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published: 2025-10-14
 
Rhodosporidium toruloides is an oleaginous yeast capable of producing a variety of biofuels and bioproducts from diverse carbon sources. Despite numerous studies showing its promise as a platform microorganism, little is known about its metabolism and physiology. In this work, we investigated the central carbon metabolism in R. toruloides IFO0880 using transcriptomics and metabolomics during growth on glucose, xylose, acetate, or soybean oil. These substrates were chosen because they can be derived from plants. Significant changes in gene expression and metabolite concentrations were observed during growth on these four substrates. We mapped these changes onto the governing metabolic pathways to better understand how R. toruloides reprograms its metabolism to enable growth on these substrates. One notable finding concerns xylose metabolism, where poor expression of xylulokinase induces a bypass leading to arabitol production. Collectively, these results further our understanding of central carbon metabolism in R. toruloides during growth on different substrates. They may also help guide the metabolic engineering and development of better models of metabolism for R. toruloides.
keywords: Conversion;Metabolomics;Transcriptomics
published: 2025-10-14
 
Efforts to engineer high-productivity crops to accumulate oils in their vegetative tissue present the possibility of expanding biodiesel production. However, processing the new crops for lipid recovery and ethanol production from cell wall saccharides is challenging and expensive. In a previous study using corn germ meal as a model substrate, we reported that liquid hot water (LHW) pretreatment enriched the lipid concentration by 2.2 to 4.2 fold. This study investigated combining oil recovery with ethanol production by extracting oil following LHW and simultaneous saccharification and co-fermentation (SSCF) of the biomass. Corn germ meal was again used to model the oil-bearing energy crops. Pretreated germ meal hydrolysate or solids (160 °C and 180 °C for 10 minutes) were fermented, and lipids were extracted from both the spent fermentation whole broth and fermentation solids, which were recovered by centrifugation and convective drying. Lipid contents in spent fermentation solids increased 3.7 to 5.7 fold compared to the beginning germ meal. The highest lipid yield achieved after fermentation was 36.0 mg lipid g−1 raw biomass; the maximum relative amount of triacylglycerol (TAG) was 50.9% of extracted oil. Although the fermentation step increased the lipid concentration of the recovered solids, it did not improve the lipid yields of pretreated biomass and detrimentally affected oil compositions by increasing the relative concentrations of free fatty acids.
keywords: Conversion;Hydrolysate;Lipidomics
published: 2024-10-28
 
This dataset contains MALDI imaging and fluorescence imaging data of 5xFAD mice and control animals. 1+2) Animal_1_5xFAD_s1 and s2 : A MATLAB file of 50 micron spatial resolution imaging of whole brain slice from a 5xFAD animal. 3) Slide28_Animal1_stitch_channels__Thioflavin S : A PNG file of the corresponding Thioflavin S- stained fluorescence image obtained post-MSI from the same section. 4) Slide28_Animal1_stitch_merged : A PNG file of the corresponding merged imaged including brightfield, Thioflavin S (GFP channel) and Hoechst staining (DAPI channel) used for image registration 5) mz_bins_use_neg.mat : A MATLAB array of the m/z channels all MSI images (whole brain slice, 50 micron spatial resolution) were binned to in order to enable comparison 6) Animal3_S18_HR.mat : A MATLAB array of high-spatial-resolution (5 micron) imaging of a 5xFAD mouse hippocampus and cortex. Due to the large dataset, 22 m/z channels are included. 7) Animal5_S18_HR.mat : A MATLAB array of high-spatial-resolution (5 micron) imaging of a wildtype mouse hippocampus and cortex 8) mz_features_22.mat : A MATLAB array of the 22 m/z channels included in the high spatial resolution imaging data
keywords: amyloid beta; 5xfad, lipids; maldi;
published: 2025-10-13
 
