The physiological function of the plant enzyme alternative oxidase has long been a topic of debate. The cyanide-resistant alternative oxidase (AOX), along with the cytochrome c oxidase (COX), catalyzes the reduction of oxygen to water in the electron transport chain of mitochondrial respiration. Although respiration via the alternative pathway (AP) results in approximately one third of the ATP production as respiration via the cytochrome pathway (CP), the AP is utilized by all plants and some fungi and animals. This “energy wasteful” pathway has been proposed to reduce oxidative stress in plant cells under a variety of stressful conditions. Virtually all previous work on the AP has been performed on laboratory-grown plants in controlled environment conditions; thus, there is little knowledge of how the AP responds to unstable conditions and multiple environmental stresses in the field. This thesis presents new methodology for studying AP respiration and the AOX protein in field-grown plants, and investigates how the AP responds to natural changes in environmental conditions in the field in several plant species grown in diverse ecosystem types. The experimental work presented here also investigates how AP activity is related to changes in total rates of respiration, and questions whether abundance of the AOX protein determines electron partitioning to the AP. AP partitioning (or relative changes in AP partitioning) varied over seasonal timescales in each of the experimental studies. Chapter 3 reports on two species of Chionochloa, a native New Zealand tussock grass growing along an altitudinal gradient. In Chapter 4, seasonal variation was studied in two tree types: Populus x canadensis, a deciduous angiosperm, and Pinus radiata, an evergreen gymnosperm. Quercus rubra trees were studied along an urban-rural gradient originating in New York City in Chapter 5. In a highly exposed and variable environment, relative changes in AP partitioning in two species of Chionochloa were correlated with the previous day’s integrated light. In Quercus rubra, the AP was instead related to temperature changes: relative AP partitioning increased in response to seasonally low temperature in trees grown at colder, more rural field sites, while at the warmer, urban sites, it increased in response to high summer temperatures. Each of these environmental conditions that were related to increases in the AP (high light, low temperatures, and heat) are potentially stressful to plants. Thus, it is possible that the increases in AP respiration observed in these studies served to oxidize excess reducing equivalents generated through stressful conditions. In Chapter 4, although AP partitioning in Populus x canadensis and Pinus radiata varied seasonally, these changes were not directly related to environmental parameters. However, AP partitioning in Populus x canadensis was clearly shown to be dependent on measurement temperature. In each of the studies presented here, changes in the AP were not related to abundance of the AOX protein. AOX protein abundance showed consistent seasonal patterns in the two deciduous angiosperms, Populus sp. and Quercus sp, and was correlated with seasonal changes in temperature in Chionochloa spp. However, the lack of correlation between protein abundance and AP partitioning indicates that the AP is subject to post-translational control and likely varies more rapidly than protein levels. In each of Chapters 3 – 5, there was no clear impact of changes in AP partitioning on rates of total respiration. As the AP produces less ATP than the CP, I hypothesized that increases in AP activity would lead to higher respiration rates in order to meet a plant’s energy demands. However, in Populus x canadensis and Quercus rubra, respiration rates remained stable during sharp increases in AP partitioning, indicating that, at least under certain conditions, increases in AP activity are accompanied by a decrease in the CP. In some of the first research studying AP partitioning in field-grown plants, this thesis illuminates possible mechanisms, functions, and implications of the AP. Over a range of plant taxa and environmental settings, this work shows that the AP does respond to stressful conditions in the wild, but that this does not result in increased respiration. Lastly, the methods presented here to study AP activity and AOX proteins in the field enable future studies to further probe the specific responses of AOX to natural stresses.