Green algae have evolved several photo-protective responses to cope with high-light stress. The present study examines the metabolic changes during photoacclimation to high-light in Chlamydomonas reinhardtii using nuclear magnetic resonance and mass spectrometry. Using principal component analysis, a clear metabolic response to highlight intensity was observed on global metabolite pools in Chlamydomonas, with major changes in the levels of amino acids and related nitrogen metabolites. Amino acid biosynthesis was induced during short-term photoacclimation presumably to alleviate excess excitation pressure in the plastid. An increase in mitochondrial metabolism through downstream photorespiratory and glyoxylate metabolism, pathways thought to act in a photo-protective capacity, was also observed. Long-term light stress resulted in a significant increase in antioxidant metabolites, ascorbate and dehydroascorbate. These results suggest that metabolism plays a direct role in coping with the imbalance in the excess excitation pressure generated during high-light stress; however, this metabolomics survey has generated additional questions about the roles of nitrogen assimilation associated metabolites in photoacclimatory responses to high-light in Chlamydomonas.

The studies in this thesis research were conducted to investigate if sensitivity to induced cortical spread depression (CSD) or the consequence of a CSD event is affected by sumatriptan induced latent sensitization. Previous studies in our lab showed persistent exposure of sumatripan to rats produced a latent state of sensitization. Using persistent sumatripan exposed rats as a model for medication overuse headache, behavior, electrical stimulation threshold to provoke a CSD event and the immunoreactivity of c-Fos in the trigeminal nucleus caudalis (TNC) were characterized. Current results showed no statistical difference of electrically induced CSD thresholds in anesthetized rats measured at day 20 in sumatripan exposed rats compared with saline treated rats. Topiramate (80 mg/kg, i.p.) used clinically for prophylaxis of migraine headache significantly increased CSD threshold in both saline and sumatriptan infused rats. CSD events appear to be associated with trigeminal vascular system activation in TNC because c-Fos expression significantly enhanced in rats with electrically stimulated CSD events. As compared to saline treated rats, sumatriptan-exposed rats demonstrated a significantly higher number of c-Fos positive cells following the electrically stimulated CSD event. Under environmental stress (bright light), sumatripan exposed rats demonstrated decreased response thresholds to periorbital and hindpaw tactile stimuli (i.e., allodynia) and enhanced c-Fos expression in TNC. A single dose of topiramate (80 mg/kg, i.p.) reversed environmental stress induced allodynia and c-Fos over-activity. Taken together, these results suggest that latent sensitization induced by persistent sumatripan exposure seems not correlated to the threshold of electrically stimulated CSD in current model. However, CSD enhanced the responses of trigeminal system in rats with sumatriptan-induced latent sensitization. The protective effects of topiramate shown in this model may be related to blocking the initiation of CSD events resulting from environmental stimulation as well as inhibiting the consequences of CSD events in primary afferents. These findings correlate with clinical observations of protective effects of topiramate for migraine prophylaxis.

The ability of Cytisus scoparius L. and Spartium junceum L. to acclimate to different levels of light and water stress was studied to understand ecological constraints on distributions of these plants. A reaction norm experimental design was used to determine growth and physiological responses of each to imposed conditions. Light treatments were stressful for both species. Increases in shade led to decreases in relative growth rates (RGR); however, light was more of a limiting factor for S. junceum because of the greater decrease in its growth rate at lowest light intensities. As light decreased, stem allocation increased in S. junceum. More differences in allocation to leaves and roots among light treatments were found in C. scoparius. Correspondingly, the greatest changes in photosynthetic characters were found in S. junceum stems and C. scoparius leaves. Differences in physiological traits did not prove to be acclimation to low light levels because quantum yield decreased when light decreased. Neither species exhibited rapid growth rates, normal allocation patterns, or proper adjustments of photosynthetic characters under light conditions below 50% full intensity. Water treatments did not cause any critical changes in growth or physiology of either species. Neither species' RGR was greatly decreased. Water availability is more of a determining factor of growth for C. scoparius because of its continued, small decline in RGR as watering frequency decreased. Spartium junceum appeared to be better adapted to the imposed water treatments with its smaller number of leaves, leaf surface area, percentage leaf weight, specific leaf area, and leaf area / stem area ratio. Spartium junceum also displayed larger root / shoot ratios in drier conditions. The magnitude of these shifts was typically greater in C. scoparius, but the RGR of this species decreased more than that of S. junceum under drier conditions. More stress was imposed on C. scoparius because its lowest water potential measurements were 40% lower than those of S. junceum and below the water potential value at turgor loss point during midday hours. When water potential decreased, Spartium junceum showed signs of acclimation because stem photosynthesis increased and leaf photosynthesis decreased while C. scoparius did not adjust its rates of photosynthesis. Water treatments did not impose enough stress to cause osmotic adjustments. Performance under these light and water treatments explained the habitat preference of each to areas where these species have become invaders along the Pacific Coast of the Unites States. / Master of Science

