The global transport and deposition of anthropogenic nitrogen (N) to downwind ecosystems are significant and continue to increase. Indeed, atmospheric deposition can be a significant source of N to many watersheds, including those in remote, unpopulated areas. Bacterial denitrification in lake sediments may ameliorate the effects of N loading by converting nitrate (NO3-) to N2 gas. Denitrification also produces nitrous oxide (N2O), a potent greenhouse gas. The ecological effects of atmospheric N inputs in terrestrial ecosystems and the pelagic zone of lakes have been well documented; however, similar research in lake sediments is lacking. This project investigates the effects N of deposition on denitrification and N2O production in lakes. Atmospheric N inputs might alter the availability of NO3- and other key resources to denitrifiers. Such altered resources could influence denitrification, N2O production, and the abundance of denitrifying bacteria in sediments. The research contrasts these responses in lakes at the ends of gradients of N deposition in Colorado and Norway. Rates of denitrification and N2O production were elevated in the sediments of lakes subject to anthropogenic N inputs. There was no evidence, however, that N deposition has altered sediment resources or the abundance of denitrifiers. Further investigation into the dynamics of nitric oxide, N2O, and N2 during denitrification found no difference between deposition regions. Regardless of atmospheric N inputs, sediments from lakes in both Norway and Colorado possess considerable capacity to remove NO3- by denitrification. Catchment-specific properties may influence the denitrifying community more strongly than the rate of atmospheric N loading. In this regard, sediments appear to be insulated from the effects of N deposition compared to the water column. Lastly, surface water N2O concentrations were greater in high-deposition lakes compared to low-deposition lakes. To understand the potential magnitude of deposition-induced N2O production, the greenhouse gas inventory methodology of Intergovernmental Panel on Climate Change was applied to available datasets. Estimated emissions from lakes are 7-371 Gg N y-1, suggesting that lakes could be an important source of N2O.

Serial femtosecond crystallography requires reliable and efficient delivery of fresh crystals across the beam of an X-ray free-electron laser over the course of an experiment. We introduce a double-flow focusing nozzle to meet this challenge, with significantly reduced sample consumption, while improving jet stability over previous generations of nozzles. We demonstrate its use to determine the first room-temperature structure of RNA polymerase II at high resolution, revealing new structural details. Moreover, the double flow-focusing nozzles were successfully tested with three other protein samples and the first room temperature structure of an extradiol ring-cleaving dioxygenase was solved by utilizing the improved operation and characteristics of these devices.

X-ray free-electron lasers provide novel opportunities to conduct single particle analysis on nanoscale particles. Coherent diffractive imaging experiments were performed at the Linac Coherent Light Source (LCLS), SLAC National Laboratory, exposing single inorganic core-shell nanoparticles to femtosecond hard-X-ray pulses. Each facetted nanoparticle consisted of a crystalline gold core and a differently shaped palladium shell. Scattered intensities were observed up to about 7 nm resolution. Analysis of the scattering patterns revealed the size distribution of the samples, which is consistent with that obtained from direct real-space imaging by electron microscopy. Scattering patterns resulting from single particles were selected and compiled into a dataset which can be valuable for algorithm developments in single particle scattering research.

Humans have dramatically increased phosphorus (P) availability in terrestrial and aquatic ecosystems. As P is often a limiting nutrient of primary production, changes in its availability can have dramatic effects on ecosystem processes. I examined the effects of calcium carbonate (CaCO3) deposition, which can lower P concentrations via coprecipitation of phosphate, on P availability in two systems: streams in the Huachuca Mountains, Arizona, and a stream, Río Mesquites, in Cuatro Ciénegas, México. Calcium carbonate forms as travertine in the former and within the microbialites of the latter. Despite these differences, CaCO3 deposition led to lowered P availability in both systems. By analyzing a three-year dataset of water chemistry from the Huachuca Mountain streams, I determined that P concentrations were negatively related to CaCO3 deposition rates. I also discovered that CaCO3 was positively correlated with nitrogen concentrations, suggesting that the stoichiometric effect of CaCO3 deposition on nutrient availability is due not only to coprecipitation of phosphate, but also to P-related constraints on biotic nitrogen uptake. Building from these observations, bioassays of nutrient limitation of periphyton growth suggest that P limitation is more prevalent in streams with active CaCO3 deposition than those without. Furthermore, when I experimentally reduced rates of CaCO3 deposition within one of the streams by partial light-exclusion, areal P uptake lengths decreased, periphyton P content and growth increased, and periphyton nutrient limitation by P decreased. In Río Mesquites, CaCO3 deposition was also associated with P limitation of microbial growth. There, I investigated the consequences of reductions in CaCO3 deposition with several methods. Calcium removal led to increased concentrations of P in the microbial biomass while light reductions decreased microbial biomass and chemical inhibition had no effect. These results suggest that CaCO3 deposition in microbialites does limit biological uptake of P, that photoautotrophs play an important role in nutrient acquisition, and, combined with other experimental observations, that sulfate reduction may support CaCO3 deposition in the microbialite communities of Río Mesquites. Overall, my results suggest that the effects of CaCO3 deposition on P availability are general and this process should be considered when managing nutrient flows across aquatic ecosystems.