Ce travail de thèse s'inscrit dans un projet régional, initié par le Parc du Lubéron et en collaboration avec le Laboratoire de Pharmacognosie et d'Ethnopharmacologie de l’Université de Marseille. Il avait pour objectif d'étudier la possibilité de domestiquer une plante sauvage, Inula montana L. (Asteraceae) connue dans la pharmacopée provençale pour ses effets anti traumatiques semblables à ceux d'Arnica montana L. et de la proposer comme nouvelle production agricole. Inula montana produit notamment des lactones sesquiterpènes, identifiées comme les métabolites secondaires responsables de ses aptitudes biologiques anti inflammatoires. Il s’est agit dans cette étude1) de déterminer les caractéristiques de croissance et de développement de la plante sauvage largement inconnue et d’identifier en conditions naturelles les facteurs les plus favorables à la production de métabolites secondaires, 2)d’étudier sa capacité à se multiplier végétativement in vitro et à former des cultures cellulaires aptes à synthétiser les molécules d’intérêt 3), de proposer un itinéraire technique applicable agronomiquement et 4) de tester les effets de divers facteurs environnementaux (fertilisation, apport de NaCl, modification du rythme circadien de l’éclairement,rayonnement UVB, ablation de feuilles, application de méthyl jasmonate ) sur la production qualitative et quantitative des lactones sesquiterpènes et des composés phénoliques. Les traits phénologiques de la plante sauvage sont impactés par l’altitude qui induit un retard dans la croissance végétative et la phase reproductrice ainsi que des modifications physiologiques et morphologiques. Les teneurs en métabolites secondaires (certaines lactones sesquiterpènes, les polyphénols totaux et flavonoïdes) varient en fonction de la saison et sont plus importantes dans le site qui présente les conditions climatiques les plus contraignantes du point de vue hydrique (sol drainant,température plus élevée et présence d’une période sèche en été). L’observation microscopique a indiqué la présence de deux types de trichomes : glandulaires (bisériés) et non glandulaires (des poils) qui sont potentiellement les structures porteuses des molécules d’intérêt. I. montana est apte à former des cals in vitro à partir d’explants racinaires, foliaires et caulinaires sur lesquels des pousses feuillées se forment. La domestication d’Inula a été réussie à partir de semences issues des plantes sauvages et en conditions agronomiques, les teneurs en lactones sesquiterpènes (costunolide, artémorine, eldarine et hydrocostunolide) et en composés phénoliques sont généralement plus élevées que chez les plantes sauvages. Les différentes contraintes appliquées pour tester les effets des facteurs environnementaux sur la production des métabolites ont montré : 1) qu’il ne peut pas être établi de corrélation entre la présence d’un stress oxydatif et une augmentation des teneurs en métabolites chez Inula 2) que l’accumulation des lactones et composés phénoliques semblent principalement favorisée lorsque la plante dispose d’un surplus de squelettes carbonés, non utilisés pour la croissance 3) enfin, les deux conditions les plus favorables à l’accumulation des métabolites chez Inula, sont : dans les feuilles, une alternance rapide de lumière et d’obscurité durant la photopériode et dans les capitules, l’application de méthyl jasmonate. Ce travail augure de bonnes perspectives en termes de valorisation d’Inula dans le secteur pharmaco-cosmétologique. Il reste à poursuivre la description du profil phytochimique de la plante et à localiser précisément les organes et/ou sous structures anatomiques concentrant les composés considérés. Ayant démontré que cette plante présente une bonne réponse à la domestication, il est également proposé de poursuivre l’étude des leviers environnementaux susceptibles d’influencer positivement et significativement le profil chimique d’Inula. / This thesis work is part of a regional project, initiated by the Luberon Park and in collaboration with the Laboratory of Pharmacognosy and Ethnopharmacology of the University of Marseille. It aimed to study the possibility of domesticating a wild plant, Inula montana L. (Asteraceae) known in the Provençal pharmacopoeia for its anti-traumatic effects similar to those of Arnica montana L. and to propose it as a new agricultural production. Inula montana produces lactones sesquiterpenes, identified as the secondary metabolites responsible for its biological anti-inflammatory properties. In this study, 1) to determine the growth and development characteristics of the largely unknown wild plant and to identify under natural conditions the most favorable factors for the production of secondary metabolites, 2) to study its characteristics. ability to multiply vegetatively in vitro and to form cell cultures able to synthesize the molecules of interest 3), to propose an agronomically applicable technical itinerary and 4) to test the effects of various environmental factors (fertilization, NaCl supply, modification circadian rhythm of illumination, UVB radiation, ablation of leaves, application of methyl jasmonate) on the qualitative and quantitative production of sesquiterpene lactones and phenolic compounds. The phenological characteristics of the wild plant are impacted by the altitude which induces a delay in the vegetative growth and the reproductive phase as well as physiological and morphological modifications. Levels of secondary metabolites (certain sesquiterpene lactones, total polyphenols and flavonoids) vary according to the season and are more important in the site which has the most water-constraining climatic conditions (draining soil, higher temperature and presence of a dry period in summer). Microscopic observation indicated the presence of two types of trichomes: glandular (biseriate) and non-glandular (hair) which are potentially the carrying structures of the molecules of interest. I. montana is able to form calli in vitro from root, foliar and shoot explants on which leafy shoots are formed. The domestication of Inula has been successful from seed from wild plants and under agronomic conditions, sesquiterpene lactone (costunolide, artemorine, eldarin and hydrocostunolide) and phenolic compounds are generally higher than in wild plants. The different constraints applied to test the effects of environmental factors on the production of metabolites have shown: 1) that there can be no correlation between the presence of oxidative stress and an increase in metabolite levels in Inula 2) that the accumulation of lactones and phenolic compounds seems mainly favored when the plant has a surplus of carbon skeletons, not used for growth; 3) finally, the two most favorable conditions for the accumulation of metabolites in Inula, are: in the leaves, a rapid alternation of light and darkness during the photoperiod and in the flower heads, the application of methyl jasmonate. This work augurs good prospects in terms of valuation of Inula in the pharmaco-cosmetological sector. It remains to continue the description of the phytochemical profile of the plant and to precisely locate the organs and / or anatomical substructures concentrating the compounds in question. Having demonstrated that this plant has a good response to domestication, it is also proposed to continue the study of environmental levers likely to positively and significantly influence the chemical profile of Inula.

