Thesis (Ph. D.)--University of Missouri-Columbia, 1998. / Typescript. Vita. Includes bibliographical references. Also available on the Internet.

Thesis (Ph. D.)--University of Missouri-Columbia, 1998. / Typescript. Vita. Includes bibliographical references. Also available on the Internet.

As one of the first land plant groups to diversify, mosses are central in understanding the origin, diversification, and early function of stomata. Unlike tracheophytes that have stomata on anatomically complex leaves and stems, mosses bear stomata exclusively on spore-bearing organs (capsules). However, stomata do not occur in all mosses and, indeed, are absence in the earliest-divergent mosses (Takakia, Andreaea, Andreaeobryum and Sphagnum), suggesting that stomata originated in mosses independently of other plants. The occurrence of structurally unique pseudostomata in Sphagnum further confounds the resolution of homology of moss stomata with those of other plants. The five studies included in this dissertation are aimed at clarifying the structure, development and evolution of moss stomata. The first study focuses on the sporophyte anatomy and stomatal ultrastructure in two structurally and phylogenetically divergent mosses, Oedipodium and Ephemerum. Oedipodium is the sister to peristomate mosses and the first extant moss with true stomata. This monospecific genus has an elaborated capsule with an extended apophysis bearing numerous long-pored stomata. In contrast, Ephemerum nests within the peristomate mosses and has a reduced capsule that lacks an apophysis and has a few round-pored stomata. Ultrastructure of stomata is similar in these two mosses and comparable to that of tracheophytes, except that the stomata of mosses are not as structurally distinct from epidermal cells as are tracheophyte stomata. Anatomical features such as the presence of a cuticle, water-conducting cells, and spongy tissues with large areas for gas exchange are more pronounced in Oedipodium sporophytes and support the role of stomata in gas exchange and water transport during development and maturation. The second study examines changes in pectin composition during development in the model moss Funaria. Stomatal movement in tracheophytes requires guard cell walls to be strong, yet flexible, because they have to undergo reversible deformation to open and close the pore. Pectins are necessary for wall flexibility and proper stomatal functioning in seed plants. In this study of Funaria, immunogold-labeling using five antibodies to pectin epitopes was conducted on guard cell walls during development to relate these features to the limited movement of stomata in moss. Movement of Funaria stomata coincides with capsule expansion when guard cell walls are thin and pectinaceous. Walls dramatically increase in thickness after pore formation and the pectin content significantly decreases in mature guard cell walls, suggesting that a decrease in flexibility is responsible for the inability to open a close previously reported in older moss guard cells. Because this was the first study to demonstrate changes in pectin composition during stomatal development in any plant, a similar study was done on Arabidopsis to identify the main types of pectins in guard cell walls. Localization of pectins in guard cell walls of Arabidopsis is similar to mosses in the stage they can move, with homogeneous walls rich in arabinan pectins that are required for wall flexibility. This study extends knowledge of pectin composition from stomata of the moss Funaria with limited stomatal movement to an angiosperm in which stomatal activity is crucial to the physiological health of the plant. The fourth study describes stomata development and internal changes in sporophyte anatomy that lead to formation of air spaces in the moss Funaria. Developing sporophytes at different stages were examined using light, fluorescence and electron microscopy; immunogold-labeling was used to investigate the presence of pectin in the newly formed cavities. Stomata in mosses do not develop from a self-generating meristemoid like in Arabidopsis, but instead they originate from a protodermal cell that differentiates directly into a guard mother cell. Epidermal cells develop from protodermal or other epidermal cells, i.e., there are no stomatal lineage ground cells. This developmental pattern is congruent with the presence of a gene ortholog of FAMA, but not SPCH and MUTE, in Physcomitrella. The final study in this dissertation focuses on the enigmatic Sphagnum. Although true stomata are absent in early-divergent mosses, Sphagnum has specialized epidermal cells, pseudostomata, that partially separate but do not open to the inside. To further understand the structure, function and evolution of pseudostomata, capsule anatomy and ultrastructure of pseudostomata were detailed. As in moss stomata, pseudostomata wall architecture and behavior facilitate capsule dehydration, shape change, and dehiscence, supporting this common function. Unlike other moss stomata, pseudostomata collapse along their ventral walls and they lack a substomatal cavity. Similarities to true stomata include two modified epidermal cells with specialized cell walls that separate by cuticle deposition and respond to drying. Pseudostomata may be interpreted as modified stomata that suppressed substomatal cavity formation, which in turn eliminated pore development. However, clarification of the homology of pseudostomata and moss stomata will require genomic studies integrated with physiological and structural data. The studies described in this dissertation significantly advance our understanding of moss stomatal development and structure, and provide a comparison point to better evaluate the evolution of stomata. Moss capsule anatomy coupled with the exclusive existence of stomata on capsules supports the concept that stomata in moss are involve in gas exchange but also facilitate drying and dispersal of spores. Changes in wall architecture coupled with a decrease in total pectin explain the inability of mature stomata to move. Development and distribution of stomata in Funaria provides evidence of a direct and less elaborated mechanism for stomatal development than described in Arabidopsis. Resolving relationships among early land plants, especially hornworts and mosses, the only bryophyte groups with stomata, is critical to understanding stomata evolution. Evaluated together, the results of this dissertation are consistent with a single origin of stomata in land plants.

