O transporte de água nas plantas é um processo complexo e que envolve a passagem por membranas celulares. Nelas existem aquaporinas, proteínas integrais de membrana, que facilitam o fluxo passivo de água e pequenos solutos. Um fator importante que influencia o transporte de água nas plantas é a condutância hidráulica radicular (LPr), que pode contribuir com até 50% de toda a resistência ao fluxo de água na planta. Várias metodologias são propostas para estimar a LPr, porém os dados obtidos são altamente variáveis, ainda que para a mesma espécie vegetal. Diante do exposto, pretende-se desenvolver e propor uma nova metodologia, o sistema hidráulico de sucção para estimativa da LPr e avaliar a contribuição das aquaporinas no transporte de água radicular em plântulas de milho. O sistema hidráulico de sucção foi construído no Laboratório de Estudos de Plantas sob Estresse (LEPSE/USP) em parceria com Laboritoire d\'Ecophysiologie des Plants sous Stress Environnementaux de Montpellier, França. Os experimentos foram conduzidos no LEPSE, onde as plântulas cresceram em hidroponia na casa de vegetação. Para avaliação da atividade de aquaporinas foi utilizado o tratamento com peróxido de hidrogênio, que em concentrações milimolares inibe os canais de água. As medidas de condutância foram realizadas sempre na mesma hora do dia, uma vez que a própria LPr e as aquaporinas apresentam ritmo circadiano. Houve redução de 53% da condutância no tratamento com peróxido em relação ao controle e correlação significativa entre a LPr e o pH, de modo que o aumento do pH provocou queda de LPr, provavelmente causada por alterações na abundância e/ou atividade de aquaporinas nas raízes de milho. Esses resultados permitem afirmar que o método de sucção foi eficaz na estimativa da condutância hidráulica, podendo ser adotado como técnica alternativa na estimativa da LPr em plantas. / The water transport in plants is a complex process which involves passage through cell membranes. In them there are aquaporins, integral membrane proteins, that facilitate passive flux of water and small solutes. An important factor influencing the water transport in plants is root hydraulic conductance (LPr), which can contribute up to 50% of the water flow resistance in the plant. Various methods are proposed for estimating LPr, but the data are highly variable, even for the same plant species. Therefore, we intend to develop and propose a new methodology, the hydraulic suction system to estimate LPr and evaluate the contribution of aquaporins in root water transport in maize seedlings. The hydraulic suction system was built at the Laboratório de Estudos de Plantas sob Estresse (LEPSE / USP) in partnership with Laboritoire d\'Ecophysiologie Plants des sous Stress Environnementaux Montpellier, France. The experiments were conducted in LEPSE where the seedlings grew in the hydroponics greenhouse. To evaluate the activity of aquaporins was used treatment with hydrogen peroxide, which in millimolar concentrations inhibits water channels. The conductance measurements were performed at the same time of day, since the LPr itself and aquaporins presents the circadian rhythm. There was a 53% reduction in conductance peroxide treatment compared to control and significant correlation between LPr and pH, so that the pH increase caused a drop LPr probably caused by alterations in the abundance and/or activity of aquaporins in maize roots. These results indicate that the suction method was efficient in estimated hydraulic conductance, may be adopted as an alternative technique to estimate the LPr in plants.

Over an extended period of time there have been plans to establish a shared facility in Lennheden to extract groundwater from the Badelunda esker to provide drinking water for the cities of Borlänge and Falun. The city of Falun is dissatisfied with the quality of its drinking water and the city of Borlänge is concerned about the risk of contamination of its existing groundwater supply at its current location. To provide a basis for a decision on this issue, the company Midvatten AB has been commissioned to perform hydrogeological investigations in the area of Lennheden. The purpose of this thesis is to design a functional groundwater model of the area between Lennheden, place of planned extraction, and Övre Tjärna, place of existing extraction, to get a better understanding of the groundwater situation in the area. A groundwater model enables simulations of different scenarios in risk assessment and contaminant transport. The aim of the thesis is that the model can be used as an aid in Midvatten’s investigations in Lennheden and that it also can be used in future projects in the area. The model has been made in Processing Modflow 5.3 and encompasses an area of 19,5 × 11 km along the Badelunda esker and the river Dalälven between Djurmo and Frostbrunnsdalen. The model has 6 layers and a cell resolution of 50 × 50 meters and 50 × 100 meters. The physical parameters of the model are comprised from different hydrological and geological investigations in the area. The Badelunda esker and the river Dalälven totally dominate the groundwater situation in the area. A large part of the work in the thesis has been focused on the calibration of the horizontal hydraulic conductivity in the esker and the vertical hydraulic conductivity in the riverbed of Dalälven. The calibration of the model implies a vertical hydraulic conductivity of 0,01 – 0,1 md-1 in the riverbed of Dalälven, depending on the riverbed thickness. Between Bäsna and Övre Tjärna, simulated and observed groundwater levels correspond well. The transport time of the water in the esker, simulated in PMPATH, also corresponds well with estimated transport times. The model can be used for simpler studies of contaminant transport. Simulation of the pumping test in Lennheden gives a good correspondence eastward in the esker from Lennheden to Övre Tjärna. Westward in the esker, from Lennheden to Bäsna, the draw down is sharper than observed. Possible reasons for the sharper draw down is an underestimation of the bulk of the esker and that the vertical hydraulic conductivity of the riverbed is set too low between Lennheden and Bäsna. In order to improve the model, the most important step should be to thoroughly investigate the river Dalälven with