Estômatos são poros responsáveis pela troca gasosa entre a folha e o meio externo. A partir da década de 80, experimentos revelaram um complexo padrão espaço temporal na abertura e fechamento dos estômatos. As experiências apontam para uma possível coordenação entre estômatos em algumas áreas da folha chamada de patches. Esse fenômeno é conhecido na literatura como patchy stomatal conductance. Frequentemente a coordenação dinâmica dos estômatos está associada à oscilações temporais na condutãncia estomática (média especial da abertura dos estômatos). Em 1997 Haefner, Buckley e Mott (HBM) publicaram uma análise numérica de um modelo dinâmico para explorar o comportamento complexo dos estômatos. O modelo é baseado em algumas características conhecidas dos estômatos e assume transporte hídrico em uma rede definida por uma geometria simples e bastante restritiva. De acordo com os autores, o modelo reproduz qualitativamente os dados experimentais. Recentemente, Ferraz e Prado mostraram que esse modelo não é capaz de reproduzir os resultados experimentais. Usando ingredientes do modelo sugerido por HBM, Ferraz e Prado sugeriram uma geometria realística de distribuição reservatórios hídricos. Embora essa configuração reproduza os patches, eles permanecem estáticos e nenhuma oscilação é observada. Sem explorar detalhes significativos, Ferraz e Prado afirmaram que a histerese na abertura estomatal poderia explicar vários aspectos dos resultados experimentais. No presente estudo comprovamos, através de uma abordagem computacional baseada em transdutores histeréticos, que a hipótese de histerese na abertura dos estômatos de fato reproduz qualitativamente os dados experimentais. Em nossa abordagem a histerese na abertura dos estômatos é emulada através de operadores chamados de histerons. A robustez da hipótese é testada usando diferentes tipos de histerons. Analisamos a correlação entre os estômatos na rede que simula a superfície da folha. Observamos que a correlação entre estômatos depende da geometria da veia. Uma análise detalhada dos parâmetros envolvidos revela uma dependência entre o período de oscilação na condutância estomática e o déficit de vapor d\'água entre a folha e o meio ambiente. Esta característica subjacente ao modelo pode inspirar novas experiências para testar a hipótese da histerese na abertura dos estômatos. / Stomata are pores on the surface of leaves responsible for controlling the exchange of gas between the plant and the environment. Experiments revealed a complex spatial-temporal pattern in the opening and closing mechanism of stomata. The main feature of the phenomenon is that stomata appear to be synchronized into clusters, known as patches. The dynamical coordination of stomata often involves oscillations in stomatal conductance. In 1997 Haefner, Buckley, and Mott (HBM) published a numerical analysis of a dynamic model to explore the complex behavior of stomata. The model is based on some known features of the stomata, and assumes that water diffuses within the leaves according to a simple geometric arrangement. According to the authors, the model reproduces qualitatively the experimental data. Recently, Ferraz and Prado showed that the computational approach of HBM is not able to reproduce the experimental results. Inspired by this model, Ferraz and Prado introduced a new geometric features that leads to static patches of stomata; however no oscillation was observed and the patches remained static. The authors suggested that hysteresis in stomatal aperture could explain several experimental aspects. We now report a further investigation of the changes suggested by Ferraz and Prado in the original model of HBM. The theoretical approach confirmed that hysteresis in the aperture mechanism of pores reproduces a variety of behaviors of stomatal conductance described in experiments. We explore the hysteresis feature through the formalism of hysteretic transducer. The robustness of the hysteretic assumption is tested by different kinds of hysteresis operators. We analyzed the correlation among stomata in the lattice. We observed that the correlation depends on the geometry of the veins. Finally, the analysis of the model reveals a dependence between the period of oscillation in the stomatal conductance time series and water vapor pressure deficits Δω - an external parameter. Further experiments might explore this underlying feature of the model.