The stomatal function and genetically based regulation of their number are interesting and important subjects of scientific research. Stomatal density on plant leaf surface is affected also by environmental factors, e.g. atmospheric CO2 concentration and irradiance (PAR). These two factors have common affect ? leaf internal CO2 concentration (Ci) in the leaf can be changed. The question might be if the Ci is the signal for expression of genes controlling stomatal development. According to the hypothesis which this thesis is based on I proposed an increase of expression of positive regulator STOMAGEN in the case when Ci is reduced. The expression of STOMAGEN was increased in plants with Ci decreased by higher PAR in contrast with plants where the Ci was decreased by adding of abscisic acid into nutrient solution. These results show no dependence of STOMAGEN expression on inner CO2 concentration.

Orientador: Ana Paula Fortuna Perez / Resumo: A família Lygodiaceae sempre teve uma taxonomia contraditória, o número de espécies aceitas no Brasil variando de duas a doze ao longo das décadas. Sendo Lygodiaceae uma família antiga, acredita-se que essas contradições possam ser causadas por híbridos. Os híbridos surgem pelo cruzamento de duas ou mais espécies e, geralmente, são primeiro identificados por suas características morfológicas intermediárias entre as espécies parentais. Normalmente, os híbridos recentes de samambaia são detectados ocorrendo junto com suas espécies parentais. Os esporos de indivíduos híbridos geralmente são mal formados e / ou abortados, portanto, inviáveis, mas os poucos esporos viáveis poderiam iniciar o processo de especiação de uma nova espécie. Considerando a sobreposição de características morfológicas, a hibridação poderia ter ocorrido entre duas espécies de Lygodium no Brasil, L. venustum e L. volubile gerando o hipotético híbrido L. × fayae. Devido à difícil identificação desses táxons, nossos estudos analisaram dois caracteres diferentes (tamanho e morfologia dos esporos e estômatos) por microscopia eletrônica de varredura, estudamos a germinação dos esporos e criamos novas descrições e uma chave de identificação às espécies aceitas para ajudar na delimitação taxonômica dos dois táxons de Lygodium e seu híbrido putativo. De acordo com nossos resultados, as espécies classificadas como L. × fayae na região sudeste do Brasil não são híbridas, mas uma variação morfológica de Lygodium venus... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The family Lygodiaceae has always had a contradictory taxonomy, the number of accepted species in Brazil varying from two to twelve through the decades. Lygodiaceae being an old family, this contradictions are believed to be caused by hybrids. Hybrids arise by the crossing of two or more species, and, usually, they are first identified by its intermediate morphological characteristics among the parental species. Normally, early fern hybrids are detected occurring along with its parental species. The spores of hybrid individuals generally are badly formed and/or aborted, thus, not viable, but the few balanced spores could starts the speciation process of a new species. Considering the overlapping of morphological characteristics, it could have occurred in two species of Lygodium in Brazil, L. venustum and L. volubile and the hypothetical hybrid L. ×fayae. Due to the difficult in identification of these taxa, our studies herein analysed two different characters (spore and stomata size and morphology) through scanning electronic microscopy, we studies the spore germination and new descriptions and an identification key were done to the accepted species to help clearing the taxonomic delimitation of the two taxa of Lygodium and their putative hybrid. According to our results, the species herein classified as L. ×fayae on the Brazilian south-east region is not a hybrid, but a morphological variation of Lygodium venustum. Additional studies on morphometric analysis with the measure... (Complete abstract click electronic access below) / Mestre