Miscanthus × giganteus (Miscanthus) is a warm-season perennial grass grown for bioenergy feedstock production. Nitrogen (N) fertilizer management is crucial for the sustainability of Miscanthus production. In our two-year study (2018 and 2019), we investigated the role of vegetation indices (VIs) in evaluating N fertilization (0 N, 56 N, 112 N, and 168 N kg ha−1) impacts on Miscanthus biomass yield and stand health. The flight campaigns were conducted early, middle, and late during the summer growing season. Among the VIs, mid-summer growing season NDRE provided the best prediction of fresh biomass (R2 = 0.87 and 0.97) and dry biomass (R2 = 0.89 and 0.97) in 2018 and 2019, respectively. The VIs generally showed that it was possible to distinguish between 0 N and 168 N treatments, but neither 0 N and 56 N kg ha−1 nor 112 N and 168 N kg ha−1 could be separated. The results from this study highlight the importance of moderate application of N (112 kg N ha−1) in improving and maintaining the stand health and biomass yield of Miscanthus over time and suggest that mid-summer growing season VIs, NDRE in particular, can be useful for assessment of Miscanthus stand health and biomass yield.
keywords: Feedstock Production;Biomass Analytics;Field Data
published: 2025-10-13
 
The oleaginous, carotenogenic yeast Rhodotorula toruloides has been increasingly explored as a platform organism for the production of terpenoids and fatty acid derivatives. Fatty alcohols, a fatty acid derivative widely used in the production of detergents and surfactants, can be produced microbially with the expression of a heterologous fatty acyl-CoA reductase. Due to its high lipid production, R. toruloides has high potential for fatty alcohol production, and in this study several metabolic engineering approaches were investigated to improve the titer of this product. Fatty acyl-CoA reductase from Marinobacter aqueolei was co-expressed with SpCas9 in R. toruloides IFO0880 and a panel of gene overexpressions and Cas9-mediated gene deletions were explored to increase the fatty alcohol production. Two overexpression targets (ACL1 and ACC1, improving cytosolic acetyl-CoA and malonyl-CoA production, respectively) and two deletion targets (the acyltransferases DGA1 and LRO1) resulted in significant (1.8 to 4.4-fold) increases to the fatty alcohol titer in culture tubes. Combinatorial exploration of these modifications in bioreactor fermentation culminated in a 3.7 g/L fatty alcohol titer in the LRO1Δ mutant. As LRO1 deletion was not found to be beneficial for fatty alcohol production in other yeasts, a lipidomic comparison of the DGA1 and LRO1 knockout mutants was performed, finding that DGA1 is the primary acyltransferase responsible for triacylglyceride production in R. toruloides, while LRO1 disruption simultaneously improved fatty alcohol production, increased diacylglyceride and triacylglyceride production, and increased glucose consumption. The fatty alcohol titer of fatty acyl-CoA reductase-expressing R. toruloides was significantly improved through the deletion of LRO1, or the deletion of DGA1 combined with overexpression of ACC1 and ACL1. Disruption of LRO1 surprisingly increased both lipid and fatty alcohol production, creating a possible avenue for future study of the lipid metabolism of this yeast.
keywords: Conversion;Genome Engineering;Genomics
published: 2025-10-13
 
CRISPR/Cas9 based genome editing has advanced our understanding of a myriad of important biological phenomena. Important challenges to multiplex genome editing in maize include assembly of large complex DNA constructs, few genotypes with efficient transformation systems, and costly/labor-intensive genotyping methods. Here we present an approach for multiplex CRISPR/Cas9 genome editing system that delivers a single compact DNA construct via biolistics to Type I embryogenic calli, followed by a novel efficient genotyping assay to identify desirable editing outcomes. We first demonstrate the creation of heritable mutations at multiple target sites within the same gene. Next, we successfully created individual and stacked mutations for multiple members of a gene family. Genome sequencing found off-target mutations are rare. Multiplex genome editing was achieved for both the highly transformable inbred line H99 and Illinois Low Protein1 (ILP1), a genotype where transformation has not previously been reported. In addition to screening transformation events for deletion alleles by PCR, we also designed PCR assays that selectively amplify deletion or insertion of a single nucleotide, the most common outcome from DNA repair of CRISPR/Cas9 breaks by non-homologous end-joining. The Indel-Selective PCR (IS-PCR) method enabled rapid tracking of multiple edited alleles in progeny populations. The ‘end to end’ pipeline presented here for multiplexed CRISPR/Cas9 mutagenesis can be applied to accelerate maize functional genomics in a broader diversity of genetic backgrounds.
keywords: gene editing; genome engineering; genotyping
published: 2025-10-10
 