Mitogen-activated protein kinases (MAPKs) belong to the most conserved signaling pathways and are found in all eukaryotes, including humans where they play important roles in various diseases and cancer. Stimulation of this signal transduction pathway by microbe-associated molecular patterns (MAMP) results in a multitude of events to regulate innate immune responses in Arabidopsis thaliana stimulating large-scale changes in gene expression. Starting from a phosphoproteomic screen in Arabidopsis thaliana wild type and mpk3, mpk4 and mpk6 mutants following microbe-associated molecular pattern (MAMP) treatment, several novel chromatin-associated proteins were identified that are differentially phosphorylated by stress-induced protein kinases. Arabidopsis Ras GTPase-activating protein SH3-domain-binding protein (AtG3BP-1) is a downstream putative substrate of the MAMP-stimulated MAPK pathway that is phosphorylated by MPK3, 4 and 6 in in vitro kinase assays. AtG3BP1 belongs to a highly conserved family of RNA-binding proteins in eukaryotes that link kinase receptormediated signaling to RNA metabolism. Here, we report the characterization of the Arabidopsis homolog of human G3BP1 in plant innate immunity. AtG3BP1 negatively regulates plant immunity and defense immune responses. Atg3bp1 mutant lines show constitutive stomata closure, expression of a number of key defense marker genes, and accumulate salicylic acid but not jasmonic acid. Furthermore, Atg3bp1 plants exhibit enhanced resistance to the biotrophic pathogen Pseudomonas syringae pv. tomato. Pathogen resistance was mediated by stomatal and apoplastic immunity in Atg3bp1. More generally, our data reinforce that AtG3BP1 is a key mediator of plant defense responses and transient expression of AtG3BP1 delivered striking disease resistance in the absence of yield penalty, highlighting a potential application of this gene in crop protection.

Stomata are cellular valves in plants that play an essential role in the regulation of gas exchange and are distributed in the epidermis of aerial organs. In Arabidopsis thaliana, stomatal production and development are coordinated by the mitogen-activated protein kinase (MAPK) signalling pathway, which modulates a variety of other processes, including cell proliferation, regulation of cytokinesis, programed cell death, and response to abiotic and biotic stress. The environment also plays a role in stomatal development, by influencing the frequency at which stomata develop in leaves. This thesis presents an analysis of stomatal development in Arabidopsis mutants in two MAPK pathways: MEKK1-MKK1/MKK2-MPK4, and MAP3K17/18-MKK3. Obtained results demonstrate the effect of stress conditions on stomatal development and specify the involvement of analysed MAPK in stomatal patterning. First, both analysed pathways modulate stomatal patterning in Arabidopsis cotyledons. Second, plant growth-promoting bacteria tested enhance stomatal density and affect guard cell morphology. Third, the sucrose or mannitol treatment increases defects in stomatal patterning. Finally, salt stress or high temperature can suppress stomatal defects in mutants of the MEKK1-MKK1/MKK2-MPK4 pathway.

<p> Plants can control water loss through their stomata in response to changing environmental conditions. Some research has been done on predicting stomatal resistance from climate variables such as solar radiation intensity, temperature, and vapour pressure deficit, but the factors involved are numerous and complex. Information on evapotranspiration from non-cultivated vegetation under field conditions is relatively scarce. This study was carried out in a subarctic coastal wetland on James Bay, an area for which little data exists. Leaf resistance measurements were collected in the field with a diffusion porometer as the 1988 growing season reached its peak. Meteorological data were also recorded. The main species examined was Carex aquatilis, a common sedge with a circumpolar distribution. As has been previously found, field resistances were lower than those reported for greenhouse-grown specimens. Attempts to derive a multivariate regression model to predict leaf resistances had a maximum explained variance under 40%. However, Principal Components Analysis suggested that cool, sunny, low-VPD conditions tend to accompany low leaf resistances. An increase in leaf resistance occurs when solar radiation intensities are low, or when the air is warm and dry. In addition, Carex aquatilis from drier areas showed less growth, and generally higher leaf resistances, than plants growing in saturated soils. Since climate models predict higher temperatures and lower soil moisture for the study area, due to carbon dioxide-induced warming, higher stomatal resistances could result. If these changes are severe enough, primary production could be reduced in the subarctic wetlands. Through the food chain, this could possibly affect species important to the economy and ecology of northern areas.</p> / Thesis / Candidate in Philosophy