regards to vertical hydraulic conductivity in the riverbed and riverbed thickness. / Under en längre tid har det funnits planer på att upprätta en ny, gemensam vattentäkt för Borlänge och Falu kommun i Badelundaåsen i Lennheden nordväst om Borlänge stad. Falu kommun är missnöjd med vattenkvaliteten i sin ytvattentäkt och Borlänge är oroad över riskerna med att ha sin nuvarande vattentäkt i Badelundaåsen lokaliserad i nära anslutning till riksväg 70 och järnvägen. Beslut i frågan ska tas under 2006 och Midvatten AB har fått i uppdrag att genomföra geohydrologiska undersökningar i och runt Lennheden som underlag till beslutet. Syftet med det här examensarbetet är att skapa en fungerande grundvattenmodell för området mellan Lennheden, plats för planerat vattenuttag, och Övre Tjärna, plats för befintligt vattenuttag, för att få en större förståelse för grundvattenmagasinet. En grundvattenmodell möjliggör simulering av olika scenarion så som föroreningstransport i grundvattnet och bestämning av skyddsområden. Målet är att modellen ska kunna användas som ett komplement i Midvattens undersökningar och även användas i framtida projekt i området. Modellen är gjord i Processing Modflow 5.3 och omfattar ett 19,5 × 11 km stort område längs Badelundaåsen och Dalälven mellan Djurmo och Frostbrunnsdalen. Modellen består av 6 lager där cellernas upplösning är 50 × 50 meter och 50 × 100 meter. De fysiska parametrar som modellen är uppbyggd av är sammanställda från olika hydrologiska och geologiska undersökningar som har utförts i området de senaste 30 åren. Badelundaåsen och Dalälven är de två komponenter som totalt dominerar grundvattensituationen i området. En stor del av arbetet har fokuserats på kalibreringen av den horisontella hydrauliska konduktiviteten i åsen och den vertikala hydrauliska konduktiviteten i Dalälvens botten. I modellen är den vertikala hydrauliska konduktiviteten i Dalälvens botten kalibrerad till intervallet 0,01 – 0,1 md-1 beroende på bottnens mäktighet. På sträckan mellan Bäsna och Övre Tjärna så fungerar modellen bra. Simulerade och observerade grundvattennivåer stämmer väl överens. Även vattnets transporttider i åsen, simulerade i PMPATH, stämmer väl överens med uppskattade transporttider. Modellen kan användas för enklare studier av ämnestransport. Vid simulering av provpumpningen i Lennheden så är överensstämmelsen god österut i åsen, från Lennheden till Övre Tjärna. Västerut i åsen, från Lennheden till Bäsna, är avsänkningen av grundvattennivån för kraftig. Möjliga orsaker till den för kraftiga avsänkningen är att åsens utbredning är underskattad och att Dalälvens botten har en för lågt ansatt vertikal hydraulisk konduktivitet längs sträckan Lennheden – Bäsna. Den klart viktigaste åtgärden för att förbättra modellen skulle vara att grundligt undersöka Dalälven, särskilt längs sträckan Djurmo – Båtsta, med avseende på dess bottens vertikala hydrauliska konduktivitet och mäktighet.

O objetivo do presente estudo foi quantificar, in vitro, a permeabilidade da dentina tratada com dois agentes anti-hiperestésicos à base de oxalato e um à base de fosfato de cálcio, por meio de testes de condutividade hidráulica e de espectroscopia de impedância eletroquímica e, ainda, determinar se espectroscopia de impedância eletroquímica pode ser considerada um método alternativo para avaliar a capacidade obstrutiva de agentes antihiperestésicos. Para o teste de condutividade hidráulica foram utilizados vinte quatro discos de dentina, obtidos de terceiros molares humanos extraídos, que foram divididos em três grupos de oito espécimes, correspondentes aos três materiais testados: gel Experimental, Sensi Kill e BisBlock. Outros dezoito discos foram selecionados para o teste de espectroscopia de impedância eletroquímica, sendo divididos em três grupos de seis espécimes, correspondentes aos três agentes anti-hiperestésicos citados anteriormente. Para ambos os testes as medidas foram realizadas nas seguintes condições: na presença de smear layer; após condicionamento com ácido fosfórico a 37% e após a aplicação dos materiais na superfície oclusal dos discos de dentina. Com base na análise estatística dos resultados, verificou-se, por meio dos dois métodos utilizados, que o gel Experimental e o BisBlock apresentaram efeitos estatisticamente semelhantes na redução da permeabilidade e foram significantemente mais eficientes que o Sensi Kill. Pode-se concluir que os agentes anti-hiperestésicos Experimental e BisBlock mostraram-se efetivos na redução da permeabilidade dentinária, pelos dois métodos de avaliação, e que o método de espectroscopia de impedância eletroquímica pode ser considerado um método alternativo para a avaliação da capacidade obstrutiva de agentes anti-hiperestésicos. / The aim of the present study was to quantify in vitro the dentin permeability of dentin treated with two potassium oxalate dentin desensitizers and one calcium phosphate agent through hydraulic conductance (Lp) and eletrochemical impedance spectroscopy (EIS) tests and, yet, to determine whether EIS may be considered a suitable alternative method to evaluate the occlusive capacity of the dentin desensitizers. For the hydraulic conductance, twenty-four dentin discs were obtained from extracted human third molars, which were divided into three groups of eight specimens, corresponding to the materials to be tested: Experimental gel, Sensi Kill and BisBlock. Other eighteen dentin discs were selected for theEIS and divided into three groups of six specimens corresponding to the three dentin desensitizers mentioned before. For both tests, the following conditions were measured: in the presence of smear layer, after 37% phosphoric acid etching and after the application of the tested materials in the occlusal surfaces of dentin discs. Based upon the statistical analysis of the results, it was demonstrated by both methods that Experimental gel and BisBlock performed statistically similarly in reducing dentin permeability and were significantly more efficient that Sensi Kill. It can be concluded that the dentin desensitizers Experimental e BisBlock were demonstrated to be effective in the reduction of dentin permeability by both experimental methods and that EIS may be considered as an alternative method for the evaluation of the obstructive capacity of dentin desensitizers.

