L’auxine est une hormone essentielle au développement des plantes, tant pour la division quel’expansion cellulaire. La transcription des gènes de réponses à l’auxine est permise par un mécanisme designalisation faisant intervenir des complexes ubiquitine ligase E3 impliquant des protéines à F-boxTIR1/AFBs, qui en présence d’auxine peuvent interagir avec les répresseurs transcriptionnels AUX/IAA pourinduire leur dégradation et activer la transcription de gènes de réponses à l’auxine. Cette transcriptionimplique des facteurs de transcription ARFs (Auxin Response Factors). Dans la feuille, un maximum d’auxinerésultant de l’activité des transporteurs d’efflux d’auxine PIN1 participe à l’initiation des dents à la marge. Lefacteur de transcription CUC2 qui permet la formation des domaines frontières intervient dans le contrôle del’orientation des transporteurs PIN1. L’auxine réprime également CUC2 limitant ainsi son expression au sinus,ce qui semble nécessaire pour la formation des dents. Les acteurs de la signalisation auxinique impliquésdans la formation des dents ne sont néanmoins pas connus chez Arabidopsis.Dans cette thèse, les profils d’expression des ARFs ont été mis en évidence dans les jeunes feuillesd’Arabidopsis au moment de la formation des dents et une cartographie fine a été établie. Trois ARFspotentiellement répresseurs ARF1, ARF3, ARF18 et trois activateurs ARF5, ARF6, ARF8 ont été identifiéscomme étant exprimés dans la zone dent/sinus. La modification de leurs profils d’expression dans des formesde feuilles modifiées par des variations d’expression de CUC2 a également été étudiée. Afin de déterminer sices ARFs sont impliqués dans la morphogénèse foliaire et plus particulièrement dans l’initiation des dents, lesimple mutant arf5-2/mpS319, et les doubles mutants arf6-2 arf8-3, arf1-5 arf3-1, arf1-5 arf18-2 et arf3-1arf18-2 ont été générés et analysés. Les feuilles matures du mutant nul arf5-2/ mpS319 présentent une tailleplus importante que celles de Col-0, suggérant que ARF5 serait impliqué dans l’expansion cellulaire. / Auxin is essential for plant development, more particularly by participating in cellular division andexpansion. Transcription of auxin response genes is allowed by the auxin signaling pathway involving the F-boxTIR1/AFBs proteins associated to ubiquitin ligase E3, which can interact with the AUX/IAA repressors in thepresence of auxin to trigger their degradation and activate transcription of early auxin response genes involvingthe Auxin Response Factors (ARF). At the leaf margin, an auxin maximum resulting from the transport of auxinmediated by the auxin efflux carrier PIN1 is required for teeth formation. The transcription factor CUC2 thatdefines boundary domains, somehow redirects PIN1 to create a convergent flux to the apex of the tooth. Auxinrepresses CUC2 thus limiting its expression to the sinus. In Arabidopsis, the actors of the auxin signalingpathway involved in leaf serration are unknown.In this thesis, the expression profiles of ARFs genes have been investigated in young leaves of Arabidopsis, morespecifically during teeth formation and a detailed map was built. Three ARFs acting as putative repressors ARF1,ARF3 and ARF18 and three activators ARF5, ARF6 and ARF8 were identified as been expressed in thesinus/teeth area. Their expression profiles were also studied in leaves exhibiting modified shapes as a result ofvariations in CUC2 expression. To determine if these ARFs are involved in leaf morphogenesis and moreparticularly in tooth initiation, the arf5-2 null mutant and the double null mutants arf6-2 arf8-3, arf1-5 arf3-1,arf1-5 arf18-2 and arf3-1 arf18-2 were generated and serration was analyzed. Length of arf5-2 mature leaves islonger than the wild type, suggesting that ARF5 could be involved in cell expansion.

The ratio of auxin to cytokinin determines de novo organogenesis in plants. Relatively little is known about the effect of genetically altered auxin signaling on in vitro organogenesis. Here, callusogenesis, shoot, and root formation were studied in loss- (LOF) and gain-of-function (GOF) alleles in two phylogenetically related Auxin Response Factors (ARFs), MONOPTEROS (MP/ARF5) and NON-PHOTOTROPHIC HYPOCOTYL 4 (NPH4/ARF7). Reduced MP activity greatly diminished shoot regeneration, and partially diminished callusogenesis and root formation. LOF in NPH4 strongly decreased callusogenesis, and mildly decreased shoot and root regeneration in particular categories of explants. By contrast, organogenesis responses were strongly increased in aerial explants carrying the GOF transgene dMP. Thus, both MP and NPH4 seem to act as positive regulators of certain organogenesis processes and the GOF dMP transgene may be of interest for stimulating organogenesis in plant species with poor regeneration properties. Also, organogenesis in vitro may reveal unknown developmental ARF functions.