Heat stress poses a serious threat to plant survival and productivity, and has a direct influence on crop yield stability. Plants response to high temperature is tightly controlled by complex genetic networks. Plants can be acclimated through gradual pre-exposure to increasing temperatures and that in turn causes higher survival in subsequent and otherwise lethal heat stress conditions. To investigate the physiological and molecular processes underlying heat acclimation and recovery, we examined changes in Arabidopsis thaliana transcriptome throughout the acclimation and the subsequent heat shock treatment. Groups of differentially expressed genes and enriched biological pathways that constitute the heat transcriptional memory were identified. The function of flavonoids in plant heat stress were further explored experimentally. In addition, we observed altered stomata density and aperture responses in heat acclimated plants, and this might be partially controlled by AGAMOUS-LIKE16 (AGL16) transcription factor and its negative regulator microRNA824 (miR824). By utilizing an automated non-invasive phenotyping facility, we have developed a protocol to record plant growth and photosynthetic performance after heat stress in wild type Arabidopsis thaliana and mutant lines at daily intervals. Through an imaging-based analysis of plants growth, we confirmed impaired thermotolerance of hsp101 compared to wild type plants by a time-series growth, morphology and chlorophyll responses. This offers a novel experimental setup for thermotolerance screenings in Arabidopsis, with defined digital markers linking the function of selected genes in heat stress responses to phenotypic traits.

In this study, the magnitude of stomatal water loss was compared between different Arabidopsis thaliana mutants. The stomatal water loss among different mutants of Populus sp. was also examined in this study. Fourteen different Arabidopsis thaliana mutants were sown and later genotyped using DNA-extraction, PCR, and gel electrophoresis. Leaf water loss analysis was made on both Arabidopsis thaliana and Populus sp. The data was then used in statistical tests, to find if there was a significant difference in rate of stomatal water loss between different genotypes. The result showed a significant difference in water loss when comparing the toc1-5 and T89 genotypes and the toc1-5 and 1119-4 genotypes in Populus sp. There was no significant difference between the 1119-4 and T89 genotypes in Populus sp. The result also showed that there was a significant difference between 25 combinations of the Arabidopsis thaliana genotypes. A conclusion was that TOC1 reduces water loss in Arabidopsis thaliana, indicating a role in stomatal clousure. Future research is needed to examine differences between genotypes among both species included in the study.

No description available.

