Orientador: Renato de Mello Prado / Abstract: Global warming is predicted to increase the intensity and duration of extreme weather events, such as droughts, heat waves, and floods, especially in tropical regions. Climate change affect growth of forage species. However, information regarding the effects of global climate change on the nutritional performance of tropical pastures is lacking, especially under field conditions. We, thus, conducted two field experiment with Panicum maximum and Stylosanthes capitata using a temperature free-air controlled enhancement system and evaluated the effects of two temperature conditions, ambient temperature and moderate warming (2°C above ambient canopy temperature), and two levels of water availability, irrigated and non-irrigated, on nutrients accumulation, nutrient use efficiency (NUE), the stoichiometric patterns of C:N:P and leaf biomass production. Both experiments was conducted using a randomized complete block design in a factorial arrangement. Our findings revealed in plants of P. maximum (C4- grass) that the N and P leaf concentration greatly decreased under water-stressed, which increased the C:N and C:P ratios, while warming increased the N:P ratio. Leaf biomass production was impaired by up to 16% under water stress and ambient temperature conditions, but the biomass production was improved by 20% under warming and irrigated conditions. Our results also showed that homeostatic instability under rainfed conditions resulted in decreased leaf biomass production, and it was ... (Complete abstract click electronic access below) / Resumo: Prevê-se que o aquecimento global aumente a intensidade e a duração dos eventos climáticos extremos, como secas, ondas de calor e inundações, especialmente nas regiões tropicais. Mudanças climáticas afetam o crescimento de espécies forrageiras. No entanto, faltam informações sobre os efeitos das mudanças climáticas globais no desempenho nutricional de pastagens tropicais, especialmente em condições de campo. Nós, assim, conduzimos dois experimento em campos com as forrageiras Panicum maximum e Stylosanthes capitata utilizando um sistema de temperatura controlada do aquecimento do ar (T-Face) e avaliou-se os efeitos de duas condições de temperatura, (temperatura ambiente) e aquecimento moderado (2°C acima da temperatura ambiente) e dois níveis de disponibilidade hídrica, (irrigada e não irrigados), no acúmulo de nutrientes, eficiência de uso de nutrientes (NUE), nos padrões estequiométricos de C:N:P e na produção de biomassa foliar. Ambos experimentos foram conduzidos utilizando um delineamento de blocos completos casualizados em arranjo fatorial. Nossos resultados revelaram que em plantas de P. maximum (pastagem C4) a concentração foliar de N e P diminuiu sob estresse hídrico, o que aumentou as relações C:N e C:P, enquanto o aquecimento aumentou a relação N:P. A produção de biomassa foliar foi prejudicada em até 16% sob condições de estresse hídrico e temperatura ambiente, mas a produção de biomassa foi melhorada em 20% sob condições de aquecimento e irrigação. Nossos resulta... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

Les changements climatiques globaux sont responsables de l'augmentation de la fréquence des épisodes de sécheresse et de fortes concentrations d'ozone troposphérique, qui peuvent se produire simultanément pour réduire la croissance des plantes en limitant la production agricole et affectant la fourniture de nourriture pour une population mondiale croissante. Ces contraintes environnementales peuvent conduire à la surproduction d'espèces réactives de l'oxygène en favorisant le stress oxydatif et provoquant un déséquilibre métabolique. Les membranes cellulaires sont les premières cibles des dommages induits par le stress et la préservation de l'intégrité cellulaire par le remodelage des lipides membranaires est essentielle pour la survie des plantes dans des conditions défavorables. L'objectif de cette étude était d'étudier les effets de la sécheresse, de l'ozone et des