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[ES] En respuesta a estreses de tipo biótico y abiótico, las plantas sintetizan proteínas de defensa y compuestos químicos de diversa naturaleza. Estos compuestos pueden actuar de manera directa, a través de propiedades antioxidantes, antifúngicas o antibacterianas, o actuar como metabolitos defensivos indirectos. Dentro de este último grupo de compuestos defensivos, cabe destacar a los compuestos fenólicos y los compuestos orgánicos volátiles (VOCs). En nuestro grupo de investigación se ha profundizado en el estudio de estos metabolitos secundarios implicados en la respuesta defensiva de las plantas. Por una parte, se identificó el ácido gentísico (GA) como una molécula señal que actúa de manera complementaria al ácido salicílico (SA) en infecciones de tipo sistémico. Además, se ha tratado de profundizar en el estudio de la biosíntesis del GA a través de la enzima salicilato 5-hidroxilasa (S5H), encargada de la conversión de SA a GA. Para ello, se ha llevado a cabo la caracterización fenotípica, molecular y química de plantas transgénicas de tomate que tienen silenciado el gen S5H mediante la técnica de RNA de interferencia (RNAi_S5H) frente a infecciones de tipo bacteriano y viroidal. Las plantas de tomate RNAi_S5H presentaron un aumento de resistencia frente a Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) y el viroide de la exocortis de los cítricos (CEVd). Del mismo modo, se llevaron a cabo análisis metabolómicos de estas plantas transgénicas RNAi_S5H tras ambas infecciones, observándose diferencias relacionadas con el metabolismo del SA, que parecen indicar que la homeostasis del SA es específica para cada interacción tomate-patógeno. Por otra parte, se identificaron algunos ésteres de (Z)-3-hexenol que eran emitidos de manera diferencial tras la infección bacteriana con la cepa avirulenta de Pst DC3000 en plantas de tomate cv. Rio Grande. Concretamente, tratamientos exógenos con el compuesto volátil butanoato de (Z)-3-hexenilo (HB) fueron capaces de inducir de manera significativa el cierre de estomas, la activación de genes defensivos y un aumento en la resistencia frente a la infección bacteriana. La eficacia de este compuesto como inductor de cierre estomático fue comprobada en diferentes cultivos agronómicos, como Arabidopsis, Medicago, Zea, Citrus y Nicotiana, confirmando su papel como un inductor de cierre estomático universal. Dado el potencial de este compuesto en agricultura, se emplearon aproximaciones genéticas, bioquímicas y farmacológicas para descifrar el mecanismo de señalización del cierre estomático mediado por HB. Una vez el volátil es percibido por los receptores de la planta, se activan diferentes componentes de la cascada de señalización defensiva, como canales permeables de Ca2+ o la producción de especies reactivas de oxígeno (ROS). Asimismo, el HB es capaz de desencadenar la activación de las proteínas quinasas activadas por mitógenos MPK3 y MPK6, induciendo el cierre estomático de una manera independiente a la síntesis y señalización mediada por ácido abscísico (ABA). Por último, la eficacia del HB fue evaluada en condiciones de campo frente a estreses tanto de tipo biótico como abiótico y en procesos de desarrollo como la maduración, proponiendo un uso del HB como un nuevo compuesto fitoprotector natural para el control de estreses de forma sostenible en agricultura. / [CA] En resposta a estressos de tipus biòtic i abiòtic, les plantes sintetitzen proteïnes de defensa i compostos químics de diversa naturalesa. Aquests compostos poden actuar de manera directa, a través de propietats antioxidants, antifúngiques o antibacterianes, o actuar com a metabòlits defensius indirectes. Dins d'aquest últim grup de compostos defensius, cal destacar als compostos fenòlics i els compostos orgànics volàtils (VOCs). En el nostre grup d'investigació s'ha aprofundit en l'estudi d'aquests metabòlits secundaris implicats en la resposta defensiva de les plantes. D'una banda, es va identificar l'àcid gentísic GA) com una molècula senyal que actua de