O objetivo do presente estudo foi quantificar, in vitro, a permeabilidade da dentina tratada com dois agentes anti-hiperestésicos à base de oxalato e um à base de fosfato de cálcio, por meio de testes de condutividade hidráulica e de espectroscopia de impedância eletroquímica e, ainda, determinar se espectroscopia de impedância eletroquímica pode ser considerada um método alternativo para avaliar a capacidade obstrutiva de agentes antihiperestésicos. Para o teste de condutividade hidráulica foram utilizados vinte quatro discos de dentina, obtidos de terceiros molares humanos extraídos, que foram divididos em três grupos de oito espécimes, correspondentes aos três materiais testados: gel Experimental, Sensi Kill e BisBlock. Outros dezoito discos foram selecionados para o teste de espectroscopia de impedância eletroquímica, sendo divididos em três grupos de seis espécimes, correspondentes aos três agentes anti-hiperestésicos citados anteriormente. Para ambos os testes as medidas foram realizadas nas seguintes condições: na presença de smear layer; após condicionamento com ácido fosfórico a 37% e após a aplicação dos materiais na superfície oclusal dos discos de dentina. Com base na análise estatística dos resultados, verificou-se, por meio dos dois métodos utilizados, que o gel Experimental e o BisBlock apresentaram efeitos estatisticamente semelhantes na redução da permeabilidade e foram significantemente mais eficientes que o Sensi Kill. Pode-se concluir que os agentes anti-hiperestésicos Experimental e BisBlock mostraram-se efetivos na redução da permeabilidade dentinária, pelos dois métodos de avaliação, e que o método de espectroscopia de impedância eletroquímica pode ser considerado um método alternativo para a avaliação da capacidade obstrutiva de agentes anti-hiperestésicos. / The aim of the present study was to quantify in vitro the dentin permeability of dentin treated with two potassium oxalate dentin desensitizers and one calcium phosphate agent through hydraulic conductance (Lp) and eletrochemical impedance spectroscopy (EIS) tests and, yet, to determine whether EIS may be considered a suitable alternative method to evaluate the occlusive capacity of the dentin desensitizers. For the hydraulic conductance, twenty-four dentin discs were obtained from extracted human third molars, which were divided into three groups of eight specimens, corresponding to the materials to be tested: Experimental gel, Sensi Kill and BisBlock. Other eighteen dentin discs were selected for theEIS and divided into three groups of six specimens corresponding to the three dentin desensitizers mentioned before. For both tests, the following conditions were measured: in the presence of smear layer, after 37% phosphoric acid etching and after the application of the tested materials in the occlusal surfaces of dentin discs. Based upon the statistical analysis of the results, it was demonstrated by both methods that Experimental gel and BisBlock performed statistically similarly in reducing dentin permeability and were significantly more efficient that Sensi Kill. It can be concluded that the dentin desensitizers Experimental e BisBlock were demonstrated to be effective in the reduction of dentin permeability by both experimental methods and that EIS may be considered as an alternative method for the evaluation of the obstructive capacity of dentin desensitizers.