contraintes combinées chez deux cultivars de haricot niébé dotés de différents degrés de tolérance à la sécheresse: EPACE-1 et IT83-D par des réponses physiologiques et par une étude portée sur la teneur et la composition des lipides de membrane, ainsi que dans l'expression de gènes liés à la biosynthèse et la dégradation des ces lipides. La sécheresse et l'ozone (120 ppb), seuls ou en combinaison, ont été appliqués aux plantes après trois semaines de germination pendant deux semaines. Après 7 et 14 jours de traitements, des paramètres physiologiques ont été déterminés. Les lipides extraits de feuilles ont été analysés par chromatographie en phase gazeuse/spectrométrie de masse. L'accumulation de transcrits (VuMGD1, VuMGD2, VuDGD1, VuDGD2, VuCLS, VuFAD7, VuFAD8, VuPLD1 et VuPAT1) a été détecté chez les feuilles par PCR en temps réel. Chez les deux cultivars, la sécheresse a limité la croissance et la photosynthèse par la fermeture des stomates, ce qui a semblé impliquer l'expression génétique de la phospholipase D (VuPLD1). L'ozone a causé des dommages foliaires et de la dégradation des galactolipides, principalement chez IT83-D. Chez ce cultivar, les dommages ont impliqué dans l'augmentation du ratio DGDG:MGDG et de l'expression de VuDGD2. De plus, une diminution partielle de la teneur en phospholipides en réponse à l'ozone a été associée à une augmentation de l'expression de VuPLD1, ce qui suggère un remplacement des phospholipides par DGDG. Contrairement, le cultivar EPACE-1 en présence d'ozone, en même temps que la dégradation des galactolipides, a montré une augmentation de la teneur en lipides de réserve et en DPG (cardiolipine). La combinaison des contraintes a conduit à des résultats similaires à ceux de la sécheresse appliquée séparément, chez les deux cultivars, ce qui reflète probablement la forte restriction de l'absorption de l'ozone en raison de la fermeture des stomates induite par la sécheresse. En conclusion, les présents résultats mettent en évidence le rôle important des lipides membranaires dans les ajustements métaboliques qui se produisent pour régler la tolérance des plantes à la sécheresse et à l'ozone / Global climate changes are responsible for the increased frequency of episodes of drought and high concentrations of tropospheric ozone, which can occur simultaneously to reduce plant growth, limiting agricultural production and affecting the food supply for an increasing world population. These environmental stresses can lead to the overproduction of reactive oxygen species, promoting oxidative stress and causing metabolic imbalance. Cell membranes are the primary targets of damage induced by stresses and the preservation of cell integrity through remodeling of membrane lipids is essential for plant survival under adverse conditions. The objective of this study was to investigate the effects of drought, ozone and the combined stresses in two cowpea cultivars with different degrees of drought tolerance: EPACE-1 and IT83-D through physiological responses and a study focused on the content and composition of membrane lipids, as well as on the expression of genes related to the biosynthesis and degradation of these lipids. Drought and ozone (120 ppb), isolated or in combination, were applied on three-week-old seedlings during two weeks. After 7 and 14 days of treatments, physiological parameters were determined. Lipids extracted from leaves were analyzed by gas chromatography/mass spectrometry. Transcript accumulation (VuMGD1, VuMGD2, VuDGD1, VuDGD2, VuCLS, VuFAD7, VuFAD8, VuPLD1 and VuPAT1) was detected in leaves by real time PCR. In both cultivars, drought inhibited the plant growth and photosynthesis through stomatal closure, which appeared to involve the gene expression of phospholipase D (VuPLD1). Ozone caused foliar injury and degradation of galactolipids, mainly in IT83-D. In this cultivar, the damage resulted in an increase in DGDG:MGDG ratio and in VuDGD2 expression. Furthermore, a partial decrease in the phospholipids content in response to