Photosynthesis is the most important process for nearly all life on earth. Photosynthetic organisms capture and transfer light energy from the sun into chemical energy which in turn provides a resource base for heterotrophic organisms. Natural light regimes are irregular and vary over magnitudes. At a certain light intensity, metabolic processes cannot keep up with the electron flow produced by the primary photoreactions, and thus reactive oxygen species (ROS) are produced. ROS are highly reactive and can damage the photosynthesis apparatus and hence plants have evolved several photoprotection mechanisms to avoid the formation of ROS. The aim of this thesis was to examine the ecological effects of variations in photoprotection in plants. In particular I wanted to study the effect on fitness and the interaction with herbivorous insects of plants with different ability in photoprotection. To study this I used wild-type and transgenic Arabidopsis thaliana plants and grew them under natural conditions in field experiments in our botanical garden in Umeå, northern Sweden. For the investigation of the plant-insect interaction, a specialist on Brassicaceae (Plutella xylostella – diamondback moth) and a generalist herbivore (Spodoptera littoralis - Egyptian cotton worm) were used. Plants that are genetically deficient in one of the photoprotection mechanisms showed reduced fitness under natural conditions. I could thus show that feedback de-excitation (FDE) is the most important photoprotection mechanism, because a lack of FDE showed the highest reduction in fitness. The comparison of field grown wild-type with FDE mutant plants, using molecular biology methods, revealed large changes in gene transcription and metabolic composition. In particular, the jasmonate pathway was upregulated in light stressed plants, especially in plants lacking FDE. Jasmonate in turn is known to be a chemical compound which induces herbivore resistance genes and other stress responses. Specialist and generalist insect herbivores responded differently in feeding (dual-choice and no-choice) and oviposition experiments with field grown plants that differed in FDE. Female diamondback moths were attracted by induced defense compounds whereas the larvae avoided these plants in feeding experiments. Generalist larvae preferred, and showed a higher survival rate, on less light-stressed plants compared to more light-stressed plants. Combining molecular biology with ecological experiments is a challenging task. To summarize my experiences, I have produced a guide for experiments on transgenic plants in common gardens. In future investigations it is important to examine natural variations in photoprotection to elucidate selection pressures on specific genes.

La photosynthèse oxygénique fournit l’énergie pour presque toutes les formes de vie sur Terre. Les premières étapes sont réalisées par quatre complexes membranaires. Deux sont des photosystèmes (PS) divisés en un core et une antenne qui, chez les plantes, est composée des protéines Lhca et Lhcb respectivement pour le PSI et le PSII. Les Lhcb les plus abondantes forment les hétérotrimères LHCII. Pour optimiser leur croissance, les organismes photosynthétiques doivent réguler précisément l’absorption de la lumière. Dans cette thèse, nous nous sommes focalisés sur un de ces mécanismes chez Arabidopsis thaliana : les transitions d’état. Cette régulation intervient en lumière faible ou fluctuante pour répondre à un déséquilibre d’excitation des photosystèmes. Elle implique la phosphorylation de Lhcb1 et Lhcb2, deux des isoformes composant le LHCII, et leur déplacement du PSII au core du PSI où elles agissent comme antenne. Récemment, l’attachement d’un LHCII additionnel à l’antenne du PSI a été rapporté, sans indice sur le rôle de la phosphorylation dans la liaison. Nous nous sommes attachés à quantifier la phosphorylation du LHCII dans les supercomplexes PSI et PSII et à déterminer les rôles respectifs de Lhcb1 et Lhcb2 dans les transitions d’état. Nous avons établi que la phosphorylation d’un seul Lhcb2 est suffisante pour l’attachement d’un LHCII au core du PSI. Nous avons ensuite isolé un complexe PSI-LHCII2 et déterminé que la liaison du second LHCII implique aussi la phosphorylation d’un seul Lhcb2. Ce travail apporte de nouvelles preuves des rôles divergents des isoformes Lhcb dans la régulation de la photosynthèse, qui sont discutées ici à la lumière de leur évolution. / Oxygenic photosynthesis directly or indirectly provides energy for almost all forms of life on earth. The first steps are performed by four membrane complexes. Two of them are photosystems (PS) organized in a core complex and an antenna system, which is composed in green organisms by Lhca and Lhcb proteins for PSI and PSII, respectively. The most abundant Lhcb proteins form the LHCII heterotrimers. To optimize