Upregulation of triacylglycerols (TAGs) in vegetative plant tissues such as leaves has the potential to drastically increase the energy density and biomass yield of bioenergy crops. In this context, constraint-based analysis has the promise to improve metabolic engineering strategies. Here we present a core metabolism model for the C4 biomass crop Sorghum bicolor (iTJC1414) along with a minimal model for photosynthetic CO2 assimilation, sucrose and TAG biosynthesis in C3 plants. Extending iTJC1414 to a four-cell diel model we simulate C4 photosynthesis in mature leaves with the principal photo-assimilatory product being replaced by TAG produced at different levels. Independent of specific pathways and per unit carbon assimilated, energy content and biosynthetic demands in reducing equivalents are about 1.3 to 1.4 times higher for TAG than for sucrose. For plant generic pathways, ATP- and NADPH-demands per CO2 assimilated are higher by 1.3- and 1.5-fold, respectively. If the photosynthetic supply in ATP and NADPH in iTJC1414 is adjusted to be balanced for sucrose as the sole photo-assimilatory product, overproduction of TAG is predicted to cause a substantial surplus in photosynthetic ATP. This means that if TAG synthesis was the sole photo-assimilatory process, there could be an energy imbalance that might impede the process. Adjusting iTJC1414 to a photo-assimilatory rate that approximates field conditions, we predict possible daily rates of TAG accumulation, dependent on varying ratios of carbon partitioning between exported assimilates and accumulated oil droplets (TAG, oleosin) and in dependence of activation of futile cycles of TAG synthesis and degradation. We find that, based on the capacity of leaves for photosynthetic synthesis of exported assimilates, mature leaves should be able to reach a 20% level of TAG per dry weight within one month if only 5% of the photosynthetic net assimilation can be allocated into oil droplets. From this we conclude that high TAG levels should be achievable if TAG synthesis is induced only during a final phase of the plant life cycle.
keywords: Feedstock Production;Modeling
published: 2025-10-10
 
Methyl methacrylate (MMA) is an important petrochemical with many applications. However, its manufacture has a large environmental footprint. Combined biological and chemical synthesis (semisynthesis) may be a promising alternative to reduce both cost and environmental impact, but strains that can produce the MMA precursor (citramalate) at low pH are required. A non-conventional yeast, Issatchenkia orientalis, may prove ideal, as it can survive extremely low pH. Here, we demonstrate the engineering of I. orientalis for citramalate production. Using sequence similarity network analysis and subsequent DNA synthesis, we selected a more active citramalate synthase gene (cimA) variant for expression in I. orientalis. We then adapted a piggyBac transposon system for I. orientalis that allowed us to simultaneously explore the effects of different cimA gene copy numbers and integration locations. A batch fermentation showed the genome-integrated-cimA strains produced 2.0 g/L citramalate in 48 h and a yield of up to 7% mol citramalate/mol consumed glucose. These results demonstrate the potential of I. orientalis as a chassis for citramalate production.
keywords: Conversion;Metabolomics
published: 2025-10-10
 
The success of a bioenergy policy relies largely on the wide adoption of perennial energy crops at the farm scale. This study uses survey data to examine potential adoption decisions by farmers in the U.S. Midwest and the causal effects of various direct and indirect influencing factors, especially heterogeneous preferences of farmers. A Bayesian network (BN) model is developed to delineate the causal relationship between farmers adoption decisions and the influencing factors. We find a dominating role of economic factors and a non-negligible impact of non-economic factors, such as the perceived environmental benefits and the extent of familiarity with perennial energy crops. To examine the effect of heterogeneity in farmer preferences, we classify the surveyed farmers into four categories based on their attitudes toward the economic, social, and environmental dimensions of perennial energy crops. We identified statistically significant between-group differences in the responses of the four types of farmers to the various influencing factors. Our findings contribute to disentangling the complicated motivations that will influence perennial energy crop adoption decisions and provide implications for more targeted policy development that need to consider the heterogeneous drivers of farmer decisions about land use.
keywords: Sustainability;Modeling
published: 2025-10-10
 