ozone was associated with an increase in VuPLD1 expression, suggesting a replacement of phospholipids by DGDG. Differently, EPACE-1 cultivar in the presence of ozone, concomitantly with the degradation of galactolipids, showed an increase in storage lipids and DPG (cardiolipin) contents. The two stresses in combination led to results similar to those observed in response to the drought treatment alone, in both cultivars, which likely reflected the severe restriction of ozone uptake resulting from drought-induced stomatal closure. In conclusion, the present results highlight the prominent role of membrane lipids in the metabolic adjustments that occur for the plant tolerance to drought and ozone

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Global climate changes are responsible for the increased frequency of episodes of drought and high concentrations of tropospheric ozone, which can occur simultaneously reducing plant growth, limiting agricultural production and affecting the food supply for an increasing world population. These environmental stresses can lead to the overproduction of reactive oxygen species, promoting oxidative stress and causing metabolic imbalance. Cell membranes are the primary targets of damage induced by stresses and the preservation of cell integrity through remodeling of membrane lipids is essential for plant survival under adverse conditions. The objective of this study was to investigate the effects of drought, ozone and the combined stresses in two cowpea cultivars with different degrees of drought tolerance: EPACE-1 (tolerant) and IT83-D (susceptible) through physiological responses and a study focused on the content and composition of membrane lipids, as well as on the expression of genes related to the biosynthesis and degradation of these lipids. Drought and ozone (120 ppb), isolated or in combination, were applied on three-week-old seedlings during two weeks. After 7 and 14 days of treatments, physiological parameters were determined. Lipids extracted from leaves were analyzed by gas chromatography/mass spectrometry. Transcript accumulation (VuMGD1, VuMGD2, VuDGD1, VuDGD2, VuCLS, VuFAD7, VuFAD8, VuPLD1 and VuPAT1) was detected in leaves by real time PCR. In both cultivars, drought inhibited the plant growth and photosynthesis through stomatal closure, which appeared to involve the gene expression of phospholipase D (VuPLD1). Ozone caused foliar injury and degradation of galactolipids, mainly in IT83-D. In this cultivar, the damage resulted in an increase in DGDG:MGDG ratio and in VuDGD2 expression. Furthermore, a partial decrease in the phospholipids content in response to ozone was associated with an increase in VuPLD1 expression, suggesting a replacement of phospholipids by DGDG. Differently, EPACE-1 cultivar in the presence of ozone, concomitantly with the degradation of galactolipids, showed an increase in storage lipids and DPG (cardiolipin) contents. The two stresses in combination led to results similar to those observed in response to the drought treatment alone, in both cultivars, which likely reflected the severe restriction of ozone uptake resulting from drought-induced stomatal closure. In conclusion, the present results highlight the prominent role of membrane lipids in the metabolic adjustments that occur for the plant tolerance to drought and ozone. / As mudanÃas climÃticas globais sÃo responsÃveis pelo aumento da frequÃncia de episÃdios de seca e de elevadas concentraÃÃes de ozÃnio troposfÃrico, que podem ocorrer simultaneamente, reduzindo o crescimento das plantas, limitando a produÃÃo agrÃcola e afetando o suprimento de alimentos para a crescente populaÃÃo mundial. Esses estresses ambientais podem levar à superproduÃÃo de espÃcies reativas de oxigÃnio, promovendo o estresse oxidativo e causando desequilÃbrio metabÃlico. As membranas celulares sÃo os primeiros alvos de danos induzidos por estresses e a preservaÃÃo da integridade celular atravÃs da remodelaÃÃo dos lipÃdios de membrana à essencial para a sobrevivÃncia