The ratio of auxin to cytokinin determines de novo organogenesis in plants. Relatively little is known about the effect of genetically altered auxin signaling on in vitro organogenesis. Here, callusogenesis, shoot, and root formation were studied in loss- (LOF) and gain-of-function (GOF) alleles in two phylogenetically related Auxin Response Factors (ARFs), MONOPTEROS (MP/ARF5) and NON-PHOTOTROPHIC HYPOCOTYL 4 (NPH4/ARF7). Reduced MP activity greatly diminished shoot regeneration, and partially diminished callusogenesis and root formation. LOF in NPH4 strongly decreased callusogenesis, and mildly decreased shoot and root regeneration in particular categories of explants. By contrast, organogenesis responses were strongly increased in aerial explants carrying the GOF transgene dMP. Thus, both MP and NPH4 seem to act as positive regulators of certain organogenesis processes and the GOF dMP transgene may be of interest for stimulating organogenesis in plant species with poor regeneration properties. Also, organogenesis in vitro may reveal unknown developmental ARF functions.

Pluvial floods are the most difficult and to date least investigated phenomena in urban hydrology. While efforts are being made to increase the knowledge base concerning this type of flooding, a large part of the difficulty lies in the nature of the precipitation. Convective storms represent most of the larger intensity short term rainfall in urban areas and is also the raintype, that is expected to increase the most in the future. The rain cells of this type have a more distinct boundary, larger intensity, a smaller extent and a shorter life span, than frontal rains. Combined with the low availability of densely spaced rain gauge networks and also low temporal resolution of measurements in 15 minutes intervals at best, makes this rain type still very difficult to analyze and even harder to predict. The resolution of cloud radar images at 2x2km and taken every 15 minutes is too coarse and the error reduction algorithms for radar based precipitation (HIPRAD) images to analysera in patterns are not sufficient by them selves to analyze the characteristics of such rainfields and the processes occurring within these fields. The spatial variation of raincells, their development and decay, the distance between them, and the velocity and direction of their movement can however be investigated employing a combination of densely spaced rain gauges and radar images to reach a more realistic representation of short-term precipitation for the use of in hydraulic models. The movement of rain fields has been investigated with two main areas of focus: The influence of direction or directional bias, often with an interest in the most crucial case referred to as the resonance effect, and in context of areal reduction of point rainfall. Most of these studies have been carried out with statistical methods and in laboratory experiments. In this study a hydraulic model was built on the terrain model of a realcity, a 28 km area in the city of Falun, to test the recently gathered information about the temporal variation of five empirical hyetographs with different peak arrival times and peak intensities, which are representative of Swedish climate. The hyetographs were produced and provided by SMHI. The empirical rain types were derived from 20 years of rain gauge observations and confirmed by radar images. For reference purposes, a standard Chicago design storm (CDS) rain was modeled as well. The simulated scenarios were modeled as a MIKE 21 hydraulic model, as a stationary scenario and in four movement directions. It was foundthat the empirical rain types produced lower inundation depth than the CDS, in a range of 20 to 50 % lower. The effect of modeling rainfall in motion produced on average only about 4-20 % lower water depths than the corresponding non-moving scenario. In a few instances, in a single evaluation point, the moving scenarios resulted in a relative water depth of a maximum of just above 1%. It was concluded that the conceptual approach of areal reduction from movement seems to be accurate and could help improve modeling rainfall in general, and specifically for cloud burst scenarios of shorter durations in urban catchments. It was also found that further investigation of the physical processes in rainfields could serve to increase the accuracy in areal reduction of precipitation for more realistic hydraulic models and in turn reduce over design. / Pluviala översvämningar är den typen, som är både svårast att reda ut och samtidigt den minst utforskade fenomenen inom urban hydrologi. Medan ansträngningar görs för att förbättra kunskapsläget, ligger den största svårigheten i nederbördens skepnad. Det är konvektiva regn som utgör de flesta av de starkare korttids regntillfällen i urbana områden och är också regntypen som förväntas att öka mest i framtiden. Regncellerna har en tydligare avgränsning, en större intensitet, mindre utsträckning, och en kortare livscykel än frontala regn. I kombination med den låga tillgängligheten av regnmätarnätverk med hög täthet i positioneringen av mätare, samt den låga tidsupplösningen av mätningar i intervaller av 15 minuter gör att konvektiva