Rice bacterial leaf streak disease caused by Xanthomonas oryzae pv. oryzicola (Xoc) is one of the most important rice bacterial diseases. Xanthomonas type III effector gene avrRxo1 is conserved in diverse Xoc strains and its homologues have been identified from several other gram-negative bacteria species such as Burkholderia and Acidovorax. In this research, we studied the protein structure of AvrRxo1 and illustrated its virulence mechanism.We determined the three-dimensional structure of the complex of AvrRxo1 and its cognate chaperone Arc1 (AvrRxo1 required chaperone 1). The AvrRxo1: Arc1 complex is structurally similar to the Zeta-epsilon family of toxin: antitoxin systems from the human bacterial pathogen Streptococcus pyogenes. AvrRxo1 and Arc1 have toxin: antitoxin-like activity in bacteria, and the toxin activity of AvrRxo1 is required for its virulence function in planta. These findings suggest that AvrRxo1 evolved from an endogenous bacterial toxin-antitoxin system.Furthermore, AvrRxo1 was shown to have virulence functions in diverse host plants including Arabidopsis thaliana. The ectopic expression of wild type avrRxo1 in Arabidopsis suppresses plant basal defense. AtVOZ (Arabidopsis vascular one zinc-finger transcription factor), which has two homologues in the Arabidopsis genome, VOZ1 and VOZ2, was identified as one of AvrRxo1 candidate interactor. The knockout of voz1/voz2 renders the plants more susceptible to the virulent pathogen Pseudomonas syringae pv. tomato (Pst) DC3000, but compromises the virulence function of AvrRxo1. The expression profiling of transgenic Arabidopsis plants expressing the avrRxo1 gene allowed us to identify Arabidopsis genes regulated by AvrRxo1 and VOZ1/2. AvrRxo1 interacts with and stabilizes VOZ2 in vivo and directly binds to the promoter region of AtCYS2 (Arabidopsis phytoCYStatin 2) to induce its expression. The overexpression of CYS2 in increased stomatal aperture size, and enhanced plant susceptibility to Pst. Therefore one of AvrRxo1 virulent functions is to regulate the expression of CYS2 by manipulating VOZ2, resulting in increased stomatal aperture. Presumably, this renders the host leaf more susceptible to colonization via the stomata. Another component of my dissertation was based on a genome-wide survey of Arabidopsis papin-like cysteine protease genes (PLCPs). The Arabidopsis genome has 31 PLCP and 7 cystatin genes, and they often worked in pairs to regulate signaling pathways in response to biotic and abiotic stress. The coordinated transcriptional regulation of all Arabidopsis PLCP and cystatin genes has never been systematically investigated. In order to unveil the mechanism of stomata-related plant immunity regulated by CYS2, we analyzed the expression patterns of 28 PLCPs and 7 cystatins in Arabidopsis in response to biotic or abiotic stress, by reprocessing and integrating microarray data from the AtGenExpress database. We also performed enzyme assays and evaluated the inhibition specificity of seven cystatins to the five most abundant PLCPs in Arabidopsis. Finally, we utilized the SVMs (support vector machines) package in R software to predict a functional network of PLCP-cystatin interplay in Arabidopsis. We identified the PLCP protein PAP4 as one of the putative targets of CYS2. The co-expression profiling indicated that the expression patterns of PAP4 and CYS2 were strongly correlated during virulent bacterial infection, and weakly correlated under drought stress. Therefore, PAP4 was determined to be a promising gene in regulating stomatal aperture size. Further research on the interplay of PAP4-CYS2 could be important for understanding AvrRxo1's virulence mechanism and regulation of plant stomatal immunity. / Ph. D.

TOO MANY MOUTHS (TMM) is a gene which has been shown to regulate cell fate and control cell division in the plant leaf epidermis. TMM encodes a leucine rich repeat-receptor like protein (LRR-RLP) which has putative function as a signal transducing factor in the cell to cell signaling pathway involved in controlling proper spacing of stomata! initials throughout the leaf. This gene has been shown to be widely conserved across species of land plants ranging from bryophytes to angiosperms. Curiously, although this gene is highly conserved, stomata! installation pathways vary considerably across these species. Leaf development in both Arabidopsis and monocot grasses has been well characterized, and contrasting stomata! development pathways have been identified between them. Dioscorea bulbifera is a basal member of the monocot family, but this species forms broad leaves similar to many eudicot species. Stomata! development in this species retains some features characteristic of grass installation pathways such as few to no secondary asymmetric divisions of meristemoids. However, new epidermal cell formation occurs by fill-in between leaf veins, more closely resembling broad leaf eudicot development. A TMM ortholog has been isolated in this species and phylogenetic analysis of the sequence may help to place the signals controlling stomata! proliferation in Dioscorea bulbifera into evolutionary perspective.