da planta sob condiÃÃes adversas. O objetivo deste trabalho foi investigar os efeitos da seca, do ozÃnio e dos estresses combinados em dois cultivares de feijÃo-de-corda dotados de diferentes graus de tolerÃncia à seca: EPACE-1 (tolerante) e IT83-D (sensÃvel) atravÃs de respostas fisiolÃgicas e de um estudo focado no teor e na composiÃÃo de lipÃdios de membrana, assim como na expressÃo de genes relacionados à biossÃntese e à degradaÃÃo desses lipÃdios. A seca e o ozÃnio (120 ppb), isolados ou combinados, foram aplicados nas plantas apÃs trÃs semanas de germinaÃÃo, durante duas semanas. ApÃs 7 e 14 dias de tratamentos, parÃmetros fisiolÃgicos foram determinados. Os lipÃdios extraÃdos de folhas foram analisados por cromatografia gasosa/espectrometria de massa. O acÃmulo de transcritos (VuMGD1, VuMGD2, VuDGD1, VuDGD2, VuCLS, VuFAD7, VuFAD8, VuPLD1 e VuPAT1) foi detectado em folhas por PCR em tempo real. Em ambos os cultivares, a seca inibiu o crescimento e a fotossÃntese atravÃs do fechamento estomÃtico, o que pareceu envolver a expressÃo gÃnica da fosfolipase D (VuPLD1). O ozÃnio causou injÃria foliar e degradaÃÃo de galactolipÃdios, principalmente em IT83-D. Neste cultivar, os danos implicaram no aumento da relaÃÃo DGDG:MGDG e da expressÃo de VuDGD2. AlÃm disso, uma diminuiÃÃo parcial no teor de fosfolipÃdios em resposta ao ozÃnio foi associada ao aumento da expressÃo de VuPLD1, sugerindo-se uma substituiÃÃo de fosfolipÃdios por DGDG. Diferentemente, o cultivar EPACE-1 em presenÃa de ozÃnio, concomitantemente com a degradaÃÃo de galactolipÃdios, mostrou um aumento nos teores de lipÃdios de reserva e de DPG (cardiolipina). A combinaÃÃo dos estresses levou a resultados semelhantes aos encontrados com a seca aplicada isoladamente, em ambos os cultivares, o que provavelmente reflete a forte restriÃÃo da absorÃÃo de ozÃnio devido ao fechamento dos estÃmatos induzido pela seca. Em conclusÃo, os presentes resultados ressaltam o proeminente papel dos lipÃdios de membrana nos processos de ajustamento metabÃlico que ocorrem para regular a tolerÃncia das plantas à seca e ao ozÃnio.

REBOUÇAS, Deborah Moura. Estudo do metabolismo de lipídios de membrana e da expressão dos genes relacionados em cultivares de Vigna unguiculata (L.) walp sob estresses abióticos combinados (seca/ozônio). 2014. 162 f. Tese (Doutorado em Bioquímica)-Universidade Federal do Ceará, Fortaleza, 2014. / Submitted by Vitor Campos (vitband@gmail.com) on 2016-09-01T23:43:36Z No. of bitstreams: 1 2014_tese_dmreboucas.pdf: 4151452 bytes, checksum: 62ddcc5c8a9f5244cb43fd01dc410a57 (MD5) / Approved for entry into archive by Jairo Viana (jairo@ufc.br) on 2016-09-05T20:07:49Z (GMT) No. of bitstreams: 1 2014_tese_dmreboucas.pdf: 4151452 bytes, checksum: 62ddcc5c8a9f5244cb43fd01dc410a57 (MD5) / Made available in DSpace on 2016-09-05T20:07:49Z (GMT). No. of bitstreams: 1 2014_tese_dmreboucas.pdf: 4151452 bytes, checksum: 62ddcc5c8a9f5244cb43fd01dc410a57 (MD5) Previous issue date: 2014 / Global climate changes are responsible for the increased frequency of episodes of drought and high concentrations of tropospheric ozone, which can occur simultaneously reducing plant growth, limiting agricultural production and affecting the food supply for an increasing world population. These environmental stresses can lead to the overproduction of reactive oxygen species, promoting oxidative stress and causing metabolic imbalance. Cell membranes are the primary targets of damage induced by stresses and the preservation of cell integrity through remodeling of membrane lipids is essential for plant survival under adverse conditions. The objective of this study was to investigate the effects of drought, ozone and the combined stresses in two cowpea cultivars with different degrees of drought tolerance: EPACE-1 (tolerant) and IT83-D (susceptible) through physiological responses and a study focused on the content and composition of membrane lipids, as well as on the expression of genes related to the biosynthesis and degradation of these