La conductivité hydraulique foliaire (KF) est une composante majeure du transport d’eau dans toute la plante. Dans les feuilles de noyer, la KF est stimulée à la lumière et est étroitement liée à l’accroissement du taux des transcrits d’aquaporines JrPIP2s. Par ailleurs, la corrélation entre la stimulation de la KF et des transcrits d’aquaporines à la lumière, n’est pas générale et dépend de l’espèce. Ici, nous étudions cette corrélation chez cinq espèces forestières (Juglans regia, Fagus sylvitica, Quercus robur, Salix alba et Populus tremula) différant par leur réponse à la lumière. Nous démontrons seulement chez le noyer (Juglans regia), la contribution des deux familles d’aquaporines PIP1s et PIP2s. Afin de mieux comprendre le rôle des JrPIP1s et JrPIP2 dans la réponse à la lumière, nous avons isolé 8 nouvelles isoformes dans les feuilles de noyer et nous avons étudié leurs profils d’expression sur une cinétique lumière. Toutes les isoformes étudiées sont accumulées à la lumière et réprimées à l’obscurité. De plus, la KF est dépendante de la qualité de lumière. Elle est réduite de 65% en absence de lumière bleue. Cette diminution serait liée à l’inhibition des transcrits d’aquaporines. Afin de caractériser les mécanismes moléculaires précoces impliqués dans la modulation de KF par la lumière, l’approche globale cDNA-AFLP a été menée sur des feuilles de noyer sous différentes conditions d’éclairement. Nous obtenons 12000 transcrits différentiels dérivés (TDFs) générés par les 128 couples d’amorces. Parmi les 187 séquences obtenues, 93 d’entre elles ont une fonction putative. Leur classification fonctionnelle montre que les gènes relatifs à la régulation cellulaire représentent environ 58% des TDFs identifiés. Les feuilles exposées à la lumière, montrent des changements dans les voies de : signalisation calcique, protéolyse, trafic vésiculaire et l’expression de divers facteurs de transcription et protéines de régulation. Pour mieux comprendre le rôle potentiel de la signalisation calcique dans la modulation de la KF par la lumière, nous avons étudié l’effet d’un inhibiteur des canaux calciques [LaCl3] et d’un antagoniste de calmoduline [W7] sur la KF et les transcrits des 10 JrPIPs. Comparées aux feuilles témoins, les inhibiteurs calciques provoquent une réduction de la KF et de la majorité des JrPIPs étudiées à la lumière. Nos résultats confirment l’implication du complexe Ca2+ /calmoduline dans la transduction du signal lumineux responsable de la stimulation de la KF et des transcrits d’aquaporines chez le noyer. / Leaf hydraulic conductance (Kleaf) takes a significant part in plant water relations. In walnut leaves, Kleaf was stimulated by light and tightly related to accumulation of JrPIP2s aquaporin transcripts. However, the light effect on Kleaf value is not systematically related to aquaporin regulation. Here we investigated the relationship between light, Kleaf and transcript levels of aquaporin in five species (Juglans regia, Fagus sylvitica, Quercus robur, Salix alba and Populus tremula) differing by the response of their Kleaf to light. Only for walnut leaves, we showed that light-increased Kleaf value is closely related to higher stimulation of both PIP1s and PIP2s aquaporins. To further investigate the involvement of aquporins (JrPIP1s and JrPIP2) in the light Kleaf modulation, 8 new full length aquaporins have been identified in walnut leaves and their expression pattern was monitored. All the aquaporin tested was turned up to be upregulated under light condition and downregulated under darkness. Moreover, we showed that the Kleaf response to light is quality-dependant, since it was reduced of 65% in the absence of blue light. Interstingly, this Kleaf reduction was correlated with a high downregulation of almost all aquaporins tested. To give an insight into the early molecular events involved in the light-induced Kleaf regulation, a large-scale transcriptomic analysis consisting of the cDNA-AFLP procedure was carried out on walnut leaves, kept at different light conditions. We obtained a total of 12,000 transcript-derived fragments (TDFs) by cDNA-AFLP with 128 primer pairs. Reliable sequences were obtained for 187 of these TDFs, and functions were attributed to 93 TDFs through BLAST searches in GenBank databases. Most of the 93 TDFs corresponded to genes encoding proteins involved in cellular regulation (58%). Leaves exposed to light showed changes in the Ca2+-signaling pathway, the ubiquitin-proteasome pathway, vesicle trafficking process and the expression of multiple transcription factors and protein regulators. To progress in understanding of a potential role for calcium signalling in light-modulated Kleaf, Kleaf values and transcript accumulation of 10 JrPIPs were monitored in leaves perfused with either a blocker of calcium channels [LaCl3] or a CaM antagonist [W7]. Compared to control, these Ca2+ -effectors led to a significant reduction in Kleaf and transcripts accumulation of almost all JrPIPs under light conditions. Our results indicate that Ca2+ /calmodulin complex may transduce the light signal required for stimulation of Kleaf and its correlated aquaporin expression.