Made available in DSpace on 2015-03-26T13:36:39Z (GMT). No. of bitstreams: 1 texto completo.pdf: 314133 bytes, checksum: 1d6a5d1c68ecfce7803cec5192e08e88 (MD5) Previous issue date: 2006-06-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Tolerance to salt stress was studied in maize seedlings (Zea mays L.) cultivars AGN 3150, BR 106, BR 201, BR 206 and SHS 4040, grown in Hoagland s nutrient solution. Treatments with NaCl 50 and 100 mM led to a decrease in the biomass production and to an increase in the relative electrolyte leakage in the roots and shoots in all cultivars. However, the largest decrease in growth and electrolyte leakage were shown by cultivar BR 106 and the smallest ones by cultivar AGN 3150, lead them to be considered as the most sensitive and the most tolerant to salt stress, respectively. The effect of NaCl 100 mM was investigated by considering some components of the antioxidative metabolism and mitochondrial respiration in roots of the seedlings. The tolerant cultivar showed increased activities of the enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione redutase (GR), as well as increase in ascorbate/dehydroascorbate ratio. The sensitive cultivar showed reduction in catalases (CAT) and GR activities as well as in ascorbate/dehydroascorbate ratio. The lipid peroxidation degree increased in roots and isolated mitochondria, only in the sensitive cultivar. In mitochondria isolated from the tolerant cultivar, NaCl treatment promoted an increase in the state 4 respiration rate and in the alternative pathway capacity. Additionally, salinity promoted a decrease in ADP/O and respiratory control ratios, indicating a lower coupling, likely due to an increase in alternative oxidase (AOX) activity. In sensitive cultivar, total respiration activity and the ADP/O ratio were strongly decreased, however the capacity of the alternative pathway was not altered. The NaCl treatment promoted decrease in the uncoupling protein (UCP) activity in the tolerant cultivar, but an increase it in the sensitive one occurred. These results suggest that the tolerant cultivar exhibits a larger efficiency in the reactive oxygen intermediates scavenging, as compared to the sensitive one. In addition, the high tolerance of cultivar AGN 3150 to the salt stress could be related to its high AOX capacity. A possible contribution of the UCP could not be confirmed from the results observed. / A sensibilidade ao estresse salino foi avaliada em plântulas de milho (Zea mays L.) das cultivares AGN 3150, BR 106, BR 201, BR 206 e SHS 4040, cultivadas em solução nutritiva de Hoagland. Os tratamentos com 50 e 100 mM de NaCl reduziram a produção de biomassa e aumentaram o extravasamento relativo de eletrólitos nas raízes e nas partes aéreas de todas as cultivares estudadas. Porém, os maiores valores de redução no crescimento e de extravasamento de eletrólitos foram apresentados pela cultivar BR 106 e os menores valores observados na cultivar AGN 3150. Portanto, entre as cultivares avaliadas, estas duas foram consideradas, respectivamente, como a mais sensível e a mais tolerante ao estresse salino. Verificou-se, então, o efeito do tratamento com 100 mM de NaCl sobre alguns componentes do metabolismo antioxidativo e sobre a respiração mitocondrial nas raízes de plântulas dessas duas cultivares. A cultivar tolerante apresentou aumento nas atividades das enzimas dismutase do superóxido (SOD), peroxidase do ascorbato (APX) e redutase da glutationa (GR), bem como aumento na razão ascorbato/desidroascorbato. A cultivar sensível apresentou redução na atividade das catalases (CAT) e GR, e também da razão ascorbato/desidroascorbato. O grau de peroxidação de lipídeos aumentou nas raízes e nas mitocôndrias isoladas, apenas na cultivar sensível. Nas mitocôndrias isoladas da cultivar tolerante, o tratamento com NaCl resultou em aumento na taxa respiratória no estado 4 e na capacidade da rota alternativa. Além disso, houve redução nas razões ADP/O e de controle respiratório, indicando um menor grau de acoplamento, possivelmente resultante da maior atividade da oxidase alternativa (AOX) na cultivar tolerante. Na cultivar sensível, a atividade respiratória total e a razão ADP/O foram fortemente reduzidas, porém a participação percentual da rota alternativa não foi alterada. O tratamento com NaCl resultou em redução na atividade da proteína desacopladora (UCP) na cultivar tolerante e em aumento desta atividade na cultivar sensível. Pelos dados obtidos, a maior tolerância da cultivar AGN 3150 está relacionada à sua maior eficiência na remoção de intermediários reativos de oxigênio, além da maior possibilidade de desvio de elétrons pela sua rota alternativa na cadeia respiratória. Entretanto, a contribuição da UCP nos mecanismos de tolerância à salinidade dessa cultivar não foi evidenciada.