El empleo de herramientas genómicas ayudará a superar dos de los retos que todavía subsisten en el campo de la mejora molecular (i.e., vía transformación): la identificación de los genes que realmente controlan los caracteres de interés agronómico y la detección de señales de regulación que permitan modular la expresión de los transgenes a nivel espacial y temporal. Entre las vías para lograr tales objetivos, destaca la mutagénesis insercional por T-DNA, que en los últimos años se ha convertido en una herramienta básica para la identificación y etiquetado de genes, así como para el análisis de su función. En efecto, la disrupción de un gen endógeno o la integración del T-DNA en la vecindad del mismo pueden ocasionar la anulación o alteración de función, dando una valiosa información sobre el papel de un cierto gen en un carácter dado. Otra aplicación de la mutagénesis insercional por T-DNA estriba en la detección de elementos de regulación mediante el empleo de los denominados "sistemas trampa" (trapping) que permiten detectar secuencias reguladoras y asignar una función a partir de datos de expresión del delator que mimetiza la expresión del gen endógeno. El aspecto más relevante de estas aproximaciones es que, tras la identificación de un cierto gen, éste queda etiquetado por el T-DNA, lo que facilita su clonación. El principal objetivo de esta Tesis Doctoral ha sido la generación una colección de líneas de inserción por T-DNA en tomate y la identificación de mutantes afectados en caracteres relacionados con el desarrollo. En concreto, se han generado más de 1200 líneas T-DNA y se han obtenido sus descendencias TG2. La caracterización de estas líneas en TG1 ha conducido a la detección de 255 mutantes (de tipo dominante, semidominante o aditivo) afectados en caracteres vegetativos y/o reproductivos. Asimismo, se ha caracterizado una pequeña muestra de progenies TG2 (en concreto 37) lo que ha permitido la identificación de 6 mutantes recesivos. / Angarita Díaz, MDP. (2009). Generación de líneas T-DNA de tomate (Solanum Lycopersicon cv.p73) e identificación de mutantes de inserción [Tesis doctoral]. Editorial Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/14718 / Palancia

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vegetable crops. Micro-Tom is a model tomato cultivar used for research due to its small size and short time to fruiting. This project evaluated I. walleriana and Micro-Tom transformed with Agrobacterium. The construct contained GFP (green fluorescent protein) and hygromycin antibiotic-resistant selectable markers, and the antisense sequence of open reading frame of INSV nucleocapsid protein (N). The N gene is expected to confer INSV resistance by RNA interference or gene silencing. The presence of transgenes was confirmed by PCR. Transgenic Impatiens was selfed for two generations. Transgenic Micro-Tom was selfed for 4 generations. Spinach was used as an INSV reservoir. Impatiens, spinach and Micro-Tom were mechanically inoculated with INSV and evaluated visually, with assay tests, ELISA testing, and PCR. Spinach was successfully infected with INSV six times of seven attempts. Impatiens and Micro-Tom had no successful inoculations of three and five attempts, respectively. / Master of Science

Made available in DSpace on 2017-07-10T17:40:42Z (GMT). No. of bitstreams: 1 Heloisa_Maria_Formentini_tese.pdf: 1264963 bytes, checksum: d416be804a86c6ba7273d6aacedf8878 (MD5) Previous issue date: 2012-08-31 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / In Brazil, the tomato is one of the vegetable species of great importance both economically and socially, however several factors are limiting its production as an example the diseases caused by nematodes of the genus Meloidogyne that limit the production in infested areas. Seeking new measures of protection and control of plant disease induced resistance is na alternative considerering that little attention has been directed to the possibility of induced resistance to root pathogens like nematodes. Therefore, this study aimed to verify the effectiveness of the chemical inducer acibenzolar-S-methyl, the biotic inductor Bacillus cereus and plant