manera complementària a l'àcid salicílic (SA) en infeccions de tipus sistèmic. A més, s'ha tractat d'aprofundir en l'estudi de la biosíntesi del GA a través d l'enzim salicilato 5-hidroxilasa (S5H), encarregada de la conversió de SA a GA. Per a això, s'ha dut a terme la caracterització fenotípica, molecular i química de plantes de transgèniques de tomaca que tenen silenciat el gen S5H mitjançant la tècnica d'RNA d'interferència (RNAi_S5H) enfront d'infeccions de tipus bacterià i viroidal. Les plantes de tomaca RNAi_S5H van presentar un augment de resistència enfront de Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) i el viroide de la exocortis dels cítrics (CEVd). De la mateixa manera, es van dur a terme anàlisi metabolómics d'aquestes plantes transgèniques RNAi_S5H després de totes dues infeccions, observant-se diferències relacionades amb el metabolisme del SA, que sembla indicar que l'homeòstasi del SA és específica per a cada interacció tomaca-patògena. D'altra banda, es van identificar alguns èsters de (Z)-3-hexenol que eren emesos de manera diferencial després de la infecció bacteriana amb el cep avirulent de Pst DC3000 en plantes de tomaca cv. Rio Gran. Concretament, tractaments exògens amb el compost volàtil butanoato de (Z)-3-hexenilo (HB) van ser capaces d'induir de manera significativa el tancament d'estomes, l'activació de gens defensius i un augment en la resistència enfront de la infecció bacteriana. L'eficàcia d'aquest compost com a inductor de tancament estomàtic va ser comprovada en diferents cultius agronòmics, com Arabidopsis, Medicago, Zea, Citrus i Nicotiana, confirmant el seu paper com un inductor de tancament estomàtic universal. Donat el potencial d'aquest compost en agricultura, es van emprar aproximacions genètiques, bioquímiques i farmacològiques per a desxifrar el mecanisme de senyalització del tancament estomàtic mediat per HB. Una vegada el volàtil és percebut pels receptors de la planta, s'activen diferents components de la cascada de senyalització defensiva, com a canals permeables de Ca2+ o la producció d'espècies reactives d'oxigen (ROS). Així mateix, el HB és capaç de desencadenar l'activació de les proteïnes cinases activades per mitógens MPK3 i MPK6, induint el tancament estomàtic d'una manera independent a la síntesi i senyalització mediada per l'àcid abscísic (ABA). Finalment, l'eficàcia del HB va ser avaluada en condicions de camp enfront d'estressos tant de tipus biòtic com abiòtic, i en processos de desenvolupament com la maduració, proposant un l'ús del HB com a nou compost fitoprotector natural per al control d'estressos de manera sostenible en agricultura. / [EN] In response to biotic and abiotic stress, plants synthesize defence proteins and chemical compounds from diverse nature. These compounds can act directly, trough antioxidant, antifungal or antibacterial properties, or indirectly as defensive metabolites. Among these group of defensive metabolites, phenolic compounds and volatile organic compounds (VOCs) present a major role. Our research group have a strong background in studying the role of plant secondary metabolites in plant defence mechanisms. On one hand, gentisic acid (GA) was first described as a signal molecule that acts complementary to salicylic acid (SA) in systemic infections. Furthermore, SA conversion to GA trough the salicylate 5-hydroxylase enzyme (S5H) has received much attention. For this purpose, S5H-silenced transgenic tomato plants (RNAi_S5H) have been phenotypically, molecularly, and chemically characterized against both, bacterial and viroidal inoculations. RNAi_S5H tomato plants resulted in enhanced resistance to both Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) and Citrus Exocortis Viroid (CEVd). Moreover, metabolomics analysis of these transgenic plants upon bacterial and viroid infections revealed differences related to SA metabolism, suggesting that SA homeostasis is specific for each tomato-pathogen interaction. On the other hand, some esters of (Z)-3-hexenol were identified to be differentially emitted by tomato cv. Rio