Dans le contexte de changement climatique, la compréhension des mécanismes régissant les pertes en eau de la vigne peut permettre d'adapter le matériel végétal pour maintenir la productivité de la vigne et la qualité du vin. L'adaptation à la sécheresse est un caractère complexe faisant intervenir des mécanismes physiologiques liés aux génotypes du greffon et du porte-greffe. Mais les effets du porte-greffe sur la régulation stomatique du greffon sont mal connus. La production par les racines de signaux chimiques tels que l'ABA et/ou hydraulique pourraient y contribuer. La réponse physiologique et moléculaire à la contrainte hydrique a été analysée sur de jeunes boutures pour 7 porte-greffes plus ou moins adaptés à la sécheresse et 2 cépages connus pour leur caractère iso ou anisohydrique. Puis 23 combinaisons greffon/porte-greffe issues de ces génotypes ont été étudiées. Une analyse métabolique sur l'accumulation de l'ABA et ses dérivés a été menée sur feuilles, racines et dans la sève xylémienne. Ces informations ont été couplées à des analyses transcriptomiques sur des gènes du métabolisme et de la signalisation de ABA, et codant des aquaporines de type PIP. L‘analyse conjointe des données physiologiques, métabolomiques et transcriptomiques ont permis d'identifier des composants moléculaires discriminant les porte-greffes selon leur fond génétique et leur adaptation à la sécheresse. Les réponses globales à la contrainte hydrique sont mieux coordonnées au sein d‘un même tissu qu‘entre racines et feuilles. A l‘échelle de la plante greffée, une prépondérance du signal hydraulique est probable. Certains gènes répondent spécifiquement aux interactions greffon/porte-greffe. / In the context of climate change, understanding the mechanisms governing the water loss of the vine is necessary to adapt the plant material to maintain the productivity of the vine and wine quality. The adaptation to drought is a complex trait involving physiological mechanisms related to scion and rootstock genotypes. But the effects of the rootstock on stomatal regulation graft are still unknown. Production by roots of chemical signals such as ABA and / or hydraulic ones be involved. Molecular and physiological responses to water stress were analyzed on young cuttings for 7 rootstocks more or less adapted to drought and 2 varieties known for their iso or anisohydric behaviour. Then 23 combinations scion / rootstock from these genotypes were investigated. Metabolic analyses for ABA and its derivatives was conducted in leaves, roots and in the xylem sap. The information was integrated with transcriptomic analyzes for genes involved in ABA metabolism and signaling, and encoding PIP aquaporins. Joint analyses of physiological data, metabolomic and transcriptomic allow the identification of the molecular components discriminating rootstocks according to their genetic background and their adaptation to drought. Global responses to water stress are better coordinated within the same tissue between roots and leaves. At the scale of the grafted plant, a preponderance of the hydraulic signal is likely. Some genes specifically respond to the scion / rootstock interactions.

Le pommier (Malus Xdomestica Borkh.) cultivé en climat à hiver doux, avec un manque d'une quantité suffisante d'heures de températures froides, présente des anomalies morphologiques et physiologiques. Sur le plan de la phénologie, il s'agit notamment d'un débourrement printanier tardif et désynchronisé entre les différents bourgeons d'un même arbre. Sur le plan agronomique, la floraison et la nouaison sont irrégulières et étalées dans le temps et conduisent à une faible production de fruits. L'objectif de ce travail de thèse est premièrement de mieux caractériser les effets des températures hivernales sur le débourrement et la croissance des bourgeons axillaires en distinguant les effets respectifs des températures hivernales et du génotype. Il s'agit ensuite de vérifier l'hypothèse que les effets des températures sur le débourrement du bourgeon s'effectue via les effets sur son statut hydrique. Les travaux ont été réalisés en France et au Brésil. En France, les expérimentations ont porté sur quatre cultivars à fort besoin (‘Granny Smith', ‘Royal Gala', ‘Starkrimson') ou faible besoin (‘Condessa') en froid, cultivés en hiver froid (conditions extérieures) et doux (serres climatisées). Nous montrons que le débourrement résulte d'une séquence d'évènements où la température hivernale joue un rôle primordial sur les mécanismes de sortie de dormance et donc sur le débourrement proprement dit, durant la période froide. Les caractéristiques propres du cultivar jouent par contre un rôle dans la croissance ultérieure des bourgeons et donc dans l'architecture finale de la pousse du pommier. Par ailleurs, la chute tardive des feuilles, caractéristique du pommier en hiver doux, a peu d'effets sur le débourrement et la croissance des bourgeons. L'analyse du statut hydrique des bourgeons a été réalisée sur le tiers distal des pousses de pommier caractérisé par une forte fréquence de ramification en climat à hiver froid. Nous montrons que, dans la période allant de l'endordormance à la phase de pré-débourrement, la conductance hydraulique à la jonction entre l'axe porteur et le bourgeon varie peu au cours de l'hiver et entre cultivars. Par ailleurs, durant cette même période le potentiel hydrique intra-bourgeon reste négatif, entre -4.35 and -2.24 MPa. Enfin, quel que soit le cultivar, nous ne montrons pas de relation entre les températures hivernales, le potentiel hydrique ou la teneur relative en eau des bourgeons, et l'aptitude au débourrement ultérieur. Ces résultats suggèrent que le bourgeon est hydrauliquement isolé de son axe porteur pendant toute la période hivernale jusqu'à quelques jours précédant le débourrement. Les expérimentations au Brésil ont porté sur le cultivar ‘Eva ‘ à faible besoin en froid, cultivés en conditions naturelles d'hiver doux. Il s'agissait de vérifier les effets possibles de la position du bourgeon le long de l'axe porteur sur sa taille et sa teneur relative en eau. Nous montrons que, tout au long de l'hiver, les bourgeons distaux sont caractérisés par une plus grande taille et une teneur relative en eau plus élevée que les bourgeons proximaux avec une forte augmentation de la teneur relative en eau une semaine avant le débourrement printanier. Le débourrement acrotone semble donc résulter d'une évolution rapide du statut hydrique du bourgeon en fin d'écodormance. L'ensemble des résultats acquis en France et au Brésil, sur des cultivars caractérisés par des besoins variables en froid hivernal, indique que l'aptitude au débourrement printanier des bourgeons de pommier est davantage lié à un « effet rameau entier » qu'au statut hydrique proprement dit des bourgeons individuels, tout au moins jusqu'à quelques jours avant le