Lipids accumulated in the vegetative tissues of cellulosic feedstocks can be a potential raw material for biodiesel and bioethanol production. In this work, bagasse of genetically engineered sorghum was subjected to liquid hot-water pretreatment at 170, 180, and 190 °C for different reaction time. Under the optimal pretreatment condition (170 °C, 20 min), the residue was enriched in glucan (57.39 ± 2.63 % w/w) and xylan (13.38 ± 0.49 % w/w). The total lipid content of the pretreated residue was 6.81% w/w, similar to that observed in untreated bagasse (6.30% w/w). Pretreatment improved the enzymatic digestibility of bagasse, allowing a recovery of 79% w/w and 86% w/w of glucose and xylose, respectively. The pretreatment and enzymatic saccharification resulted in a 2-fold increase in total lipid in enzymatic residue compared to the original bagasse. Thus, pretreatment and enzymatic hydrolysis enabled high sugar recovery while concentrating triglycerides and free fatty acids in the residue.
keywords: Conversion;Feedstock Production;Feedstock Bioprocessing
published: 2025-10-10
 
Mitochondrion is generally considered as the most promising subcellular organelle for compartmentalization engineering. Much progress has been made in reconstituting whole metabolic pathways in the mitochondria of yeast to harness the precursor pools (i.e., pyruvate and acetyl-CoA), bypass competing pathways, and minimize transportation limitations. However, only a few mitochondrial targeting sequences (MTSs) have been characterized (i.e., MTS of COX4), limiting the application of compartmentalization engineering for multigene biosynthetic pathways in the mitochondria of yeast. In the present study, based on the mitochondrial proteome, a total of 20 MTSs were cloned and the efficiency of these MTSs in targeting heterologous proteins, including the Escherichia coli FabI and enhanced green fluorescence protein (EGFP) into the mitochondria was evaluated by growth complementation and confocal microscopy. After systematic characterization, six of the well-performed MTSs were chosen for the colocalization of complete biosynthetic pathways into the mitochondria. As proof of concept, the full α-santalene biosynthetic pathway consisting of 10 expression cassettes capable of converting acetyl-coA to α-santalene was compartmentalized into the mitochondria, leading to a 3.7-fold improvement in the production of α-santalene. The newly characterized MTSs should contribute to the expanded metabolic engineering and synthetic biology toolbox for yeast mitochondrial compartmentalization engineering.
keywords: Conversion;Metabolic Engineering
published: 2025-10-10
 
Glucose and xylose are the major sugars present in cellulosic hydrolysates. The cellulosic sugars can be used for the production of platform chemicals. In this study, productions of lipid and ethanol by yeasts were compared for concentrated bioenergy sorghum syrup. Bioenergy sorghum was hydrothermally pretreated at 50% w/w solids in a continuous industrial reactor and sequentially mechanically refined using a burr mill to improve biomass accessibility for hydrolysis. Fed-batch enzymatic hydrolysis was conducted with 50% w/v solids loading and cellulase cocktail (50 FPU/g biomass) to achieve 230 g/L sugar concentration. Various strains of Rhodosporidium toruloides were evaluated for converting sugars into lipids, and strain Y-6987 had the highest lipid titer (9.2 g/L). The lipid titer was improved to 19.0 g/L by implementing a two-stage culture scheme, where the first stage was optimized for yeast growth and the second for lipid production. For ethanol production, the engineered Saccharomyces cerevisiae SR8ΔADH6 was used to coferment glucose and xylose. Ethanol fermentation was optimized for media nutrients (YP, YNB/urea, and urea), cellulosic sugar concentration, and sulfite conditioning to maximize the ethanol concentration from sorghum syrups. Fermentation of 70% v/v concentrated hydrolysate conditioned with sulfite produces 50.1 g/L ethanol from 141 g/L of sugars.
keywords: Conversion;Feedstock Bioprocessing
published: 2025-10-10
 
The presence of free fatty acids interferes with the conversion of plant oils to biodiesel. Four strong and weak base resins were evaluated for the removal of free fatty acids (FFA) from oil. Amberlite FPA 51 showed the highest adsorption capacity of FFA. A resin concentration above 3% could enable a higher percentage FFA adsorption. The adsorption process fitted a pseudo-first-order kinetic model and achieved equilibrium in approximately 8 h. A full factorial design was used to optimize the resin and FFA concentrations at a fixed temperature (40° C). A ratio of resin to fatty acid concentrations above 1.875 was sufficient for 70% adsorption and the amount adsorbed continued to increase with further added resin. A two-step washing of resin using hexane and ethanol recovered approximately 67.55% ± 4.05% of the initially added fatty acid. The resin that was used was regenerated with 5% NaOH and re-used for a minimum of three consecutive cycles. However, the adsorption capacity diminished to 75% of the initial cycle in cycles 2 and 3. Thus, the work presents a resin-based process for deacidification of oil to reduce fatty acid content of oil for biodiesel production.
keywords: Conversion;Feedstock Bioprocessing
published: 2025-10-10
 
