Matching Items (86)
Description
The oceans play an essential role in global biogeochemical cycles and in regulating climate. The biological carbon pump, the photosynthetic fixation of carbon dioxide by phytoplankton and subsequent sequestration of organic carbon into deep water, combined with the physical carbon pump, make the oceans the only long-term net sink for anthropogenic carbon dioxide. A full understanding of the workings of the biological carbon pump requires a knowledge of the role of different taxonomic groups of phytoplankton (protists and cyanobacteria) to organic carbon export. However, this has been difficult due to the degraded nature of particles sinking into particle traps, the main tools employed by oceanographers to collect sinking particulate matter in the ocean. In this study DNA-based molecular methods, including denaturing gradient gel electrophoresis, cloning and sequencing, and taxon-specific quantitative PCR, allowed for the first time for the identification of which protists and cyanobacteria contributed to the material collected by the traps in relation to their presence in the euphotic zone. I conducted this study at two time-series stations in the subtropical North Atlantic Ocean, one north of the Canary Islands, and one located south of Bermuda. The Bermuda study allowed me to investigate seasonal and interannual changes in the contribution of the plankton community to particle flux. I could also show that small unarmored taxa, including representatives of prasinophytes and cyanobacteria, constituted a significant fraction of sequences recovered from sediment trap material. Prasinophyte sequences alone could account for up to 13% of the clone library sequences of trap material during bloom periods. These observations contradict a long-standing paradigm in biological oceanography that only large taxa with mineral shells are capable of sinking while smaller, unarmored cells are recycled in the euphotic zone through the microbial loop. Climate change and a subsequent warming of the surface ocean may lead to a shift in the protist community toward smaller cell size in the future, but in light of these findings these changes may not necessarily lead to a reduction in the strength of the biological carbon pump.
ContributorsAmacher, Jessica (Author) / Neuer, Susanne (Thesis advisor) / Garcia-Pichel, Ferran (Committee member) / Lomas, Michael (Committee member) / Wojciechowski, Martin (Committee member) / Stout, Valerie (Committee member) / Arizona State University (Publisher)
Created2011
Description
Many studies over the past two decades examined the link between climate patterns and discharge, but few have attempted to study the effects of the El Niño Southern Oscillation (ENSO) on localized and watershed specific processes such as nutrient loading in the Southwestern United States. The Multivariate ENSO Index (MEI) is used to describe the state of the ENSO, with positive (negative) values referring to an El Niño condition (La Niña condition). This study examined the connection between the MEI and precipitation, discharge, and total nitrogen (TN) and total phosphorus (TP) concentrations in the Upper Salt River Watershed in Arizona. Unrestricted regression models (UMs) and restricted regression models (RMs) were used to investigate the relationship between the discharges in Tonto Creek and the Salt River as functions of the magnitude of the MEI, precipitation, and season (winter/summer). The results suggest that in addition to precipitation, the MEI/season relationship is an important factor for predicting discharge. Additionally, high discharge events were associated with high magnitude ENSO events, both El Niño and La Niña. An UM including discharge and season, and a RM (restricting the seasonal factor to zero), were applied to TN and TP concentrations in the Salt River. Discharge and seasonality were significant factors describing the variability in TN in the Salt River while discharge alone was the significant factor describing TP. TN and TP in Roosevelt Lake were evaluated as functions of both discharge and MEI. Some significant correlations were found but internal nutrient cycling as well as seasonal stratification of the water column of the lake likely masks the true relationships. Based on these results, the MEI is a useful predictor of discharge, as well as nutrient loading in the Salt River Watershed through the Salt River and Tonto Creek. A predictive model investigating the effect of ENSO on nutrient loading through discharge can illustrate the effects of large scale climate patterns on smaller systems.
