycopersicum L.), and sweet orange, with the genes encoding the PR-8 and CDR-1 proteins isolated from Citrus sinensis. The cotyledons from in vitro germinated Micro-Tom tomato seeds were used as explants source for genetic transformation experiments with PR-8 and CDR-1 genes. These explants were subcultured until the appearance of regenerating shoots and after transgenic plants, which were acclimatized and taken to a greenhouse. The transgenic plants were confirmed by PCR and the number of gene insertions by Southern blot. The plants were grown until T1 generation was obtained. Simultaneously genetic transformation experiments were performed with epicotyl segments from \'Hamlin\' sweet orange seeds germinated in vitro with CDR-1 gene in order to obtain transgenic plants and their characterization. Simultaneously, the standard curve construction was performed by qPCR analysis for identification of Pseudomonas syringae pv. tomato. Thirteen transgenic plants of Micro-Tom tomato with PR-8 gene and three with CDR-1 gene were obtained. The transformation efficiencies were around 0,38 to 1,98%. Three transgenic plants of Micro-Tom tomato with PR-8 gene were characterized by southern blot, and the number of inserts ranged from 1 to 3. Nineteen transgenic \'Hamlin\' sweet orange plants with CDR-1 gene were obtained through genetic transformation experiments, and the transformation efficiency was 2,06 to 5,96%. One plant was characterized, by Southern blot and has one DNA copy number in the plant genome.

Tomato (Solanum lycopersicum L.) consumption has increased every year due to the fruit attractiveness, several utilizations, and beneficial effects for human health. However, tomato fruits can accumulate a Cd concentration that exceeds the safety threshold for human consumption of vegetables, even when plants are grown in soil with acceptable Cd level. Cd is a non-essential, hazardous element to biological systems, triggering several diseases in humans. In plants, Cd disturbs the antioxidant machinery, changes the nutritional status, and impairs the photoassimilate production and/or partitioning, hence reducing fruit yield and quality. However, distinct tomato accessions can present contrasting tolerance degree to Cd toxicity, as detected by our group in previous studies. The use of these accessions is a powerful approach to identify strategies employed by plants to cope with Cdinduced challenges, and the acknowledgement of such strategies can be potentially used in breeding and biotechnological programs to improve fruit yield and quality in crops that were cultivated in contaminated fields. The set of studies that compose the present thesis aimed (i) to identify the main mechanisms for the contrasting tolerance degree to Cd-induced toxicity in tomato accessions after short and long-term Cd exposure; (ii) to evaluate the relationship among tolerance degree and fruits attributes in plants that were grown in Cd-containing soil, and (iii) to determine the transgenerational effects of Cd-induced stress. In the first experiment, nine tomato accessions with a varied tolerance degree, which was based on biomass accumulation, to Cd exposure were grown in hydroponic solution containing CdCl2 35 μM for 6 days. Avoidance of high Mg concentration in roots was identified as a plant strategy to mitigate Cd toxicity by preventing formation of root hairs. Regarding the mode of action of Cd toxicity, Mn excess in leaves, in addition to the high Cd concentration per se, seems to be coupled to leaf damages that are enhanced by the increased Zn and B concentrations in the photosynthetic tissues. In the second experiment, tolerant (Yoshimatsu) and sensitive (Tropic Two Orders) genotypes were grown in Cd-containing soil, in order to evaluated production parameters. After plant exposure to Cd, the tolerant genotype presented an increased fruit diameter, height and weight, when compared to the control plants. In both cultivars, Cd concentration varied according to the following descending order: roots = leaf blade > (floral receptacle, peduncle and sepals) > stem = fruit peel = fruit pulp. Moreover, data suggested that floral receptacle and its related-structures acted as a barrier to the Cd transportation to the fruits, but it was not enough to avoid Cd reaching the fruits. Furthermore, Cd exposure provoked remarkable reductions in the Mg concentration in roots of sensitive and tolerant genotypes, revealing that both tomato cultivars are able to employ this mechanism for plant acclimation to long-term Cd exposure. Considering such information, it is possible that, under the short-term Cd exposure, tolerant accessions activate this mechanism either early or faster than sensitive genotypes. In addition, positive transgenerational effects on seed germination and vigor of the tolerant genotype were triggered by the plant-mother cultivation in Cd-containing media, despite of the increased chromosomal abnormality. This work reported new insights