extracts of rosemary (Rosmarinus officinalis) and turmeric (Curcuma longa) in the induced resistance in susceptible and resistant tomato in plants infected with Meloidogyne incognita race 3. Two experiments were conducted simultaneously both followed the 2 x 6 factorial design with two tomato genotypes, one susceptible to M. incognita (Santa Clara) and a resistant (Ivety) and six treatments: ASM (125 mg i.a L -1 ), Bacillus cereus (6.10 7 CFU mL -1 ), rosemary 10%, turmeric 10%, water and a control (no inoculum and no spraying in the aerial part), with five replicates. Each vase with a capacity of 2 L, were filled with a mixture of soil, sand and compost previously autoclaved and homogenized in the ratio 2:2:1 and were transplanted to each one three tomato seedlings susceptible and resistant. The treatments were sprayed in the aerial part in tomato plants in all vases except the absolute control. At 72 h after the first application of treatments was carried out the inoculation of 407/100 cm 3 of J2 and eggs per vase. In the first experiment, using destructive samples of tomato treated and inoculated with M. incognita were determined the response of both genotypes to the treatments applied to the enzymatic activity of peroxidase, polyphenol oxidase, chitinase, β-1,3 glucanase and phenylalanine ammonia-lyase from roots of tomato plants that were macerated and homogenized to withdrawals in time 0 h, 24 h, 48 h, 96 h and 120 h after the first application of the treatments. In the second experiment, the variables analyzed to determine the effect of treatments on nematode population were the number of root-knots, juveniles and eggs in the soil accomplished at 56 days after the first application of the treatments that were reapplied every seven days during this period. From the results obtained it was concluded that there was a reduction in the number of root-knots in the roots of tomato plants showing no difference between the two genotypes for plants that received the treatments with acibenzolar-S-methyl, turmeric, rosemary and B. cereus. There was a reduction in the formation of root-knots in susceptible cultivar, confirming their potential in protecting the genotypes used against the attack of M. incognita. For the enzimatic activity peroxidase was the enzyme strongly associated with resistance with the highest activity in resistant genotype if compared to susceptible regardless of inducer treatment. In susceptible tomato B. cereus stood out in the induction of chitinase and peroxidase whereas for the resistant tomato rosemary induced peroxidase and polyphenol oxidase and rosemary extracts and turmeric induced chitinase enzyme to the susceptible genotype / No Brasil o tomate é uma das espécies de hortaliças de grande importância tanto no ponto de vista econômico quanto social, no entanto vários fatores são limitantes para sua produção como exemplo as doenças causadas por fitonematoides principalmente as espécies do gênero Meloidogyne que inviabilizam a produção nas áreas infestadas. Buscando novas medidas de proteção e controle de doenças de plantas a indução de resistência é uma alternativa haja vista que pouca atenção tem sido direcionada a possibilidade de indução de resistência à patógenos do sistema radicular como os fitonematoides. Assim este trabalho teve como objetivo verificar a eficácia do indutor químico acibenzolar-S-metil, do indutor biótico Bacillus cereus e de extratos vegetais de alecrim (Rosmarinus officinalis) e cúrcuma (Curcuma longa) na indução de resistência em tomateiros suscetível e resistente infectados com Meloidogyne incógnita raça 3. Foram conduzidos dois experimentos simultaneamente ambos seguiram o esquema fatorial 2 x 6 com dois genótipos de tomateiro um suscetível à M. incognita (Santa Clara) e um resistente (Ivety) e seis tratamentos: ASM (125 mg i.a L -1 ), Bacillus cereus (6.10 7 UFC mL -1 ), alecrim 10%, cúrcuma 10%, água e uma testemunha absoluta (sem inóculo e sem pulverização na parte aérea), com cinco repetições. Cada vaso, com capacidade para 2 L, foram preenchidos com a mistura de solo, areia e composto orgânico previamente autoclavados e homogeneizados na proporção 2:2:1 e para