ContributorsSversvold, Darren (Author) / Neuer, Susanne (Thesis advisor) / Elser, James (Committee member) / Fenichel, Eli (Committee member) / Arizona State University (Publisher)
Created2012
Description
Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is widely accepted as the world's most abundant enzyme and represents the primary entry point for inorganic carbon into the biosphere. Rubisco's slow carboxylation rate of ribulose-1,5-bisphosphate (RuBP) and its susceptibility to inhibition has led some to term it the "bottle neck" of photosynthesis. In order to ensure that Rubisco remains uninhibited, plants require the catalytic chaperone Rubisco activase. Activase is a member of the AAA+ superfamily, ATPases associated with various cellular activities, and uses ATP hydrolysis as the driving force behind a conformational movement that returns activity to inhibited Rubisco active sites. A high resolution activase structure will be an essential tool for examining Rubisco/activase interactions as well as understanding the activase self-association phenomenon. Rubisco activase has long eluded crystallization, likely due to its infamous self-association (polydispersity). Therefore, a limited proteolysis approach was taken to identify soluble activase subdomains as potential crystallization targets. This process involves using proteolytic enzymes to cleave a protein into a few pieces and has previously proven successful in identifying crystallizable protein fragments. Limited proteolysis, utilizing two different proteolytic enzymes (alpha-chymotrypsin and trypsin), identified two tobacco activase products. The fragments that were identified appear to represent most of what is considered to be the AAA+ C-terminal all alpha-domain and some of the AAA+ N-terminal alpha beta alpha-domain. Identified fragments were cloned using the pET151/dTOPO. The project then moved towards cloning and recombinant protein expression in E. coli. NtAbeta(248-383) and NtAbeta(253-354) were successfully cloned, expressed, purified, and characterized through various biophysical techniques. A thermofluor assay of NtAbeta(248-383) revealed a melting temperature of about 30°C, indicating lower thermal stability compared with full-length activase at 43°C. Size exclusion chromatography suggested that NtAbeta(248-383) is monomeric. Circular dichroism was used to identify the secondary structure; a plurality of alpha-helices. NtAbeta(248-383) and NtAbeta(253-354) were subjected to crystallization trials.
ContributorsConrad, Alan (Author) / Wachter, Rebekka (Thesis advisor) / Moore, Thomas (Committee member) / Redding, Kevin (Committee member) / Arizona State University (Publisher)
Created2012
Description
The heliobacterial reaction center (HbRC) is widely considered the simplest and most primitive photosynthetic reaction center (RC) still in existence. Despite the simplicity of the HbRC, many aspects of the electron transfer mechanism remain unknown or under debate. Improving our understanding of the structure and function of the HbRC is important in determining its role in the evolution of photosynthetic RCs. In this work, the function and properties of the iron-sulfur cluster FX and quinones of the HbRC were investigated, as these are the characteristic terminal electron acceptors used by Type-I and Type-II RCs, respectively. In Chapter 3, I develop a system to directly detect quinone double reduction activity using reverse-phase high pressure liquid chromatography (RP-HPLC), showing that Photosystem I (PSI) can reduce PQ to PQH2. In Chapter 4, I use RP-HPLC to characterize the HbRC, showing a surprisingly small antenna size and confirming the presence of menaquinone (MQ) in the isolated HbRC. The terminal electron acceptor FX was characterized spectroscopically and electrochemically in Chapter 5. I used three new systems to reduce FX in the HbRC, using EPR to confirm a S=3/2 ground-state for the reduced cluster. The midpoint potential of FX determined through thin film voltammetry was -372 mV, showing the cluster is much less reducing than previously expected. In Chapter 7, I show light-driven reduction of menaquinone in heliobacterial membrane samples using only mild chemical reductants. Finally, I discuss the evolutionary implications of these findings in Chapter 7.
ContributorsCowgill, John (Author) / Redding, Kevin (Thesis advisor) / Jones, Anne (Committee member) / Fromme, Petra (Committee member) / Arizona State University (Publisher)
Created2012
Description
Phosphorus (P), an essential nutrient for growth of all organisms, is often in limited biological supply for herbivore consumers compared to other elements, such as carbon (C). Ecological stoichiometry studies have assessed responses of filter-feeding zooplankton from the genus Daphnia to single and multi-species food resources that are P-limited, finding decreased growth as a result to changes in metabolic processes and feeding behavior. Conversely, recent laboratory studies have shown that P-rich algal food resources also result in decreased growth rates for Daphnia, though the possible mechanisms behind this maladaptive response is understudied. Moreover, no published study tests the existence of the “stoichiometric knife edge” hypothesis for low C:P under field conditions. To address this lack of information, I measured growth rate as well as respiration and ingestion rates for D. magna, D. pulicaria, and D. pulex that were fed natural lake seston experimentally enriched with different levels of PO43-. I found heterogeneous effects of high dietary P across Daphnia species. Growth rate responses for D. magna were strong and indicated a negative effect of high-P, most likely as a result to decreased ingestion rates that were observed. The seston treatments did not elicit significant growth rate responses for D. pulex and D. pulicaria, but significant responses to respiration rates were observed for all species. Consumer body stoichiometry, differences in seston C:P for each experiment, or differential assimilation by producer types may be driving these results. My study suggests that the stoichiometric knife edge documented in laboratory studies under low C:P conditions may not operate to the same degree when natural seston is the food source; diet diversity may be driving complex nuances for consumer performance that were previously overlooked.