Grande plants upon infection with the avirulent strain of the bacterium Pst DC3000. Particularly, treatments with the volatile (Z)-3-hexenyl butyrate (HB) resulted in significant stomatal closure, defence genes induction and enhanced resistance to the bacteria. Moreover, the efficacy of this compound as a stomata closer was tested in different agronomic crop as Arabidopsis, Medicago, Zea, Citrus y Nicotiana plants, postulating HB as a new universal stomata closer. Due to its potent properties, the signalling pathway of the HB-mediated stomata closure has been deciphered by using different genetic, biochemical, and pharmacological approaches. The perception of this volatile by plant receptors appeared to initiate different defence signalling events, including the activation of Ca2+ permeable channels or reactive oxygen species (ROS) burst. Moreover, HB triggered the activation of the mitogen-activated protein kinases MPK3 and MPK6, inducing stomatal closure independently of abscisic acid (ABA) biosynthesis and signalling. Additionally, HB efficacy has been also tested in field conditions and against both biotic and abiotic stresses, and also during ripening, proposing HB as a new natural phytoprotector for the sustainable control of stresses in agriculture. / This work was funded by Grant AICO/2017/048 from the Generalitat Valenciana and by Grant INNVAL10/18/005 from the Agència Valenciana de la Innovació (Spain). / Payá Montes, C. (2023). Secondary Metabolites in Plant Defence Mechanisms [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/193041

From the Proceedings of the 1977 Meetings of the Arizona Section - American Water Resources Assn. and the Hydrology Section - Arizona Academy of Science - April 15-16, 1977, Las Vegas, Nevada / Relative water content (RWC), water potential (P), and gas exchange were measured on saltcedar at the Bernardo, New Mexico, lysimeter site. RWC and s were closely correlated; but, water potential measurements, taken with a pressure bomb, were more convenient and reliable. RWC and r decreased sharply from sunup until about 0900, when minimum values of about -26 bars T or 80% RWC were reached. Water status then remained constant or improved slightly through late afternoon. Transpiration rates typically remained high until about noon and then began a steady, gradual decrease that continued throughout the afternoon. The data suggest that water stress may be a factor in initiating stomatal closure; however, transpiration continued to decline despite a constant or improved leaf water status. Maximum net photosynthetic rates occurred by 0900, and depressions throughout the remainder of the day were largely accounted for by increased leaf temperatures. Afternoon depressions in transpiration and photosynthesis occurred in twigs held at constant temperature and relative humidity, suggesting that a diurnal rhythm may be involved in control of gas exchange. Water status of plants growing on the lysimeters was comparable to that of plants in adjacent natural stands; gas exchange rates were slightly higher for the lysimeter-grown plants.

Made available in DSpace on 2016-06-02T19:31:38Z (GMT). No. of bitstreams: 1 1456.pdf: 806888 bytes, checksum: c1a017c0d739e9db5205b7784a9e4a47 (MD5) Previous issue date: 2007-06-06 / Universidade Federal de Sao Carlos / Young plants of Coffea arabica grafted over C. canephora were submitted to six watering cycles (WC) during 35 days. Each WC was accomplished until net photosynthesis (PN) close to zero or leaf water potential (&#936;) close to -2.0 MPa in Acauã (AC), Mundo- Novo (MN), and Obatã (OB) cultivars. After 2 WC (about 10 days) gs dropped from 0.15 to 0.01 mol m-2 s-1 in all cultivars changing the pattern of leaf gas exchange in subsequent WC. From 3 to 6 WC the cultivars showed high oscillations of water use efficiency or substomatal CO2 concentration (Ci) besides negative PN at the end of WC. After 3 WC, gs increased faster than PN after watering and Ci increased indicating some damage in photosynthetic machinery. It happened strongly in OB, which