cada vaso foram transplantados três mudas de tomateiro suscetível e resistente. Os tratamentos foram pulverizados na parte aérea dos tomateiros em todos os vasos com exceção da testemunha absoluta. Às 72 h após a primeira aplicação dos tratamentos foi realizada a inoculação de 407/100 cm 3 de J2 e ovos por vaso. No primeiro experimento, utilizando amostras destrutivas de tomateiros tratados e inoculados com M. incognita foram determinadas a resposta dos dois genótipos aos tratamentos aplicados para a atividade enzimática das enzimas peroxidase, polifenoloxidase, quitinase, β-1,3 glucanase e fenilalanina amônia-liase a partir do macerado homogeneizado das raízes dos tomateiros para o tempo de coleta 0 h, 24 h, 48 h, 96 h e 120 h após a aplicação dos tratamentos. No segundo experimento, as variáveis analisadas para determinar o efeito dos tratamentos sobre a população do nematoide foram o número de galhas, juvenis e ovos presentes no solo realizado aos 56 dias após a primeira aplicação dos tratamentos que foram reaplicados a cada sete dias durante este período. A partir dos resultados obtidos concluiu-se que houve uma redução no número de galhas no sistema radicular dos tomateiros não apresentando diferença entre os dois genótipos para as plantas que receberam os tratamentos com acibenzolar-S-metil, cúrcuma, alecrim e Bacillus cereus. Houve uma redução na formação de galhas na cultivar suscetível, confirmando seu potencial na proteção dos genótipos utilizados contra o ataque do M. incognita. Para a atividade enzimática a peroxidase foi a enzima que esteve fortemente associada à resistência com a atividade superior no genótipo resistente em relação ao suscetível independentemente do tratamento indutor. No tomateiro suscetível o B. cereus destacou-se na indução de peroxidase e quitinase enquanto que para o tomateiro resistente o alecrim induziu peroxidase e polifenoloxidase e os extratos de alecrim e cúrcuma induziram a enzima quitinase para o genótipo suscetível

Some microorganisms are known to form mutualistic symbiosis with plant roots and by their impact they can improve some plant parameters. These symbiotic microorganisms, which are able to improve some plant parameters, include especially mycorrhizal fungi, plant growth promoting bacteria and some saprotrophic mycoparasitical fungi. Mechanisms of changes of these parameters, as influenced by symbiotic microorganisms, are known only in part and nowadays are being actively researched. Aims of this work were to find out if selected microbial treatments influence selected growth, physiological and yield parameters of plants and contents of selected substances in fruits. Within this work were made three pot greenhouse experiments (experiments 1, 2 and 3) and three pilot greenhouse experiments (experiments 4, 5 and 6), performed on tomato (Solanum lycopersicum) and cucumber (Cucumis sativus) plants. Plants were grown in hydroponics using a carrier of rockwool and they were watered by nutrient solution. As microbial treatments for plants in experiments have been used a mixture of arbuscular mycorrhizal fungi (AM), mixture of plant growth promoting bacteria (PGPB), saprotrophic mycoparasitical fungus Trichoderma harzianum (Th) and various mutual combinations of these treatments. There have been observed...

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Agradecemos a compreensão. on 2017-01-11T17:23:55Z (GMT) / Submitted by LUCIANA GOMES DE ALMEIDA (luciana@biodinamica.org.br) on 2017-02-05T16:18:18Z No. of bitstreams: 1 Luciana Dissertação Mestrado definitivo 090117.pdf: 1935780 bytes, checksum: 6a96f8cbad659a357aa0c5cee25806e4 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2017-02-08T15:48:59Z (GMT) No. of bitstreams: 1 almeida_lg_me_bot.pdf: 1935780 bytes, checksum: 6a96f8cbad659a357aa0c5cee25806e4 (MD5) / Made available in DSpace on 2017-02-08T15:48:59Z (GMT). No. of bitstreams: 1 almeida_lg_me_bot.pdf: 1935780 bytes, checksum: 6a96f8cbad659a357aa0c5cee25806e4 (MD5) Previous issue date: 2016-11-10 / Objetivou-se com esta pesquisa estudar a curva de crescimento e absorção de nutrientes do tomateiro ‘Débora Victory’, cultivado em sistema orgânico a campo, em