ContributorsCurrier, Courtney M (Author) / Currier, James (Thesis advisor) / Harrison, Jon (Committee member) / Neuer, Susanne (Committee member) / Arizona State University (Publisher)
Created2015
Description
Microscopic algae have been investigated extensively by researchers for decades for their ability to bioremediate wastewater and flue gas while producing valuable biomass for use as feed, fuel, fertilizer, nutraceutical, and other specialty products. Reports of the exciting commercial potential of this diverse group of organisms started appearing in the literature as early as the 1940’s. However, nearly 80 years later, relatively few successful commercial microalgae installations exist and algae have not yet reached agricultural commodity status. This dissertation examines three major bottlenecks to commercial microalgae production including lack of an efficient and economical cultivation strategy, poor management of volatile waste nutrients, and costly harvesting and post processing strategies. A chapter is devoted to each of these three areas to gain a better understanding of each bottleneck as well as strategies for overcoming them.
The first chapter demonstrates the capability of two strains of Scenedesmus acutus to grow in ultra-high-density (>10 g L-1 dry weight biomass) cultures in flat panel photobioreactors for year-round production in the desert Southwest with record volumetric biomass productivity. The advantages and efficiency of high-density cultivation are discussed. The second chapter focuses on uptake and utilization of the volatile components of wastewater: ammonia and carbon dioxide. Scenedesmus acutus was cultured on wastewater from both municipal and agricultural origin and was shown to perform significantly better on flue gas as compared to commercial grade CO2 and just as well on waste nutrients as the commonly used BG-11 laboratory culture media, all while producing up to 50% lipids of the dry weight biomass suitable for use in biodiesel. The third chapter evaluates the feasibility of using gravity sedimentation for the harvesting of the difficult-to-separate Scenedesmus acutus green algae biomass followed by microfluidization to disrupt the cells. Lipid-extracted biomass was then studied as a fertilizer for plants and shown to have similar performance to a commercially available 4-6-6 fertilizer. Based on the work from these three chapters, a summary of modifications are suggested to help current and future microalgae companies be more competitive in the marketplace with traditional agricultural commodities.
The first chapter demonstrates the capability of two strains of Scenedesmus acutus to grow in ultra-high-density (>10 g L-1 dry weight biomass) cultures in flat panel photobioreactors for year-round production in the desert Southwest with record volumetric biomass productivity. The advantages and efficiency of high-density cultivation are discussed. The second chapter focuses on uptake and utilization of the volatile components of wastewater: ammonia and carbon dioxide. Scenedesmus acutus was cultured on wastewater from both municipal and agricultural origin and was shown to perform significantly better on flue gas as compared to commercial grade CO2 and just as well on waste nutrients as the commonly used BG-11 laboratory culture media, all while producing up to 50% lipids of the dry weight biomass suitable for use in biodiesel. The third chapter evaluates the feasibility of using gravity sedimentation for the harvesting of the difficult-to-separate Scenedesmus acutus green algae biomass followed by microfluidization to disrupt the cells. Lipid-extracted biomass was then studied as a fertilizer for plants and shown to have similar performance to a commercially available 4-6-6 fertilizer. Based on the work from these three chapters, a summary of modifications are suggested to help current and future microalgae companies be more competitive in the marketplace with traditional agricultural commodities.