Plant architecture influences the microenvironment throughout the canopy layer. Plants with a more erect leaf architecture allow for an increase in planting densities and allow more light to reach lower canopy leaves. This is predicted to increase crop carbon assimilation. Frictional resistance to wind reduces air movement in the lower canopy, resulting in higher humidity. By increasing the proportion of canopy photosynthesis in the more humid lower canopy, gains in the efficiency of water use might be expected, although this may be slightly offset by the more open erectophile form canopy. An anatomical feature in members of the Poaceae family that impacts leaf angle is the articulated junction of the sheath and blade, which also bares the ligule and auricles. Mutants, which lack ligules and auricles, show no articulation at this junction, resulting in leaves that are near vertical. In maize, these phenotypes termed liguleless result from null mutations of genes: ZmLG1 (Zm00001eb67740) and ZmLG2 (Zm00001eb147220). In sorghum, SbiRTx430.06G264300 (SbLG1) and SbiRTx430.03G392300 (SbLG2) are annotated as the respective maize homologues. A hair-pin element designed to down-regulate both SbLG1 and SbLG2 was introduced into the grain sorghum genotype RTx430. Derived transgenic events harbouring the hair-pin failed to develop ligules and displayed reduced leaf angles to the vertical, but less vertical than in null mutations. Under field settings, plots sown with these sorghum events having an erect architecture phenotype displayed an increase in photosynthesis in lower canopy levels, which led to increases in above-ground biomass and seed yield, without an increase in water use.
keywords: Genome Engineering; Photosynthesis; Sorghum; Water Use Efficiency
published: 2025-10-10
 
This zip file contains a UNIX-format DayCent model executable, input files, automation code, and associated directory structure necessary to re-produce the DayCent analysis underlying the manuscript. The main script “autodaycent.py” (written for Python 2.7) opens an interactive command line routine that facilitates: Calibrating the DayCent pine growth model; Initializing DayCent for a set of case studies sites; Executing an ensemble of model runs representing case study site reforestation, grassland restoration, or conversion to switchgrass cultivation; and Results analysis & generation of manuscript Fig. 3. Note that the interactive analysis code requires that all input files to be contained in the directory structure as uploaded, without modification. Executable versions of the DayCent model compatible with other operating systems are available upon request.
keywords: Feedstock Production;Modeling
suppressed by curator
 