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / There are many studies showing that the salinity may directly affect the nitrate uptake, from their osmotic effect, nature of the salt and its ionic composition. However, little is known about the mechanisms related to the salt ability to inhibit the nitrate acquisition indirectly. This study was carried with aim to induce inhibition of NO3- influx in cowpea roots of indirect form triggered by a negative feedback mechanism, caused by the increase in the pool of free amino acids in the tissue, induced by salt stress.For this, were done three isolated studies and continuous.The exogenous glutamine application promoted an increase in the free amino acids content.The presence of glutamine decreased significantly the nitrate acquisition.The free ammonium can also be listed as a key-compound in the role of nitrate influx regulation, since use of the MSX (Methionine sulfoximine) promoted the increase of NH4+ content and also reduced nitrate influx, but, in a lesser degree when compared to treatment with AZA (Azaserine). Salt stress caused a reduction in NO3- influx by decrease in the growth of plants induced by salt. The data indicated which this reduction in the influx is triggered by increase of amino acids content, mainly the glutamine, that is main likely compound to act as signal in the N-feedback regulation. / Existem muitos estudos mostrando que a salinidade pode afetar a absorÃÃo de nitrato de forma direta, a partir do seu efeito osmÃtico, da natureza do sal e de sua composiÃÃo iÃnica. Entretanto, pouco se sabe sobre os mecanismos relacionados com a capacidade do sal em inibir a aquisiÃÃo de nitrato de forma indireta. O presente estudo teve como objetivo induzir a inibiÃÃo do influxo de NO3- em raÃzes de feijÃo-caupà de forma indireta desencadeada por um mecanismo de feedback negativo, provocado pelo aumento no pool de aminoÃcidos livres no tecido, induzido por estresse salino. Para isso, foram realizados trÃs estudos isolados e contÃnuos. A aplicaÃÃo de glutamina exÃgena promoveu um aumento no conteÃdo de aminoÃcidos livres. A presenÃa de glutamina reduziu significativamente a aquisiÃÃo de nitrato. O amÃnio livre tambÃm pode ser listado como um composto-chave no papel da regulaÃÃo do influxo de nitrato, pois a utilizaÃÃo do MSO (Metionina sulfoximina) promoveu o aumento do conteÃdo de NH4+ e tambÃm reduziu o influxo de nitrato, porÃm em menor grau quando comparado ao o tratamento com AZA (Azaserina). O estresse salino causou uma reduÃÃo no influxo de NO3-, pela diminuiÃÃo no crescimento das plantas induzida pelo sal. Adicionalmente, estes dados indicaram que esta reduÃÃo no influxo està ligada ao aumento do teor de aminoÃcidos, principalmente a glutamina, que à o principal componente para atuar como sinal na regulaÃÃo por N-feedback.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / In this work were studied diverse physiological mechanisms, as the osmotic adjustment, photosynthesis, including gas exchange and chlorophyll fluorescence parameters, as well the oxidative responses in physic nuts submitted to different abiotic stresses as: salinity, drought and high temperature. The first experiment aimed to study the effects of increase of the NaCl concentrations (0, 25, 50, 75 and 100 mM) in the ions accumulate (Na+, Cl- and K+) and some growth variables, as well evaluate the water state and the principals solutes (organic and inorganic) involved on the osmotic adjustment of physic nuts plants under this stressful condition. Physic nuts plants showed sensibility to salt stress, presenting a reduction by 50% in dry matter from by 47 mM of NaCl concentration during 15-d. This sensibility should be due the leaf Na+ and Cl- high accumulation, associated a strong reduction in the K+ concentration, induced by high Na+ content. On the other hand, physic nuts plants were able osmotic adjust to salinity due a severe decrease on the osmotic potential and increase of leaf water state, principally in the higher NaCl levels. Of the solutes studied, was observed that salt ions (Na+ and Cl-) contributed with the most of the osmotic adjustment, while that the K+ contribution was decreased strongly by NaCl. The glycinebetaine compared to proline was more important to the osmotic adjustment of physic nuts leaves, as in the absence as in presence of different NaCl levels in the nutrient solution. The second experiment evaluated the resistance of photosynthetic apparatus of physic nuts plants submitted to different time of exposure (7-d and 14-d of treatment and 3-d of recovery) to salt stress (100 mM of NaCl).The changes caused on photochemistry activity and leaf gas exchange were evaluate by Na+ and Cl- accumulation and decrease of K+/Na+ ratio in the leaves. After 7-d of treatment was observed a major action of osmotic effects. However, after 14-d of treatment, the ionic effects caused by Na+ and Cl- excessive accumulation and by K+/Na+ ratio strong