Digital Commons Network^™

Chloroplast Division Protein Arc3 Regulates Chloroplast Ftsz-Ring Assembly And Positioning In Arabidopsis Through Interaction With Ftsz2, Min Zhang, Aaron J. Schmitz, Deena K. Kadirjan-Kalbach, Allan D. Terbush, Katherine W. Osteryoung

Department of Agronomy and Horticulture: Faculty Publications

Chloroplast division is initiated by assembly of a mid-chloroplast FtsZ (Z) ring comprising two cytoskeletal proteins, FtsZ1 and FtsZ2. The division-site regulators ACCUMULATION AND REPLICATION OF CHLOROPLASTS3 (ARC3), MinD1, and MinE1 restrict division to the mid-plastid, but their roles are poorly understood. Using genetic analyses in Arabidopsis thaliana, we show that ARC3 mediates division-site placement by inhibiting Z-ring assembly, and MinD1 and MinE1 function through ARC3. ftsZ1 null mutants exhibited some mid-plastid FtsZ2 rings and constrictions, whereas neither constrictions nor FtsZ1 rings were observed in mutants lacking FtsZ2, suggesting FtsZ2 is the primary determinant of Z-ring assembly in vivo. arc3 …

Resource Allocation For Maximizing Prediction Accuracy And Genetic Gain Of Genomic Selection In Plant Breeding: A Simulation Experiment, Aaron Lorenz

Department of Agronomy and Horticulture: Faculty Publications

Allocating resources between population size and replication affects both genetic gain through phenotypic selection and quantitative trait loci detection power and effect estimation accuracy for marker-assisted selection (MAS). It is well known that because alleles are replicated across individuals in quantitative trait loci mapping and MAS, more resources should be allocated to increasing population size compared with phenotypic selection. Genomic selection is a form of MAS using all marker information simultaneously to predict individual genetic values for complex traits and has widely been found superior to MAS. No studies have explicitly investigated how resource allocation decisions affect success of genomic …

Evaluation Of Public Sweet Sorghum A-Lines For Use In Hybrid Production, Jeffrey F. Pedersen, Scott E. Sattler, William F. Anderson

Department of Agronomy and Horticulture: Faculty Publications

A fundamental need for commercialization of sweet sorghum [Sorghum bicolor (L.) Moench] as a bioenergy crop is an adequate seed supply, which will require development of hybrid varieties using dwarf seed-parent lines. A set of six public sweet sorghum A-lines (Dwarf Kansas Sourless, KS9, N36, N38, N39, and N4692) were crossed with a set of six public sweet sorghum cultivars (Brawley, Kansas Collier, Dale, Sugar Drip, Waconia, and Wray). Grain, fiber, and sugar yields were determined, and conversion formulas were applied to estimate ethanol yields. Hybrids were grown in fields at Ithaca, NE, USA, in 1983– 1984 fertilized with …

Yield Gap Analysis—Rationale, Methods And Applications—Introduction To The Special Issue, M.K. Van Ittersum, Kenneth Cassman

Department of Agronomy and Horticulture: Faculty Publications

Yield gap analysis is an increasingly popular concept. It is a powerful method to reveal and understand the biophysical opportunities to meet the projected increase in demand for agricultural products towards 2050, and to support decision making on research, policies, development and investment that is needed. This Special Issue presents the state-of-the-art about concepts, methods and applications of yield gap analysis. The methodological papers emphasize the need for agronomically sound and relevant analyses, from local to global scales. The fourteen papers provide examples of applications to different crops, climate zones and production conditions, at various spatial extents and with different …

Agricultural Innovation To Protect The Environment, Jeffrey Sayer, Kenneth Cassman

Department of Agronomy and Horticulture: Faculty Publications

In a world of 9.5 billion people, global demand for food, fiber, and biofuels has to be met with minimal possible increases in land, water, fossil fuels, and the minerals used to produce fertilizers (1–4). The problem is debated at three levels: first, that agriculture will not be able to produce enough because it will come up against both biophysical and environmental limits that restrict yields (3, 5, 6); second, that the need to expand and intensify agriculture will destroy the broader environmental values of forests, wetlands, marine systems, and their associated biodiversity (7–9); and third, that there are institutional …