ContributorsWray, Joshua (Author) / Dempster, Thomas (Thesis advisor) / Roberson, Robert (Thesis advisor) / Bingham, Scott (Committee member) / Neuer, Susanne (Committee member) / Arizona State University (Publisher)
Created2019
Description
Quagga mussels are an aquatic invasive species capable of causing economic and ecological damage. Despite the quagga mussels’ ability to rapidly spread, two watersheds, the Salt River system and the Verde River system of Arizona, both had no quagga mussel detections for 8 years. The main factor thought to deter quagga mussels was the stratification of the two watersheds during the summer, resulting in high temperatures in the epilimnion and low dissolved oxygen in the hypolimnion. In 2015, Canyon Lake, a reservoir of the Salt River watershed, tested positive for quagga mussel veligers. In this study, I used Landsat 7 and Landsat 8 satellite data to determine if changes in the surface temperature have caused a change to the reservoir allowing quagga mussel contamination. I used a location in the center of the lake with a root mean squared error (RMSE) of 0.80 and a correlation coefficient (R^2) of 0.82, but I did not detect any significant variations in surface temperatures from recent years. I also measured 21 locations on Canyon Lake to determine if the locations in Canyon Lake were able to harbor quagga mussels. I found that summer stratification caused hypolimnion dissolved oxygen levels to drop well below the quagga mussel threshold of 2mg/L. Surface temperatures, however were not high enough throughout the lake to prevent quagga mussels from inhabiting the epilimnion. It is likely that a lack of substrate in the epilimnion have forced any quagga mussel inhabitants in Canyon Lake to specific locations that were not necessarily near the point of quagga veliger detection sampling. The research suggests that while Canyon Lake may have been difficult for quagga mussels to infest, once they become established in the proper locations, where they can survive through the summer, quagga mussels are likely to become more prevalent.
ContributorsLau, Theresa (Author) / Fox, Peter (Thesis advisor) / Neuer, Susanne (Committee member) / Abbaszadegan, Morteza (Committee member) / Arizona State University (Publisher)
Created2018
Description
Time-resolved serial femtosecond crystallography is an emerging method that allows for structural discovery to be performed on biomacromolecules during their dynamic trajectory through a reaction pathway after activation. This is performed by triggering a reaction on an ensemble of molecules in nano- or microcrystals and then using femtosecond X-ray laser pulses produced by an X-ray free electron laser to collect near-instantaneous data on the crystal. A full data set can be collected by merging a sufficient number of these patterns together and multiple data sets can be collected at different points along the reaction pathway by manipulating the delay time between reaction initiation and the probing X-rays. In this way, these ‘snapshot’ structures can be viewed in series to make a molecular movie, allowing for atomic visualization of a molecule in action and, thereby, a structural basis for the mechanism and function of a given biomacromolecule.
This dissertation presents results towards this end, including the successful implementations of the first diffusive mixing chemoactivated reactions and ultrafast dynamics in the femtosecond regime. The primary focus is on photosynthetic membrane proteins and enzymatic drug targets, in pursuit of strategies for sustainable energy and medical advancement by gaining understanding of the structure-function relationships evolved in nature. In particular, photosystem I, photosystem II, the complex of photosystem I and ferredoxin, and 3-deoxy-D-manno-2-octulosonate-8-phosphate synthase are reported on, from purification and isolation, to crystallogenesis, to experimental design and data collection and subsequent interpretation of results and novel insights gained.
This dissertation presents results towards this end, including the successful implementations of the first diffusive mixing chemoactivated reactions and ultrafast dynamics in the femtosecond regime. The primary focus is on photosynthetic membrane proteins and enzymatic drug targets, in pursuit of strategies for sustainable energy and medical advancement by gaining understanding of the structure-function relationships evolved in nature. In particular, photosystem I, photosystem II, the complex of photosystem I and ferredoxin, and 3-deoxy-D-manno-2-octulosonate-8-phosphate synthase are reported on, from purification and isolation, to crystallogenesis, to experimental design and data collection and subsequent interpretation of results and novel insights gained.