about the effects of Cd exposure on tomato development, tolerance mechanisms, fruit quality and yield of tomato, as well as Cd distribution in the plants. / O consumo de tomate (Solanum lycopersicum L.) tem aumentado a cada ano devido a atratividade dos frutos, suas diversas utilizações e efeitos benéficos para a saúde humana. No entanto, os frutos de tomate podem acumular uma concentração de cádmio (Cd) que excede o limiar de segurança para o consumo humano, mesmo quando as plantas são cultivadas em solo com níveis aceitáveis de Cd. Cádmio e um elemento não-essencial, extremamente perigoso para os sistemas biológicos, desencadeando varias doenças em seres humanos. Nas plantas, o Cd perturba a maquinaria antioxidante, altera o estado nutricional e prejudica a produção e /ou o particionamento de fotoassimilados, frequentemente reduzindo a produtividade e qualidade de frutos. No entanto, diferentes acessos de tomateiros podem apresentar contrastantes graus de tolerância a toxicidade gerada pela exposição ao Cd, como detectado em estudos anteriores de nosso grupo. O uso desses acessos e uma abordagem poderosa para identificar as estratégias empregadas pelas plantas para lidar com os desafios induzidos pelo Cd; e o conhecimento de tais estratégias pode ser potencialmente utilizado em programas biotecnológicos e de melhoramento genético. Deste modo, o conjunto de estudos que compõem a presente tese objetivou (i) identificar os principais mecanismos que suportam o grau de tolerância contrastante a toxicidade induzida por Cd em acessos de tomate após exposição a curto e longo prazos a este metal pesado; (ii) avaliar a relação entre o grau de tolerância e os atributos físico-químico de frutos oriundos de tomateiros cultivados em solo contendo Cd, e (iii) determinar os efeitos transgeracionais do estresse induzido por Cd. No primeiro experimento, nove acessos de tomateiro com graus variados de tolerância a exposição ao Cd, baseado na acumulação de biomassa, foram cultivados em solução hidropônica contendo 35 μM de CdCl2 durante 6 dias. O impedimento de elevada concentração de magnésio (Mg) em raízes foi identificado como possível estratégia da planta para mitigar a toxicidade de Cd, por meio da evitação da formação de pelos radiculares. Em relação ao modo de ação da toxicidade induzida por Cd, o excesso de Mn, em adição a elevada concentração de Cd, parece estar acoplado aos danos foliares que são acentuados ainda mais pelas altas concentrações de zinco (Zn) e boro (B) nos tecidos fotossintéticos de plantas sob exposição ao Cd. No segundo experimento, os genótipos tolerantes (Yoshimatsu) e sensíveis (Tropic Two Orders) foram cultivados em solo contendo Cd, a fim de avaliar os parâmetros de produção. O genótipo tolerante apresentou frutos com maior diâmetro, altura e peso após o cultivo em solo contendo Cd, quando comparado as plantas controle. Em ambas as cultivares, a concentração de Cd variou de acordo com a seguinte ordem descendente: raízes = folíolos> (receptáculo floral, pedúnculo e sépalas) > caule = casca de fruta = polpa de fruta. Alem disso, dados sugerem que o receptáculo floral e suas estruturas atuaram como uma barreira ao transporte de Cd para os frutos, entretanto, ela não foi suficiente para evitar que o Cd atingisse os frutos. Em adição, a exposição ao Cd provocou notáveis reduções na concentração de Mg nas raízes de genótipos sensíveis e tolerantes, revelando que a aclimatação das plantas depende do baixo status de Mg em tecidos radiculares. Desde que ambas as cultivares são capazes de empregar este mecanismo, os dados sugerem que, durante a exposição a curto prazo ao Cd, acessos tolerantes são capazes de ativa-lo ou mais cedo ou mais rápido do que acessos sensíveis. Ademais, efeitos transgeracionais positivos na germinação e vigor das sementes do genótipo tolerante foram desencadeados pelo cultivo planta-mãe em solo com Cd, apesar do aumento de anormalidades cromossômicas. Este trabalho reportou novos conhecimentos sobre os efeitos da exposição ao Cd sobre o desenvolvimento do tomateiro, mecanismos de tolerância, qualidade e rendimento de frutos, bem como a distribuição de Cd dentro da planta.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / To verify the variability interference of the estimates of the values used to determinate the plot size with simulations of different numbers of plants per plot and groupings of crops, a five-experiment work was carried out. One in the winter/spring of 2004, other in the summer/autumn of 2005 for the zucchini and three experiments with tomato in the winter/spring of 2007, accomplished at Department of Phytotechny of UFSM, RS. In the experiments with zucchini, plastic tunnel was used, with drip irrigation, black mulching of PEDB, three lines with 24 plants per row, spacing among plants 0,8m and among lines 1,2m. For the tomato, the same plastic tunnels were used in two experiments and a plastic greenhouse in the third, that one with dimensions of 24m