New Technologies Reduce Greenhouse Gas Emissions From Nitrogenous Fertilizer In China, Wei-Feng Feng, Zheng-Xia Dou, Pan He, Xiao-Tang Ju, David Powlson, Dave Chadwick, David Norse, Yue-Lai Lu, Ying Zhang, Liang Wu, Xin-Ping Chen, Kenneth Cassman, Fu-Suo Zhang

Department of Agronomy and Horticulture: Faculty Publications

Synthetic nitrogen (N) fertilizer has played a key role in enhancing food production and keeping half of the world’s population adequately fed. However, decades of N fertilizer overuse in many parts of the world have contributed to soil, water, and air pollution; reducing excessive N losses and emissions is a central environmental challenge in the 21st century. China’s participation is essential to global efforts in reducing N-related greenhouse gas (GHG) emissions because China is the largest producer and consumer of fertilizer N. To evaluate the impact of China’s use of N fertilizer, we quantify the carbon footprint of China’s N …

Testing The Fuel Efficiency Of Tractors With Continuously Variable And Standard Geared Transmissions, Christopher N. Howard, Michael F. Kocher, Roger M. Hoy, Erin E. Blankenship

Department of Agricultural and Biological Systems Engineering: Faculty Publications

A John Deere 8295R IVT tractor with a continuously variable transmission (CVT) and a John Deere 8295R PowerShift (PST) tractor with a standard geared transmission (GT) were tested for fuel consumption at three different travel speeds with six different load levels applied per speed. The JD 8295R PST tractor was tested both at full throttle (FT) and shifted up two gears and throttled back (SUTB) to achieve the same travel speed as at full throttle. For each travel speed with each transmission mode, fuel consumption was determined to be linearly related to drawbar power. Linear regression analyses were performed, and …

Co2 Uptake And Ecophysiological Parameters Of The Grain Crops Of Midcontinent North America: Estimates From Flux Tower Measurements, Tagir G. Gilmanov, Bruce K. Wylie, Larry L. Tieszen, Tilden P. Meyers, Vern S. Baron, Carl J. Bernacchi, David P. Billesbach, George G. Burba, Marc L. Fischer, Aaron J. Glenn, Niall P. Hanan, Jerry L. Hatfield, Mark W. Heuer, Steven E. Hollinger, Daniel M. Howard, Roser Matamala, John H. Prueger, Mario Tenuta, David G. Young

Department of Agricultural and Biological Systems Engineering: Faculty Publications

We analyzed net CO₂ exchange data from 13 flux tower sites with 27 site-years of measurements over maize and wheat fields across midcontinent North America. A numerically robust “light-soil temperature- VPD”-based method was used to partition the data into photosynthetic assimilation and ecosystem respiration components. Year-round ecosystem-scale eco-physiological parameters of apparent quantum yield, photosynthetic capacity, convexity of the light response, respiration rate parameters, ecological light-use efficiency, and the curvature of the VPD-response of photosynthesis for maize and wheat crops were numerically identified and interpolated/extrapolated. This allowed us to gap-fill CO₂ exchange components and calculate annual totals and budgets. …

Seepage-Induced Streambank Erosion And Instability: In Situ Constant-Head Experiments, T. L. Midgley, G. A. Fox, G. V. Wilson, D. M. Heeren, E. J. Langendoen, A. Simon

Department of Agricultural and Biological Systems Engineering: Faculty Publications

The effects of seepage on streambank erosion and failure are less understood compared to fluvial processes, especially the linkage between surface water and groundwater mechanisms. Field data are needed to validate laboratory seepage erosion and instability conclusions and to understand how engineering tools and methods may be applied to field conditions. An innovative trench injection system was utilized to provide a constant head on a near-streambank groundwater system when filled with stream water. This research was performed on a streambank of Dry Creek, a deeply incised stream with near-vertical banks located in Mississippi. Experiments included installing a trench (2.8 m …

In Situ Soil Pipeflow Experiments On Contrasting Streambank Soils, T. L. Midgley, G. A. Fox, G. V. Wilson, R. M. Felice, D. M. Heeren

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Soil piping has been attributed as a potential mechanism of instability of embankments and streambanks. Limited field work has been conducted on quantifying and modeling pipeflow and internal erosion processes in the field with either natural or artificially created soil pipes. This research utilized an innovative constant-head trench system to conduct constant-head soil pipe experiments in two contrasting streambanks: Dry Creek in northern Mississippi and Cow Creek in northern Oklahoma. Experiments included open pipes, in which the soil pipe was directly connected to the constant- head trench and open at the streambank face, and clogged pipes, which involved plugging the …