reduction in the leaves, caused permanent damages on the photosynthesis of physic nuts plants due as the stomatal limitations as non-stomatal ones. The third experiment aimed to study the comparative effects between the salt stress (50 mM of NaCl) and water stress (induced for PEG 6000), both with osmotic potential of â0.22 MPa on the photosynthesis, water relations and growth of physic nuts plants. The water stress effects induced for PEG in the leaf growth, electrolyte leakage and leaf gas exchange were more deleterious than by NaCl ones. In the both stresses was observed decrease in the leaf CO2 assimilation due the stomatal and non-stomatal limitations. However, the chlorophyll fluorescence parameters did not affect. The fourth experiment aimed to evaluate the relative contribution of organic and inorganic solutes on the osmotic adjustment of leaves and roots physic nuts plants in different water restriction levels. Of the solutes studied, the K+ and soluble sugar were the most involved in the osmotic adjustment as in the leaves as in roots. Others solutes as, Na+, Cl-, total amino acids and glycibetaine, also presented a effective role in the reduction of osmotic potential in both organs. On the other hand, the leaf proline content, although has increased significantly, was not sufficient to promote an effective participation of this amino acid in the osmotic adjustment of physic nuts plants. The same experiment aimed to observe the isolated and combined effects of water stress and high temperature on the photosynthesis and evaluate the oxidative defenses system in physic nuts plants. The photosynthetic apparatus was more sensitive to water stress than heat ones, been that the combination of them caused deleterious effects yet large in this complex. Additionally, the oxidative damages also were more marked in the combined stress. In general, the data shown that physic nuts plants, although present ability to adjust osmotically to salinity and drought, have their photosynthetic apparatus very affected in this stressful conditions. Even as, the defense system against oxidative damages appears has not been efficient in plants exposure at the drought and heat isolated and combined stresses / In this work were studied diverse physiological mechanisms, as the osmotic adjustment, photosynthesis, including gas exchange and chlorophyll fluorescence parameters, as well the oxidative responses in physic nuts submitted to different abiotic stresses as: salinity, drought and high temperature. The first experiment aimed to study the effects of increase of the NaCl concentrations (0, 25, 50, 75 and 100 mM) in the ions accumulate (Na+, Cl- and K+) and some growth variables, as well evaluate the water state and the principals solutes (organic and inorganic) involved on the osmotic adjustment of physic nuts plants under this stressful condition. Physic nuts plants showed sensibility to salt stress, presenting a reduction by 50% in dry matter from by 47 mM of NaCl concentration during 15-d. This sensibility should be due the leaf Na+ and Cl- high accumulation, associated a strong reduction in the K+concentration, induced by high Na+ content. On the other hand, physic nuts plants were able osmotic adjust to salinity due a severe decrease on the osmotic potential and increase of leaf water state, principally in the higher NaCl levels. Of the solutes studied, was observed that salt ions (Na+ and Cl-) contributed with the most of the osmotic adjustment, while that the K+contribution was decreased strongly by NaCl. The glycinebetaine compared to proline was more important to the osmotic adjustment of physic nuts leaves, as in the absence as in presence of different NaCl levels in the nutrient solution. The second experiment evaluated the resistance of photosynthetic apparatus of physic nuts plants submitted to different time of exposure (7-d and 14-d of treatment and 3-d of recovery) to salt stress (100 mM of NaCl).The changes caused on photochemistry activity and leaf gas exchange were evaluate by Na+ and Cl- accumulation and decrease of K+/Na+ ratio in the leaves. After 7-d of treatment was observed a major action of osmotic effects. However, after 14-d of treatment, the ionic effects caused by Na+ and Cl- excessive accumulation and by K+/Na+ ratio strong reduction in the leaves, caused permanent damages on the photosynthesis of physic nuts plants due as the stomatal limitations as non-stomatal ones. The third experiment aimed to study the comparative effects between the salt stress (50 mM of NaCl) and water stress (induced for xv PEG 6000), both with osmotic potential of â0.22 MPa on the photosynthesis, water relations and growth of physic nuts plants. T e water stress effects induced for PEG in the leaf growth, electrolyte leakage and leaf gas exchange were more deleterious than by NaCl ones. In the both stresses was observed decrease in