long and 10m wide in 8 cultivation rows. In the five experiments, each plant was considered as a basic unit (UB) of the plot size. The sequence of two, three and four plants in the crop line formed the plots of two, three and four UB of size (X). For each estimate in each experiment, analyses were accomplished considering only considered individual and combined harvests. For each one of the five experiments, the estimates of the average, of the variance, of the variation coefficient, of the index relative information and of the relative coefficient had been obtained. The Bartlett test was applied among the variances of the individual and combined harvests in each size of simulated plot and among the plot sizes in each individual and combined harvest to verify the homogeneity among the variances within each harvest in different plot sizes. For both cultures, there were some increases in the estimates of the average and in the variance of the fitomass of fruits with the increase of the size of the plot and/or the number of grouped harvests. For the zucchini, the variances, among the grouped harvests, were homogeneous, considering the grouping of three harvests, while for the tomato only in the groups of six harvests. The relative information index and the relative coefficient present more appropriate values, independently of each culture, considering the groups of harvests. The arrangement of two groups of combined harvests, joined with plots of three plants for Italian zucchini and of four plants for tomato, reduce the variability among plots. / Para verificar qual a interferência da variabilidade das estimativas dos valores usados para determinação do tamanho de parcela com simulações de diferentes números de plantas por parcela e agrupamentos de colheitas, realizou-se um trabalho com cinco experimentos. Um no inverno/primavera de 2004, outro no verão/outono de 2005 para a abobrinha italiana e três experimentos com tomate no inverno/primavera de 2007, realizados no Departamento de Fitotecnia da UFSM, RS. Nos experimentos com abobrinha italiana, foram utilizados túnel plástico, com irrigação por gotejamento, camalhões com mulching preto de PEDB, três linhas de cultivo, 24 plantas por linha, espaçamento entre plantas 0,8m e entre filas 1,2m. Já para a cultura do tomate, foram utilizados os mesmos túneis plásticos em dois experimentos e uma estufa plástica no terceiro, essa com dimensões de 24m de comprimento por 10m de largura em oito linhas de cultivo. Nos cinco experimentos, cada planta foi considerada como sendo a unidade básica (UB) do tamanho de parcela. A seqüência de duas, três e quatro plantas na linha de cultivo formou as parcelas de duas, três e quatro UB de tamanho (X). Foram realizadas, para cada estimativa em cada experimento, análises considerando-se apenas colheitas individuais e colheitas agrupadas. Para cada um dos cinco experimentos, foram obtidas as estimativas da média, da variância, do coeficiente de variação, do índice de informação relativa e do coeficiente relativo. Aplicou-se o teste de Bartlett entre as variâncias das colheitas individuais e agrupadas em cada tamanho de parcela simulado e entre os tamanhos de parcela em cada colheita individual e agrupadas para verificar a homogeneidade entre as variâncias dentro de cada colheita em diferentes tamanhos de parcela. Tanto para a cultura da abobrinha italiana quanto para a do tomate, há acréscimos nas estimativas da média e da variância da fitomassa de frutos com o aumento do tamanho da parcela e/ou o número de colheitas agrupadas. Para a abobrinha italiana, as variâncias entre as colheitas agrupadas apresentaram-se homogêneas a partir do agrupamento de três colheitas, enquanto que para o tomate somente nos agrupamentos de seis colheitas. O índice de informação relativa e o coeficiente relativo apresentam valores mais adequados, independente da cultura, a partir dos agrupamentos de colheitas. O arranjo de dois grupos de colheitas agrupadas, combinados com parcelas de três plantas para abobrinha italiana e de quatro plantas para tomate, reduzem a variabilidade entre parcelas.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The aim of this work were determine the optimum plot size and the uniformity assay size for estimating the optimum plot size in order to evaluate the fresh fitomass of lettuce plants and the mass of fruits of pepper, tomato, snap bean and zucchini. Production data were collected in uniformity assay with lettuce, pepper, tomato, snap bean and zucchini conducted at the Departamento de Fitotecnia da Universidade Federal de Santa Maria. Inside the planting row were planned different uniformity assay sizes. For each uniformity assay size planned were performed 3.000 resamples with replacement and were estimated for each sample the coefficient of spatial autocorrelation of the first order (p), the variance (s2), the mean (m) and the optimum plot size (Xo) by the method of maximum curvature of the coefficient of variation