In this study, leaf anatomies of five species of Limonium Mill. (L. echioides, L. globuliferum, L. tamaricoides, L. anatolicum and L. Sinuatum) are studied. Taca belonging to five different sections of the genus Limonium as / L. echioides of the section Schizyhymenium, L. globuliferum of the section Sphaerostachys, L. tamaricoides of the section Limonium, L. anatolicum of the section Sarcophyllum and L. sinuatum of the section Pteroclados are used in this study. Throughout the species studied, L. tamaricoides and L. anatolicum are endemic to Turkey. In order to examine anatomy of leaves, the paraffin sectioning and hand cross sectioning methods are used. Avarage stomata length, width and number per 210.68&micro / m x 263.27&micro / m area of the leaves are examined and the stomata types due to epidermal cells are defined. Additionally, the vascular bundles, upper and lower epiderms of leaves of each species are defined. It is determined that except the species L. globuliferum and L. tamaricoides, the stomata density per 210.68&micro / m x 263.27&micro / m area of the leaves are approximately same for upper and lower epidermis, but in these species, the stomata numbers at upper epidermis is higher than the stomata numbers in lower epidermis, which might be related with the altitude, microclimate and habitat of these two species. It is also found that except the shoot leaf of L. sinuatum, the phloem is closer to lower epiderm than xylem in vascular bundles. But in shoot leaf of L. sinuatum, the phloem circularly covers the xylem and makes a closed circle around.

Células estomáticas e reações metabólicas de plantas foram modelados por meio da teoria dos grafos neste trabalho; a distância entre estômatos vizinhos na folha foi adotada como parâmetro utilizado para a conectividade em redes onde os estômatos foram definidos como nodos. A direção da formação de produtos e substratos em reações metabólicas determinou a conectividade nas redes metabólicas, onde cada metabólito foi definido como um nodo. As redes de estômatos foram capazes de gerar uma grande quantidade de informação geométrica associada à distância entre os estômatos. Estas medidas se mostraram uma poderosa ferramenta para avaliar a plasticidade fenotípica em folhas de plantas. A adaptação de plantas a condições ambientais extremas, como altas taxas de umidade e grandes variações no tempo de exposição à luz, puderam ser quantificadas por parâmetros de redes. Parâmetros topológicos globais das redes metabólicas mostraram que elas possuem propriedades estatísticas e topológicas de redes livre escala, como nos seres vivos em geral. Entretanto, alguns parâmetros topológicos locais das redes como a medida hub-score, geram vetores de características que, se comparados entre plantas, geram informação filogenética. Além disso, nós comprovamos que é possível construir modelos que sugerem uma organização geral para o metabolismo, por meio de algorítmos de conectividade hierárquica. O algorítmo de k-cores foi usado para gerar camadas de conectividade nas redes metabólicas. A atribuição química dos metabólitos ao longo das camadas k-core, mostra que a hierarquia de conexões está associada a especialização do metabolismo. Isto sugere que o algorítimo também gera informação sobre a evolução da maquinaria metabólica. Portanto, o modelo para conectar elementos de uma rede metabólica adotado neste trabalho, traz informações naturais sobre as plantas, o que sugere que exista parâmetros físicos das reações metabólicas representados pelo modelo. / Stomatic cells and metabolic reactions were modeled by graph theory in this work. The distance between stomata was adopted as connectivity parameter in the networks where stomata were defined as nodes. The direction of formation from products and substrates in the metabolic reactions, determined the connectivity from the metabolic networks, where each metabolite was defined as a node. The networks of stomata were able to generate a large amount of geometric information based at distance between the stomata. These measures represented a powerful tool to evaluate the phenotypic plasticity in leaves of plants. Global topological parameters from plant the metabolic networks revealed that plant metabolic networks has the topology of free scale networks, as in living beings in general. However, some local topological parameters of the networks such as the hub-score, can be organized as characteristic vectors with differential phylogenetic information. In addition, we have shown that it is possible to construct models that suggest a general organization for the metabolism through algorithms with iterative percolation from network connectivity. The k-cores algorithm was used to generate layers of connectivity in the metabolic networks. The chemical assignment of the metabolites along the k-core layers shows that the hierarchy of connections is associated with specialization of metabolism. This suggests that the algorithm also generates information about the evolution of the metabolic machinery. Therefore, the model used to connect elements of the metabolic networks adopted in this work, brings natural information about the plants, which suggests that there are physical parameters of the metabolic reactions represented by the model.