showed progressive reduction of PN after 3 WC and the lowest values of &#936; (-2.0 MPa) at the end of 6 WC. The survivorships for cultivars were 80% till 3 WC and 70% after 6 WC. The gas exchange pattern alteration could indicate the potential hardening of AC, MN and OB cultivars. After field plantation of hardened and control (without perivous hardening) Mundo-Novo and Obatã cultivars, with 180-days-old, it was observed significant (p<0.05) greater accumulation of leaf, stem and root biomass in hardened in control plants. Stem diameter, height and the number of leaf and branch were also higher (p<0.05) in previous hardened individuals. Greater differences in crown structure and biomass between control and hardened plants were observed mainly after the first period of low water availability (210 days under field conditions). The higher leaf number and leaf area in more branched crowns could carbon assimilation and grain production. For practical proposes, 3 WC before growing under field conditions are sufficient for changing consistently the leaf gas exchange pattern in 3 cultivars studied, keeping photosynthetic machinery free of damage and high survivorship. Previous hardening proportioned less interrupted vegetative development after transplantation under field conditions in both cultivars. / Plantas jovens de Coffea arabica enxertadas sobre C. canephora foram submetidas a 6 ciclos de suspensão de rega (CR) durante 35 dias. Cada CR foi acompanhado até que a fotossíntese líquida (PN) fosse próxima de zero ou até que o potencial hídrico foliar (&#936;) fosse próximo de -2.0 MPa nos cultivares Acauã (AC), Mundo-Novo (MN) e Obatã (OB). Após 2 CR (cerca de 10 dias) os valores médios de condutância estomática (gs) diminuíram de 0,15 para 0,01 mol m-2 s-1 em todos os cultivares, mudando o padrão de trocas gasosas nos subseqüentes CR. De 3 a 6 CR os cultivares apresentaram maiores oscilações da eficiência do uso da água e da concentração substomática de CO2, além de valores negativos de PN ao final dos CR. Após 3 CR ocorreu um aumento proporcionalmente maior em gs do que em PN após rega, com um aumento correspondente de Ci, indicando possíveis danos no aparato fotossintético. Este evento ocorreu de forma mais clara em OB, o qual demonstrou redução progressiva de PN após 3 CR e os menores valores de &#936; (-2,0 MPa) ao final de 6 CR. A sobrevivência dos cultivares foi de 80% em 3 CR e 70% em 6 CR. As alterações dos padrões de trocas gasosas nos cultivares AC, MN e OB podem indicar que os exemplares foram potencialmente rustificados. Após o plantio de MN e OB rustificados e controle (sem prévia rustificação) em campo, com 180 dias de idade, foi observado no tratamento rustificado valores significativamente (p<0,05) maiores do acúmulo de biomassa e do número de componentes estruturais da copa. A rustificação prévia proporcionou maior acumulo de biomassa de folhas, de caule e de raízes que nas plantas controle em ambos os cultivares. O diâmetro do caule, a altura, o número de folhas e o número de ramos também foram significativamente maiores (p<0,05) nos indivíduos previamente rustificados. As maiores diferenças da estrutura da copa e da biomassa acumulada entre plantas controle e rustificadas foram encontradas principalmente após o primeiro período de rezudida disponibilidade hídrica (210 dias em condições de campo). O maior número de folhas e a maior área foliar em copas mais ramificadas podem proporcionar um aumento da assimilação de carbono por planta e maior produção de grãos nas plantas rustificadas. Em termos práticos, 3 CR antes do crescimento em campo são suficientes para mudar consistentemente o padrão de trocas gasosas foliares nos 3 cultivares estudados, mantendo o aparato fotossintético livre de danos e proporcionando maior sobrevivência. A prática de rustificação prévia antes do plantio deve ser adotada nos cultivares estudados, pois o custo é reduzido e o desenvolvimento vegetativo inicial é fortemente favorecido após o transplante sob condições de campo.