model (CMCV). Posteriorly, were also calculated the minimum values, percentile 2,5%, mean, percentile 97,5% and maximum values of statistics p, s2, m and Xo. For Xo statistic, even was calculated the amplitude of the confidence interval of 95%. Finally, were determined the uniformity assay size, starting from the smaller uniformity assay size planned (three basic units) and considering as uniformity assay size the number of basic units from which the amplitude of the confidence interval of 95% of optimum plot size was minor or equal to two basic experimental units. The uniformity assay size influences the estimation of the optimum plot size for evaluate the fresh fitomass of lettuce plants and the mass of pepper, tomato, snap bean and zucchini fruits. Uniformity assay with 27 basic units (27 plants) are enough for estimate the optimum plot size for evaluate the fresh fitomass of lettuce plants with a confidence interval of 95% minor or equal to two basic experimental units. Uniformity assay with pepper with 29 basic units (29 plants) in plastic greenhouse, with tomato with 12 basic units (12 plants) in plastic tunnel and with snap bean with 21 basic units (42 plants) in plastic tunnel, are enough for estimate the optimum plot size for evaluate the mass of fruits with a confidence interval of 95% minor or equal to two basic experimental units. Uniformity assay with snap bean with 18 basic units (36 plants) and with zucchini with ten basic units (ten plants) in plastic greenhouse are enough for estimate the optimum plot size for evaluate the mass of fruits with a confidence interval of 95% minor or equal to three basic experimental units. It is recommended the use of parcels with six basic experimental units for evaluate the fresh fitomass of lettuce plants and parcels with seven, five, six and eight basic experimental units, respectively, for evaluate the mass of fruits of pepper, tomato, snap bean and zucchini. / O objetivo deste trabalho foi determinar o tamanho de parcela e o tamanho de ensaio de uniformidade para estimar o tamanho de parcela a fim de avaliar a fitomassa fresca de plantas de alface e a massa de frutos de pimentão, de tomateiro, de feijão-vagem e de abobrinha italiana. Dados de produção de alface, de pimentão, de tomateiro, de feijão-vagem e de abobrinha italiana foram coletados em ensaios de uniformidade realizados no Departamento de Fitotecnia da Universidade Federal de Santa Maria. Dentro de cada linha de cultivo foram planejados diferentes tamanhos de ensaios de uniformidade. Para cada tamanho de ensaio de uniformidade planejado foram realizadas 3.000 reamostragens com reposição e estimados para cada amostra o coeficiente de autocorrelação espacial de primeira ordem (p), a variância (s2), a média (m) e o tamanho de parcela (Xo) pelo método da curvatura máxima do modelo do coeficiente de variação (CMCV). Posteriormente, também foram calculados os valores mínimo, percentil 2,5%, média, percentil 97,5% e máximo das estatísticas p, s2, m, e Xo. Ainda, para a estatística Xo, calculou-se a amplitude do intervalo de confiança de 95%. Finalmente, determinou-se o tamanho de ensaio de uniformidade, partindo-se do menor tamanho de ensaio planejado (três unidades básicas) e considerando como tamanho de ensaio de uniformidade o número de unidades básicas a partir do qual a amplitude do intervalo de confiança de 95% foi menor ou igual a duas unidades experimentais básicas. O tamanho do ensaio de uniformidade influencia a estimativa do tamanho de parcela para avaliar a fitomassa fresca de plantas de alface e a massa de frutos de pimentão, de tomateiro, de feijão-vagem e de abobrinha italiana. Ensaios de uniformidade com 27 unidades básicas (27 plantas) são suficientes para estimar o tamanho de parcela para avaliar a fitomassa fresca de plantas de alface com amplitude do intervalo de confiança de 95% menor ou igual a duas unidades experimentais básicas. Ensaios de uniformidade de pimentão com 29 unidades básicas (29 plantas) em estufa plástica, de tomateiro com 12 unidades básicas (12 plantas) em túnel plástico e de feijão-vagem com 21 unidades básicas (42 plantas) em túnel plástico, são suficientes para estimar o tamanho de parcela para avaliar a massa de frutos com amplitude do intervalo de confiança de 95% menor ou igual a duas unidades experimentais básicas. Ensaios de uniformidade de feijão-vagem com 18 unidades básicas (36 plantas) e de abobrinha italiana com dez unidades básicas (dez plantas) em estufa plástica são suficientes para estimar o tamanho de parcela para avaliar a massa de frutos com amplitude do intervalo de confiança de 95% menor ou igual a três unidades experimentais básicas. Recomenda-se o uso de parcelas com seis unidades experimentais básicas para avaliar a fitomassa fresca de plantas de alface e com sete, cinco, seis e oito unidades experimentais básicas, respectivamente, para avaliar a massa de frutos de pimentão, de tomateiro, de feijão-vagem e de abobrinha italiana.