débourrement effectif. La pousse annuelle de pommier apparait donc comme une unité morphologique et physiologique intégrée qui, dans un contexte climatique donné, conditionne le statut hydrique de chaque bourgeon et son aptitude au débourrement. / The apple tree (Malus Xdomestica Borkh.) presents morphological and physiological anomalies when grown in mild winter climates with insufficient winter chilling to overcome winter dormancy. Symptoms are typically delayed and erratic budburst, entailing desynchronized flowering and fruit-set and poor agronomic performances. This thesis aimed at gaining more insights on the following issues. Firstly, what are the effects of winter temperatures on axillary burdburst and bud outgrowth, and what are the respective effects of winter temperatures and cultivar?, and secondly, is there a link between the temperature-dependent budburst and bud water status? Works were done in France and Brazil. In France, experiments were carried out in controlled conditions on four apple cultivars characterized by either high chilling (‘Granny Smith', ‘Royal Gala', ‘Starkrimson') or low chilling (‘Condessa') requirements and were submitted to outdoor-cold and greenhouse-mild winter temperatures. We showed that the actual shoot architecture and budburst resulted from an ordered sequence of events with a pivotal role of winter temperatures on the dormancy completion of individual lateral buds. Endogenous factors related to the cultivar branching pattern overtook the temperature effect on the lateral bud outgrowth. Furthermore, the delayed senescence and subsequent leaf persistence during winter, characterizing the apple tree in the mild winter temperature conditions, had only a weak effect on the topological distribution of budburst and lateral outgrowth. The analyses of bud water status were done on distal buds only, characterized by high budburst frequency in cold winter conditions. We showed that, from endodormancy to the pre-budburst stage, xylem conductance at the stem-to-bud junction did not show consistent changes across cultivars and winter temperature treatments. Bud water potential had negative values, between -4.35 and -2.24 MPa, depending on cultivars and winter temperature treatments. Moreover, whatever the cultivar, there were no significant trends across dates for the effects of winter temperatures on bud water potential and relative water content without a consistent relationship with actual spring budburst frequency. These results suggested that lateral buds were hydraulically isolated from the parent stem during winter until a few days before budburst. The other set of experiments was carried out in Brazil, under mild winter conditions, on the low chilling apple cultivar ‘Eva'. The objectives were to gain more insights on the effect of the position of the over-wintering lateral bud along the whole-parent shoot on bud size and water content. Results highlighted that distal buds were larger and had a higher water content than proximal buds with a strong increase of water content a week before spring budburst. It was concluded that the acrotonic pattern of budburst was mainly established during ecodormancy. As a whole, we showed that spring budburst seemed more related to a whole-shoot effect than to the water status of the individual bud during winter dormancy. Our study substantiated the importance of the whole shoot as an integrated morphological and physiological unit in driving budburst and further growth.

La performance d'une plante repose en partie sur sa capacité à capturer l'énergie lumineuse via la croissance foliaire. La littérature souligne deux limitations majeures de la croissance, de nature métabolique ou hydraulique. Nous testons ici l'hypothèse originale que l'importance relative de ces deux limitations est structurée par l'ontogenèse de la feuille chez Arabidopsis thaliana. Nous montrons que la disponibilité en carbone restreint la croissance des jeunes feuilles, tandis qu'une compétition hydraulique entre croissance et transpiration s'accroît au cours de l'ontogenèse. La mise en place de cette limitation hydraulique s'explique par une dégradation de la capacité du xylème et probablement des aquaporines à approvisionner la feuille en eau, malgré une diminution ontogénétique de la transpiration. Cette dernière est la conséquence de l'acquisition progressive de la sensibilité des stomates aux signaux de fermeture, notamment l'obscurité et l'acide abscissique (ABA), hormone induite par la sécheresse. Enfin, nous mettons en évidence une nouvelle composante de la sensibilité stomatique à l'ABA, conservée chez des mutants décrits comme insensibles à cette hormone : l'ABA induit une diminution de la conductance hydraulique foliaire qui abaisse le potentiel hydrique foliaire et in fine la conductance stomatique. Ce mécanisme chez les feuilles développées contribuerait sous stress hydrique à rediriger le flux d'eau vers les feuilles en croissance. Plus généralement, le contrôle des stomates par des mécanismes hydrauliques induits par l'ABA pourrait être une composante majeure de l'ajustement entre offre et demande en eau chez les plantes soumises à un stress hydrique. / In plants, leaf growth is the central process allowing energy capture and space colonization. The literature suggests that leaf growth is predominantly determined by metabolic and hydraulic limitations. Here, we test the original hypothesis that the relative importance of metabolics and hydraulics on the control of leaf growth is organized according to leaf ontogeny in Arabidopsis thaliana. We show that leaf carbon balance limits growth of the young leaves which therefore grow at a slower rate in the nighttime, while a hydraulic limitation gradually establishes in the daytime, when growth and transpiration competes for water. This gradual hydraulic limitation is underlain by a deterioration of leaf venation and probably aquaporins capacity to supply water to the leaf, despite an ontogenetic decrease in transpiration. This decline in transpiration occurs because stomata acquire throughout leaf ontogeny their sensitivity to the major closure signals, including darkness and abscisic acid (ABA), a hormone induced by drought. Finally, we discover a novel component of stomatal sensitivity to ABA, conserved in mutants described as insensitive to ABA in isolated epidermis: ABA induces a decrease in leaf hydraulic conductance which lowers leaf water potential and stomatal conductance according to a hydraulic cascade. Decreasing leaf hydraulic conductance through ABA action in fully expanded leaves would contribute to redirect water flow to the young leaves under water stress. More generally, controlling stomata by ABA-induced hydraulic mechanisms could be a crucial component of the coordination between water supply and water demand in plants under water challenging conditions.