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The objective of this study was to determine the response of plant transpiration and leaf growth to the fraction of transpirable soil water (FTSW) in potato and cassava clones. Three experiments were conducted with potato, two of them were in Spring, with plantings on 08/20/2008 (BAT1) and on 10/20/2008 (BAT2), and one experiment in Fall, with planting on 03/25/2009 (BAT3). Two experiments were conducted with cassava, with plantings on 09/11/2009 (MAN1) and on 09/08/2010 (MAN2). The experiments were conducted in 8 liter pots filled with soil inside a plastic house. Two potato clones (Macaca and SMINIA 793101- 3) and two cassava clones (Fécula Branca and Fepagro RS 13) were used. Soil water, represented by the FTSW, transpiration and leaf growth were measured on a daily basis during the period of soil drying. The threshold FTSW that starts to affect transpiration, which is an indicator of stomata closure, was 0.38, 0.47 and 0.28 for the clone Macaca and 0.48, 0.51 and 0.32 for the clone SMINIA793101-3 in BAT1, BAT2 and BAT3, respectively, whereas leaf growth decreased at a greater threshold FTSW for Macaca . These results indicate that the clone SMINIA793101-3 is more tolerant to soil water deficit than Macaca . For cassava, the threshold FTSW for transpiration was 0.45 in the clone Fécula Branca and 0.50 in the clone Fepagro RS 13 . The onset of reduction in leaf growth was at a FTSW of 0.51 in the clone Fécula Branca and 0.49 in the clone Fepagro RS 13 .When data were separated in days with low and high vapor pressure deficit at 3PM, threshold FTSW for transpiration and leaf growth changed in both species and within each species. These results confirm the genetic variability available in each species and are a reminder on the complex genotype and environment relationships that characterize agricultural crops. / O objetivo deste trabalho foi determinar a resposta da planta em termos de transpiração e de crescimento foliar em clones de batata e de mandioca ao conteúdo de água disponível no solo, representado pela fração de água transpirável no solo (FATS). Foram realizados três experimentos com a cultura da batata, sendo dois na primavera (cultivo de safra), com plantios em 20/08/2008 (BAT1) e 20/10/2008 (BAT2) e um no outono (cultivo de safrinha), com plantio em 25/03/2009 (BAT3), e dois experimentos com a cultura da mandioca com plantio em 11/09/2009 (MAN1) e 08/09/2010 (MAN2). Os experimentos foram conduzidos em vasos de 8 litros preenchidos com solo e dispostos no interior de um abrigo plástico telado. Os clones de batata utilizados foram Macaca e SMINIA793101-3 e os clones de mandioca foram Fécula Branca e Fepagro RS 13. A água disponível, representada pela FATS, a transpiração e o crescimento foliar foram medidos diariamente durante o período de imposição da deficiência hídrica em cada experimento. Para a cultura da batata a FATS crítica que começa a afetar a transpiração, indicativo do início do fechamento estomático, foi de 0,38, 0,47 e 0,28 no clone Macaca e 0,48, 0,51 e 0,32 no clone SMINIA793101-3, para os experimentos BAT1, BAT2 e BAT3, respectivamente, enquanto o crescimento foliar começou a ser reduzido com um valor de FATS crítica maior para o clone Macaca, indicando que o clone SMINIA793101-3 é mais tolerante ao déficit hídrico no solo que o clone Macaca. Para a cultura da mandioca, a FATS crítica em que a transpiração começa a ser reduzida foi de 0,45 para o clone Fécula Branca e 0,50 para o clone Fepagro RS 13. O crescimento foliar começou a ser reduzido quando a FATS atingiu valores de 0,51 no clone Fécula Branca e 0,49 no clone Fepagro RS 13. Quando os dados foram separados em dias com baixa e alta demanda evaporativa do ar, representada pelo déficit de pressão de vapor do ar às 15 horas, houve diferença na FATS crítica que começa a afetar a transpiração e o crescimento foliar para as duas espécies e dentro de cada espécie. Estes resultados confirmam a variabilidade genética disponível em cada espécie e são um lembrete da complexa relação entre genótipo e ambiente que caracteriza as culturas agrícolas.

