Theses and Dissertations from UMD
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Item SPATIO-TEMPORAL ANALYSIS OF PHOTOTROPISM IN ARABIDOPSIS SEEDLINGS(2019) Pritchard, Candace; Murphy, Angus S; Plant Science and Landscape Architecture (PSLA); Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Optimization of light capture during seedling development is a major determinant of plant fitness. As seedlings emerge from the soil, the processes of photomorphogenesis and phototropism optimize deployment of structures that capture light for photosynthesis. Photomorphogenesis produces hypocotyl thickening, cotyledon expansion, and chloroplast maturation. Concurrent phototropic responses initiated by blue light position the expanding cotyledons to maximize photosynthesis. The mechanisms underlying both processes have been explored for more than 140 years, but are still not fully understood. This dissertation seeks to provide a better understanding of phototropism by exploring the timing and localization of the constituent mechanisms downstream of the well-characterized perception of blue light by the PHOTOTROPIN photoreceptors. The experiments described herein characterize temporally and spatially distinct processes involved in asymmetric auxin accumulations that lead to differential hypocotyl elongation. To better identify the link between early perception and later auxin transport and elongation events, an open-air system was used to remove seedling hindrance and provide better spatio-temporal resolution. These experiments confirmed the more rapid bending conferred by loss of the ATP Binding Cassette class B (ABCB) 19 auxin efflux transporter and loss of differential elongation in the mid hypocotyl elongation zone in higher order pinformed mutants. However, apart from the enhancement of phototropic bending observed in abcb19 and pin4 mutants, no auxin transport mutants tested showed alterations in early phototropic responses, and no mutant exhibited a delay in the onset of phototropic bending. Recently identified CBC1 and CBC2 (CONVERGENCE OF BLUE LIGHT (BL) AND CO2 1/2) have been shown to act in downstream signaling during phot1-mediated regulation of stomatal conductance. Similarly, during phototropism cbc1cbc2 double mutants show early defects in phot1-mediated phototropism. Further, CBC1 and CBC2 have been shown to regulate S-type anion channels. Analysis of S-type anion channel mutants also reveals defects in early bending responses. These results point to blue light-dependent regulation of anion channel activity having an important role during the earliest stages of phototropism.Item SEASONAL NITROGEN CYCLING AND HOMEOSTASIS IN POPULUS: SOURCE-SINK COMMUNICATION(2018) Li, Gen; Coleman, Gary D; Plant Science and Landscape Architecture (PSLA); Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Nitrogen (N) is an essential nutrient for plant growth, development and reproduction. Seasonal N cycling is an adaption to nutrient limitation and a feature of the perennial lifestyle of trees. Poplar (Populus) is a model system used to study forest tree genetics and molecular biology, including seasonal N cycling. The accumulation of Bark storage proteins (BSP) is a central feature of seasonal N cycling in poplar, yet our understanding of the contribution of the BSP storage pool to N remobilization during growth and mechanisms that regulate BSP accumulation, catabolism and N remobilization is limited. The research presented in this dissertation is directed towards advancing knowledge of the regulation of seasonal N cycling in poplar using a combination of experimental approaches. The role of the N storage pool to N remobilization was examined through N sink manipulations and the specific role of BSP storage was investigated by N source reduction via RNAi mediated knockdown of BSP accumulation. From this it was found that N remobilization from BSP significantly contributes to shoot growth following dormancy and initial shoot growth is source limited. Poplar bark transcriptome analyses during regrowth following dormancy revealed an enrichment for up-regulated genes associated with auxin transport and signaling. Based on the transcriptome analysis experiments that manipulate auxin production or polar auxin transport were performed and the results indicate that BSP catabolism and N remobilization likely involves polar auxin transport from expanding buds and shoots and auxin-mediated regulation of protease gene expression. Analysis of DNA microarrays of bark RNA during short-day (SD) induction of BSP gene expression was used to identify putative regulatory factors that may play a role of BSP accumulation. The transcriptome analyses indicated that SD represses the expression of genes involved in ethylene production as well as a reduction in bark ethylene biosynthesis. Additionally, treatment of excised stems with ACC or ethephon repressed BSP gene expression while AVG induced BSP gene expression. This repression was reduced in ethylene-insensitive poplars expressing Arabidopsis dominant gain-of-function allele etr1-1. Furthermore, transient expression of ERF12 and ERF41, two transcription factors with the greatest induction in SD treatment, in transgenic tobacco stably transformed with BSPA promoter fused with GUS resulted in enhanced GUS activity suggesting ERF12 and ERF41 may act as positive regulators of BSP gene expression. Since glutamine plays a pivotal role in N partitioning during N storage and remobilization, the possible role of the PII glutamine sensor was studied and it was found that the transcript levels of PII increased in bark during SD-induced leaf senescence and BSP accumulation. PII knockdown poplars using RNAi showed reduced glutamine-induced BSP gene expression. Moreover, glutamine-induced BSP gene expression was also inhibited by 2-OG, a PII-NAGK interaction antagonist, suggesting a possible role of PII-glutamine sensing in BSP accumulation. Taken together, this study provides important insights into the mechanism of seasonal N accumulation and remobilization in poplar.Item Shaping the Dicot Fruit: Molecular and Genomic Approaches to Fruit Development(2016) Hawkins, Charles; Liu, Zhongchi; Cell Biology & Molecular Genetics; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)The fruit is one of the most complex and important structures produced by flowering plants, and understanding the development and maturation process of fruits in different angiosperm species with diverse fruit structures is of