[Truncated abstract. Please see the pdf format for the complete text. Also, formulae and special characters can only be approximated here. Please see the pdf version for an accurate reproduction.] This thesis examines the water use, ecophysiology and hydraulic architecture of Eucalyptus marginata (jarrah) growing on bauxite mine rehabilitation sites in the jarrah forest of south-western Australia. The principal objective was to characterise the key environment and plant-based influences on tree water use, and to better understand the dynamics of water use over a range of spatial and temporal scales in this drought-prone ecosystem. A novel sap flow measurement system (based on the use of the heat pulse method) was developed so that a large number of trees could be monitored concurrently in the field. A validation experiment using potted jarrah saplings showed that rates of sap flow (transpiration) obtained using this system agreed with those obtained gravimetrically. Notably, diurnal patterns of transpiration were measured accurately and with precision using the newly developed heat ratio method. Field studies showed that water stress and water use by jarrah saplings on rehabilitation sites were strongly seasonal: being greatest in summer when it was warm and dry, and least in winter when it was cool and wet. At different times, water use was influenced by soil water availability, vapour pressure deficit (VPD) and plant hydraulic conductance. In some areas, there was evidence of a rapid decline in transpiration in response to dry soil conditions. At the end of summer, most saplings on rehabilitation sites were not water stressed, whereas water status in the forest was poor for small saplings but improved with increasing size. It has been recognised that mature jarrah trees avoid drought by having deep root systems, however, it appears that saplings on rehabilitation sites may have not yet developed functional deep roots, and as such, they may be heavily reliant on moisture stored in surface soil horizons. Simple predictive models of tree water use revealed that stand water use was 74 % of annual rainfall at a high density (leaf area index, LAI = 3.1), high rainfall (1200 mm yr-1) site, and 12 % of rainfall at a low density (LAI = 0.4), low rainfall (600 mm yr-1) site, and that water use increased with stand growth. A controlled field experiment confirmed that: (1) sapling transpiration was restricted as root-zone water availability declined, irrespective of VPD; (2) transpiration was correlated with VPD when water was abundant; and (3) transpiration was limited by soil-to-leaf hydraulic conductance when water was abundant and VPD was high (> 2 kPa). Specifically, transpiration was regulated by stomatal conductance. Large stomatal apertures could sustain high transpiration rates, but stomata were sensitive to hydraulic perturbations caused by soil water deficits and/or high evaporative demand. No other physiological mechanisms conferred immediate resistance to drought. Empirical observations were agreeably linked with a current theory suggesting that stomata regulate transpiration and plant water potential in order to prevent hydraulic dysfunction following a reduction in soil-to-leaf hydraulic conductance. Moreover, it was clear that plant hydraulic capacity determined the pattern and extent of stomatal regulation. Differences in hydraulic capacity across a gradient in water availability were a reflection of differences in root-to-leaf hydraulic conductance, and were possibly related to differences in xylem structure. Saplings on rehabilitation sites had greater hydraulic conductance (by 50 %) and greater leaf-specific rates of transpiration at the high rainfall site (1.5 kg m-2 day1) than at the low rainfall site (0.8 kg m-2 day1) under near optimal conditions. Also, rehabilitation-grown saplings had significantly greater leaf area, leaf area to sapwood area ratios and hydraulic conductance (by 30-50 %) compared to forest-grown saplings, a strong indication that soils in rehabilitation sites contained more water than soils in the forest. Results suggested that: (1) the hydraulic structure and function of saplings growing under the same climatic conditions was determined by soil water availability; (2) drought reduced stomatal conductance and transpiration by reducing whole-tree hydraulic conductance; and (3) saplings growing on open rehabilitation sites utilised more abundant water, light and nutrients than saplings growing in the forest understorey. These findings support a paradigm that trees evolve hydraulic equipment and physiological characteristics suited to the most efficient use of water from a particular spatial and temporal niche in the soil environment.