ContributorsCoe, Jesse (Author) / Fromme, Petra (Thesis advisor) / Sayres, Scott (Thesis advisor) / Mujica, Vladimiro (Committee member) / Redding, Kevin (Committee member) / Arizona State University (Publisher)
Created2018
Description
Safe, readily available, and reliable sources of water are an essential component of any municipality’s infrastructure. Phoenix, Arizona, a southwestern city, has among the highest per capita water use in the United States, making it essential to carefully manage its reservoirs. Generally, municipal water bodies are monitored through field sampling. However, this approach is limited spatially and temporally in addition to being costly. In this study, the application of remotely sensed reflectance data from Landsat 7’s Enhanced Thematic Mapper Plus (ETM+) and Landsat 8’s Operational Land Imager (OLI) along with data generated through field-sampling is used to gain a better understanding of the seasonal development of algal communities and levels of suspended particulates in the three main terminal reservoirs supplying water to the Phoenix metro area: Bartlett Lake, Lake Pleasant, and Saguaro Lake. Algal abundances, particularly the abundance of filamentous cyanobacteria, increased with warmer temperatures in all three reservoirs and reached the highest comparative abundance in Bartlett Lake. Prymnesiophytes (the class of algae to which the toxin-producing golden algae belong) tended to peak between June and August, with one notable peak occurring in Saguaro Lake in August 2017 during which time a fish-kill was observed. In the cooler months algal abundance was comparatively lower in all three lakes, with a more even distribution of abundance across algae classes. In-situ data from March 2017 to March 2018 were compared with algal communities sampled approximately ten years ago in each reservoir to understand any possible long-term changes. The findings show that the algal communities in the reservoirs are relatively stable, particularly those of the filamentous cyanobacteria, chlorophytes, and prymnesiophytes with some notable exceptions, such as the abundance of diatoms, which increased in Bartlett Lake and Lake Pleasant. When in-situ data were compared with Landsat-derived reflectance data, two-band combinations were found to be the best-estimators of chlorophyll-a concentration (as a proxy for algal biomass) and total suspended sediment concentration. The ratio of the reflectance value of the red band and the blue band produced reasonable estimates for the in-situ parameters in Bartlett Lake. The ratio of the reflectance value of the green band and the blue band produced reasonable estimates for the in-situ parameters in Saguaro Lake. However, even the best performing two-band algorithm did not produce any significant correlation between reflectance and in-situ data in Lake Pleasant. Overall, remotely-sensed observations can significantly improve our understanding of the water quality as measured by algae abundance and particulate loading in Arizona Reservoirs, especially when applied over long timescales.
ContributorsRussell, Jazmine Barkley (Author) / Neuer, Susanne (Thesis advisor) / Fox, Peter (Committee member) / Myint, Soe (Committee member) / Arizona State University (Publisher)
Created2018
Description
Over the last century, X-ray crystallography has been established as the most successful technique for unravelling the structure-function relationship in molecules. For integral membrane proteins, growing well-ordered large crystals is a challenge and hence, there is room for improving current methods of macromolecular crystallography and for exploring complimentary techniques. Since protein function is deeply associated with its structural dynamics, static position of atoms in a macromolecule are insufficient to unlock the mechanism.
The availability of X-ray free electron lasers presents an opportunity to study micron-sized crystals that could be triggered (using light, small molecules or physical conditions) to capture macromolecules in action. This method of ‘Time-resolved serial crystallography’ answers key biological questions by capturing snapshots of conformational changes associated with multi-step reactions. This dissertation describes approaches for studying structures of large membrane protein complexes. Both macro and micro-seeding techniques have been implemented for improving crystal quality and obtaining high-resolution structures. Well-diffracting 15-20 micron crystals of active Photosystem II were used to perform time-resolved studies with fixed-target Roadrunner sample delivery system. By employing continuous diffraction obtained up to 2 A, significant progress can be made towards understanding the process of water oxidation.
Structure of Photosystem I was solved to 2.3 A by X-ray crystallography and to medium resolution of 4.8 A using Cryogenic electron microscopy. Using complimentary techniques to study macromolecules provides an insight into differences among methods in structural biology. This helps in overcoming limitations of one specific technique and contributes in greater knowledge of the molecule under study.
The availability of X-ray free electron lasers presents an opportunity to study micron-sized crystals that could be triggered (using light, small molecules or physical conditions) to capture macromolecules in action. This method of ‘Time-resolved serial crystallography’ answers key biological questions by capturing snapshots of conformational changes associated with multi-step reactions. This dissertation describes approaches for studying structures of large membrane protein complexes. Both macro and micro-seeding techniques have been implemented for improving crystal quality and obtaining high-resolution structures. Well-diffracting 15-20 micron crystals of active Photosystem II were used to perform time-resolved studies with fixed-target Roadrunner sample delivery system. By employing continuous diffraction obtained up to 2 A, significant progress can be made towards understanding the process of water oxidation.
Structure of Photosystem I was solved to 2.3 A by X-ray crystallography and to medium resolution of 4.8 A using Cryogenic electron microscopy. Using complimentary techniques to study macromolecules provides an insight into differences among methods in structural biology. This helps in overcoming limitations of one specific technique and contributes in greater knowledge of the molecule under study.
ContributorsRoy Chowdhury, Shatabdi (Author) / Fromme, Petra (Thesis advisor) / Ros, Alexandra (Committee member) / Redding, Kevin (Committee member) / Arizona State University (Publisher)
Created2018