lipids. Drought and ozone (120 ppb), isolated or in combination, were applied on three-week-old seedlings during two weeks. After 7 and 14 days of treatments, physiological parameters were determined. Lipids extracted from leaves were analyzed by gas chromatography/mass spectrometry. Transcript accumulation (VuMGD1, VuMGD2, VuDGD1, VuDGD2, VuCLS, VuFAD7, VuFAD8, VuPLD1 and VuPAT1) was detected in leaves by real time PCR. In both cultivars, drought inhibited the plant growth and photosynthesis through stomatal closure, which appeared to involve the gene expression of phospholipase D (VuPLD1). Ozone caused foliar injury and degradation of galactolipids, mainly in IT83-D. In this cultivar, the damage resulted in an increase in DGDG:MGDG ratio and in VuDGD2 expression. Furthermore, a partial decrease in the phospholipids content in response to ozone was associated with an increase in VuPLD1 expression, suggesting a replacement of phospholipids by DGDG. Differently, EPACE-1 cultivar in the presence of ozone, concomitantly with the degradation of galactolipids, showed an increase in storage lipids and DPG (cardiolipin) contents. The two stresses in combination led to results similar to those observed in response to the drought treatment alone, in both cultivars, which likely reflected the severe restriction of ozone uptake resulting from drought-induced stomatal closure. In conclusion, the present results highlight the prominent role of membrane lipids in the metabolic adjustments that occur for the plant tolerance to drought and ozone. / As mudanças climáticas globais são responsáveis pelo aumento da frequência de episódios de seca e de elevadas concentrações de ozônio troposférico, que podem ocorrer simultaneamente, reduzindo o crescimento das plantas, limitando a produção agrícola e afetando o suprimento de alimentos para a crescente população mundial. Esses estresses ambientais podem levar à superprodução de espécies reativas de oxigênio, promovendo o estresse oxidativo e causando desequilíbrio metabólico. As membranas celulares são os primeiros alvos de danos induzidos por estresses e a preservação da integridade celular através da remodelação dos lipídios de membrana é essencial para a sobrevivência da planta sob condições adversas. O objetivo deste trabalho foi investigar os efeitos da seca, do ozônio e dos estresses combinados em dois cultivares de feijão-de-corda dotados de diferentes graus de tolerância à seca: EPACE-1 (tolerante) e IT83-D (sensível) através de respostas fisiológicas e de um estudo focado no teor e na composição de lipídios de membrana, assim como na expressão de genes relacionados à biossíntese e à degradação desses lipídios. A seca e o ozônio (120 ppb), isolados ou combinados, foram aplicados nas plantas após três semanas de germinação, durante duas semanas. Após 7 e 14 dias de tratamentos, parâmetros fisiológicos foram determinados. Os lipídios extraídos de folhas foram analisados por cromatografia gasosa/espectrometria de massa. O acúmulo de transcritos (VuMGD1, VuMGD2, VuDGD1, VuDGD2, VuCLS, VuFAD7, VuFAD8, VuPLD1 e VuPAT1) foi detectado em folhas por PCR em tempo real. Em ambos os cultivares, a seca inibiu o crescimento e a fotossíntese através do fechamento estomático, o que pareceu envolver a expressão gênica da fosfolipase D (VuPLD1). O ozônio causou injúria foliar e degradação de galactolipídios, principalmente em IT83-D. Neste cultivar, os danos implicaram no aumento da relação DGDG:MGDG e da expressão de VuDGD2. Além disso, uma diminuição parcial no teor de fosfolipídios em resposta ao ozônio foi associada ao aumento da expressão de VuPLD1, sugerindo-se uma substituição de fosfolipídios por DGDG. Diferentemente, o cultivar EPACE-1 em presença de ozônio, concomitantemente com a degradação de galactolipídios, mostrou um aumento nos teores de lipídios de reserva e de DPG (cardiolipina). A combinação dos estresses levou a resultados semelhantes aos encontrados com a seca aplicada isoladamente, em ambos os cultivares, o que provavelmente reflete a forte restrição da absorção de ozônio devido ao fechamento dos estômatos induzido pela seca. Em conclusão, os presentes resultados ressaltam o proeminente papel dos lipídios de membrana nos processos de ajustamento metabólico que ocorrem para regular a tolerância das plantas à seca e ao ozônio.