growth, photosynthetic organisms have to precisely regulate their light harvesting. In this thesis, we focused on one of these mechanisms in Arabidopsis thaliana: state transitions. This regulation occurs in low or fluctuating light to answer for the imbalance of excitation between photosystems. It involves the phosphorylation of Lhcb1 and Lhcb2, two of the isoforms composing LHCII. The phosphorylated LHCII trimers detach from PSII and bind to PSI core and act as an antenna for it. Recently the attachment of additional LHCII trimers to PSI on its antenna side has been reported, with no clue of the role of phosphorylation in this binding. We set on quantifying the LHCII phosphorylation of purified PSI and PSII supercomplexes to determine the respective roles of the Lhcb1 and Lhcb2 isoforms in state transitions. We established that the phosphorylation of a single Lhcb2 protein is sufficient for the binding of PSI core. We then isolated a PSI-LHCII2 supercomplex, and determined that the binding of the additional trimer also involves the phosphorylation of a single Lhcb2 isoform per trimer. This work brings new evidences for the divergent roles of the Lhcb isoforms in light harvesting and its regulation, which are discussed here in the light of their evolution.

Plants absorb light for photosynthesis, but not all is used. Excess light energy may lead to photoinhibition of photosynthesis and irreversible photooxidative damage. Plants have evolved mechanisms for energy dissipation under high light stress. One such response may involve production of ELIP. It is of interest to know what signal(s) may be involved in ELIP expression. My hypothesis is that redox status of the chloroplast photosynthetic electron transport Chain (PETC) and/or chlororespiration may induce ELIP expression. Using the Arabidopsis thaliana immutans (im) chlororespiratory mutant, this hypothesis was tested. Etiolated seedlings of this variegated mutant were subjected to various light intensities over 0-24 hr period and ELIP mRNA levels were analyzed. These were compared with the wild type plants treated in the same manner. It was found that mature thylakoids may not be required for ELIP expression, and that both photoreceptor-dependent and independent components may be involved in ELIP expression.

Environmental stress conditions can drastically affect plant growth and productivity. In contrast to soil moisture or salinity that can gradually change over a period of days or weeks, changes in light intensity or temperature can occur very rapidly, sometimes over the course of minutes or seconds. So, in our study we have taken an metabolomics approach to identify the rapid response of plants to light stress. In the first part we have focused on the ultrafast (0-90 sec) metabolic response of local tissues to light stress and in the second part we analyzed the metabolic response associated with rapid systemic signaling (0-12 min). Analysis of the rapid response of Arabidopsis to light stress has revealed 111 metabolites that significantly alter in their level during the first 90 sec of light stress exposure. We further show that the levels of free and total glutathione accumulate rapidly during light stress in Arabidopsis and that the accumulation of total glutathione during light stress is dependent on an increase in nitric oxide (NO) levels. We further suggest that the increase in precursors for glutathione biosynthesis could be linked to alterations in photorespiration, and that phosphoenolpyruvate could represent a major energy and carbon source for rapid metabolic responses. Taken together, our analysis could be used as an initial road map for the identification of different pathways that could be used to augment the rapid response of plants to abiotic stress. In addition, it highlights the important role of glutathione in initial stage of light stress response. Light-induced rapid systemic signaling and systemic acquired acclimation (SAA) are thought to play an important role in the response of plants to different abiotic stresses. Although molecular and metabolic responses to light stress have been extensively studied in local leaves, and to a lesser degree in systemic leaves, very little is known about the metabolic responses that occur in the different tissues that connect the local to the systemic leaves. These could be important in defining the specificity of the systemic response as well as in supporting the propagation of different systemic signals, such as the reactive oxygen species (ROS) wave. Here we report that local application of light stress to one rosette leaf resulted in a metabolic response that encompassed local, systemic and transport tissues (tissues that connect the local and systemic tissues), demonstrating a high degree of physical and metabolic continuity between different tissues throughout the plant. We further show that the response of many of the systemically altered metabolites could be associated with the function of the ROS wave, and that the level of eight different metabolites is altered in a similar way in all tissues tested (local, systemic, and transport tissues). These compounds could define a core metabolic signature for light stress that propagates from the local to the systemic leaves. Taken together, our findings suggest that metabolic changes occurring in cells that connect the local and systemic tissues could play an important role in mediating rapid systemic signaling and systemic acquired acclimation to light stress.