Les derniers acteurs de la voie de signalisation à l’éthylène sont des facteurs de transcription appelés ERF (Ethylene Response Factors). La connaissance de leur rôle spécifique dans la régulation des processus développementaux dépendant de l’éthylène reste limitée. Les travaux présentés dans la thèse concernent la caractérisation fonctionnelle du gène Sl-ERF.B3, un membre de cette grande famille de régulateurs transcriptonnels dans la tomate (Solanum lycopersicum). Utilisant une stratégie répresseur dominant ; il est montré en particulier que ce gène intervient dans la mise en place de la réponse à l’éthylène et dans le contrôle de la maturation du fruit. L’expression d’une construction ERF.B3-SRDX, une version chimérique de Sl-ERF.B3 fusionné à un domaine répresseur de type EAR, entraine des phénotypes pléotropiques aussi bien dans la signalisation de l’éthylène que dans le développement des parties végétatives et des organes reproducteurs. Ainsi, une altération de la triple réponse à l’éthylène est constatée chez les lignées transgéniques et au stade adulte, les plantes présentent des phénotypes d’épinastie des feuilles, de sénescence prématurée des fleurs et d’abscission accélérée des fruits. L’ensemble de ces observations est corrélée avec une modification de l’expression de gènes impliqués dans la biosynthèse et la réponse à l’éthylène. Ces données suggèrent que ERF.B3 intervient dans un mécanisme de rétro-control de la réponse à l’éthylène en agissant à la fois sur les gènes de biosynthèse et de signalisation de l’hormone. Au niveau du fruit, la sur-expression d’ERF.B3-SRDX entraine une modification du processus de maturation avec un retard notable de l’avènement de l’acquisition de la compétence à murir. Cependant, une fois la maturation initiée, elle s’accompagne d’une forte production d’éthylène et d’une accélération du ramollissement du fruit. A l’inverse, l’accumulation de pigment est inhibée par altération de la voie de biosynthèse des caroténoïdes. Ces données phénotypiques sont corrélées avec le niveau d’expression des gènes clés impliqués dans ces processus. Les résultats indiquent que dans les lignées transgéniques, il y a découplage de certaines caractéristiques de la maturation du fruit et permettent de mettre en lumière le rôle d’ERF.B3 dans la régulation des processus de développement dépendant de l’éthylène chez la tomate. / Ethylene Response Factors (ERFs) are known to be the last transcription factors of the ethylene transduction pathway. Their specific role in ethylene-dependent developmental processes remains poorly understood. This work demonstrated a specific role of Sl- ERF.B3, a member of the ERF gene family in tomato (Solanum lycopersicum), in mediating ethylene response and fruit ripening through a dominant repressor strategy. ERF.B3-SRDX dominant repressor etiolated seedlings displayed partial constitutive ethylene-response in the absence of ethylene and adult plants exhibited typical ethylenerelated alterations such as leaf epinasty, premature flower senescence and accelerated fruit abscission. The multiple symptoms related to enhanced ethylene sensitivity correlate with the altered expression of ethylene biosynthesis and signaling genes, suggesting the involvement of Sl-ERF.B3 in a feedback mechanism regulating components of ethylene production and response. In addition, over-expression of ERF.B3-SRDX in tomato results in alterations in both fruit morphology and ripening process. The attainment of competence to ripen is dramatically delayed in ERF.B3-SRDX fruits but once ripening proceeds it is associated with high climacteric ethylene production and enhanced fruit softening while pigment accumulation is strongly reduced. Moreover, a number of genes involved in the fruit ripening process showed expression pattern deviating from that of wild type. These data suggest a putative role of Sl-ERF.B3 in the transcriptional network underlying the ripening process and uncover a mean for uncoupling some of the main features of fruit ripening such as fruit softening and pigment accumulation. Overall, the study highlighted the importance of an ERF gene in ethylene-mediated developmental processes such as plant growth and fruit ripening.