the leaf CO2 assimilation due the stomatal and nonstomatal limitations. However, the chlorophyll fluorescence parameters did not affect. The fourth experiment aimed to evaluate the relative contribution of organic and inorganic solutes on the osmotic adjustment of leaves and roots physic nuts plants in different water restriction levels. Of the solutes studied, the K+ and soluble sugar were the most involved in the osmotic adjustment as in the leaves as in roots. Others solutes as, Na+, Cl-, total amino acids and glycibetaine, also presented a effective role in the reduction of osmotic potential in both organs. On the other hand, the leaf proline content, although has increased significantly, was not sufficient to promote an effective participation of this amino acid in the osmotic adjustment of physic nuts plants. The same experiment aimed to observe the isolated and combined effects of water stress and high temperature on the photosynthesis and evaluate the oxidative defenses system in physic nuts plants. The photosynthetic apparatus was more sensitive to water stress than heat ones, been that the combination of them caused deleterious effects yet large in this complex. Additionally, the oxidative damages also were more marked in the combined stress. In general, the data shown that physic nuts plants, although present ability to adjust osmotically to salinity and drought, have their photosynthetic apparatus very affected in this stressful conditions. Even as, the defense system against oxidative damages appears has not been efficient in plants exposure at the drought and heat isolated and combined stresses

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / A salinizaÃÃo dos solos tem se constituÃdo um problema sÃrio para a agricultura, principalmente nas regiÃes Ãridas e semi-Ãridas em todo o mundo. O excesso de sais no solo afeta tanto as Ãreas irrigadas como as nÃo irrigadas, limitando, portanto o crescimento e a produtividade das culturas. O presente trabalho teve por objetivo avaliar os efeitos da inoculaÃÃo com fungo micorrÃzico arbuscular e da adubaÃÃo orgÃnica no desenvolvimento do sabià (Mimosa caesalpiniaefolia Benth), sob estresse salino. Portanto, foi instalado um experimento em casa de vegetaÃÃo do Departamento de CiÃncias do Solo da Universidade Federal do CearÃ, no Campus do Pici em Fortaleza. O delineamento experimental adotado foi o inteiramente casualizado, em esquema fatorial, 2 (presenÃa e ausÃncia de fungos micorrÃzicos arbusculares) X 2 (presenÃa e ausÃncia de composto orgÃnico) X 5 nÃveis de condutividade elÃtrica da Ãgua de irrigaÃÃo (0,7, 1,2, 2,2, 3,2 e 4,2 dS m1), com 4 repetiÃÃes. Ao final de 60 dias apÃs a germinaÃÃo das plantas foi coletado o experimento, onde determinou-se os fatores de crescimento, as variÃveis microbiolÃgicas, o pH do solo apÃs o cultivo, os teores minerais (N, P, K, Ca, Mg, Na, Cl) e as relaÃÃes Raiz/Parte AÃrea (R/PA) e SÃdio/PotÃssio (Na/K). A salinidade promoveu maior reduÃÃo na produÃÃo de matÃria seca das plantas nÃo micorrizadas e nÃo adubadas. A inoculaÃÃo com fungo micorrÃzico arbuscular aumentou a produÃÃo de matÃria seca das plantas de sabiÃ. As plantas adubadas com matÃria orgÃnica apresentaram maior desenvolvimento, quando comparadas com as plantas nÃo adubadas. As plantas micorrizadas apresentaram maiores conteÃdos de nutrientes, quando comparadas com as plantas nÃo micorrizadas. / Salt affected soils management is an important subject to be studied, especially when irrigated arid and semi-arid regions are considered, as soil salt excess may limit growth and plant productivity. The objective of this dissertation was to evaluate the effect of mycorrhizal arbuscular fungi (AMF) inoculation on growth of sabià plants (Mimosa caesalpiniaefolia Benth.), cultivated under salt stress in the presence or absence of organic compost. Thus, an experiment was conducted under green-house conditions to evaluate the potential of inoculated and fertilized plant establishment submitted to salt stress, in a complete randomized factorial design 2 (inoculated or non-inoculated plants) X 2 (fertilized or non-fertilized with organic compost) X 5 levels of irrigation water electrical conductivity (0.7, 1.2, 2.2, 3.2 and 4.2 dS mâ), with 4 replications. After 60 days from germination, plants were harvested and then and analyzed to evaluate growth factors, microbiological variables, soil pH, shoot-root and sodium/potassium ratios and mineral nutrient content (N, P. Ca, Mg, Na and Cl). Salinity decreased growth and plant biomass of non-mycorrhizal fertilized plants. Inoculation with AMF increased plant dry matter production when compared with non-inoculated plants under salt stress. When compared with non-fertilized plants, organic compost fertilization increased plant growth, Nutrient contents were always much higher in inoculated plants and in organic compost fertilized plants.