Use Of A Surrogate To Evaluate The Impact Of Tillage On The Transport Of Steroid Hormones From Manure-Amended Agricultural Fields, Sagor Biswas, William L. Kranz, Charles A. Shapiro, Mitiku Mamo, Shannon L. Bartelt-Hunt, Daniel D. Snow, David P. Shelton, David D. Tarkalson, Terry L. Mader, Simon J. Van Donk, Tian C. Zhang

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Beef feedlot manure distributed to row crop production areas is a potential surface water contaminant source of the steroid hormones commonly used in beef cattle production. This article reports on research conducted at the University of Nebraska Haskell Agricultural Laboratory near Concord, Nebraska, in July 2009. Manure, collected from beef feedlot pens, was stockpiled for ten months prior to application to a row crop field. Previous research identified that the detection frequency of steroid hormones in beef manure varies greatly. Thus, a surrogate (17α-ethynylestradiol, EE2) was applied at a rate of 75 g ha^-1 to ensure detectable concentrations in …

Runoff Nutrient Transport As Affected By Land Application Method, Swine Growth Stage, And Runoff Rate, J. E. Gilley, Shannon L. Bartelt-Hunt, S. J. Lamb, X. Li, D. B. Marx, D. D. Snow, D. B. Parker, B. L. Woodbury

Department of Agricultural and Biological Systems Engineering: Faculty Publications

This study was conducted to measure the effects of slurry application method, swine growth stage, and flow rate on runoff nutrient transport. Swine slurry was obtained from production units containing grower pigs, finisher pigs, or sows and gilts. The swine slurry was applied using broadcast, disk, or injection methods at a rate required to meet annual nitrogen requirements for corn. Three 30 min simulated rainfall events, separated by 24 h intervals, were applied to the experimental plots at an intensity of 70 mm h^-1. Inflow was applied at the top of each plot in four successive increments after …

Hourly And Daily Single And Basal Evapotranspiration Crop Coefficients As A Function Of Growing Degree Days, Days After Emergence, Leaf Area Index, Fractional Green Canopy Cover, And Plant Phenology For Soybean, Suat Irmak, Lameck O. Odhiambo, James Specht, Koffi Djaman

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Hourly evapotranspiration (ET) crop coefficients (K_c) are needed to optimize the effectiveness and efficiency of high-frequency micro- and sprinkler irrigation practices involving the application of water multiple times a day. However, not much is known about the daily and seasonal patterns and magnitudes in hourly K_c values for soybean. In addition, locally developed K_c values are necessary for more robust within-season irrigation management, crop ET estimation, and water balance analyses. Hourly and daily Kc functions were developed for soybean in south-central Nebraska through extensive field research. Actual crop evapotranspiration (ET_a) was measured using a …

Maize Evapotranspiration, Yield Production Functions, Biomass, Grain Yield, Harvest Index, And Yield Response Factors Under Full And Limited Irrigation, Koffi Djaman, Suat Irmak, William R. Rathje, Derrel L. Martin, Dean E. Eisenhauer

Department of Agricultural and Biological Systems Engineering: Faculty Publications

South-central Nebraska is one of the most extensively irrigated areas in the U.S., with over 65,000 active irrigation wells, and maize is the major agronomical crop produced. Maize production in this region requires supplementary irrigation for maximum productivity. Effective on-farm implementation of full and limited irrigation practices for potential improvements of crop productivity requires knowledge of locally developed crop yield response to water functions. In this study, the effects of full and limited irrigation practices on maize (Zea mays L.) plant height, leaf area index (LAI), grain yield and biomass production, actual crop evapotranspiration (ET_a), yield production functions, …

Use Of A Surrogate To Evaluate The Impact Of Tillage On The Transport Of Steroid Hormones From Manure-Amended Agricultural Fields, Sagor Biswas, William Kranz, Charles A. Shapiro, Mitiku Mamo, Shannon L. Bartelt-Hunt, Daniel D. Snow, David P. Shelton, David D. Tarkalson, Terry L. Mader, Simon J. Van Donk, Tian C. Zhang

