Dataset Search Results

The photosynthetic capacity or the CO2-saturated photosynthetic rate (Vmax), chlorophyll, and nitrogen are closely linked leaf traits that determine C4 crop photosynthesis and yield. Accurate, timely, rapid, and non-destructive approaches to predict leaf photosynthetic traits from hyperspectral reflectance are urgently needed for high-throughput crop monitoring to ensure food and bioenergy security. Therefore, this study thoroughly evaluated the state-of-the-art physically based radiative transfer models (RTMs), data-driven partial least squares regression (PLSR), and generalized PLSR (gPLSR) models to estimate leaf traits from leaf-clip hyperspectral reflectance, which was collected from maize (Zea mays L.) bioenergy plots with diverse genotypes, growth stages, treatments with nitrogen fertilizers, and ozone stresses in three growing seasons. The results show that leaf RTMs considering bidirectional effects can give accurate estimates of chlorophyll content (Pearson correlation r=0.95), while gPLSR enabled retrieval of leaf nitrogen concentration (r=0.85). Using PLSR with field measurements for training, the cross-validation indicates that Vmax can be well predicted from spectra (r=0.81). The integration of chlorophyll content (strongly related to visible spectra) and nitrogen concentration (linked to shortwave infrared signals) can provide better predictions of Vmax (r=0.71) than only using either chlorophyll or nitrogen individually. This study highlights that leaf chlorophyll content and nitrogen concentration have key and unique contributions to Vmax prediction.

This dateset contains data files necessary to replicate figures from "Idealized Particle-Resolved Large-Eddy Simulations to Evaluate the Impact of Emissions Spatial Heterogeneity on CCN Activity" submitted to Atmospheric Chemistry and Physics. Within the compressed folder data.zip are two subdirectories, "processed_data" and "spatial-het". The "processed_data" directory contains netCDF files which contain a subset of simulation output used in figure generation. The "spatial-het" subdirectory contains a .csv file with spatial heterogeneity values computed via an exact algorithm of the spatial heterogeneity metric described by Mohebalhojeh et al. 2025. The subdirectory "sh-patterns" contains .csv files for each emissions scenario. Each entry corresponds to a single grid cell over a domain of dimension 100x100 (lateral resolution of the computational domain employed in this paper). Within scripts.zip are python notebooks for generating figures. Additional python modules are included which contain helper functions for notebooks. Furthermore, a Fortran version of the spatial heterogeneity metric is included alongside shells scripts for creating a python environment in which the code can be compiled and convert into a Python module. Note that the create_env.sh and compile_nsh.sh scripts must be run prior to executing cells in notebooks to make use of the spatial heterogeneity subroutines. <b>*Note*:</b> New in this V2: Following an initial review, an additional figure was requested, which required updates to both data.zip (adding one new NC file: no-heterogeneity_met-vars_subset.nc) and scripts.zip (a minor addition to a Python notebook). A README in PDF format has also been uploaded to provide a summary of the dataset.

During the transformation of wild-type (WT) Arabidopsis thaliana, a T-DNA containing OLEOSIN-GFP (OLE1-GFP) was inserted by happenstance within the GBSS1 gene, resulting in significant reduction in amylose and increase in leaf oil content in the transgenic line (OG). The synergistic effect on oil accumulation of combining gbss1 with the expression of OLE1-GFP was confirmed by transforming an independent gbss1 mutant (GABI_914G01) with OLE1-GFP. The resulting OLE1-GFP/gbss1 transgenic lines showed higher leaf oil content than the individual OLE1-GFP/WT or single gbss1 mutant lines. Further stacking of the lipogenic factors WRINKLED1, Diacylglycerol O-Acyltransferase (DGAT1), and Cys-OLEOSIN1 (an engineered sesame OLEOSIN1) in OG significantly elevated its oil content in mature leaves to 2.3% of dry weight, which is 15 times higher than that in WT Arabidopsis. Inducible expression of the same lipogenic factors was shown to be an effective strategy for triacylglycerol (TAG) accumulation without incurring growth, development, and yield penalties.

Conversion of corn fiber to ethanol in the dry grind process can increase ethanol yields, improve coproduct quality and contribute to process sustainability. This work investigates the use of two physio-chemical pretreatments on corn fiber and effect of cellulase enzyme dosage to improve ethanol yields. Fiber separated after liquefaction of corn was pretreated using (1) hot water pretreatment (160°C for 5, 10 or 20 min); and (2) wet disk milling and converted to ethanol. The conversion efficiencies of hot water pretreated fiber were higher than untreated fiber, with highest increase in conversion (10.4%) achieved for 5-minute residence time at 160 °C. Disk milling was not effective in increasing conversion compared to other treatments. Hydrolysis and fermentation of untreated fiber with excess cellulase enzymes resulted in 33.3% higher conversion compared to untreated fiber. Note: in “Table1_Treatments.csv”, NA = Not applicable.