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / A reduÃÃo da atividade fotossintÃtica em plantas sob estresse salino ocorre devido à restriÃÃo do influxo de CO2 no mesofilo foliar, causada em parte pelo fechamento estomÃtico. Nessas condiÃÃes, a reduÃÃo da fotossÃntese pode levar a danos oxidativos, os quais sÃo atribuÃdos, principalmente, a produÃÃo excessiva de perÃxido de hidrogÃnio devido o aumento da atividade fotorrespiratÃria. O aumento da pressÃo de CO2 no meio externo pode favorecer seu influxo no mesofilo, favorecendo a fotossÃntese e atenuando os danos oxidativos sob salinidade. Neste sentido, na presente pesquisa foi avaliada a eficiÃncia fotossintÃtica e danos oxidativos em plantas de cajueiro cultivadas sob salinidade quando expostas a condiÃÃes normais e elevadas de CO2. Para tanto, plÃntulas jovens (30 dias pÃs-plantio) foram cultivadas na ausÃncia (controle) ou presenÃa de salinidade (NaCl 100 mM), por oito dias, em casa de vegetaÃÃo, e em seguida foram expostas a pressÃes de CO2 atmosfÃrica (380 ppm) e elevada (760 ppm), em cÃmara de crescimento sob condiÃÃes controladas, durante 7 dias. Os resultados demonstram que o cultivo em altas pressÃes de CO2, em um curto perÃodo, induziu a iniciaÃÃo de um processo de aclimataÃÃo em plÃntulas de cajueiro. Essa aclimataÃÃo inicial està associada a reduzidas taxas de assimilaÃÃo do carbono e elevados conteÃdos de carboidratos. Durante o cultivo salino, em CO2 ambiente, as mudas apresentaram respostas tÃpicas a plantas estressadas, com maiores nÃveis de TBARS, H2O2, superior relaÃÃo Na+/K+, atividade da APX e, induÃÃo do mecanismo antioxidativo nÃo enzimÃtico, com maiores acÃmulos de prolina e antocianinas. Enquanto que o cultivo em alto CO2 refletiu em respostas conflitantes, principalmente, quanto Ãs alteraÃÃes oxidativas. As plÃntulas submetidas à combinaÃÃo da salinidade e de altas pressÃes de CO2 estimularam a peroxidaÃÃo lipÃdica, a dissipaÃÃo nÃo- fotoquÃmica (NPQ), o acÃmulo de aminoÃcidos, flavonÃides e antocianinas, superiores ao cultivo salino em CO2 ambiente. Por outro lado o conteÃdo de H2O2 e, variÃveis relacionadas a fotorrespiraÃÃo, como a atividade da GO e o conteÃdo de glioxilato reduziram. Esses resultados sugerem que altas pressÃes de CO2 induziram danos aditivos à salinidade, reflexo de uma interaÃÃo que se mostrou complexa, durante um perÃodo inicial de aclimataÃÃo. Adicionalmente, pelo cajueiro ser um modelo nativo e contrastante, mais estudos serÃo necessÃrios, principalmente avaliando seu comportamento ao longo do tempo, em um perÃodo mais extenso. / The reduction of photosynthetic activity in plants under salt stress occurs due to the restriction of the influx of CO2 in the leaf mesophyll, caused in part by stomatal closure. Under these conditions, the reduction of the photosynthesis can lead to oxidative damage, which are assigned, mainly, the excessive production of hydrogen peroxide due to increased activity photorespiratory. Thus, the increase of CO2 pressure in the external environment may favor the influx in the mesophyll, favoring photosynthesis and attenuating the oxidative damage under salinity. In this sense, the present study was to evaluate the photosynthetic efficiency and oxidative damage in cashew plants grown under salinity when exposed to normal and elevated CO2. To this end, young seedlings (30 days after planting) were cultured in the absence (control) or presence of NaCl (100 mM NaCl) for eight days, in a greenhouse, and then were exposed to atmospheric pressure of CO2 (380 ppm) and elevated (760 ppm) in a growth chamber under controlled conditions, for seven days. The results show that cultivation at high pressures of CO2, in a short period, induced the initiation of a process of acclimation of cashew seedlings. This initial acclimation is associated with reduced rates of carbon assimilation and high carbohydrate contents. During cultivation saline, CO2 ambient, the seedlings showed typical responses to salt stress, with higher levels of TBARS, H2O2, higher Na + / K +, APX activity and induction of antioxidant non-enzymatic mechanism, as proline and anthocyanin. Whereas the culture in high CO2, reflected in conflicting responses, mainly related to oxidative changes. Seedlings subjected to a combination of salinity and high pressures of CO2 stimulated lipid peroxidation, non-photochemical dissipation (NPQ), the accumulation of amino acids, flavonoids and anthocyanins, higher cultivation in saline CO2 environment. On the other hand, have reduced the content of H2O2 and variables associated with photorespiration, as GO activity and content of glyoxylate. These results suggest that high pressures of CO2 induced damage additives to salinity, reflecting a complex interaction that was shown during an initial period of acclimation. In addition, the cashew to be a model native and contrasting, more studies are needed, especially be evaluating behavior over time, in a longer period