Submitted by Gabriela Lopes (gmachadolopesufpel@gmail.com) on 2016-09-14T18:22:14Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Tese Juliano Dutra Schmitz.pdf: 9582623 bytes, checksum: f579b0458e2ab6f4ae5579715de842ea (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2016-09-15T19:46:43Z (GMT) No. of bitstreams: 2 Tese Juliano Dutra Schmitz.pdf: 9582623 bytes, checksum: f579b0458e2ab6f4ae5579715de842ea (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-09-15T19:46:43Z (GMT). No. of bitstreams: 2 Tese Juliano Dutra Schmitz.pdf: 9582623 bytes, checksum: f579b0458e2ab6f4ae5579715de842ea (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2014-12-03 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / A macieira (Malus X domestica Borkh.) apresenta anomalias fisiológicas quando cultivada em regiões de inverno ameno, onde o frio hibernal é insuficiente para superação da dormência. Assim, na presente tese foram estudados três temas de pesquisa. No tema 1 estudou-se a distribuição e fenologia da brotação e crescimento inicial da ramificação primaveril; No tema 2 estudou-se a brotação primaveril através da determinação do status hídrico de gemas laterais e da condutância hidráulica do xilema. O tema 3 realizou-se a análise do comportamento da brotação primaveril de uma cultivar de baixo exigência em frio cultivada em inverno ameno. Para isso, foram montados dois dispositivos experimentais: Experimento 1: realizado em Montpellier/França, onde foram estudadas quatro cultivares de macieira, com diferentes requerimentos em frio (‘Condessa’, ‘Granny Smith’, ‘Royal Gala’ e ‘Starkrimson’), submetidas a dois regimes térmicos (inverno frio, condições naturais de Montpellier; e inverno ameno, temperatura controlada em casa-de-vegetação). Experimento 2: realizado com a cultivar de baixo requerimento em frio ‘Eva’ sob regime térmico hibernal ameno (condição natural de Capão do leão/Brasil). A partir do experimento 1, dois artigos foram redigidos . Conclui-se a partir dos resultados obtidos (artigos 1 e 2) que as temperaturas hibernais têm o principal efeito na distribuição da ramificação ao longo do eixo principal e no tempo para brotação; a presença de folha das plantas submetidas ao regime térmico de inverno ameno não afeta a distribu ição de ramos prolépticos vegetativos; a cultivar exerce efeito no crescimento da ramificação. Com relação ao status hídrico, conclui-se que durante o inverno (período de dormência) as gemas laterais permanecem hidraulicamente isoladas do eixo principal; assim como o potencial de brotação está relacionado a um efeito ramo inteiro (todo eixo principal) do que ao potencial individual de cada gema lateral. Através do experimento 2, um artigo foi elaborado, tendo por objetivo testar a hipótese que a posição em que a gema lateral está localizada sobre o eixo principal têm efeito na brotação primaveril, no conteúdo de água e tamanho das mesmas. Pode-se concluir deste estudo que uma semana antes a brotação, as gemas localizadas na zona distal possuem maior potencial de crescimento (maior frequência de brotação e menor tempo médio para brotação), além de apresentarem maior umidade ponderal e tamanho. / The apple tree (Malus X domestica Borkh.) presents morphological and physiological anomalies when grown in mild winter climates with insufficient winter chilling to overcome winter dormancy. Symptoms are typically delayed and erratic budburst, entailing desynchronized flowering and fruitset and poor agronomic performances. This thesis aimed at gaining more insights on the following issues. Firstly, what are the effects of winter temperatures on axillary burdburst and bud outgrowth, and what are the respective effects of winter temperatures and cultivar?, and secondly, is there a link between the temperature-dependent budburst and bud water status? Works were done in France and Brazil. In France, experiments were carried out in controlled conditions on four apple cultivars characterized by either high chilling (‘Granny Smith’, ‘Royal Gala’, ‘Starkrimson’) or low chilling (‘Condessa’) requirements and were submitted to outdoor-cold and greenhouse-mild winter temperatures. We showed that the actual shoot architecture and budburst resulted from an ordered sequence of events with a pivotal role of winter temperatures on the dormancy completion of individual lateral buds. Endogenous factors related to the cultivar branching pattern overtook the temperature effect on the lateral bud outgrowth. Furthermore, the delayed senescence and subsequent leaf persistence during winter, characterizing the apple tree in the mild winter temperature conditions, had only a weak effect on the topological distribution of budburst and lateral outgrowth. The analyses of bud water status were done on distal buds only, characterized by high budburst frequency in cold winter conditions. We showed that, from endodormancy to the pre-budburst stage, xylem conductance at the stem-to-bud junction did not show consistent changes across cultivars and winter temperature treatments. Bud water potential had negative values, between -4.35 and -2.24 MPa, depending on cultivars and winter temperature treatments. Moreover, whatever the cultivar, there were no significant trends across dates for the effects of winter temperatures on bud water potential and relative water content without a consistent relationship with actual spring budburst frequency. These results suggested that lateral buds were hydraulically isolated from the parent stem during winter until a few days before budburst. The other set of experiments was carried out in Brazil, under mild winter conditions, on the low chilling apple cultivar ‘Eva’. The objectives were to gain more insights on the effect of the position of the over-wintering lateral bud along the whole-parent shoot on bud size and water content. Results highlighted that distal buds were larger and had a higher water content than proximal buds with a strong increase of water content a week before spring budburst. It was concluded that the acrotonic pattern of budburst was mainly established during ecodormancy. As a whole, we showed that spring budburst seemed more related to a whole-shoot effect than to the water status of the individual bud during winter dormancy. Our study substantiated the importance of the whole shoot as an integrated morphological and physiological unit in driving budburst and further growth.