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The lower oxygen limit (LOL) in plants marks the oxygen (O2) level where the metabolism shifts from being predominantly aerobic to anaerobic; recent work has shown that respiratory-based indicators of this metabolic shift are well-correlated with changes in chlorophyll a fluorescence signals. The physiological and biochemical changes at the root of this relationship have not been well-described in the literature. The processes involved are spatially separated: chlorophyll fluorescence is associated with the lightdependent reactions and emanates from the chloroplasts whereas aerobic respiration and fermentation occurs in the mitochondria and cytosol, respectively. Evidences outlined in this thesis are used to suggest the mechanistic link between these three regions of the cell is a fluid exchange of cellular reductant. When mitochondrial respiration is inhibited as a result of inadequate O2, used as a terminal electron acceptor, glycolytic reductant in the form of NADH accumulates in the cytosol. Reductant imbalances between the cytosol and organelles can be adjusted indirectly using translocators. Excess chloroplastic reductant is used to reduce the plastoquinone (PQ) pool via NADPH-dehydrogenase, a component of the chlororespiratory pathway, effectively decreasing the photochemical quenching (qP) capacity thereby inducing a switch from minimum fluorescence (Fo) to a higher relative fluorescence (F) value where qP < 1. Subjecting dark-adapted photosystems to low-intensity light increased Fo to a slightly higher F value due to a lightinduced reduction of the oxidized PQ pool when the O2 was above the LOL, but decreased F as a result of a PSI-driven oxidation of the already over-reduced PQ pool when the O2 was below the LOL. Low O2 was also shown to increase violaxanthin deepoxidation and non-photochemical quenching (qN), likely a reflection of the overreduced state of the photosystems and associated pH decrease. Dynamic controlled atmosphere (DCA) is a fluorescence-based controlled atmosphere (CA) system that sets the optimum atmosphere for fruits and vegetables based on a product’s fluorescence response. Experiments in this thesis on the relationship between O2, temperature, light, metabolism, pigmentation and chlorophyll fluorescence were used to interpret the physiology behind fluorescence changes, suggest improved DCA techniques and outline potentially profitable avenues for future research.

Thesis (M.S.)--Oregon State University, 1993. / Typescript (photocopy). Includes bibliographical references (leaves 37-42). Also available on the World Wide Web.

Chlorophyll fluorescence imaging has now become a versatile and standard tool in fundamental and applied plant research. This method captures time series images of the chlorophyll fluorescence emission of whole leaves or plants upon various illuminations, typically combination of actinic light and saturating flashes. Several conventional chlorophyll fluorescence parameters have been recognized that have physiological interpretation and are useful for, e.g., assessment of plant health status and early detection of biotic and abiotic stresses. Chlorophyll florescence imaging enabled us to probe the performance of plants by visualizing physiologically relevant fluorescence parameters reporting physiology and biochemistry of the plant leaves. Sometimes there is a need to find the most contrasting fluorescence features/parameters in order to quantify stress response at very early stage of the stress treatment. The conventional fluorescence utilizes well defined single image such as F0, Fp, Fm, Fs or arithmetic combinations of basic images such as Fv/Fm, PSII, NPQ, qP. Therefore, although conventional fluorescence parameters have physiological interpretation, they may not be representing highly contrasting image sets. In order to find the effect of stress treatments at very early stage, advanced statistical techniques, based on classifiers and feature selection methods, have been developed to select highly contrasting chlorophyll fluorescence images out of hundreds of captured images. We combined sets of highly performing images resulting in images with very high contrast, the so called combinatorial imaging. The application of advanced statistical methods on chlorophyll fluorescence imaging data allows us to succeed in tasks, where conventional approaches do not work. This thesis aims to explore the application of conventional chlorophyll fluorescence parameters as well as advanced statistical techniques of classifiers and feature selection methods for high-throughput screening. We demonstrate the applicability of the technique in discriminating three species of the same family Lamiaceae at a very early stage of their growth. Further, we show that chlorophyll fluorescence imaging can be used for measuring cold and drought tolerance of Arabidopsis thaliana and tomato plants, respectively, in a simulated high ? throughput screening.