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Beef feedlot manure distributed to row crop production areas is a potential surface water contaminant source of the steroid hormones commonly used in beef cattle production. This article reports on research conducted at the University of Nebraska Haskell Agricultural Laboratory near Concord, Nebraska, in July 2009. Manure, collected from beef feedlot pens, was stockpiled for ten months prior to application to a row crop field. Previous research identified that the detection frequency of steroid hormones in beef manure varies greatly. Thus, a surrogate (17α-ethynylestradiol, EE2) was applied at a rate of 75 g ha^-1 to ensure detectable concentrations in …

Transport Of Steroid Hormones In The Vadose Zone After Land Application Of Beef Cattle Manure, Simon Van Donk, Sagor Biswas, William L. Kranz, Daniel D. Snow, Shannon L. Bartelt-Hunt, Terry L. Mader, Charles A. Shapiro, David P. Shelton, David D. Tarkalson, Tian .. Zhang, Steve Ensley

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Few studies have been conducted to investigate the potential contamination of groundwater from manureborne steroid hormones. The objective of this study was to monitor leaching of steroid hormones at the bottom of 2.4 m deep percolation lysimeters installed in plots treated with beef cattle manure. Soil samples were also collected from surrounding field plots. Treatments consisted of application of manure from two treatment methods (stockpiling and composting) or no manure application. Laboratory analyses of manure, soil, and leachate samples used liquid chromatography and tandem mass spectrometry to identify 17 steroid hormones and metabolites. Progesterone, estrone, β-zearalenol, and 4-androstenedione were detected …

Development And Evaluation Of Ordinary Least Squares Regression Models For Predicting Irrigated And Rainfed Maize And Soybean Yields, Vivek Sharma, Daran R. Rudnick, Suat Irmak

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Understanding the relationships between climatic variables and soil physical and chemical properties with crop yields on large scales is critical for evaluating crop productivity to make better assessments of local and regional food security, policy, land and water resource allocation, and management decisions. In this study, ordinary least squares(OLS) regression models were developed to predict irrigated and rainfed maize and soybean yields at the county level as a

function of explanatory variables [precipitation (P), actual crop evapotranspiration (ETa), organic matter content (OMC), cation exchange capacity (CEC), clay content (CC), and available soil water capacity (ASW)] of the dominant soil type …

Water Productivity Of Corn And Dry Bean Rotation On Very Fine Sandy Loam Soil In Western Nebraska, William E. Spurgeon, C. Dean Yonts

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Subsurface drip irrigation (SDI) was used to irrigate corn and dry beans in a crop rotation study for four years from 2005 to 2008 to evaluate the accumulative effect of irrigation amount on yield and irrigation water use efficiency (I_WUE). This study was conducted on a very fine sandy loam soil using a short season corn hybrid typical for the area. Four irrigation treatments based on providing a fraction of the estimated crop evapotranspiration (ET_c) were evaluated. The treatments were designed to replace a certain percentage of ET_c and were denoted as I₁₂₅, …

Surface- And Hydrogel-Mediated Delivery Of Nucleic Acid Nanoparticles, Angela K. Pannier, Tatiana Segura

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Gene expression within a cell population can be directly altered through gene delivery approaches. Traditionally for nonviral delivery, plasmids or siRNA molecules, encoding or targeting the gene of interest, are packaged within nanoparticles. These nanoparticles are then delivered to the media surrounding cells seeded onto tissue culture plastic; this technique is termed bolus delivery. Although bolus delivery is widely utilized to screen for efficient delivery vehicles and to study gene function in vitro, this delivery strategy may not result in efficient gene transfer for all cell types or may not identify those delivery vehicles that will be efficient in vivo. …

Implementation Of A ”Rapid Design Challenge” In A Cross-Disciplinary Senior Capstone Course And Evaluation Of Device Performance, Abby M. Kelly, Austin Lammers, David D. Jones, Richard Stowell, Roger Hoy, Evan Curtis, Angela K. Pannier

Department of Agricultural and Biological Systems Engineering: Faculty Publications

The senior capstone experience within the Department of Biological Systems Engineering at the University of Nebraska-Lincoln is a two-semester, two-course sequence intended to give senior students realistic design experience, working with real projects, real clients, faculty consultants, and teammates to produce a deliverable that meets the client’s needs. Students within this course sequence come from two different degree programs (agricultural engineering and biological systems engineering) and within each degree program from a variety of “emphasis areas”, e.g. biomedical engineering, environmental engineering, machine design. While the goal of this course is for students to experience a “real” design project, we felt …

Acetyl-Coa Carboxylases Are Checkpoints In Adipocyte Differentiation, Elizabeth Lyn Cordonier, Daniel Camara Teixeira, Zhonghi Han, Angela K. Pannier, Janos Zempleni

Department of Agricultural and Biological Systems Engineering: Faculty Publications

No abstract provided.

