Regulation of woody plant development by gibberellin catabolic and signaling genes

Gibberellin (GA) is a growth promoting hormone implicated in regulating a diversity of plant processes. This dissertation examines the role of GA metabolic and signaling genes in woody plant growth and development. Transgenic modifications, expression analysis, physiological/biochemical assays, biometric measurements and histological analysis were used to understand the regulatory roles these genes play in the model woody plant, Populus. Our results highlight the importance of GA regulatory genes in woody perennial growth, including: phenology, wood formation, phenotypic plasticity, and growth/survival under field conditions. We characterize two putative Populus orthologs of the SHORT INTERNODES (SHI) gene from Arabidopsis, a negative regulator of GA signaling. RNAi-mediated suppression of Populus SHI-like genes increased several growth-related traits, including extent of xylem proliferation, in a dose-dependent manner. Three Populus genes, sharing sequence homology to the positive regulator of GA signaling gene PHOTOPERIOD-RESPONSIVE 1 (PHOR1) from Solanum, are up-regulated in GA-deficient and insensitive plants suggesting a conserved role in GA signaling. We demonstrate that Populus PHOR1-like genes have overlapping and divergent function(s). Two PHOR1-like genes are highly expressed in roots, predominantly affect root growth (e.g., morphology, starch quantity and gravitropism), and induced by short-days (SD). The other PHOR1-like gene is ubiquitously expressed with a generalized function in root and shoot development. The effects of GA catabolic and signaling genes on important traits (e.g., adaptive and productivity traits) were studied in a multi-year field trial. Transgenics overexpressing GA 2-oxidase (GA2ox) and DELLA genes showed tremendous variation in growth, form, foliage, and phenology (i.e., vegetative and reproductive). Observed gradients in trait modifications were correlated to transgene expression levels, in a manner suggesting a dose-dependent relationship. We explore GA2ox and DELLA genes involvement in mediating growth responses to immediate short-term drought stress, and SD photoperiods, signaling prolonged periods of stress (e.g., winter bud dormancy). GA2ox and DELLA genes show substantial up-regulation in response to drought and SDs. Transgenics overexpressing homologs of these genes subjected to drought and SD photoperiods show hypersensitive growth restraint and increased stress resistances. These results suggest growth cessation (i.e., dormancy) in response to adverse conditions is mediated by GA regulatory genes.

Speciation with gene-flow in eastern North American red oaks (Quercus section Lobatae)

As foundational species, oaks (Quercus : Fagaceae) support the activities of both humans and wildlife. However, many oaks in North America are declining, a crisis exacerbated by the previous disappearance of other hard mast-producing trees. In addition, the economic demands placed on this drought-tolerant group may intensify if climate change extirpates other, relatively mesophytic species. Genetic tools can help address these management challenges. To this end, we developed a suite of 27 microsatellite markers, of which 22 are derived from expressed sequence tags (ESTs). Many of these markers bear significant homology to known genes and may be able to directly assay functional genetic variation. Markers obtained from enriched microsatellite libraries, on the other hand, are typically located in heterochromatic regions and should reflect demographic processes. Considered jointly, genic and genomic microsatellites can elucidate patterns of gene-flow and natural selection, which are fundamental to both an organism's evolutionary ecology and conservation biology. To this end, we employed the developed markers in an FST-based genome scan to detect the signature of divergent selection among the red oaks (Quercus section Lobatae). Three candidate genes with putative roles in stress responses demonstrated patterns of diversity consistent with adaptation to heterogeneous selective pressures. These genes may be important in both local genetic adaptation within species and divergence among them. Next, we used an isolation-with-migration model to quantify levels of gene-flow among four red oaks species during speciation. Both speciation in allopatry and speciation with gene-flow were found to be major drivers of red oak biodiversity. Loci playing a key role in speciation are also likely to be ecologically important within species