No description available.

Made available in DSpace on 2015-03-26T13:36:40Z (GMT). No. of bitstreams: 1 texto completo.pdf: 450533 bytes, checksum: d14de51f177a16c047cb046fafb1e533 (MD5) Previous issue date: 2009-02-18 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Mutations in GUN (Genome UNcoupled) genes have helped to understand the pathways of the chloroplast signal transduction that control the expression of nuclear photosynthetic genes. In this context, the gun4 genotype has been characterized in biochemical and molecular aspects. However, little is known whether this mutant produces a phenotype physiological aspects. In order to complement the phenotypic characterization of gun4 genotype, the aim of this study was evaluate the photosynthetic responses of gun4 plants of Arabidopsis thaliana under different levels of light and under light stress. The gun4 plants, in the absence of light stress showed reduced levels of chlorophyll a and b, minimal (Fo) and maximal (Fm) fluorescence in dark-adapted leaves, and quantum yield of unregulated energy dissipation of photosystem II (ΦNO) in 20%, 26%, 18% e 32%, respectively. Also, the effective quantum yield of the FSII (ΦFSII) and photochemical quenching (qL) were 11% and 20% higher, respectively. Higher values of ETR (electrons transport rate), ΦFSII and qL were also found in gun4 under different light intensities, showing that gun4 can maximize the transfer of light energy absorbed in the light harvest complex of FSII for the photosynthetic process. The rate of net carbon assimilation (A) and stomatal conductance (gs) were 43% and 61% lower in gun4 respectively, despite no difference in the flow of absorption of 14CO2 was observed, suggesting that the lower magnitude of photosynthesis is due to resistance to influx of CO2. After 14 hours of light stress, there was no difference in A, and the values of the electron transport rate (ETR) and ΦFSII were reduced by 35% and 50% respectively, both in gun4 and wild type (WT) plants. However, the quantum yield potential of FSII (Fv/Fm) decreased significantly only in WT, suggesting that gun4 may have a greater ability to minimize photoinhibitory effects. These results provide additional evidences for the higher non-photochemical quenching (qN) and quantum yield of regulated energy dissipation of the gun4 FSII (ΦNPQ). Together, the photochemical phenotypes of gun4 may explain the increase (48%) in the photochemical global network connectance after 28 hours of stress, demonstrating the greater ability of this genotype to adjust its photosynthetic apparatus in response to light stress. Finally, the results found in this study demonstrated that gun4 transfers and dissipates more efficiently the excess of light energy absorbed under light stress condition, despite a strong reduction in chlorophyll content. The results suggest the existence of a mechanism for the adjustment of the photosystem in order to offset the reduction of light absorption, indicating the importance of this mutant to explore the mechanisms that control the plasticity of the photochemical protein complex. / Mutações em genes denominados GUN (Genome UNcoupled) têm auxiliado na compreensão das rotas de transdução de sinais do cloroplasto que controlam a expressão de genes fotossintéticos nucleares. Nesse sentido, o genótipo gun4 tem sido caracterizado em aspectos bioquímicos e moleculares, no entanto pouco se sabe sobre aspectos fisiológicos desse mutante. Diante disso, de forma a complementar essa caracterização fenotípica, o objetivo desse trabalho foi avaliar as respostas fotossintéticas do mutante gun4 de Arabidopsis thaliana sob diferentes níveis de luz. Plantas gun4, na ausência de estresse luminoso, apresentaram níveis de clorofila a e b, fluorescências mínima (Fo) e máxima (Fm) de folhas adaptadas ao escuro e rendimento quântico de dissipação não regulada de energia do fotossistema II (ΦNO) reduzidos em 20%, 26%, 18% e 32%, respectivamente, enquanto que o rendimento quântico efetivo do FSII (ΦFSII) e o coeficiente de extinção fotoquímico (qL) foram 11% e 20% maiores, respectivamente. Valores maiores de ETR (taxa de transporte de elétrons), ΦFSII e qL também foram observados em gun4 sob diferentes intensidades luminosas, demonstrando que este fenótipo transfere de forma mais eficiente a energia luminosa absorvida no complexo coletor de luz do FSII para o processo fotossintético. A taxa de assimilação líquida de carbono (A) e a condutância estomática (gs) foram 43% e 61% menores