As anthropogenic nutrient loads threaten the health of the Chesapeake Bay, lotic processes throughout its headwaters may buffer increased nitrogen inputs by converting them to stable forms, ultimately through denitrification to N2 gas. However, the temporal environmental factors controlling baseflow nitrogen retention are poorly understood, particularly temperature, shading, and dissolved organic matter dynamics. This study therefore attempts to elucidate the effects of these environmental variables on nitrogen cycling within the Fair Hill Natural Resources Management Area (Fair Hill), a forested watershed within the Piedmont physiographic province of the Chesapeake Bay. As expected, groundwater and allochthonous organic matter inputs set the foundation for lotic biogeochemistry at Fair Hill, creating a nutrient-limited, heterotrophic reach. Within this setting, three temporal "hot-moments" of in-stream nutrient processing were observed: the release of ammonium and phosphate during the warm - but shaded - growing season; nitrate uptake during autumnal leaf-fall; and a unique spike of nitrate uptake and respiration-induced degradation of labile organic matter during a drought. Consequently, the baseflow capacity of this headwater stream to buffer nutrient exports to the Chesapeake Bay constantly varies throughout the year in response to light availability, temperature, and in-stream organic matter dynamics. / Master of Science / Throughout the Chesapeake Bay watershed, ecological processes known as nitrogen retention can naturally remove nitrogen pollution from small streams (a.k.a. headwater streams), and hence the Chesapeake Bay watershed. However, in-stream nitrogen retention varies throughout the year due to seasonal changes in temperature, shading (as leaves grow in the spring or fall off in the fall), and the amount and type of organic matter in the stream. This study examines how these three variables (temperature, shading, and dissolved organic matter dynamics) affect nitrogen retention in a headwater, forested stream within the Fair Hill Natural Resources Management Area (Fair Hill) located in the Piedmont region of the Chesapeake Bay watershed. As expected, groundwater and organic matter inputs set the foundation for in-stream conditions at Fair Hill, creating an environment with low concentrations of nitrate and phosphate (thus causing the stream to be nutrient-limited), while also creating a heterotrophic environment, which is an environment where more oxygen is consumed by microbes than produced by algae and plants. Additionally, three seasonal patterns regarding in-stream nutrient dynamics were observed at Fair Hill. Firstly, in-stream ammonium and phosphate concentrations increased during the warm - but shaded - growing season. Secondly, in-stream nitrate concentrations decreased when leaves fell in the fall. Thirdly, during a drought, in-stream nitrate removal increased while in-stream organic matter became more degraded. Consequently, in-stream nutrient retention at Fair Hill varies constantly throughout the year in response to light availability, temperature, and in-stream organic matter dynamics.

Black Cottonwood (Populus trichocarpa) is a fast-growing, economically important tree species. P. trichocarpa was the first tree to have its genome fully sequenced and is considered the model organism for genomic research in trees. Of the macronutrients in plants, Nitrogen (N) is required in the greatest amounts and is generally the limiting nutrient in terrestrial ecosystems. Inorganic N-transport is performed by four families of transporter proteins, AMT1 and AMT2 for ammonium (NH4+) and NRT1 and NRT2 for nitrate (NO3-). I have created phylogenetic reconstructions of each of these transporter families in 22 members of Viridiplantae whose genomes have been fully sequenced. Based on these phylogenies, I have introduced a new classification system for the transporter families that better represents the evolutionary and functional relatedness of the proteins. These phylogenies were supplemented with topology predictions, subcellular localization predictions, and in silico expression profiling in order to suggest functional characterization of the groups. This facilitated candidate gene selection for NH4+ and NO3- uptake transporters from P. trichocarpa. Expression profiling was performed on two of these candidates. Results suggest that PtAMT1-1 may be a high-affinity, root-localized NH4+ transporter. In contrast, PtNRT2-6 is a high-affinity NO3- transporter localized to the dormant bud, but its physiological functions remain largely enigmatic. Flux profiles of NH4+, NO3-, and H+ in the first 1.4 cm of root tips of three-week-old P. trichocarpa seedlings and cuttings were measured using the Microelectrode Ion Flux mEasurement (MIFE) system to demonstrate the activity of AMTs and NRTs under nutrient-abundant and nutrient-deficient conditions. I found mainly N-efflux from roots of cuttings while seedling roots exhibited N-uptake. This is the first report of such a difference. This highlights an unexpected but clear physiological difference between seedling and cutting roots, which are frequently used in experimental setups. / Graduate / 0817 / 0369 / 0715 / neilvonw@gmail.com