Equisetum giganteum L., a giant horsetail, is one of the largest living members of an ancient group of non-flowering plants with a history extending back 377 million years. Its hollow upright stems grow to over 5 m in height. Equisetum giganteum occupies a wide range of habitats in southern South America. Colonies of this horsetail occupy large areas of the Atacama river valleys, including those with sufficiently high groundwater salinity to significantly reduce floristic diversity. The purpose of this research was to study the ecophysiological and biomechanical properties that allow E. giganteum to successfully colonize a range of habitats, varying in salinity and exposure. Stem ecophysiological behavior was measured via steady state porometry (stomatal conductance), thermocouple psychrometry (water potential), chlorophyll fluorescence, and ion specific electrodes (xylem fluid solutes). Stem biomechanical properties were measured via a 3-point bending apparatus and cross sectional imaging. Equisetum giganteum stems exhibit mechanical characteristics of semi-self-supporting plants, requiring mutual support or support of other vegetation when they grow tall. The mean elastic moduli (4.3 Chile, 4.0 Argentina) of E. giganteum in South America is by far the largest measured in any living horsetail. Stomatal behavior of E. giganteum is consistent with that of typical C3 vascular plants, although absolute values of maximum late morning stomatal conductance are very low in comparison to typical plants from mesic habitats. The internode stomata exhibit strong light response. However, the environmental sensitivity of stomatal conductance appeared less in young developing stems, possibly due to higher cuticular conductance. Exclusion of sodium (Na) and preferential accumulation of potassium (K) at the root level appears to be the key mechanism of salinity tolerance in E. giganteum. Overall stomatal conductance and chlorophyll fluorescence were little affected by salinity, ranging from very low levels up to half strength seawater. This suggests a high degree of salinity stress tolerance. The capacity of E. giganteum to adapt to a wide variety of environments in southern South America has allowed it to thrive despite tremendous environmental changes during their long tenure on Earth.

Les diatomées constituent le principal groupe du phytoplancton dans les océans, contribuant à près de 20% de la production primaire globale. Dans leur environnement très variable, les diatomées sont particulièrement efficaces dans leur capacité à ajuster leur activité photosynthétique en dissipant sous forme de chaleur l’énergie lumineuse absorbée en excès, par un processus appelé le « Non-Photochemical Quenching of chlorophyll fluorescence », (NPQ). Chez la diatomée modèle, Phaeodactylum tricornutum, il a été montré que LHCX1, une protéine proche des antennes photosynthétiques, est impliquée dans le NPQ. Par des approches intrégrées de génétique, biologie moléculaire, biochimie, imagerie des cinétiques de fluorescence et spectroscopie ultrarapide, j’ai étudié le rôle de la famille des LHCX chez P. tricornutum. J’ai tout d’abord pu corréler une expression différentielle des 4 gènes LHCX de P. tricornutum avec différentes dynamiques de NPQ et activités photosynthétiques, dans différentes conditions de lumiére et nutriments. En localisant les LHCX dans les differents complexes photosynthétiques et les différents sites de dissipation d’énergie, j’ai pu proposer un modèle de régulation dynamique du NPQ impliquant à court terme principalement LHCX1 au niveau des centres réactionnels, et une autre isoforme, possiblement LHCX3, au niveau des antennes lors d’un stress lumineux prolongé. Enfin, par le criblage d’une série de mutants potentiellement dérégulés dans leur contenu en LHCXs, j’ai pu identifier des lignées avec un NPQ altéré qui pourront constituer des nouveaux outils de recherche. Dans l’ensemble ce travail de thèse a permis de mettre en évidence la diversification fonctionnelle et l’importance de la famille des LHCX dans la fine modulation des capacités de collecte de lumière et de photoprotection, expliquant sans doute en partie le succès des diatomées dans leur environnement très fluctuant. / Diatoms dominate phytoplanktonic communities in contemporary oceans, contributing to 20% of global primary productivity. In their extremely variable environment, diatoms are especially efficient in adjusting their photosynthetic activity by dissipating as heat the light energy absorbed in excess, through a process called “Non-Photochemical Quenching of chlorophyll fluorescence”, (NPQ). In the model diatom Phaeodactylum tricornutum, it has been shown that LHCX1, a photosynthetic antenna-related gene, is involved in the NPQ process. Through integrated approaches of genetics, molecular biology, biochemistry, study of the kinetics of chlorophyll fluorescence yields and ultrafast spectroscopy, I studied the role of the LHCX family in the photoprotection activity of P. tricornutum. I first correlated a differential regulation of the 4 P. tricornutum LHCX genes with different dynamics of NPQ and photosynthetic activity, in different light and nutrient conditions. By localizing the LHCXs in fractioned photosynthetic complexes and the different sites of energy dissipation, I was able to propose a model of dynamic regulation of NPQ capacity involving mainly the LHCX1 in the reaction centers, during short-term high light responses. During prolonged high light stress, the quenching occurs mainly in the antennas, potentially mediated by the LHCX3 isoform. Finally, using photosynthetic parameters, I screened a series of transgenic lines putatively deregulated in their LHCX amount, and I identified lines with altered NPQ, which could represent novel investigation tools. Altogether, this work highlighted the functional diversification and the importance of the LHCX protein family in the fine-tuning of light harvesting and photoprotection capacity, possibly contributing to explain diatoms success in their highly fluctuating environment.