Analyse fonctionnelle de la protéine Enhancer of Zeste, SlEZ2, chez la tomate, Solanum lycopersicumLes protéines Polycomb, initialement découvertes chez la drosophile, ont récemment caractérisées chez les plantes où elles remplissent des fonctions essentielles au cours du développement de la plante. Chez la drosophile, les protéines polycomb (PcG) agissent sous forme de trois complexes multi-protéiques : PRC1, PRC2 et PhoRC. Seulement, deux de ces complexes ont été identifiés chez les plantes : un orthologue fonctionnel du complexe PRC1 (PRC1-like) et PRC2. Le complexe PRC2 maintien la chromatine dans un état condensé et intervient dans le contrôle du développement des fleurs, des graines, des fruits et des feuilles. Chez la tomate Solanum lycopersicum, le complexe PRC2 est composé de trois protéines polycomb : SlEMF2 (EMbryotic Flower), SlFIE (Fertilization Independent Endosperm) and SlE(Z) (Enhancer of Zeste). Les protéines SlE(Z) portent l’activité histone méthyl transférase qui permet la mise en place de la marque répressive H3K27me3. Chez la plante modèle, Arabidopsis thaliana, cette marque joue un rôle essentiel au cours du développement de la plante Afin d’étudier le rôle du complexe PRC2 dans le développement du fruit et de la plante de tomate, et plus particulièrement de la protéine SlE(Z), nous avons identifié trois gènes codant les protéines SlE(Z) : SlEZ1, SlEZ2 et SlEZ3. Au laboratoire, il a récemment été montré que la protéine SlEZ1 intervient au cours du développement floral (How Kit et al., 2010). L’objectif de ce travail est de déterminer la fonction de la protéine SlEZ2 au cours du développement du fruit et de la plante de tomate. Pour cela, nous avons analysé des plantes transgéniques sous exprimant le gène SlEZ2, orthologue au gène CURLY LEAF d’A. thaliana, par stratégie RNAi. Ce travail indique que la protéine SlEZ2 est impliquée dans la croissance de la plante de tomate, ainsi que dans le développement des feuilles, des fleurs et des fruits. Les plantes transgéniques présentent des phénotypes pléiotropes tels que des fleurs et des feuilles modifiées, un fort taux d’avortement des fruits, des fruits de texture et de couleur altérées ainsi qu’une réduction de la taille des plantes. De plus, nous avons identifiés quatre gènes ciblés par la protéine SlEZ2 dont l’expression est dérégulée dans les feuilles. Il s’agit de deux gènes à MADS box, TAG1 et TAGL1, ainsi que de deux gènes KNOX, LeT6 et TKN4. / Functional analysis SlEZ2, a tomato Enhancer of zeste proteinPolycomb proteins, first discovered in Drosophila, have been identified in plants and play essential functions in plant development. In Drosophila, polycomb proteins (PcG) acts as a complex and three have been identified: PRC1, PRC2 and PhoRC. However, only two polycomb complexes have been identified in plants: like-PCR1 and PRC2. The PCR2 complex maintain chromatin in a closed state and control flower, seed, fruit and leaf development.In tomato Solanum lycopersicum, PRC2 is composed by three polycomb proteins SlEMF2 (EMbryotic Flower), SlFIE (Fertilization Independent Endosperm) and SlE(Z) (Enhancer of Zeste)(Enhancer of Zeste). SlE(Z) proteins have a methyltransferase activity that puts in place an repressive epigenetic mark a trimethylation of lysine 27 histone 3. In plant model, Arabidopsis thaliana, this mark plays an essential role in plant development but little is known about PRC2 role in plant and fruit development of tomato. In order to unravel the function of the E(z) protein in the control of tomato fruit and plant development, we have characterized three E(z) encoding genes, namely SlEz1, SlEz2 and SlEZ3. In a recent work, we reported that SlEZ1 protein plays a role in flower development (How Kit at al., 2010). The aim of this present study was to determine the function of the SlEZ2 protein in plant and fruit development. We present our results focusing on RNAi transgenic plants which underexpressed SlEZ2 gene, homologue of Curly Leaf Arabidopsis gene. This analysis indicates that SlEZ2 protein is implicated in tomato plant growth and affects also leaf, flower and fruit development. Phenotypes include abnormal flowers and leafs, fruit development abortion, altered fruit colour and texture and plant of reduced size. Moreover, we characterize four target genes of SlEZ2 genes in leaves which present a deregulated expression : TAG1, TAGL1, LeT6 and TKN4.