published: 2024-01-04
 
This is a collection of 31 quasi-linear convective system (QLCS) mesovortices (MVs) that were manually identified and analyzed using the lowest elevation scan of the nearest relevant Weather Surveillance Radar–1988 Doppler (WSR-88D) during the two years (springs of 2022 and 2023) of the Propagation, Evolution, and Rotation in Linear Storms (PERiLS) field campaign. Throughout the two years of PERiLS, a total of nine intensive observing periods (IOPs) occurred (see https://catalog.eol.ucar.edu/perils_2022/missions and https://catalog.eol.ucar.edu/perils_2023/missions for exact IOP dates/times). However, only six of these IOPs (specifically, IOPs 2, 3, and 4 from both years) are included in this dataset. The inclusion criteria were based on the presence of strictly QLCS MVs within the C-band On Wheels (COW) domain, one of the research radars deployed in the field for the PERiLS project. Further details on how MVs were identified are provided below. This analysis was completed using the Gibson Ridge radar-viewing software (GR2Analyst). Each MV had to be produced by a QLCS, defined as a continuous area of 35 dBZ radar reflectivity over at least 100 km when viewed from the lowest elevation scan. The MVs analyzed also had to pass through/near the COW’s domain at some point during their lifetimes to allow for additional analysis using the COW data. Tornadic (TOR), wind-damaging (WD), and non-damaging (ND) MVs were analyzed. ND MVs were ones that usually had a tornado warning placed on them but did not produce any damage and persisted for five or more radar scans; this was done to target the strongest MVs that forecasters thought could be tornadic. The QLCS MVs were identified using objective criteria, which included the existence of a circulation with a maximum differential velocity (dV; i.e., the difference between the maximum outbound and minimum inbound velocities at a constant range) of at least 20 kt over a distance ≤ 7 km. The following radar-based characteristics were catalogued for each QLCS MV at the lowest elevation angle of the nearest WSR-88D: latitude and longitude locations of the MV, the genesis to decay time of the MV, the maximum dV across the MV, the maximum rotational velocity (Vrot; i.e., dV divided by two), diameter of the MV, the range from the radar of the MV center, and the height above radar level of the MV center. In the Excel sheet, there are a total of 37 sheets. 32 of the 37 sheets are for each MV that was examined. One of those MVs (sheet titled 'EFU_tor_iop3') was not included in the final count of MVs (31). This MV produced an EFU tornado and only tornadoes that were given ratings were used to calculate MV statistics. The 31 MV sheets that were used to calculate MV statistics are labeled following the convention 'mv#_iop#_qlcs'. ‘mv#’ is the unique number that was assigned to each MV for clear identification, 'iop#' is the IOP in which the MV occurred, 'qlcs' denotes that the MV was produced by a QLCS, and the 2023 IOPs are denoted by ‘_2023’ after ‘qlcs’ in the sheet name. In these sheets, there are notes on what was visually seen in the radar data, damage associated with each MV (using the National Centers for Environmental Information (NCEI) database), and the characteristics of the MV at each time step of its lifetime. The yellow rows in each of the sheets indicate the last row of data included in the pretornadic, predamaging (wind damage), and pre-nondamaging statistics. The orange boxes in the notes column indicate any reports that were in NCEI but not in GR2Analyst. There are also sheets that examine pretornadic and predamaging diameter trends, box and whisker plot statistics of the overall characteristics of the different types of MVs, and the overall characteristics of each MV, with one Excel sheet (‘combined_qlcs_mvs’) examining the characteristics of each MV over its entire lifetime and one Excel sheet (‘combined_qlcs_mvs_before_report’) examining the characteristics of each MV before it first produced damage or had a tornado warning placed on it.
keywords: quasi-linear convective system; QLCS; tornado; radar; mesovortex; PERiLS; low-level rotation; tornadic; nontornadic; wind-damaging; Propagation, Evolution, and Rotation in Linear Storms; tornado warning; C-band On Wheels
published: 2025-10-10
 
β‐Carotene is a natural pigment and health‐promoting metabolite, and has been widely used in the nutraceutical, feed, and cosmetic industries. Here, we engineered a GRAS yeast Saccharomyces cerevisiae to produce β‐carotene from xylose, the second most abundant and inedible sugar component of lignocellulose biomass. Specifically, a β‐carotene biosynthetic pathway containing crtYB, crtI, and crtE from Xanthophyllomyces dendrorhous was introduced into a xylose‐fermenting S. cerevisiae. The resulting strain produced β‐carotene from xylose at a titer threefold higher than from glucose. Interestingly, overexpression of tHMG1, which has been reported as a critical genetic perturbation to enhance metabolic fluxes in the mevalonate pathway and β‐carotene production in yeast when glucose is used, did not further improve the production of β‐carotene from xylose. Through fermentation profiling, metabolites analysis, and transcriptional studies, we found the advantages of using xylose as a carbon source, instead of glucose, for β‐carotene production to be a more respiratory feature of xylose consumption, a larger cytosolic acetyl‐CoA pool, and an upregulated expression level of rate‐limiting genes in the β‐carotene‐producing pathway, including ACS1 and HMG1. As a result, 772.8 mg/L of β‐carotene was obtained in a fed‐batch bioreactor culture with xylose feeding. Considering the inevitable large scale production of xylose when cellulosic biomass‐based bioeconomy is implemented, our results suggest xylose utilization is a promising strategy for overproduction of carotenoids and other isoprenoids in engineered S. cerevisiae.
keywords: Conversion;Genome Engineering
published: 2025-10-10
 