Master of Science / Department of Agronomy / P. V. Vara Prasad / Grain sorghum (Sorghum bicolor L. Moench) is the fourth most important cereal crop grown throughout the semi-arid regions of the world. It is a staple food crop in Africa and Asia, while it is an important feed crop in the United States (US). More recently it is increasingly becoming important as a potential bioenergy feedstock crop around the world. The state of Kansas is the largest producer of grain sorghum in the US and contributes 40% of the total production. Drought is one of the major environmental factors limiting sorghum production in the semi-arid regions of the US, Asia and Africa. It is estimated that global crop losses due to drought stress exceed $10 billion annually. In crop production, drought stress can be classified into pre- or post-flowering. Even though the world collections of sorghum contain over 35,000 accessions, the genetic base currently used in breeding programs is very small (about 3%). Thus, it is important to identify diverse breeding lines for crop improvement. The diversity (association) panel consisting of 300 sorghum lines from all over the world was assembled for trait evaluation and association mapping. In this research these lines were grouped into the five major races (Figure 1) and 10 intermediate races of sorghum. The objectives of the research are to: (i) quantify the performance of the diversity panel under field conditions in Kansas, (ii) identify critical physiological traits affected by drought at both pre- and post-flowering stages of sorghum development, (iii) identify the most sensitive stage to drought stress during the reproductive phase of sorghum development and, (iv) test the feasibility of using a chlorophyll fluorescence assay (CVA) as a tool for identifying stay-green lines in grain sorghum during early stages of crop development. Field experiments were conducted in 2006 and 2007 in two locations in Kansas (Manhattan and Hays) under rain fed and irrigated conditions for the association panel. Objectives (iii) and (iv) were achieved with controlled environment experiments conducted in the greenhouse at the agronomy department, Kansas State University in 2006 and 2007. Results showed that there was large genetic variability among and within different races in the diversity panel for growth, physiological traits and yield components. Some genotypes showed yield stability across the different environments that were investigated. Drought significantly decreased seed number and harvest index across genotypes and races. In grain sorghum the period prior to flowering (panicle initiation) was the most sensitive stage to drought stress, in terms of its effect on seed-set, during reproductive development. A cell viability assay showed that there were significant differences in the loss of cell viability between leaf sample of stay green and non-stay green genotypes when leaf samples are collected in the morning and subjected to high respiratory demand. Therefore the chlorophyll fluorescence assay has potential as a tool for stay green trait screening at early stages of growth in grain sorghum.