Stomatal development and its regulation by environmental conditions (light, CO2 concentration) and physiological processes in the leaf of higher plants were investigated. The study was based on the assumptions that stomatal development should be regulated by signals coming from both external environment and leaf interior, and that the signal should be transduced from cotyledons to leaves. Transgenerational effect in stomatal development was also studied. Molecular and physiological approaches were applied to reveal the relationship between leaf environment, stomatal development, stomatal function and leaf physiology.

Como organismos multicelulares complexos, as plantas requerem um extraordinário nível de coordenação entre as células para que se desenvolvam ordenadamente. Para a coordenação dessas atividades, as células devem freqüentemente ser aptas a se comunicar umas com as outras a certas distâncias. Os principais meios de comunicação intercelular são os hormônios e mensageiros químicos que carreiam as informações entre as células e assim coordenam seu crescimento e desenvolvimento. O ácido abscísico (ABA) é o hormônio envolvido nos processos fisiológicos quando as plantas são expostas a alguns tipos de estresse, como deficiência hídrica. Relativamente grandes quantidades de ABA são rapidamente sintetizadas em folhas em resposta ao estresse hídrico, onde o principal papel parece ser a regulação da abertura e fechamento estomático. No início do estresse hídrico, o ABA carreado pelo fluxo xilemático é sintetizado nas raízes que estão em contato direto com o solo seco. Devido a esse transporte poder ocorrer antes que o baixo potencial de água no solo cause qualquer mudança mensurável no estado hídrico das folhas, acredita-se que o ABA seja um sinal das raízes que ajuda a reduzir a taxa de transpiração nas folhas pelo fechamento estomático. O principal objetivo do trabalho foi determinar se o ABA sintetizado nas raízes de Lycopersicon esculentum é crucial na regulação do comportamento estomático ou se o ABA produzido pelas folhas poderia agir como sinal hormonal que permitisse a sobrevivência da planta em condições de deficiência hídrica. De forma similar, Lycopersicon pennellii, uma espécie tolerante ao estresse hídrico, foi estudada com o objetivo de determinar a possível função dos sinais químicos das raízes ou de outro mecanismo de tolerância a seca. Para tanto, plantas com 30 dias de idade obtidas de sementes de L. esculentum cv Lukullus, L. pennellii e um mutante chamando notabilis foram enxertadas umas nas outras e crescidas em casa de vegetação. Após dois meses, medidas de condutância estomática, transpiração e potencial da água na folha foram realizadas em três condições de água no solo: bem irrigado, estresse hídrico e reirrigação. Os valores de condutância estomática e transpiração mostraram que o comportamento estomático foi determinado pelo genótipo da parte aérea das plantas enxertadas. Também verificou-se que a tolerância a seca em L. pennellii não foi determinada exclusivamente por caracteres morfo-anatômicos da parte aérea. / Multicellular plants are complex organisms and their orderly development requires an extraordinary measure of coordination between cells. In order to coordinate their activities, cells must be able to communicate with each other, often at some distance. The principal means of intercellular communication are the hormones, chemical messengers that carry information between cells and thus coordinate their growth and development. Abscisic acid (ABA) is the hormone involved in physiological processes when plants are exposed to some sort of stress: water deficiency, for example. Relatively large amounts of ABA are rapidly synthesized in leaves in response to water stress, where it appears to play a major role in regulating stomatal opening and closure. As water stress begins, some of the ABA carried out by the xylem stream is synthesized in roots that are in direct contact with the drying soil. Because this transport can occur before the low water potential of the soil causes any measurable change in the water status of the leaves, ABA is believed to be a root signal that helps reduce the transpiration rate in the leaves by closing stomata. The main objective of the work was to determine if ABA synthesized in the roots of Lycopersicon esculentum is crucial in regulation stomata behavior or ABA produced by the leaves could act as hormone signal to improve plant survival under water shortage conditions. In a similar way, Lycopersicon pennellii, a water stress tolerant specie, was studied in order to determine the possible role of chemical root signals or any other mechanism. For that, 30 d-old plants obtained from seeds of L. esculentum cv Lukullus, L. pennellii and a mutant named notabilis were grafted to each other and grown in greenhouse conditions. Two months later, measurements of stomatal conductance, transpiration and leaf water potential were carried out. These measurements were done under three soil water conditions: plenty, stress and reirrigation. The values of stomatal conductance and transpiration showed that stomatal behavior was influenced by the genotype of the aerial part of grafted plants. It was also verified that the drought tolerance of L. pennellii was not only determined by morpho-anatomical characteristics of plant shoot.