The nonconventional yeast Issatchenkia orientalis has emerged as a potential platform microorganism for production of organic acids due to its ability to grow robustly under highly acidic conditions. However, lack of efficient genetic tools remains a major bottleneck in metabolic engineering of this organism. Here we report that the autonomously replicating sequence (ARS) from Saccharomyces cerevisiae (ScARS) was functional for plasmid replication in I. orientalis, and the resulting episomal plasmid enabled efficient genome editing by the CRISPR/Cas9 system. The optimized CRISPR/Cas9-based system employed a fusion RPR1′-tRNA promoter for single guide RNA (sgRNA) expression and could attain greater than 97% gene disruption efficiency for various gene targets. Additionally, we demonstrated multiplexed gene deletion with disruption efficiencies of 90% and 47% for double gene and triple gene knockouts, respectively. This genome editing tool can be used for rapid strain development and metabolic engineering of this organism for production of biofuels and chemicals.
keywords: Conversion;Genomics;Genome Engineering;Transcriptomics
published: 2025-10-09
 
Aging-related yield decline in Miscanthus × giganteus (miscanthus) remains a major constraint to sustainable biomass production. This study evaluated how nitrogen (N) management and soil fertility influence yield-component traits and productivity in aging miscanthus. Trials were conducted at two sites established in 2008 at the University of Illinois Energy Farm, Urbana, IL. (i) The Sun Grant trial received 0, 60, and 120 kg N ha−1 annually until 2015. Starting 2021, half of each plot received 60 or 120 kg N ha−1, resulting in six legacy-contemporary treatments: 0N–0N, 0N–120N, 60N–0N, 60N–60N, 120N–0N, 120N–120N. (ii) The Energy Farm trial remained unfertilized until 2014, when one half of each plot received 56 kg N ha−1, forming two treatments: 0N–0N, 0N–56N. Sun Grant trial results showed N fertilization increased tiller density (tillers m−2) and tiller weight (g tiller−1) in juvenile to early-mature miscanthus (2011–2015). After N withdrawal, both traits declined (20 % and 40 %), though legacy effects persisted in tiller weight in the aging stands (2020–2023). Contemporary N had little effect on tiller density but increased tiller weight by 34 %–77 %, resulting in 23 %–106 % higher machine-harvested biomass yield in 0–120N, 60-60N, and 120-120N plots. At the Energy Farm trial, 0N–56N plots yielded 59 %–108 % more biomass than 0N–0N. Soil total N increased (Sun Grant: 47 % by 2020; Energy Farm: 58 % by 2023), while Mehlich-3 P (42 %–44 %) and K (21 %–46 %) declined. These findings identify tiller weight as a key determinant of biomass yield in aging miscanthus and highlight the need for P and K management for long-term productivity.
keywords: miscanthus; nitrogen; soil
published: 2025-10-08
 
Rubisco activase (Rca) facilitates the release of sugar‐phosphate inhibitors from the active sites of Rubisco and thereby plays a central role in initiating and sustaining Rubisco activation. In Arabidopsis, alternative splicing of a single Rca gene results in two Rca isoforms, Rca‐α and Rca‐β. Redox modulation of Rca‐α regulates the function of Rca‐α and Rca‐β acting together to control Rubisco activation. Although Arabidopsis Rca‐α alone less effectively activates Rubisco in vitro , it is not known how CO2 assimilation and plant growth are impacted. Here, we show that two independent transgenic Arabidopsis lines expressing Rca‐α in the absence of Rca‐β (“Rca‐α only” lines) grew more slowly in various light conditions, especially under low light or fluctuating light intensity, and in a short day photoperiod compared to wildtype. Photosynthetic induction was slower in the Rca‐α only lines, and they maintained a lower rate of CO2 assimilation during both photoperiod types. Our findings suggest Rca oligomers composed of Rca‐α only are less effective in initiating and sustaining the activation of Rubisco than when Rca‐β is also present. Currently there are no examples of any plant species that naturally express Rca‐α only but numerous examples of species expressing Rca‐β only. That Rca‐α exists in most plant species, including many C3 and C4 food and bioenergy crops, implies its presence is adaptive under some circumstances.
keywords: Feedstock Production;Biomass Analytics;Phenomics
published: 2025-10-07
 