Au sein de l’unité mixte de recherche 619 de l’Institut National de Recherche Agronomique, le groupe « Organogénèse du Fruit et Endoréduplication » étudie les acteurs moléculaires prenant part au contrôle du cycle cellulaire dans le fruit de tomate. L’objet de la présente thèse est l’étude de l’inhibiteur du cycle cellulaire Kip-Related Protein, et son rôle durant le développement du fruit. Identification de motifs protéiques fonctionnels chez l’Inhibiteur de Kinase Cycline-Dependent SlKRP1 chez Solanum lycopersicum : Leur rôle dans les interactions avec des partenaires du cycle cellulaire Les Kip-related proteins (KRPs) jouent un rôle majeur dans la régulation du cycle cellulaire. Il a été montré qu’ils inhibent les complexes CDK/Cyclin et ainsi bloquent la progression du cycle cellulaire. Malgré leur manque d’homologie avec leurs homologues animaux au delà de leur motif de liaison CDK/Cyclin, localisé à l’extrémité C-terminal de la protéine dans les séquences de plante, des études antérieurs ont montré la présence de motifs conservés spécifiques aux plantes chez certaines KRPs. Nous n’avons cependant que peu d’information concernant leur fonction. Nous montrons ici que les KRPs sont distribués en deux sous groupes phylogénétiques, et que chaque sous-groupe dispose de courts motifs spécifiques conservés. Les KRPs du sous-groupe 1 disposent ainsi de six motifs conservés entre eux. Utilisant SlKRP1, qui appartient au sous-groupe 1, nous avons identifié des motifs responsables de la localisation de la protéine et de ses interactions protéine-protéine. Nous montrons que le motif 2 est responsable de l’interaction avec CSN5, une sous-unité du complexe signalosome, et que le motif 5 a un effet redondant avec le motif 3 pour ce qui est de la localisation sub-cellulaire de la protéine. Nous montrons de plus que SlKRP1 est capable de guider SlCDKA1 et SlCycD3;1 vers le noyau, et ce même en l’absence du motif de liaison CDK/Cycline précédemment référencé. Ce nouveau site d’interaction est probablement localisé dans la partie centrale de la séquence de SlKRP1. Ces résultats apportent de nouveaux indices quant au rôle de la partie encore méconnue de cette protéine. La surexpression de SlKRP1 dans le mésocarpe de tomate détruit la proportionnalité entre endoréduplication et taille cellulaire Le fruit est un organe spécialisé résultant du développement de l’ovaire après pollinisation et fertilisation, et qui offre un environnement adéquat pour la maturation des graines et leur dispersion. De part leur importance en nutrition humaine et leur importance économique, les espèces à fruit charnu ont été le sujet d’étude développementales principalement orientée vers la formation de l’ovaire, la mise à fruit et la maturation du fruit. La phase de croissance du fruit a été beaucoup moins étudiée, bien que la division cellulaire et la croissance cellulaire prenant place durant cette période soient cruciales à la détermination de la taille finale du fruit, ainsi que de sa masse et sa forme. Le développement du mésocarpe du fruit de tomate se déroule par la succession d’une phase de division cellulaire suivie d’une phase d’expansion cellulaire associée à l’endoréduplication, menant à la formation de cellules géantes (jusqu’à 0,5mm) avec des niveaux de ploïdie pouvant atteindre 256C. Bien qu’une relation évidente entre endoréduplication et croissance cellulaire ait été montrée par de nombreux exemples chez les plantes, le rôle exact de l’endoréduplication n’a toujours pas été élucidé, étant donné que la plupart des expériences induisant une modification du niveau d’endoréduplication dans la plante affectaient aussi la division cellulaire. Nous avons étudié la cinétique du dévelopement du mésocarpe de tomate au niveau morphologique et cytologique et avons étudié l’effet de la diminution du niveau d’endoréduplication sur le dévelopement du fruit en sur-exprimant l’inhibiteur du cycle cellulaire Kip-Related Protein 1 (SlKRP1) spécifiquement dans les cellules en croissance du mésocarpe de tomate. Nous montrons une proportionnalité directe entre endoréduplication et taille cellulaire durant le développement normal du fruit, ce qui nous a permis de construire un modèle de développement du mésocarpe définissant l’épaisseur du péricarpe en ne prenant en compte que le nombre de divisions cellulaires et le nombre de tours d’endoréduplication. De façon surprenante, les mésocarpes de tomate affectés dans leur niveau d’endoréduplication par la sur-expression de SlKRP1 ne sont pas affectés