regn fortfarande är svåra att analysera och ännu svårare att förutse. Upplösningen av molnradar bilder av 2x2 km som tas varje 15:de minut är för grova och algoritmer för felreducering av bilder från radarbaserad nederbördsdata (HIPRAD) för analys av regn mönster är inte tillräckligt noggranna, för sig, för att kunna analysera egenskaperna av sådana regnfält och de processerna som karakteriserar dessa. Den spatiala variationen inom regnceller, deras utveckling och förfall, avståndet mellan dem samt riktningen och hastigheten kan ändå undersökas med hjälp av kombinationen av regnmätarnätverk och radar bilder för att uppnå mer realistiska korttids nederbördsscenarier för användning i hydrauliska model. Studier, som har undersökt regn i rörelse har varit fokuserade på två huvudområden: Betydelsen av riktningen, i vilken regnet rör sig, där den största effekten som denna riktningsbias kan uppnå, har döpts resonans effekt och i samband med ytreducering (areal reduction) av punkt nederbörd. De flesta av dessa studier har genomförts med hjälp av statistiska metoder och laboratorieexperiment. I denna studie skapades en hydraulisk modell baserad på en realistisk terräng av ett existerade urbant område, en yta på 28 km i Falun, för att testa den nyligen utvärderade informationen om temporala intensitets fördelningen som representerar det svenska klimatet. Regndatat producerades och tillhandahölls av SMHI och representerar en mätserie från regnmätare över en period av 20 år. Som referens modellerades även ett Chicago regn (CDS). Med hjälp av en MIKE21 hydraulisk modell, simulerades ett stationärt scenario och fyra rörelseriktningar för varje empirisk hyetograf. Resultaten visade att de empiriska regntyperna skapade översvämningar med 20-50% lägre vattendjup än CDS regnet. Att modellera rörelsen resulterade i 4-20% lägre vattennivåer jämfört med respektive stationär scenario. I några enstaka tillfällen, i en av evalueringspunkterna, skapade de rörliga scenarierna större resultat, med lite över 1% i det största fallet. Det drogs slutsatsen att konceptet av areal reduction genom molnrörelse verkar vara korrekt och skulle kunna hjälpa att förbättra sättet att modellera regn generellt, men också specifikt för skyfalls scenarier med korta varaktigheter över urbana avrinningsområden. Man kom ytterligare till slutsatsen att framtida studier i samband med de fysiska processerna i regnceller skulle kunna användas för att höja noggrannheten av ytreducering av nederbörd för mer realistiska hydrauliska modeller, som i sin tur kunde minska överdesign.

L'auxine est une hormone végétale qui coordonne plusieurs processus de développement des plantes à travers la régulation d'un ensemble spécifique de gènes. Les Auxin Response Factors (ARF) sont des régulateurs transcriptionnels qui modulent l'expression de gènes de réponse à l’auxine. Des données récentes montrent que les membres de la famille des ARF sont impliqués dans la régulation du développement des fruits de la nouaison à la maturation. L'objectif principal de la thèse est d’étudier la part qui revient aux ARF dans le contrôle du développement et de la maturation des fruits et d’en comprendre les mécanismes d’action. L’analyse des données d’expression disponibles dans les bases de données a révélé que, parmi tous les ARF de tomates, SlARF2 affiche le plu haut niveau d'expression dans le fruit avec un profil distinctif d’expression associé à la maturation. Nous avons alors entrepris la caractérisation fonctionnelle de SlARF2 afin d’explorer son rôle dans le développement et la maturation des fruits. Deux paralogues, SlARF2A et SlARF2B, ont été identifiés dans le génome de la tomate. Nous avons montré que l’expression de SlARF2A dans le fruit est régulée par l'éthylène tandis que celle de SlARF2B est induite par l'auxine. La sous-expression de SlARF2A, comme celle de SlARF2B, entraine un retard de maturation alors que l’inhibition simultanée des deux paralogues conduit à une inhibition plus sévère de la maturation suggérant une redondance fonctionnelle entre les deux paralogues lors de la maturation des fruits. Les fruits présentant une sous-expression des gènes SlARF2 produisent de faibles quantités d'éthylène, montrent une faible accumulation de pigments et une plus grande fermeté. Le traitement avec de l'éthylène exogène ne peut pas inverser les phénotypes de défaut de maturation suggérant que SlARF2 pourrait agir en aval de la voie de signalisation de l'éthylène. L'expression des gènes clés de biosynthèse et de signalisation de l'éthylène est fortement perturbée dans les lignées sous-exprimant SlARF2 et les gènes majeurs qui contrôlent le processus de maturation (RIN, CNR, NOR, TAGL1) sont sensiblement sous-régulés. Les données suggèrent que SlARF2 est essentiel pour la maturation des fruits et qu’il pourrait agir au croisement des voies de signalisation de l'auxine et de l'éthylène. Dans le but de mieux comprendre les mécanismes moléculaires par lesquels les ARF régulent l'expression des gènes de réponse à l'auxine, nous avons étudié l'interaction des SlARFs avec des partenaires protéiques ciblés, principalement les co-répresseurs de type Aux/IAA et Topless (TPL) décrits comme les acteurs clés