Yarrowia lipolytica was found natively to produce erythritol, mannitol, and arabitol during growth on glucose, fructose, mannose, and glycerol. Osmotic stress is known to increase sugar alcohol production, and was found to significantly increase erythritol production during growth on glycerol. To better understand erythritol production from glycerol, since it was the most promising sugar alcohol, we measured the expression of key genes and intracellular metabolites. Osmotic stress increased the expression of several key genes in the glycerol catabolic pathway and the pentose phosphate pathway. Analysis of intracellular metabolites revealed that amino acids, sugar alcohols, and polyamines are produced at higher levels in response to osmotic stress. Heterologous overexpression of the sugar alcohol phosphatase increased erythritol production and glycerol utilization in Y. lipolytica. We further increased erythritol production by increasing the expression of native glycerol kinase (GK), and transketolase (TKL). These data show the growth and titers produced.
keywords: Conversion;Genome Engineering
published: 2025-10-07
 
This dataset accompanies the research paper "Nine circles of elastic brittle fracture: A series of challenge problems to assess fracture models" by Kamarei, Zeng, Dolbow, and Lopez-Pamies, CMAME (2026). The dataset contains computational mesh files, figure data, and supporting materials for nine standardized challenge problems designed to assess fracture models in isotropic elastic brittle materials under quasi-static loading conditions. The nine challenge problems span the complete range of fracture nucleation and propagation mechanisms: strength nucleation (uniaxial tension, biaxial tension, torsion), Griffith nucleation (pure-shear, double cantilever beam, trousers), and strength-Griffith mediated nucleation (single edge notch, indentation, poker-chip). The dataset includes: High-quality mesh files for all nine challenge problems Figure data and results for both linear elastic (soda-lime glass) and non-linear elastic (PU elastomer) brittle materials
keywords: Fracture mechanics; Finite element analysis; Phase-field modeling; Strength; Toughness; Fracture nucleation; Fracture propagation; Experimental validation
published: 2025-10-03
 
Plant cell wall hydrolysates contain not only sugars but also substantial amounts of acetate, a fermentation inhibitor that hinders bioconversion of lignocellulose. Despite the toxic and non-consumable nature of acetate during glucose metabolism, we demonstrate that acetate can be rapidly co-consumed with xylose by engineered Saccharomyces cerevisiae. The co-consumption leads to a metabolic re-configuration that boosts the synthesis of acetyl-CoA derived bioproducts, including triacetic acid lactone (TAL) and vitamin A, in engineered strains. Notably, by co-feeding xylose and acetate, an engineered strain produces 23.91 g/L TAL with a productivity of 0.29 g/L/h in bioreactor fermentation. This strain also completely converts a hemicellulose hydrolysate of switchgrass into 3.55 g/L TAL. These findings establish a versatile strategy that not only transforms an inhibitor into a valuable substrate but also expands the capacity of acetyl-CoA supply in S. cerevisiae for efficient bioconversion of cellulosic biomass.
keywords: Conversion;Genome Engineering
published: 2021-05-17
 
Please cite as: Wuebbles, D., J. Angel, K. Petersen, and A.M. Lemke, (Eds.), 2021: An Assessment of the Impacts of Climate Change in Illinois. The Nature Conservancy, Illinois, USA. https://doi.org/10.13012/B2IDB-1260194_V1 Climate change is a major environmental challenge that is likely to affect many aspects of life in Illinois, ranging from human and environmental health to the economy. Illinois is already experiencing impacts from the changing climate and, as climate change progresses and temperatures continue to rise, these impacts are expected to increase over time. This assessment takes an in-depth look at how the climate is changing now in Illinois, and how it is projected to change in the future, to provide greater clarity on how climate change could affect urban and rural communities in the state. Beyond providing an overview of anticipated climate changes, the report explores predicted effects on hydrology, agriculture, human health, and native ecosystems.
keywords: Climate change; Illinois; Public health; Agriculture; Environment; Water; Hydrology; Ecosystems
published: 2025-09-04
 
This dataset contains the following to replicate figures from "TChem-atm (v2.0.0): Scalable Performance-Portable Multiphase Atmospheric Chemistry" submitted to Geophysical Model Development (GMD). It contains (1) the simulation inputs, outputs and analysis notebook for recreating the PartMC-CAMP and PartMC-TChem-atm comparison and (2) scripts, timing results and analysis tools for recreating the performance evaluation. Users can either inspect the raw output to verify the results of the manuscript or rerun simulations using the provided inputs. Additionally, modifiying the inputs allows for for further exploration of both model simulation and performance characteristics.
keywords: Atmospheric chemistry; Aerosols; Numerical solvers; Particle-resolved modeling; GPUs
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