em gun4, respectivamente. Porém, não houve diferença no fluxo de absorção de 14CO2, sugerindo que a menor magnitude de A seja devido a uma restrição difusional ao influxo de CO2. Após 14 h de estresse, não houve diferença em A e os valores da taxa de transporte de elétrons (ETR) e ΦFSII foram reduzidos em 35% e 50%, respectivamente, tanto nas plantas gun4 como no tipo selvagem (WT). Entretanto, o rendimento quântico potencial do FSII (Fv/Fm) decresceu significativamente apenas em WT, sugerindo que gun4 possa minimizar, mais eficientemente, efeitos fotoinibitórios. Isto se evidencia, adicionalmente, nos maiores valores de coeficiente de extinção não fotoquímico (qN) e rendimento quântico de dissipação regulada de energia do FSII (ΦNPQ). Em conjunto, esses fenótipos fotoquímicos de gun4 podem explicar o maior aumento (48%) na conectância global da rede fotoquímica após 28 h de estresse, demonstrando a maior capacidade deste genótipo em ajustar seu aparato fotossintético em resposta ao estresse luminoso. Por fim, os resultados encontrados neste trabalho demonstraram a alta capacidade do genótipo gun4 em dissipar o excesso de energia luminosa absorvida sob estresse e, apesar da forte redução no teor de clorofila, transferir esta mais eficientemente. Isso sugere a existência de um mecanismo de adaptação dos fotossistemas de forma a compensar a redução na absorção de luz, indicando um valor importante deste mutante para explorar os mecanismos que controlam a plasticidade dos complexos protéicos fotoquímicos.

Si bien se ha realizado una intensa selección genética a favor de frutos de girasol con mayor valor agronómico (mayor peso y contenido de materia grasa, resistencia a enfermedades, etc.) sorprende la escasez de información referida a los eventos anatómicos que controlan su crecimiento y madura-ción. La dinámica y el grado de superposición de los procesos de división y aumento de volumen celular y diferenciación de tejidos del ovario y pericarpo, y del óvulo, saco embrionario y semilla, que definen el peso y la estructura final que alcanza el fruto de girasol, no han sido estudiados. Delimitar temporal o fenológicamente dichos procesos resulta información indis-pensable para plantear y comparar trabajos en que se inves-tiguen las relaciones e interacciones entre factores genéticos y ambientales sobre el peso y la estructura final que alcanza el fruto de girasol. Estos parámetros inciden sobre el rendimiento del cultivo y la facilidad con que la cáscara se separa de la pepa, la que puede evaluarse determinando la aptitud al descascarado (AD) de los frutos, durante su indus-trialización. Los objetivos de esta tesis fueron: 1) establecer un modelo de desarrollo histogénico del fruto de girasol en una escala temporal y fenológica, en dos híbridos de similar base genética que diferían en su AD; 2) Analizar las modificaciones que se producen en el patrón de desarrollo de los frutos de tres posiciones del capítulo, al reducir la radiación incidente sobre el cultivo en un 80% mediante sombreado durante la preantesis (Pre-A) y postantesis temprana (Post-A) y 3) Eva-luar el efecto que el genotipo y las modificaciones generadas por los tratamientos de estrés lumínico tienen sobre la AD y parámetros subyacentes de los frutos. El modelo de desarrollo de los frutos fue similar entre híbridos. Entre los estadios reproductivos R2 y R4 se produjo la diferenciación del saco embrionario y de los granos de polen. La división celular en la pared del ovario (futuro pericarpo) fue disminuyendo a partir de R2, no observándose células en división en R4, momento en que quedó fijado el tamaño potencial del pericarpo. El ta-maño final del pericarpo se estableció al completarse la escle-rificación y acumulación de materia seca del mismo, entre 10 y 13 días después de la antesis (DDA). El tamaño potencial del embrión se fijó 18 DDA cuando ya había transcurrido la etapa inicial del período de rápida acumulación de materia seca del embrión. Este período se extendió desde 11 DDA hasta, aproximadamente, 29 a 32 DDA cuando se registró el máximo peso del embrión y la madurez fisiológica (MF) del fruto. El sombreado de Pre-A redujo el período de acumulación de materia seca del pericarpo de los frutos de la posición interna del capítulo. El sombreado de Post-A también modificó la dinámica de crecimiento del pericarpo. Así, el peso del pericarpo (PP) de los frutos medios e internos se mantuvo constante desde el inicio del tratamiento hasta 4-7 días después de finalizado el