Como organismos multicelulares complexos, as plantas requerem um extraordinário nível de coordenação entre as células para que se desenvolvam ordenadamente. Para a coordenação dessas atividades, as células devem freqüentemente ser aptas a se comunicar umas com as outras a certas distâncias. Os principais meios de comunicação intercelular são os hormônios e mensageiros químicos que carreiam as informações entre as células e assim coordenam seu crescimento e desenvolvimento. O ácido abscísico (ABA) é o hormônio envolvido nos processos fisiológicos quando as plantas são expostas a alguns tipos de estresse, como deficiência hídrica. Relativamente grandes quantidades de ABA são rapidamente sintetizadas em folhas em resposta ao estresse hídrico, onde o principal papel parece ser a regulação da abertura e fechamento estomático. No início do estresse hídrico, o ABA carreado pelo fluxo xilemático é sintetizado nas raízes que estão em contato direto com o solo seco. Devido a esse transporte poder ocorrer antes que o baixo potencial de água no solo cause qualquer mudança mensurável no estado hídrico das folhas, acredita-se que o ABA seja um sinal das raízes que ajuda a reduzir a taxa de transpiração nas folhas pelo fechamento estomático. O principal objetivo do trabalho foi determinar se o ABA sintetizado nas raízes de Lycopersicon esculentum é crucial na regulação do comportamento estomático ou se o ABA produzido pelas folhas poderia agir como sinal hormonal que permitisse a sobrevivência da planta em condições de deficiência hídrica. De forma similar, Lycopersicon pennellii, uma espécie tolerante ao estresse hídrico, foi estudada com o objetivo de determinar a possível função dos sinais químicos das raízes ou de outro mecanismo de tolerância a seca. Para tanto, plantas com 30 dias de idade obtidas de sementes de L. esculentum cv Lukullus, L. pennellii e um mutante chamando notabilis foram enxertadas umas nas outras e crescidas em casa de vegetação. Após dois meses, medidas de condutância estomática, transpiração e potencial da água na folha foram realizadas em três condições de água no solo: bem irrigado, estresse hídrico e reirrigação. Os valores de condutância estomática e transpiração mostraram que o comportamento estomático foi determinado pelo genótipo da parte aérea das plantas enxertadas. Também verificou-se que a tolerância a seca em L. pennellii não foi determinada exclusivamente por caracteres morfo-anatômicos da parte aérea. / Multicellular plants are complex organisms and their orderly development requires an extraordinary measure of coordination between cells. In order to coordinate their activities, cells must be able to communicate with each other, often at some distance. The principal means of intercellular communication are the hormones, chemical messengers that carry information between cells and thus coordinate their growth and development. Abscisic acid (ABA) is the hormone involved in physiological processes when plants are exposed to some sort of stress: water deficiency, for example. Relatively large amounts of ABA are rapidly synthesized in leaves in response to water stress, where it appears to play a major role in regulating stomatal opening and closure. As water stress begins, some of the ABA carried out by the xylem stream is synthesized in roots that are in direct contact with the drying soil. Because this transport can occur before the low water potential of the soil causes any measurable change in the water status of the leaves, ABA is believed to be a root signal that helps reduce the transpiration rate in the leaves by closing stomata. The main objective of the work was to determine if ABA synthesized in the roots of Lycopersicon esculentum is crucial in regulation stomata behavior or ABA produced by the leaves could act as hormone signal to improve plant survival under water shortage conditions. In a similar way, Lycopersicon pennellii, a water stress tolerant specie, was studied in order to determine the possible role of chemical root signals or any other mechanism. For that, 30 d-old plants obtained from seeds of L. esculentum cv Lukullus, L. pennellii and a mutant named notabilis were grafted to each other and grown in greenhouse conditions. Two months later, measurements of stomatal conductance, transpiration and leaf water potential were carried out. These measurements were done under three soil water conditions: plenty, stress and reirrigation. The values of stomatal conductance and transpiration showed that stomatal behavior was influenced by the genotype of the aerial part of grafted plants. It was also verified that the drought tolerance of L. pennellii was not only determined by morpho-anatomical characteristics of plant shoot.