immense interest. In the work presented here, molecular genetics and genomic analysis are used to explore the processes that form the fruit in two species: The model organism Arabidopsis and the diploid strawberry Fragaria vesca. One important basic question concerns the molecular genetic basis of fruit patterning. A long-standing model of Arabidopsis fruit (the gynoecium) patterning holds that auxin produced at the apex diffuses downward, forming a gradient that provides apical-basal positional information to specify different tissue types along the gynoecium’s length. The proposed gradient, however, has never been observed and the model appears inconsistent with a number of observations. I present a new, alternative model, wherein auxin acts to establish the adaxial-abaxial domains of the carpel primordia, which then ensures proper development of the final gynoecium. A second project utilizes genomics to identify genes that regulate fruit color by analyzing the genome sequences of Fragaria vesca, a species of wild strawberry. Shared and distinct SNPs among three F. vesca accessions were identified, providing a foundation for locating candidate mutations underlying phenotypic variations among different F. vesca accessions. Through systematic analysis of relevant SNP variants, a candidate SNP in FveMYB10 was identified that may underlie the fruit color in the yellow-fruited accessions, which was subsequently confirmed by functional assays. Our lab has previously generated extensive RNA-sequencing data that depict genome-scale gene expression profiles in F. vesca fruit and flower tissues at different developmental stages. To enhance the accessibility of this dataset, the web-based eFP software was adapted for this dataset, allowing visualization of gene expression in any tissues by user-initiated queries. Together, this thesis work proposes a well-supported new model of fruit patterning in Arabidopsis and provides further resources for F. vesca, including genome-wide variant lists and the ability to visualize gene expression. This work will facilitate future work linking traits of economic importance to specific genes and gaining novel insights into fruit patterning and development.Item Influence of Nitrogen and Sink Competition on Shoot Growth of Poplar(2016) Egekwu, Chioma; Coleman, Gary D; Plant Science and Landscape Architecture (PSLA); Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Terrestrial and oceanic biomass carbon sinks help reduce anthropogenic CO2 emissions and mitigate the long-term effect of increasing atmospheric CO2. Woody plants have large carbon pools because of their long residence time, however N availability can negatively impact tree responses to elevated CO2. Seasonal cycling of internal N in trees is a component that contributes to fitness especially in N limited environments. It involves resorption from senescing leaves of deciduous trees and storage as vegetative storage proteins (VSP) in perennial organs. Populus is a model organism for tree biology that efficiently recycles N. Bark storage proteins (BSP) are the most abundant VSP that serves as seasonal N reserves. Here I show how poplar growth is influenced by N availability and how growth is influenced by shoot competition for stored N reserves. I also provide data that indicates that auxin mediates BSP catabolism during renewed shoot growth. Understanding the components of N accumulation, remobilization and utilization can provide insights leading to increasing N use efficiency (NUE) of perennial plants.Item The role of auxin on the evolution of embryo development and axis formation in land plants(2005-03-10) Poli, DorothyBelle; Cooke, Todd J.; Cell Biology & Molecular Genetics; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)ABSTRACT Title of Dissertation: THE ROLE OF AUXIN ON THE EVOLUTION OF EMBRYO DEVELOPMENT AND AXIS FORMATION IN LAND PLANTS DorothyBelle Poli, Doctor of Philosophy, 2005 Dissertation directed by: Professor Todd J. Cooke, Cell Biology and Molecular Genetics This thesis examined the role of auxin in the evolution of land plants. Several approaches were used to study how auxin regulates the development in the bryophyte sporophytes. The altered growth of isolated young sporophytes exposed to applied auxin (indole-3-acetic acid) or an auxin antagonist (p-chlorophenoxyisobutyric acid) suggested that endogenous auxin regulates the rates of axial growth in all bryophyte divisions. In the hornwort Phaeoceros personii, auxin moved at very low fluxes, was insensitive to an auxin-transport inhibitor (N-[1-naphthyl]phthalamic acid), and exhibited a polarity ratio close to 1.0, implying that auxin moves by simple diffusion. The liverwort Pellia epiphylla exhibited somewhat higher auxin fluxes, which were sensitive to transport inhibitors but lacked any measurable polarity. Thus, auxin movement in liverwort sporophytes appears to result from facilitated diffusion. In the moss Polytrichum ohioensis, auxin movement was predominantly basipetal in young sporophytes and occurred at high fluxes exceeding those measured in maize coleoptiles. In older sporophytes, acropetal auxin flux had increased beyond the level measured for basipetal flux in the specimens observed in several, but not all, seasons. The evidence from both inhibitor treatments and isolated tissues is consistent with the interpretation that the cortex carries out basipetal transport in both younger and older sporophytes, whereas the central vascular tissues carries out basipetal transport in younger sporophytes and acropetal flux in older sporophytes. Given the significant differences in fall rainfall in the collection years, the purported sensitivity of vascular tissue development may account for the seasonal variation observed in these experiments. Auxin regulators and polar transport were also used to study the regulation of the embryogenesis of the fern Marsilea vestita. Auxin biosynthesis inhibitors affected initial cell proliferation resulting in the formation of aborted embryos, p-chlorophenoxyisobutyric acid delayed growth and development in all stages of embryogenesis while -naphthaleneacetic acid mediated rapid cell proliferation that caused enlarged disorganized embryos. Polar auxin transport inhibitors caused no significant abnormalities, which suggested a limited role for polar transport in fern embryogenesis. In conclusion, this evidence suggests that auxin is ultimately involved in the establishment of the body plans in all land plant sporophytes.