Appliqués à forte dose, les rayonnements UV-C sont délétères, alors que administrés à faibles doses, ces memes rayonnements stimulent des réponses bénéfiques. Ce phénomène est connu sous le nom d’hormesis et la dose bénéfique est qualifiée d’hormique. L’application des faibles doses d’UV-C sur les fruits et les légumes en post-récolte a permis donc d’augmenter leur résistance vis à vis des pathogènes (Charles et al, 2008), d’améliorer leur qualité nutritionnelle (Mercier et al, 2001) et d’améliorer leur rendement et leur croissance (Siddiqui et al, 2011). Ces travaux ont été réalisés sur des espèces différentes. Dans le cadre de ma thèse, nous avons traité une seule espèce de laitues romaines var Claudius après récolte avec une dose non délétère d’UV-C (0.85kJ.m-2) puis nous avons évalué son effet sur leur aspect de résistance aux champignons Botrytis cinerea (BC87) et Sclerotinia minor (SM), sur leurs valeurs nutritionnelles au cours du stockage et sur leurs réponses à la contrainte saline du milieu. L’analyse de tous les résultats obtenus,a mis en évidence que la dose d’UV-C choisie diminue la sensibilité de la laitue romaine à ces deux agents pathogènes (Ouhibi et al. 2014a), améliore sa valeur nutritionnelle en augmentant la concentration des composées phénoliques et de l’acide ascorbique et acquière aux plantes provenant de grains prétraitées aux UV-C un meilleur potentiel d'adaptation à la contrainte saline (Ouhibi et al, 2014 b). Le traitement des plantes avec une dose d’UV-C non délétère permet d’agir de façon bénéfique et simultanément sur différents processus physiologiques. / Applied in high doses, UV-C radiations are harmful, while administered at low doses, these same radiations stimulate beneficial answers. This phenomenon is known as hormesis and the beneficial dose is qualified hormic. The application of low doses of UV-C on fruits and vegetables in post harvest enhances resistance against pathogens (Charles et al, 2008), improve their nutritional quality (Mercier et al, 2001) and their performance to grow (Siddiqui et al, 2011). This works were carried out on different species. In my thesis, we treated a single species of romaine lettuce var claudius after harvest with a non-harmful dose of UV-C (0.85kJ.m-2) and we evaluated its effect on resistance to Botrytis cinerea (BC87) and Sclerotinia minor (SM), in their nutritional value during storage and their responses to salt stress. The analysis of all the results obtained showed that the UV-C dose decreases the sensitivity of romaine lettuce to these two pathogens (Ouhibi et al. 2014a), improve nutritional value by increasing the content in phenolic compound, in ascorbic acid and acquire to plants from seeds pre-treated with UV-C greater potential for adaptation to salt stress (Ouhibi et al, 2014 b).

>Magister Scientiae - MSc / The production of reactive oxygen species (ROS) is prominent in all aerobic metabolisms including plants. For this reason, the redox homeostasis of the production and scavenging of these intermediates is imperative for growth, development and survival during unfavourable conditions. In this study, a putative glutathione peroxidase gene (Glyma17g34110) from Glycine max (soybean) was identified and analyzed. The successful characterisation of Glyma17g34110 provided evidence of it being a glutathione peroxidase using glutathione as its preferred electron donor and substrate. Furthermore, it is known that antioxidant enzymes such as GPX exist in various tissues, performing a diverse set of functions. By a bioinformatic analysis of Glyma17g34110 and its promoter region, it was indicated that Glyma17g34110 could be a putative chloroplast protein that could play an important role in photosynthesis.One of the major factors affecting plant growth and development worldwide is abiotic stresses such as salinity. In the presence of salinity the production of harmful ROS is increased, resulting in detrimental reactions with important biological features (DNA, protein and lipid membranes), leading to cell death. The analysis of Glyma17g34110 under salt stress revealed that it is a salt sensitive gene and thus, the down-regulation of Glyma17g34110 could be due to the lack of known defence and response cis-acting elements present in the promoter region. Furthermore, it was proven in previous studies that the application of exogenous nitric oxide (NO) increases the activity of antioxidant enzymes. In this thesis it was observed that the presence of exogenously applied NO increased the expression of Glyma17g34110 tremendously in all soybean tissues (leaves, roots and nodules) investigated.Studies have found numerous cis-acting elements to be NO responsive, however, none of these elements were found in the promoter region upstream of glyma17g34110. This suggests that novel cis-acting elements could be present in the promoter region of Glyma17g34110.Thus, increasing the expression of Glyma17g34110 during salinity in the presence of NO, as well as the identification of these novel cis-acting elements, could lead to the enhancement of the defence mechanisms against ROS, which could lead to increasing plant tolerance to stress.