Pappelsorten (Populus) für den Anbau in Kurzumtriebsplantagen sollten neben einer hohen Biomasseproduktion und der Vermehrbarkeit über Steckhölzer auch Trockentoleranz aufweisen. Trockenheit stellt einen wichtigen abiotischen Stressor dar, der das Wachstum der Pflanze, ihre Vitalität und ihre Produktivität negativ beeinflussen kann. Polyploide Sorten von Getreidepflanzen (Triticum), krautigen Pflanzen (Lonicera, Spathiphyllum und Nicotiana) aber auch Baumarten (Betula) zeigen eine höhere Toleranz gegenüber Stress, wie z.B. Trockenstress, im Vergleich zu Sorten mit niedrigerem Ploidiegrad. Hybridaspen Populus tremula (L.) x P. tremuloides (Michx.) der Sektion Populus können auch auf Grenzertragsböden ökonomische Zuwächse verzeichnen, auf denen andere Baumarten ausfallen. Den Hybridaspen fehlt allerdings die Vermehrbarkeit über Steckhölzer. Anderen Pappelarten der Sektionen Tacamahaca und Aigeiros wie beispielsweise P. nigra (L.) and P. trichocarpa (Torr. & Gray) x P. deltoides (Bartram ex Marsh) zeigen diese entscheidende Eigenschaft. Da Kreuzungen zwischen den Sektionen Populus und Tacamahaca oder Aigeiros auf natürlichem Weg schwierig sind, wurde die Methode der somatischen Hybridisierung eingesetzt. Der Klon P. tremula x P. tremuloides ('Münden 2') wurden als ein Fusionspartner und einer der Klone P. x canescens (INRA clone No. 717 1‑B4), P. nigra oder P. trichocarpa x P. deltoides (B19) als zweiter Fusionspartner in Hybridisierungsexperimenten verwendet. Blattmorphologisch und durch Mikrosatellitenanalysen konnten die erhaltenen Fusionsprodukte dem Ausgangsklon P. tremula x P. tremuloides ('Münden 2') zugeordnet werden, wiesen dabei aber einen tetraploiden Chromosomensatz auf. In der vorliegenden Dissertation war das zentrale Ziel, Trockenstressreaktionen tetraploider Hybridaspenlinien (HAL) im Vergleich zur diploiden Ausgangslinie P. tremula x P. tremuloides ('Münden 2') zu untersuchen. Dazu wurden die folgenden Fragen aufgestellt. (i) Sind die tetraploiden HAL dem diploiden Ausgangsklon unter Trockenheit überlegen z.B. hinsichtlich des Wasserverbrauchs und des Verwelkungs- und Vertrocknungszeitpunktes der Blätter? (ii) Unterscheiden sich die tetraploiden HAL von dem diploiden Ausgangklon in morphologischen Ausprägungen wie beispielsweise der Stomatamorphologie, der Höhe, der Biomasse und der Gesamtblattfläche? (iii) Ist die höhere Trockentoleranz der tetraploiden HAL durch den erhöhten Ploidiegrad verursacht? (iv) Zeigen die tetraploiden HAL unter Trockenstress früh physiologische Veränderungen im Kohlenhydratgehalt, dem relativen Blattwassergehalt und der stomatären Leitfähigkeit? (v) Gibt es genetische Modifikationen abgesehen von der Tetraploidie wie z.B. Duplikationen und Deletionen, die eventuell Einfluss auf das Trockensressverhalten haben könnten? In einem ersten Experiment wurden vier tetraploide HAL untersucht. Zwei HAL zeigten einen geringeren Wasserverbrauch und drei HAL eine spätere Verwelkung als der diploide Ausgangsklon. Außerdem entwickelten die tetraploiden HAL eine geringere Stomatadichte, größere Stomata und eine vergleichbare oder geringere Höhe, Biomasse und Gesamtblattfläche im Vergleich zum diploiden Ausgangsklon. Das zweite Experiment wurde mit vier diploiden und 16 weiteren tetraploiden HAL durchgeführt. Auch die diploiden Linien stammten aus Protoplastenfusions-experimenten. Die tetraploiden HAL wiesen im Mittel eine höhere Überlebensrate als die diploiden HAL auf, entwickelten aber durchdschnittlich ein geringeres Höhenwachstum. Unter Berücksichtigung der Höhe verwelkten 11 tetraploide, aber keine diploide HAL später als der diploide Ausgangsklon. In einem dritten Experiment wurden zwei Linien aus