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A COOPERATIVE RELATIONSHIP BETWEEN CYANOBACTERIA AND E. COLI FOR THE PRODUCTION OF BIOFUELS Marguerite Christian Program: NSF EPSCoR Mentor: Christie Howard Department: Biochemistry and Molecular Biology University of Nevada, Reno E. coli can be engineered to produce resources such as biodiesel and bioethanol, however, the growth media costs account for 30-40% of the total cost, making it less efficient than other methods. The goal of this research is to produce biofuels at a drastically lowered cost. One way science can reduce the cost is to genetically engineer photosynthetic bacteria like cyanobacteria to produce biofuels from CO2. However, in a number of cases cyanobacteria has been shown to be less productive in making biofuels than E. coli, presumably due to lack of transport proteins or internal toxicity issues. As an alternative approach, the University of Nevada, Reno undergraduate iGEM (International Genetically Machine) team is using cyanobacteria to address the problem of high media cost by transforming the cyanobacteria Synechocystis PCC 6803 to produce high levels of glucose to be released into the media and feed our genetically engineered E. coli. By co-culturing these two organisms, direct carbon transfer from CO2 to ethanol and C12 fatty acids can be achieved. To achieve this goal, the cyanobacteria needs to produce glucose in high concentrations as the carbon source. To raise glucose concentrations, glycogen production via the agp gene will be knocked out, leading to a higher sucrose production. The bacteria will then be engineered to overexpress the gene encoding invertase, which converts sucrose to glucose and fructose. To increase output of these products, overexpression of glf, the gene encoding a transporter of the glucose and fructose, will also be engineered into cyanobacteria. My research will focus on designing the construct to knock out the agp gene. The ultimate goal of the iGEM team is to use genetically engineered cyanobacteria to sustain E. coli for the purpose of producing biofuel products at a significantly lowered cost. CATERPILLAR-PARASITOID INTERACTION DIVERSITY ALONG AN ALTITUDINAL GRADIENT WITHIN THE SIERRA NEVADA AND EASTERN SLOPE OF THE ANDES MOUNTAINS Kirsha Frederickson Program: GURA Mentor: Lee Dyer Major: Biology University of Nevada, Reno It is a major goal of ecological studies to quantify diversity of trophic interactions and to understand how these interactions are related to major global issues of climate change, biodiversity loss, and ecosystem services. As a follow-up to a similar study conducted by the PI (K. Fredrickson) and other students, we will continue to test the hypothesis that altitudinal gradients in interaction diversity are consistent across large latitudinal gradients. Standardized methods of plot sampling will be used to estimate the caterpillar-parasitoid abundances and diversity in the Sierra Nevada, along altitudinal gradients. A total of 100 plots were deployed in summer 2010, and another 100 plots will be deployed this summer. The plant genus, Alnus, is the focal plant in the study; all caterpillars reared will be collected off of this plant. 9
NUTRIENT CONCENTRATIONS IN O-HORIZON LEACHATE Rachel Funk Program: NSF REU Mentor: Wally Miller Major: Environmental Sciences Eureka College O Horizon leachate from Sierran forests can be an important contributor of inorganic nutrients in soil, overland and/or litter interflow, and water quality. The nutrient content and leachability from litter (Oi) and duff (Oe) materials is therefore important, as is the duration of extraction (leaching) event. The effect of material and extraction time on leachate nutrient concentration and amount was investigated for three O horizon materials (Oi, Oe, and Oi+Oe) from seven sites across the Sierras. The extractions were performed over 4 extraction intervals (10 min, 30 min, 60 min and 120 min) with 3 replications for a 3x4x3 study design. The results will show how nutrients are discharged from the o-horizon, including which layers serve as sources of nutrients and how concentrations are affected by extraction time. HYDROTHERMAL CARBONIZATION OF WOODY BIOMASS Brian Gallaspy Program: NSF EPSCoR Mentors: Kent Hoekman, Wei Yan Department: Chemical Engineering University of Nevada, Reno Hydrothermal carbonization (HTC) is a process by which lignocellulosic (woody) biomass is converted into a solid fuel having a higher energy density than the starting feedstock. Broadly, lignocellulosic biomass is composed of three components: cellulose, hemi-cellulose and lignin. The HTC reaction is carried out by placing the feedstock in hot, pressurized water. Under these conditions, the hemicellulose readily breaks down into simple sugars that are dissolved into the water. The recovered solid product has a higher C/O ratio, and thereby a higher energy density. Under optimal HTC conditions, the biochar product has similar properties to a low-rank coal. One implication of this is that the HTC process could be used to produce a renewable solid fuel that could replace or supplement coal in existing power plants with little to no modification of the equipment used. This project aims to investigate how the reaction conditions affect the product. The reaction is carried out in a specially designed twochamber reactor which allows the water to be heated to the desired reaction conditions in the lower chamber, while the biomass waits in the upper chamber. The purpose of the two-chamber reactor is to allow for fine control of temperature conditions. Once the desired temperature is reached, a valve is opened and the biomass falls into the water and the reaction begins. The conditions being varied are: temperature (200 °C, 230 °C and 260 °C), reaction hold time (1, 3 and 5 minutes), and particle size (14-28 mesh, 10-14 mesh and 8-10 mesh). The product is then pelletized, and characterized using calorimetry along with a number of other physical tests including strength, attrition and hydrophobicity. 10
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