Itápolis-SP e estudar os teores de N, P e K de seis biofertilizantes ao longo do processo de fermentação. Para avaliação do híbrido Débora Victory, os tratamentos foram constituídos por nove épocas de amostragem, aos 2, 32, 46, 60, 77, 91, 109, 122 e 137 dias após o transplante (DAT) realizado em 01/04/2014. As plantas atingiram acúmulo estimado de massa seca e fresca total na última coleta de 550,2 e 9.528,2 g planta-1, respectivamente. A ordem decrescente do acúmulo total de nutrientes foi K > N > Ca > P > S > Mg, com valores estimados de 22,6; 10,4; 5,0; 2,3; 2,1 e 1,6 g planta-1, respectivamente. Na última coleta, o maior acúmulo de N, P, K ocorreu nos frutos e de Ca, S e Mg na parte vegetativa (caule+folhas+ inflorescências). O estudo da composição dos biofertilizantes foi constituído por seis biofertilizantes, elaborados com a mesma base para a fermentação (composta de 1 kg de farelo de arroz, 250 mL de melaço, 250 mL de microorganismos, marca EMBIOTIC, misturados em 15 litros de água), diferindo entre si pelo acréscimo de um ingrediente: T1 = torta de mamona (2 kg); T2 = farinha de sangue (2 kg); T3 = pó de casco e chifres (2 kg); T4 = farinha de ossos (2 kg); T5 = farinha de sangue (1 kg); T6 = farinha de ossos (1 kg) e cinco momentos de avaliação, aos 3, 7, 15, 22 e 30 dias após o preparo (DAP) dos biofertilizantes. Em cada momento foram analisados os teores de N, P e K e a condutividade elétrica. O biofertilizante a base de pó de casco e chifres apresentou maior teor de N do que os outros, alcançando o teor de 6,8 g L-1 aos 30 DAP. Os biofertilizantes a base de torta de mamona e pó de casco e chifres apresentaram maior teor de P, com médias de 3,2 e 2,6 g L-1, respectivamente, do que os de farinha de ossos (1 e 2 kg). O biofertilizante a base de torta de mamona apresentou maior teor de K, média de 2,5 g L-1, do que todos os demais, que foram iguais entre si. Independente do tipo de biofertilizante, os teores de K não se alteraram, os de N e a condutividade elétrica aumentaram e os de P reduziram ao longo do tempo de fermentação até os 30 DAP. / The objective of this research was to study the growth curve and absorption of nutrients in tomato 'Debora Victory', grown in organic system field in Itápolis-SP and to study the contents of N, P and K of six biofertilizers during the process fermentation. To evaluate the Débora Victory hybrid, the treatments consisted of nine sampling times at 2, 32, 46, 60, 77, 91, 109, 122 and 137 days after transplanting (DAT) held on 01.04.2014. The plants had estimated total dry and fresh weight, in the last sampling, of 550.2 and 9528.2 g plant-1, respectively. The descending order of total nutrient accumulation was K> N> Ca> P> S> Mg, with estimates of 22.6; 10.4; 5.0; 2.3; 2.1 and 1.6 g plant-1, respectively. In the last sampling, the greater accumulation of N, P, K occurred in fruits and Ca, Mg and S in the vegetative parts (stem + leaves + inflorescences). The experiment consisted of six biofertilizers, prepared on the same basis for fermentation (composed of 1 kg of rice bran, 250 ml of molasses, 250 ml of microorganisms, brand EMBIOTIC, mixed in 15 liters of water), differing from each other by an ingredient addition: T1 = castor bean cake (2 kg), T2 = blood meal (2 kg); T3 = hoof and horn powder (2 kg), T4 = bone meal (2 kg), T5 = blood meal (1 kg); T6 = bone meal (1 kg)) and five dates of evaluation: 3, 7, 15, 22 and 30 days after preparation (DAP) of biofertilizers. The following variables were analyzed: contents of N, P and K and the electrical conductivity. The hoof and horn powder biofertilizer showed higher N content than the others, reaching a maximum of 6.8 g L-1 at 30 DAP. The castor bean cake and hoof and horn powder, showed higher P contents, with average of 3.2 and 2.6 g L-1, respectively, than bone meal (1 and 2 kg) biofertilizer. The castor bean cake biofertilizer showed higher K content, average of 2.5 g L-1, than the others that did not differ each other. Regardless of the type of biofertilizer, the K content did not change, the N content and the electrical conductivity increased and the P content reduced during the time of fermentation until 30 DAP.