mismo. Luego de ello, el PP se incrementó durante 3 a 4 días. En MF, la reducción en el PP (18 a 52%) y en el espesor (20-33%) de la capa media (ECM) del mismo fue similar entre tratamientos de sombreado. El sombreado de Pre-A redujo el número de estratos (NTE) e incrementó el grosor de las paredes celulares de la CM del pericarpo en la posición media e interna del capítulo. Por el contrario, el tratamiento de Post-A redujo el número de estratos esclerificados de la CM (NEECM) y el espesor y el contenido de celulosa, hemicelulosa y lignina de las paredes celulares del pericarpo en las tres posiciones del capítulo. En MF, el peso de los embriones (PE) de los frutos del sombreado de Pre-A, fue inferior al tratamiento control (Con) y el de los frutos de Post-A fue igual al de los frutos del control. Ello estuvo asociado a la reducción del número de células de los cotiledones de los frutos en las tres posiciones del capítulo, la duración del periodo de llenado de los embriones (PLLE) de los frutos externos, la tasa de crecimiento de los embriones (TCE) de los frutos medios y la TCE y el PLLE de los frutos en la posición interna en el sombreado de Pre-A. El sombreado de Post-A solo prolongó la duración de la fase de lento crecimiento del embrión (fase lag) de los frutos medios e internos. La reducción en el rendimiento por planta fue similar en ambos tratamientos de sombreado, variando, con respecto al Con, los componentes del rendimiento a través de los cuales se realizó el ajuste. Las plantas sombreadas en Pre-A presentaron el mismo número de frutos llenos, pero el peso individual de los mismos fue inferior al Con. Por el contrario, en el sombreado de Post-A se redujo el número de frutos llenos por planta, mientras que su peso individual fue similar al Con. La heterogeneidad entre los frutos de las distintas posiciones del capítulo no fueron modi-ficadas por los tratamientos de sombreado, y estuvieron asociadas a diferencias, en el número de células de los cotile-dones, excepto entre los frutos medios e internos del trata-miento de Pre-A, y en la TCE, excepto entre los frutos externos y medios del tratamiento de Post-A. Las variaciones en la AD de los frutos resultaron tanto consecuencia de dife-rencias entre híbridos como de las condiciones de manejo del cultivo (sombreado). Así, la frecuencia de radios de parén-quima por mm de sección transversal de pericarpo, que determinó las diferencias en la AD entre híbridos, se mantuvo, salvo ligeras variantes, constante entre localidades. La menor AD observada en el sombreado de Post-A, con respecto al tratamiento Con y al sombreado de Pre-A, estuvo asociada a la reducción en el grosor de las paredes celulares del pericarpo. Los resultados de esta tesis permiten avanzar en el marco actual de conocimiento sobre los procesos involucrados en la determinación de la estructura y peso final que alcanza el fruto de girasol, componente que incide sobre el rendi-miento del cultivo y la calidad de los subproductos obtenidos durante la industrialización de sus frutos. Además, constitu-yen el primer aporte que analiza los efectos del estrés lumínico (sombreado) sobre los cambios morfo-anatómicos asociados al crecimiento y desarrollo del pericarpo y/o embrión de girasol, así como, sobre las variaciones en la AD de sus frutos. Palabras clave: girasol, Helianthus annuus L., tamaño potencial del pericarpo, tamaño final del fruto, aptitud al descascarado, estrés lumínico. / In spite of the intense genetic selection to obtain sunflower fruits with a higher agronomic value (higher weight and oil content, resistance to diseases, etc.), there is a surprising lack of information on the developmental events that control their growth and maturity. No one has studied the dynamics and overlapping degree of the cell division and volume increase processes and the differentiation of ovary, pericarp, ovule, embryo sac and seed tissues which define the final weight and structure of the sunflower fruit. Understanding the timing and phenological definition of these processes is essential to proposing and comparing research on the relations and interactions between genetic and environmental factors determining final weight and structure (volume, weight and anatomic structure) of the sunflower fruit. These last variables have an impact on the crop yield and on the ability of the hull to separate from the seed, which can be assessed by establishing the dehulling ability (DA) of fruits during processing. The objectives of this thesis are: 1) to