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O feijÃo-de-corda (Vigna unguiculata) à uma das principais leguminosas cultivadas no Brasil. Entretanto, devido à presenÃa de fatores biÃticos e abiÃticos sÃo geradas perdas significativas na produtividade desta cultura. O VÃrus do Mosaico Severo do Caupi (CPSMV) tem atacado com grande frequÃncia os cultivos brasileiros. AlÃm disso, devido o feijÃo-de-corda ser cultivado em regiÃes onde os Ãndices pluviomÃtricos sÃo baixos, a deficiÃncia hÃdrica à bastante comum. Esse estresse promove impacto negativo substancial no crescimento e desenvolvimento das plantas, resultando em diminuiÃÃo na produÃÃo de biomassa. Em condiÃÃes de campo, os estresses mencionados acima podem ocorrer tanto individualmente quanto simultaneamente. Portanto, o presente estudo teve como objetivo estudar as respostas do feijÃo-de-corda (genÃtipo CE-31, sin. PitiÃba) submetido ao estresse hÃdrico, infectado com o CPSMV e sob pressÃo dos dois estresses combinados. Sob essas condiÃÃes, a cinÃtica enzimÃtica de proteÃnas antioxidantes (superÃxido dismutase, catalase e ascorbato peroxidase) e PR-proteÃnas (glucanase, quitinase e guaiacol peroxidase), bem como a dosagem de perÃxido de hidrogÃnio e quantificaÃÃo do CPSMV em folhas de feijÃo-de-corda foram realizadas. AlÃm disso, proteÃnas diferencialmente acumuladas em folhas de feijÃo-de-corda submetidas à esses estresses foram quantificadas e identificadas por LC-MS/MS e bancos de dados disponÃveis. A infecÃÃo viral nÃo alterou a maioria dos parÃmetros fisiolÃgicos. Jà a deficiÃncia hÃdrica contribuiu para a diminuiÃÃo da fotossÃntese lÃquida, transpiraÃÃo e condutÃncia estomÃtica. Atà 6 dias apÃs inoculaÃÃo do CPSMV, o vÃrus nÃo agravou os efeitos negativos da seca. Os resultados da cinÃtica enzimÃtica revelaram que as respostas sÃo alteradas de formas diferentes frente aos estresses quando impostos individualmente ou simultaneamente e que a presenÃa do estresse hÃdrico favoreceu a infecÃÃo viral apÃs 2 dias de inoculaÃÃo. A anÃlise proteÃmica revelou 117 proteÃnas diferencialmente acumuladas pertencentes a diferentes categorias de acordo com as funÃÃes fisiolÃgicas: energia e metabolismo, fotossÃntese, metabolismo de proteÃnas, homeostase redox, fatores de regulaÃÃo e processamento de RNA, resposta ao estresse, defesa de plantas e outras. Estes resultados sÃo importantes para a compreensÃo dos mecanismos celulares que atuam no feijÃo-de-corda, genÃtipo CE-31, quando submetido ao estresse hÃdrico, à infecÃÃo pelo CPSMV e a ambos os estresses de forma combinada. / Cowpea (Vigna unguiculata) is a legume (Fabaceae) with high nutritional value grown in Brazil. However, biotic and abiotic factors are responsible for causing significant losses to the growth and yield of leguminous plants. Cowpea mosaic severe virus (CPSMV) is one of the more severe problems affecting cowpea production in Brazil. Also, cowpea are mainly grown in regions where rainfall is low, thus water deficit is quite common. This stress promotes substantial negative impact on growth and development of plants, resulting in decreased of biomass production. Under field conditions, the stresses mentioned above can occur both individually and simultaneously. Therefore, this work aimed to study the responses of cowpea (CE-31 genotype, syn. Pitiuba) subjected to water deficit, infected with CPSMV and under pressure of the two combined stresses. Under these conditions, enzyme kinetics of antioxidant protein (superoxide dismutase, catalase and ascorbate peroxidase) and PR-proteins (glucanase, chitinase and guaiacol peroxidase) as well as hydrogen peroxide dosage and quantification of CPSMV in cowpea leaves were performed. In addition, differentially accumulated