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.

No description available.

Les plantes présentent une incroyable diversité de tailles, formes et couleurs, étroitement liée à certaines de leurs fonctions biologiques telles que la photosynthèse, la reproduction, etc. De ce fait, la façon dont ces organismes multicellulaires acquièrent des formes complexes est une question clé en biologie du développement. La morphologie des organes végétaux résulte en effet de la modulation, à l’échelle cellulaire, de patrons d’expression génétique, de croissance et de différenciation. Bien que la morphogénèse ait été largement étudiée d’un point de vue moléculaire, nous ne savons toujours pas comment ces réseaux génétiques sont traduits en formes biologiques. Le but de ce projet de recherche est donc d’étudier le développement des organes latéraux (feuilles juvéniles, feuilles caulinaires et organes floraux, id sépales, pétales et anthères) chez l’espèce modèle Arabidopsis thaliana. Afin d’approcher la question du rôle des interactions complexes entre cellules et organes lors du développement, nous nous intéressons à la variabilité entre les organes, mais aussi à la variabilité cellulaire intrinsèque de chaque organe. Nous avons donc testé (1) si la diversité de formes observées chez les organes latéraux résulte de modulations d’un programme développemental commun; (2) si la croissance et le développement des organes latéraux est un phénomène stochastique ou dépend de mécanismes sous-jacents spécifiques. Pour ce faire, nous utilisons une approche multidisciplinaire basée sur la génétique, la microscopie confocale et l’analyse d’images 3D pour extraire les patrons de croissance inhérents aux différents organes. Les résultats de la première étude (Chapitre 2) montrent que la forme des organes dépend de l’équilibre entre croissance et différentiation, dont la régulation précise permet l'acquisition de fonctions hautement spécialisées. La feuille caulinaire, par exemple, présente un retard de différenciation qui permet une activité morphogénétique prolongée et une redistribution de la croissance. À travers la suppression transitoire de la croissance lors des premiers stades de développement, la trajectoire développementale de la feuille caulinaire permet sa double fonction, à la fois protectrice et photosynthétique.\par La deuxième étude (Chapitre 3), quant-à-elle, s’intéresse aux comportements des cellules individuelles, dont la croissance, bien que contrôlée par des informations positionnelles, est souvent hétérogène. Cette variabilité résulte de la différenciation de cellules spécialisés, les stomates, qui suivent un programme de développement spécifique. Le comportement autonome de ces cellules, asynchrone, est la principale source de variabilité dans des tissus dont la croissance est autrement homogènes. Dans l’ensemble, cette thèse a permis de mettre en lumière l’importance de la temporalité lors du développement des organes végétaux. Que ce soit à l’échelle de l’organe, du tissu ou de la cellule, la modulation et la synchronisation de la croissance et de la différentiation sont nécessaires à l’acquisition des formes stéréotypiques des organes et à leur complexité fonctionnelle. / Plants display an incredible diversity of sizes, shapes, and colors, closely linked to some of their biological functions, such as photosynthesis, reproduction, etc. How these multicellular organisms acquire complex shapes is, therefore, a key question in developmental biology. The morphology of plant organs results from cell-level modulation of patterns of gene expression, growth, and differentiation. Although morphogenesis has been extensively studied from a molecular point of view, how genetic networks are translated into biological forms is still unclear. Thus, the aim of this research project is to study the development of lateral organs (rosette leaves, cauline leaves, and floral organs, i.e. sepals, petals, and anthers) in the model species Arabidopsis thaliana. To address the question of the role of complex cell-organ interactions during development, we are interested not only in variability between organs but also in the intrinsic cellular variability of each organ. We, therefore, tested (1) whether the diversity of shapes observed in lateral organs results from modulations of a common developmental program; (2) whether the growth and development of lateral organs is a stochastic phenomenon or depends on specific underlying mechanisms. To this end, we are using a multidisciplinary approach based on genetics, confocal microscopy, and 3D image analysis to extract the growth patterns inherent in the different organs. The results of the first study (Chapter 2) show that organ shape depends on the balance between growth and differentiation, which fine regulation enables the acquisition of highly specialized functions. The cauline leaf, for example, shows a delay in differentiation that allows for prolonged morphogenetic activity and growth redistribution. Through the transient growth suppression at early stages, the cauline leaf developmental trajectory allows for its dual function, from protection to photosynthesis. The second study (Chapter 3) focuses on the behavior of individual cells, whose growth, although controlled by positional information, is often heterogeneous. This variability results from the differentiation of specialized cells, the stomata, which follow a specific developmental program. The autonomous, asynchronous behavior of these cells is the main source of variability in tissues whose growth is otherwise homogeneous. Overall, this thesis has shed light on the importance of timing in plant organ development. Whether at the organ, tissue, or cell level, modulation and synchronization of growth and differentiation are necessary for the acquisition of stereotypic organ shapes and functional complexity.