Network Analysis Of Endogenous Gene Expression Profiles After Polyethyleneimine-Mediated Dna Delivery, Timothy M. Martin, Sarah A. Plautz, Angela K. Pannier

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Background—DNA delivery systems, which transport exogenous DNA to cells, have applications that include gene therapy, tissue engineering and medical devices. Although the cationic nonviral DNA carrier polyethyleneimine (PEI) has been widely studied, the molecular factors and pathways underlying PEI-mediated DNA transfer remain largely unknown, preventing the design of more efficient delivery systems.

Methods—HEK 293 T cells were treated with polyplexes formed with PEI and pEGFPLuc encoding for green fluorescent protein (GFP). Transfected cells expressing GFP were flowseparated from treated, untransfected cells. Gene expression profiles were obtained using Affymetrix HG-U133 2.0 microarrays and differentially expressed genes were identified using R/ Bioconductor. …

Avhrr-Ndvi-Based Crop Coefficients For Analyzing Long-Term Trends In Evapotranspiration In Relation To Changing Climate In The U.S. High Plains, Denis Mutiibwa, Suat Irmak

Department of Agricultural and Biological Systems Engineering: Faculty Publications

Studies in regions of extensive irrigation practices have revealed a significant influence of evaporative cooling on regional temperatures as a result of surface energy redistribution during evaporation. In the U.S. High Plains, maximum temperatures during the last quarter of the 20th century have been decreasing. We investigated the trends in evapotranspiration (ET or latent heat) fluxes originating from increasing irrigation practices in the High Plains region from 1981 to 2008. We estimated actual ET (ETc) over the entire High Plains from the spatial crop coefficients (Kc) and spatial reference (potential) ET (ETref). We proposed …

Changing Room Cues Reduces The Effects Of Proactive Interference In Clark’S Nutcrackers, Nucifraga Columbiana, Jody L. Lewis, Alan C. Kamil, Kate E. Webbink

Avian Cognition Papers

To determine what factors are important for minimizing interference effects in spatial memory, Clark’s Nutcrackers, Nucifraga columbiana were tested for their spatial memory for two serial lists of locations per day. In this experiment two unique landmark sets were either different between List 1 and List 2 or the same. We found that Nutcrackers were most susceptible to interference when the landmark sets were the same. This study suggests that repeatedly testing animal memory in the same room, with the same cues, can hamper recall due to interference.

Differences In Relative Hippocampus Volume And Number Of Hippocampus Neurons Among Five Corvid Species, Kristy L. Gould, Karl E. Gilbertson, Andrew J. Hrvol, Joseph C. Nelson, Abigail L. Seyfer, Rose M. Brantner, Alan C. Kamil

Avian Cognition Papers

The relative size of the avian hippocampus (Hp) has been shown to be related to spatial memory and food storing in two avian families, the parids and corvids. Basil et al. [Brain Behav Evol 1996;47: 156-164] examined North American food-storing birds in the corvid family and found that Clark’s nutcrackers had a larger relative Hp than pinyon jays and Western scrub jays. These results correlated with the nutcracker’s better performance on most spatial memory tasks and their strong reliance on stored food in the wild. However, Pravosudov and de Kort [Brain Behav Evol 67 (2006), 1-9] raised questions …

Effect Of Salts On The Co-Fermentation Of Glucose And Xylose By A Genetically Engineered Strain Of Saccharomyces Cerevisiae, Elizabeth Casey, Nathan S. Mosier, Jiri Adamec, Zachary Stockdale, Nancy Ho, Miroslav Sedlak

Department of Biochemistry: Faculty Publications

Background: A challenge currently facing the cellulosic biofuel industry is the efficient fermentation of both C5 and C6 sugars in the presence of inhibitors. To overcome this challenge, microorganisms that are capable of mixed-sugar fermentation need to be further developed for increased inhibitor tolerance. However, this requires an understanding of the physiological impact of inhibitors on the microorganism. This paper investigates the effect of salts on Saccharomyces cerevisiae 424A(LNH-ST), a yeast strain capable of effectively co-fermenting glucose and xylose.