dans la répression des gènes dépendant de la signalisation auxinique. Une fois les gènes codant pour les membres de la famille TPL de tomate isolés, une approche double hybride dans la levure a permis d’établir des cartes exhaustives d'interactions protéine-protéine entre les membres des ARFs et des Aux/IAA d’une part et les ARFs et les TPL d’autre part. L'étude a révélé que les Aux/IAA interagissent préférentiellement avec les SlARF activateurs et qu’à l’inverse les Sl-TPL interagissent uniquement avec les SlARF répresseurs. Les données favorisent l'hypothèse que les ARF activateurs recrutent les Sl-TPL via leur interaction avec les Aux/IAA, tandis que les ARF répresseurs peuvent interagir directement avec les Sl-TPL. Les études d’interactions ont permis également d’identifier de nouveaux partenaires comme les protéines VRN5 et LHP1, composantes des complexes Polycomb PRC impliqués dans la repression par voie épigénétique de la transcription par modification de l'état de méthylation des histones. Au total, le travail de thèse apporte un nouvel éclairage sur le rôle et les mécanismes d'action des ARF et identifie SlARF2 comme un nouvel élément du réseau de régulation contrôlant le processus de maturation des fruits chez la tomate. / The plant hormone auxin coordinates plant development through the regulation of a specific set of auxin-regulated genes and Auxin Response Factors (ARFs) are transcriptional regulators modulating the expression of auxin-response genes. Recent data demonstrated that members of this gene family are able to regulate fruit set and fruit ripening. ARFs are known to act in concert with Aux/IAA to control auxin-dependent transcriptional activity of target genes. However, little is known about other partners of ARFs. The main objective of the thesis research project was to gain more insight on the involvement of ARFs in fruit development and ripening and to uncover their interaction with other protein partners beside Aux/IAAs. Mining the tomato expression databases publicly available revealed that among all tomato ARFs, SlARF2 displays the highest expression levels in fruit with a marked ripening-associated pattern of expression. This prompted us to uncover the physiological significance of SlARF2 and in particular to investigate its role in fruit development and ripening. Two paralogs, SlARF2A and SlARF2B, were identified in the tomato genome and transactivation assay in a single cell system revealed that the two SlARF2 proteins are nuclear localized and act as repressors of auxin-responsive genes. In fruit tissues, SlARF2A is ethylene-regulated while SlARF2B is auxin-induced. Knock-down of SlARF2A or SlARF2B results in altered ripening with spiky fruit phenotype, whereas simultaneous down-regulation of SlARF2A and SlARF2B leads to more severe ripening inhibition suggesting a functional redundancy among the two SlARF2 paralogs during fruit ripening. Double knock-down fruits produce less climacteric ethylene and show delayed pigment accumulation and higher firmness. Exogenous ethylene treatment cannot reverse the ripening defect phenotypes suggesting that SlARF2 may act downstream of ethylene signaling. The expression of key ethylene biosynthesis and signaling genes is dramatically disturbed in SlARF2 down-regulated fruit and major regulators of the ripening process, like RIN, CNR, NOR, TAGL1, are under-expressed. The data support the notion that SlARF2 is instrumental to fruit ripening and may act at the crossroads of auxin and ethylene signaling. Altogether, while ethylene is known as a key hormone of climacteric fruit ripening, the ripening phenotypes associated with SlARF2 down-regulation bring unprecedented evidence supporting the role of auxin in the control of this developmental process. To further extend our knowledge of the molecular mechanism by which ARFs regulate the expression of auxin-responsive genes we sought to investigate interactions SlARF and putative partners, mainly Aux/IAAs and Topless co-reppressors (TPLs) reported to be key players in gene repression dependent on auxin signaling. To this end, genes encoding all members of the tomato TPL family were isolated and using a yeast-two-hybrid approach comprehensive protein-protein interaction maps were constructed. The study revealed that Aux/IAA interact preferentially with activator SlARFs while Sl-TPLs interact only with repressor SlARFs. The data support the hypothesis that activator ARFs recruit Sl-TPLs co-repressors via Aux/IAAs as intermediates, while repressor ARFs can physically interact with Sl-TPLs. Further investigation indicated that SlARFs and Sl-TPLs can interact with polycomb complex PRC1 PRC2 components, VRN5 and LHP1, known to be essential players of epigenetic repression of gene transcription through the modification of histones methylation status. These data establish a potential link between ARFs and epigenetic regulation and thereby open new and original perspectives in understanding the mode of action of ARFs. Altogether, the thesis work provides new insight on the role of ARFs and their underlying action mechanisms, and defines SlARF2 as a new component of the regulatory network controlling the ripening process in tomato.