Contrôle de l'oxydation et de la dégradation du pool de vitamine C chez la tomate et impact sur la qualité du fruit et la tolérance au stress. Le métabolisme de l’ascorbate et plus principalement le statut redox du pool d’ascorbate sont impliqués dans la tolérance au stress et dans les processus primaires de croissance et de développement de la plante. La teneur et le statut redox de l’ascorbate chez les plantes sont régulés par (i) ses voies de biosynthèse, (ii) par le cycle ascorbate-glutathion permettant le recyclage des formes semi-oxydées et oxydées de l’ascorbate et (iii) par sa dégradation, l’ensemble de ces processus étant sous le contrôle de l’environnement. Au cours de ce travail de thèse, des méthodes de transgénèse nous ont permis d’identifier, chez différents génotypes de tomate à petit et gros fruits, les bouleversements physiologiques et métaboliques permettant de compenser des modifications de l’activité des enzymes monodéhydroascorbate réductase (impliqué dans le cycle ascorbate-glutathion) et ascorbate oxydase. Nous avons observé d’importantes modifications phénotypiques altérant le rendement en fruits de la plante sous conditions de culture pouvant générer un stress et également en condition normale de culture. Des liens entre l’activité des enzymes précités avec le métabolisme des sucres, la photosynthèse et la conductance stomatique sont révélés. Le déséquilibre entre les activités oxydantes et réductrices de ces enzymes constitue la première étape vers une dégradation de l’ascorbate. Le taux de dégradation se révèle très faible à la lumière, tandis qu’à l’obscurité une forte accumulation des produits de dégradation l’oxalate, le thréonate ainsi que l’oxalyl-thréonate est observé dans les feuilles de tomate. Enfin, l’activité de l’enzyme MDHAR est corrélée au taux de dégradation à l’obscurité. Les travaux de cette thèse mettent en avant l’importance du statut redox du couple ascorbate / monodéhydroascorbate dans les processus de croissance cellulaire et entre dans la régulation du rendement chez la tomate, et influe la dégradation de l’ascorbate. / Ascorbate metabolism and particularly ascorbate redox status are involved in stress tolerance and growth processes of plant cells. The concentration of ascorbate and its redox status are under control of (i) its biosynthetic pathways, (ii) the ascorbate-glutathione cycle allowing recycling of semi-oxidized and oxidized forms of ascorbate and (iii) its degradation rate. These processes are under environmental control. Transgenic lines modified for the activity of monodehydroascorbate reductase (involved in ascorbate-glutathione cycle) and ascorbate oxidase were generated in cherry and large-fruited genotypes of tomato. Physiological and metabolic modifications related to the modification of these enzyme activities were studied. We observed large phenotypic alterations that affected fruit yield under both stress conditions and normal growth conditions. Links between ascorbate recycling and sugar metabolism, photosynthesis and stomatal conductance were also revealed. An imbalance between the oxidizing and reducing activities of these enzymes is the first step leading to ascorbate degradation. We have shown that the degradation rate was very low under light, whereas under darkness the degradation compounds oxalate, threonate and oxalyl-threonate accumulated in tomato leaves. Also, the degradation rate is correlated with MDHAR activity. These results highlight the crucial role of the redox status of the ascorbate / monodehydroascorbate couple in growth processes and yield stability in tomato, and the impact on ascorbate degradation.

This work was carried out in collaboration with the Department of Adaptive Biotechnologies, Global Change Research Centre AS CR. It deals with the quantitative analysis of photosynthetic cell cultures. It uses images captured by a confocal fluorescent microscope to the automatic determining the number of cells in the sample. The work consists of a theoretical analysis, which briefly describes fluorescence and confocal microscopy. It also concisely introduces a microscope Leica TCS SP8 X, which I used to scan data. One capture is devoted to the theory of digital image processing. The second part deskribes the development of algorithm for processing 3D data and simplified algorithm for processing 2D data and its program implementations in a programming environment MATLAB R2013b. Grafical user interface is explained in detail. Done measurement are presented at the conclusion. It mentions compiled sample preparation protocol. The results of the program are compared with manual counting. Number of cells per 1 ml are determined by created program in samples of cell cultures Chenopodium rubrum (Cr) and Solanum lycopersicum (To).