proteins from cowpea leaves subjected to these stresses were identified and quantified by LC-MS/MS. Viral infection did not affect the majority of physiological parameters. Already water deficit contributed to the decrease in net photosynthesis, transpiration and stomatal conductance. At 6 days after CPSMV inoculation, the virus did not worsen the negative effects of drought. The results revealed that the enzyme kinetics of the responses was changed across different ways when the stress imposed individually or simultaneously and the presence of water deficit favored viral infection after 2 days of CPSMV inoculation. The proteomic analysis revealed 117 differentially accumulated proteins belonging to different categories according to the physiological functions: energy and metabolism, photosynthesis, protein metabolism, redox homeostasis, regulation factors and RNA processing, response to stress, plant defense and others. These results are important for understanding the cellular mechanisms that operate in cowpea (CE-31 genotype), when subjected to water deficit, CPSMV infection and both combined stresses.

La carotte présente un grand intérêt nutritionnel comme source alimentaire en caroténoïdes. Pourtant, la connaissance des mécanismes d’accumulation de ces composés est un enjeu majeur. Si le déterminisme génétique a été relativement bien étudié, l’impact de stress sur l’accumulation des caroténoïdes chez la carotte reste méconnu. Ce travail de thèse vise donc à déterminer (i) l’impact de stress biotique et abiotique appliqués individuellement ou en combinaison sur les teneurs en caroténoïdes dans les feuilles et racines de carotte,(ii) les mécanismes de régulation pouvant expliquer ces variations (iii) si le métabolisme secondaire est affecté spécifiquement, indépendamment du métabolisme primaire.Les résultats mettent en évidence un effet négatif des différentes conditions de stress, en particulier appliquées en combinaison, sur les teneurs en caroténoïdes dans les feuilles et les racines de carotte, mais dépendent du génotype. La régulation transcriptionnelle au niveau de la voie de biosynthèse des caroténoïdes ne peut expliquer qu’en partie les variations de teneurs. Les variations de teneurs en chlorophylles des feuilles et sucres des racines sont corrélées à celles des teneurs en caroténoïdes, suggérant des mécanismes communs de régulation.Ce travail montre que l’impact de stress en culture, et en particulier leur combinaison, est une composante importante de l’élaboration de la qualité nutritionnelle. Les travaux doivent être poursuivis afin d’établir un schéma plus précis de la régulation de l’accumulation des caroténoïdes chez la carotte. / Carrot presents a high nutritional interest as a carotenoid intake source. However, knowledge about accumulation mechanisms of these compounds is a major issue. While genetic determinism was relatively well studied, the impact of stresses on carotenoid accumulation in carrot remains unknown. This thesis work aims to determine (i) the impact of biotic and abiotic stresses applied individually or in combination on carotenoid contents in carrot leaves and roots, (ii) the regulation mechanisms which could explainthese variations and (iii) if secondary metabolism is specifically affected independently from primarymetabolism. Results bring to light a negative effect of the different stress conditions, particularly applied in combination, on carotenoid contents in carrot leaves and roots but depend on genotypes. Transcriptional regulation based on carotenoid biosynthetic genes can only partially explain contentvariations. Chlorophyll content variations in leaves and sugar content variations in roots are correlated to those of carotenoids suggesting common regulation mechanisms. This work shows that the impact of stress on culture, and particularly in combination, is an important determinism of nutritional quality. Further works need to be performed to establish a more precise regulation network pattern of carotenoid accumulation in carrot.