establish a histogenic development model, on both a time and a phenological scale, of the fruit of two sunflower hybrids with similar genetic background but with different fruit DA; 2) to analyze the modifications of the fruit development pattern, in three positions of the capitulum, with 80% reduction of incident radiation (shading) on the crop during preanthesis (Pre-A) and early postanthesis (Post-A); and 3) to assess the effect of genotype and light stress on the fruit DA and underlying parameters. The fruit development model was similar between hybrids. Differentiation of the embryo sac and the pollen grains took place between stages R2 and R4. The ovary wall (future pericarp) cell division decreased after R2, with no dividing cells found at R4, when the potential size of the pericarp was determined. The final size of the pericarp was established between 10 and 13 days after anthesis (DAA) upon completion of its sclerification and accumulation of dry matter. The embryo potential size was established by 18 DAA, when the initial stage of rapid dry matter accumulation of the embryo had already passed. The last stage lasted from the 11 DAA until about 29 to 32 DAA when the maximum weight of the embryo (MF) was recorded. Pre-A shading decreased the dry matter accumulation period of the pericarp in the capitulum central position. Post-A shading also modified the pericarp growth dynamics. In this way, the pericarp weight (PW) of the mid and central fruits remained constant during the treatment and for 4-7 days after it was completed. After that the PW increased during 3 to 4 days. At MF, the PW reduction (18 to 52%) and thickness (20-33%) of the middle layer (ML) of the pericarp was similar in both shading treatments. In the Pre-A treatment, the number of strata decreased and the thickness of the cell wall of the ML increased in the capitulum mid and central positions. On the contrary, the Post-A treatment decreased the number of sclerified strata of the ML, as well as the thickness and the content of cellulose, hemicellulose and lignin of the pericarp cell wall of the fruits from the three positions on the capitulum. At PM, the weight of the embryos (EW) from the Pre-A shade was lower than the EW of the Con and Post-A treatment fruits. The EW reduction was associated with the reduction in the cotyledon cells number at the three capitulum positions, the duration of the embryo filling period (EFP) of the peripheral fruits, the embryo growth rate (EGR) of mid fruits and the EGR and the EEP of central fruits from Pre-A shading. The Post-A shading extended the slow growth phase duration of the embryo (lag phase) of the central and mid fruits only. The reduction of yield per plant, with respect to Con, was similar in both shading treatments. The yield components responsible for the adjustments depended on the treatment. Pre-A shaded plants produced the same number of filled fruits but with a lower weight per fruit. On the contrary, in Post-A shading the number of filled fruits decreased while their individual weight was similar to Con. The heterogeneity among fruits in the different capitulum positions was not modified by the shading treatments, and it was associated to differences in cotyledons cell number, except for the mid and central fruits of the Pre-A treatment, and to EGR, except between peripheral and mid fruits of the Post-A treatment. Variations in the DA were both a consequence of the differences between hybrids and the crop management conditions (shading). Thus the frequency of parenchyma radii by mm of cross section of the pericarp, that determined the DA differences among hybrids, remained the same, except for slight variants, throughout locations. The lowest EW/PW ratio observed in Post-A shading with respect to Con treatment and the Pre-A shading, resulted from the PW reduction and not from EW, which was associated with a reduction in cell wall thickness and in the DA.The results of this thesis are the first contribution that analyzed the effects of light stress (shading) on the morphological and anatomical changes associated to growth and development of the sunflower pericarp and embryo. They improve our understanding of the processes involved in the determination of the structure and finalweight of the sunflower fruit, a component that impacts crop yield and also the quality of the byproducts of fruit processing. Keywords: sunflower, Helianthus annuus L., pericarp potential size, fruit final size, dehulling ability, light stress.