Results: In this study, we show that salts can be significant inhibitors of S. cerevisiae. All 6 pairs of anions …

Functional Convergence Of Structurally Distinct Thioesterases From Cyanobacteria And Plants Involved In Phylloquinone Biosynthesis, Fabienne Furt, William J. Allen, Joshua R. Widhalm, Peter Madzelan, Robert C. Rizzo, Gilles J. Basset, Mark A. Wilson

Department of Biochemistry: Faculty Publications

The synthesis of phylloquinone (vitamin K₁) in photosynthetic organisms requires a thioesterase that hydrolyzes 1,4-dihydroxy- 2-naphthoyl-CoA (DHNA-CoA) to release 1,4- dihydroxy-2-naphthoate (DHNA). Cyanobacteria and plants contain distantly related hotdog-fold thioesterases that catalyze this reaction, although the structural basis of these convergent enzymatic activities is unknown. To investigate this, the crystal structures of hotdog-fold DHNA-CoA thioesterases from the cyanobacterium Synechocystis (Slr0204) and the flowering plant Arabidopsis thaliana (AtDHNAT1) were determined. These enzymes form distinct homotetramers and use different active sites to catalyze hydrolysis of DHNACoA, similar to the 4-hydroxybenzoyl-CoA (4-HBA-CoA) thioesterases from Pseudomonas and Arthrobacter. Like the 4-HBA-CoA …

Food Webs And Food Sovereignty: Research Agenda For Sustainability, Charles A. Francis, Michelle Miller, Molly Anderson, Nancy Creamer, Michelle Wander, Jacob Park, Thomas Green, Brent Mccown

Department of Agronomy and Horticulture: Faculty Publications

Future food production will be constrained by the scarcity of fossil fuel and fresh water as well as increasing intensity and unpredictability of weather events and climate changes. The assurance of food security and equity for many consumers is complicated by concentration of ownership of land and other production resources, as well as a global corporate food systems model that is driven by profit at the expense of people and the environment. To assess potential alternatives to the contemporary global food chain, well focused research is needed on local food production and food webs where small- and midscale family farms …

Rapid Detection And Quantification Of Triacylglycerol By Hplc–Elsd In Chlamydomonas Reinhardtii And Chlorella Strains, Naoko Kobayashi, Eric A. Noel, Austin Barnes, Julian Rosenberg, Concetta C. Dirusso, Paul N. Black, George A. Oyler

Department of Biochemistry: Faculty Publications

Triacylglycerol (TAG) analysis and quantification are commonly performed by first obtaining a purified TAG fraction from a total neutral lipid extract using thinlayer chromatography (TLC), and then analyzing the fatty

acid composition of the purified TAG fraction by gas chromatography (GC). This process is time-consuming, labor

intensive and is not suitable for analysis of small sample sizes or large numbers. A rapid and efficient method for monitoring oil accumulation in algae using high performance liquid chromatography for separation of all lipid classes combined with detection by evaporative light scattering (HPLC–ELSD) was developed and compared to the conventional TLC/GC method. TAG …

Probing Arabidopsis Chloroplast Diacylglycerol Pools By Selectively Targeting Bacterial Diacylglycerol Kinase To Suborganellar Membranes, Bagyalakshmi Muthan, Rebecca Roston, John E. Froehlich, Christoph Benning

Department of Biochemistry: Faculty Publications

Diacylglycerol (DAG) is an intermediate in metabolism of both triacylglycerols and membrane lipids. Probing the steady-state pools of DAG and understanding how they contribute to the synthesis of different lipids is important when designing plants with altered lipid metabolism. However, traditional methods of assaying DAG pools are difficult, because its abundance is low and because fractionation of subcellular membranes affects DAG pools. To manipulate and probe DAG pools in an in vivo context, we generated multiple stable transgenic lines of Arabidopsis (Arabidopsis thaliana) that target an Escherichia coli DAGkinase (DAGK) to each leaflet of each chloroplast envelope membrane. …