The Canterbury plains are frequently exposed to summer drought and climate predictions forecast that the severity and frequency of summer drought will increase. The most commonly used pasture grass, perennial ryegrass (Lolium perenne L.), is drought sensitive. One possible method to maintain sward dry matter (DM) production under water stress is to use an alternative grass species such as tall fescue (Festuca arundinacea Schreb.). The objective of this research was to compare summer DM production of monoculture swards of perennial ryegrass and tall fescue under various seasonal drought regimes to study physiological and biochemical drought responses of each species. Data were collected over a period of two-summer seasons, Season One (2006-2007) and Season Two (2007-2008) in an automated rain shelter at Lincoln, Canterbury. Drought treatments included exposure of plants to a spring or autumn drought or a four-weekly "irrigated" drought regime. DM yields of the two species were similar under each watering regime. The control treatment, under non-limiting conditions, has the highest accumulated yield in both Season One and Two for ryegrass (17.1 and 15.7 t DM ha⁻¹) and tall fescue (18.8 and 16.0 t DM ⁻¹) respectively. Spring and autumn drought treatments were similar for the two species in accumulated yield in either season, however the exposure to drought stress returned yields lower than the control. Consistently, the lowest-yielding treatment was the four-weekly irrigated drought, which resulted in an average yield across species in Season One of 10.1 t DM ha⁻¹ and 8.35 t DM ha⁻¹ in Season Two. Growth rates of the swards were calculated using accumulated DM production against accumulated thermal time using a base temperature of 3°C for both species. The control treatments showed a strong linear relationship for both species in both seasons, though Season Two showed a period of approximately 390 °Cd of no growth. Spring growth was similar for all treatments until October when both the spring drought and four-weekly irrigated treatment deviated from the control as water stress commenced. Growth also ceased under autumn drought later in the season. The physiological drought responses between species and among treatments differed. Tall fescue under control conditions had the highest photosynthesis rates of 20.5 µmol CO₂ m⁻² s⁻¹,or 22% higher than ryegrass, whereas the four-weekly irrigated treatment showed no inter-species differences. Differences were also found for other gas exchange parameters. Physiological water use efficiency (phys WUE) in ryegrass was 15% greater than tall fescue in Season Two. Photosynthesis and gas exchange rates against leaf water potential showed declining gas flow in both species across all treatments in response to drying soil conditions and across all irrigation treatments. The osmo-protectant proline was 22% higher in concentration in ryegrass than in tall fescue in Season Two and increased in drought stressed treatments in both seasons. Water stress was found to reduce total chlorophyll concentrations in all treatments and in tall fescue, while little change occurred in the chlorophyll a:b ratio. In conclusion, the findings from this thesis suggest similar DM responses for the two species under drought. The findings suggests that tall fescue performs more as a "water user" under drought conditions, compared with perennial ryegrass, which is more a "water saver." Resonses to the changing environment to a point, before "shuttting up shop" through lower stomatal conductance.