dem zweiten Experiment ausgewählt, die vergleichbares Höhenwachstum zeigten, aber später verwelkten als der diploide Ausgangklon. Es wurden physiologische Merkmale wie der Wasserverbrauch und der Verwelkungszeitpunkt der Blätter beobachtet und der relative Blattwassergehalt, die stomatäre Leitfähigkeit und der Kohlenhydratgehalt in einer Zeitreihe untersucht. Geringerer Wasserverbrauch im Vergleich zum diploiden Ausgangsklon konnte in beiden tetraploiden HAL gezeigt werden. Diese Beobachtung deckt sich mit dem höheren relativen Bodenwassergehalt der tetraploiden HAL im Vergleich zum diploiden Ausgangsklon. Insgesamt spricht das für geringeren Trockenstress in den tetraploiden HAL. Auf der anderen Seite sollte ein geringerer Wasserverbrauch auch eine geringere stomatäre Leitfähigkeit nach sich ziehen. Dies war in den tetraploiden HAL nicht durchgehend der Fall. Relativ zur Kontrolle konnten die tetraploiden Linien aber eine höhere stomatäre Leitfähigkeit bei Bodenwassergehalten aufrechterhalten, bei denen die diploide Ausgangslinie bereits ihre Stomata geschlossen hatte. Dies weist ebenfalls auf eine höhere Trockentoleranz der tetraploiden HAL hin. Genetische Modifikationen wie beispielsweise Duplikationen und Deletionen wurden mit Hilfe von “copy number variation”(“Kopienzahlvariation”)-Analysen an drei tetraploiden HAL und der diploiden Ausgangslinie durchgeführt. Die drei tetraploiden Linien zeigten vergleichbares Höhenwachstum, aber spätere Verwelkung als die diploide Ausgangslinie. Bei dieser Analyse konnten nur drei Segmente detektiert werden, die bei allen drei tetraploiden HAL im Vergleich zum diploiden Ausgangsklon verändert vorlagen. Die putativen Gene der detektierten Segmente wiesen keine Annotation auf. Eine der drei tetraploiden HAL zeigte eine hohe Anzahl von 90 Segmenten, die hier aber weder in dem diploiden Ausgangsklon noch in den beiden anderen tetraploiden Linien erhöht vorlagen. Diese Linie zeigte zwar eine höhere Trockentoleranz als der diploide Ausgangsklon, aber nicht gegenüber der anderen untersuchten tetraploiden HAL (Experiment 3). Beide tetraploiden HAL waren bezüglich des Wasserverbrauchs, des Verwelkungszeitpunktes und der stomatären Leitfähigkeit ähnlich. Dies lässt vermuten, dass die Trockentoleranz durch die Tetraploidie und nicht durch weitere genetische Modifikationen bedingt ist. Trotzdem ist eine weiterführende funktionelle Untersuchung der drei Segmente, die in allen tetraploiden HAL verändert auftraten, sinnvoll, da auch hier ein trockenrelevantes Gen verändert sein könnte. Um eine Relevanz der putativen Gene bezüglich Trockenheit zu testen, könnte zukünftig das Verhalten von Knock-out-Mutanten, denen die entsprechende Sequenz fehlt, unter Trockenstress untersucht werden. Abschließend ist zu bemerken, dass neben den hier untersuchten HAL mehrere hundert Fusionsprodukte in einem Verbundprojekt genetisch charakterisiert wurden. Die Heterofusionsfrequenz war äußerst gering und die Hybriden waren bereits in der in vitro Phase nicht überlebensfähig. Geringe Heterofusionsfrequenz wurde beteits in anderen Arten beobachtet (Cyclamen, Brassicaceae, Fabaceae, Poaceae, Solanaceae). Da in dieser Dissertation nur Homofusionslinien verwendet wurden, wurde die Vermehrbarkeit über Steckhölzer nicht getestet, sondern der Fokus auf das Trockenstressverhalten der tetraploiden Homofusionslinien gelegt. Zusammenfassend zeigen die Ergebnisse, dass veränderte morphologische Ausprägungen und eine höhere Trockentoleranz in den HAL mit erhöhten Ploidiegrad auftraten. Dies prädestiniert die tetraploiden Linien für Standorte, an denen Wasser als limitierender Faktor auftritt und hohe Ausfallraten durch Dürre erwartet werden.