au niveau de la taille des cellules ou du fruit, ni dans leur contenu métabolique. Nos résultats démontrent pour la première fois qu’alors que le niveau de ploïdie est étroitement lié avec la taille des cellules et du fruit, l’endoréduplication n’est pas responsable de la croissance cellulaire du mésocarpe de tomate. / Within the Joint Research Unit 619 of the National Institute of Agronomic Research (INRA), the group "Organogenesis of the Fruit and endoreduplication" examines the molecular players involved in cell cycle control in tomato fruit. The purpose of this thesis is the study of the cell cycle inhibitor Kip-Related Protein and its role during fruit development. Identification of protein motifs in the functional inhibitor of Cyclin-Dependent Kinase in Solanum lycopersicum SlKRP1: Their role in interactions with partners in the cell cycle The Kip-related proteins (KRPs) play a major role in the regulation of cell cycle. It has been shown to inhibit the CDK / Cyclin and thus block cell cycle progression. Despite their lack of homology with their counterparts in animals beyond their binding motif CDK / Cyclin, located at the C-terminal protein sequences in the plant, previous studies have shown the presence of conserved motifs plant specific in some KRPs, but there is little information about their function. We show here that the KRPs are distributed into two phylogenetic groups, and that each subgroup has specific short conserved motifs. The KRPs from subgroup 1 have six conserved motifs. Using SlKRP1, which belongs to subgroup 1, we have identified the motifs responsible for the localization of the protein and protein-protein interactions. We demonstrate that the pattern 2 is responsible for the interaction with CSN5, a subunit of the signalosome complex, and that the motif 5 is redundant with motif 3 with respect to the sub-cellular localization of the protein. We also show that SlKRP1 is capable of guiding SlCDKA1 and SlCycD3; 1 to the nucleus, even in the absence of CDK / cyclin binding motif previously referenced. This new site of interaction is probably located in the central part of the sequence of SlKRP1. These results provide new clues about the role of the little-known part of this protein. Overexpression of SlKRP1 in tomato mesocarp disrupts the proportionality between endoreduplication and cell size The fruit is a specialized organ which results from the ovary after pollination and fertilization, and provides a suitable environment for seed maturation and dispersal. Because of their importance in human nutrition and economic importance, fleshy fruit species have been the subject of study mainly focused on the developmental formation of the ovary, fruit set and fruit ripening. The stage of fruit growth has been much less studied, although cell division and cell growth taking place during this period are crucial to determining the final size of the fruit, as well as its mass and shape. The development of tomato fruit mesocarp occurs by the estate of a phase of cell division followed by a phase of cell expansion associated with endoreduplication, leading to the formation of giant cells (up to 0.5 mm) with ploidy levels of up to 256C. Although a clear relationship between endoreduplication and cell growth has been shown by many examples in plants, the exact role of endoreduplication has still not been elucidated, since most of the experiments leading to a change in the level of endoreduplication in plants also affected cell division. We studied the kinetics of the development of tomato mesocarp morphologically and cytologically and studied the effect of the reduced level of endoreduplication in the development of the fruit over-expressing the cell cycle inhibitor Kip-Related Protein 1 (SlKRP1) specifically in the growing cells of the tomato mesocarp. We show a direct proportionality between endoreduplication and cell size during normal development of the fruit, which allowed us to build a model for development of mesocarp defining the thickness of the pericarp by taking into account the number of cell divisions and the number of rounds of endoreduplication. Surprisingly, the tomato mesocarps affected in their level of endoreduplication by over-expression of SlKRP1 are not affected in terms of cell size and fruit, or on their metabolic content. Our results demonstrate for the first time that while the level of ploidy is closely linked with cell size and fruit, endoreduplication is not responsible for the cell growth of tomato mesocarp.