Effet de la salinité pendant la croissance de plants de riz sur la biosynthèse de la 2-acétyl-1-pyrroline (2AP), composé aromatique clé des variétés de riz aromatique,le rôle de la 1-pyrroline dans la biosynthèse de la 2AP ainsi que l'origine du groupe acétyle ont été étudiés. La variété aromatique de Camargue (France), Aychade, a été cultivée soit dans un milieu liquide soit dans des pots en serre. Dans le milieu de culture liquide, une augmentation significative de la teneur en 2AP dans les feuilles a été observée quelle que soit la concentration en NaCl (de 1,7 à 170 mM). Une corrélation positive a été mise en évidence entre la teneur en sel et en 2AP. A de plus fortes concentrations en NaCl, il a également pu être montré qu'il y avait une relation entre la concentration en proline, un précurseur aminé majeur de la 2AP, et la teneur en 2AP. Inversement, les changements en acide -aminobutyrique (GABA), un des métabolites compétitif de la biosynthèse de la 2AP, n'ont pas été corrélés avec ceux de la 2AP. Dans les expérimentations menées en serre, la variété Aychade a été cultivée dans une solution saline à 30 mM, correspondant à une conductivité électrique de 3800 ± 400 S.cm-1, valeur considérée comme suffisante pour induire un stress salin sur le plant de riz. Cette solution saline a été appliquée à des dates et pour des durées différentes en fonction du cycle de développement de la plante. A la phase végétative, une relation positive a été observée dans les feuilles entre le traitement salin et la teneur en 2AP ainsi qu'en proline. Il est intéressant de noter que tous les grains provenant des plants traités avec la solution saline contenaient des teneurs nettement plus élevées en 2AP par rapport au contrôle. La synthèse de la 2AP la plus importante s'est produite lorsque les plants ont été soumis à un traitement salin pendant l'ensemble des phases végétative et reproductive. Cependant, dans le dernier cas, le rendement en grain était beaucoup plus faible. En supplémentant des cals de riz avec de la 1-pyrroline, il a été démontré que cette imine cyclique était un facteur limitant pour la biosynthèse de la 2AP non seulement dans la variété aromatique, mais également dans la non aromatique. Les essais conduits avec le glucose et les acides gras marqués 13C ont montré que ces molécules pourraient être précurseurs du groupe acétyle de la molécule de 2AP. / The effect of salinity during rice plant growth on the biosynthesis of 2-acetyl-1-pyrroline (2AP), characteristic flavour compound of fragrant rice cultivars was studied. Additionally role of 1-pyrroline in 2AP biosynthesis together with source of acetyl group were investigated. Aychade, a fragrant rice was cultivated either in a liquid medium adapted for seedlings or in pots in a greenhouse. A significant increase in 2AP levels in the leaves was observed whatever the concentration of NaCl (1.7 to 170 mM NaCl) in the culture medium. A positive correlation was found between the level of salt and that of 2AP. There was a relationship between the concentration of proline, a major amino precursor of 2AP and 2AP level at highest concentrations of NaCl. Conversely the changes in -aminobutyric acid (GABA), a competitive metabolite in 2AP biosynthesis were not correlated with those of 2AP. In greenhouse experiments, Aychade cultivar was grown under one level of salt solution, 30 mM that corresponds to EC of 3800 ±400 S.cm-1, value considered sufficient to induce salt stress in rice. Timing and duration of application of salt solution varied according to the growth stages. At vegetative phase a positive relation was observed between salt treatment and 2AP as well as proline levels in the leaves. Interestingly grains from all salt treated plants contained significantly higher levels of 2AP than those from control. The highest synthesis occurred when plants were subjected to salt treatment during whole vegetative and reproductive phases. However in the latter case crop yield decreased significantly. Through supplementation of rice callus with 1-pyrroline it was demonstrated that this cyclic imine was limiting factor for the 2AP biosynthesis not only in a fragrant variety but also in a non fragrant variety. Experiments with 13C labelled glucose and fatty acids showed that glucose and fatty acids could be precursors of acetyl group of 2AP.

>Magister Scientiae - MSc / Abiotic stress, mainly in the form of extreme temperatures, drought and salinity has caused major crop losses worldwide, putting a severe strain on agriculture. Salinity severely limits plant growth and productivity and affects all aspects of the plant’s development including the most crucial stage; germination. This study investigated the effect of salt (NaCl) stress on Sorghum bicolor seedlings and the role of exogenously applied calcium (Ca2+) to ameliorate the effects of salt stress during germination. Sorghum seeds were germinated in the presence and absence of various NaCl (100, 200 and 300 mM) and Ca2+ (5, 15 and 35 mM) concentrations. Several assays including physiological (germination and growth assays), biochemical (osmolytes and oxidative stress markers), anatomical (epidermal and xylem layers) and expression profiles of key genes [antioxidant (SbSOD, SbAPX2 and SbCAT3), Salt Overly Sensitive (SbSOS1, 2 and 3) pathway enzymes and the vacuolar Na+/H+ exchanger antiporter2 (SbNHX2)] were investigated. Salt stress delayed germination and negatively affected growth as observed by the reduced root and shoot length and decreased fresh and dry weight. There was an increase in proline content and oxidative stress markers (H2O2 and MDA) under salt stress. Oxidative stress resulted in damage to the epidermal and xylem layers as observed on Scanning Electron Microscopy (SEM) images. Quantitative real-time polymerase chain reaction revealed that salt stress induced the expression of SbAPX2, SbCAT3 and SbSOS1 genes, whereas SbSOD4A, SbSOS2, SbSOS3 and SbNHX2 genes were not affected by salt. Exogenous application of Ca2+ counteracted the harmful effects of salt stress by improving germination efficiency, promoting seedling growth, reducing oxidative damage and the Na+/K+ ratio, indicating the protective effect. Ca2+ also effectively protected the epidermis and xylem layers from the severe damage caused by salt stress. In the presence of Ca2+ the expression of SbAPX2 and SbCAT3 was reduced except for the SbNHX2 gene, which increased by 65-fold compared to the control. The results obtained suggests that sorghum is able to respond to salt stress by inducing osmolytes, the antioxidant defence system as well as the SOS pathway. Furthermore, 5 mM Ca2+ was determined as the optimum Ca2+ concentration required to enhance sorghum’s tolerance to salt stress.