The optimal flowering time for bioenergy crop miscanthus is essential for environmental adaptability and biomass accumulation. However, little is known about how genes controlling flowering in other grasses contribute to flowering regulation in miscanthus. Here, we report on the sequence characterization and gene expression of Miscanthus sinensisGhd8, a transcription factor encoding a HAP3/NF-YB DNA-binding domain, which has been identified as a major quantitative trait locus in rice, with pleiotropic effects on grain yield, heading date and plant height. In M. sinensis, we identified two homoeologous loci, MsiGhd8A located on chromosome 13 and MsiGhd8B on chromosome 7, with one on each of this paleo-allotetraploid species’ subgenomes. A total of 46 alleles and 28 predicted protein sequence types were identified in 12 wild-collected accessions. Several variants of MsiGhd8 showed a geographic and latitudinal distribution. Quantitative real-time PCR revealed that MsiGhd8 expressed under both long days and short days, and MsiGhd8B showed a significantly higher expression than MsiGhd8A. The comparison between flowering time and gene expression indicated that MsiGhd8B affected flowering time in response to day length for some accessions. This study provides insight into the conserved function of Ghd8 in the Poaceae, and is an important initial step in elucidating the flowering regulatory network of Miscanthus.

DayCent MUVP version (Methanogenesis, UV litter degradation and Photosynthesis). DAYCENT is the daily time-step version of the CENTURY biogeochemical model (Parton et al., 1994). DAYCENT simulates fluxes of C and N among the atmosphere, vegetation, and soil (Del Grosso et al., 2001a; Parton et al., 1998). Key submodels include soil water content and temperature by layer, plant production and allocation of net primary production (NPP), decomposition of litter and soil organic matter, mineralization of nutrients, N gas emissions from nitrification and denitrification, and CH4 oxidation in non-saturated soils.

In this study, different process schemes were designed and evaluated for biodiesel production from engineered cane lipids with uncertain fatty acid compositions. Four different process schemes were compared under (i) thermal glycerolysis and (ii) enzymatic glycerolysis approaches. These schemes were based on the biodiesel yield and economic indicators such as the net present value (NPV) and the minimum selling price (MSP) of biodiesel. A scheme with polar lipid separation under thermal glycerolysis resulted in the maximum NPV ($96.5 million) and minimum MSP ($1107/ton biodiesel), respectively. Through local sensitivity analysis, it was concluded that the cane lipid percentage is the most significant factor influencing process economics. A conjoint analysis of the lipid procurement price and cane lipid percent suggested that 15% cane lipids with a low lipid procurement price ($0.536/kg) results in a positive NPV. When the cane lipid price is higher (>$0.80/kg), a 20% lipid content should be considered to achieve a positive NPV. At 20% cane lipids, the worst-case and best-case scenarios were evaluated by analyzing the interplay of the three most important parameters, The best-case scenario revealed that the minimum NPV under any process scheme could yield more than $100 million (or MSP: $0.80/L), and the worst-case analysis showed that losses incurred by the plant could be as high as $80 million (MSP: $1.36/L). A Monte Carlo simulation indicated that there is a 70% chance of the plant being profitable (NPV > 0).

This repository provides the data and code used to reproduce key plots from the manuscript and to extend discussions that were only briefly covered therein. All MATLAB scripts were developed and tested in MATLAB R2024a. All Python scripts were developed and tested in Python 3.11.2. * <b> NOTE:</b> New in this V3: 1. 2 new MATLAB files (ChiralPointGroups.m and THz_current_estimation.m), ChiralPointGroups.pdf (a compiled version of ChiralPointGroups.m) and theoretical model code (theoretical_model.zip) are added. More information can be found in the readme. 2. Updated and renamed "publication_data.zip" (in V2) to "data_and_analysis.zip" 3. Change License from CC BY to "Other license". Licensing Terms: Data (all .mat files) is under CC BY and Code is released under MIT license. Therefore, V3 is bound to this new license. V2 is still under CC BY. <b>→ Data and analysis code (data_and_analysis.zip):</b> The dataset is organized into five subfolders. Each subfolder corresponds to a unique combination of experimental conditions, including: • Magnetic field orientation (B ∥ c or B ⟂ c) • Scan parameter (magnetic field or temperature) • Pump laser polarization (linear s, linear p, or circular) • Detection polarization (linear s) Each folder contains: • The raw time-domain data files (.mat) • Oscillator parameters extracted via linear prediction algorithm (.mat) • MATLAB scripts (.m) that generate plots of the raw data, processed fits, and amplified modes. Each script should be run within its corresponding folder to ensure proper loading of the associated data files. Folder summary: 1. B_parallel_c_linear_spump_sprobe_field: B ∥ c, s-polarized pump, s-polarized THz detection, magnetic field dependence 2. B_parallel_c_linear_spump_sprobe_temperature: B ∥ c, s-polarized pump, s-polarized THz detection, temperature dependence 3. B_perp_c_linear_spump_sprobe_field: B ⟂ c, s-polarized pump, s-polarized THz detection, magnetic field dependence 4. B_perp_c_linear_spump_sprobe_temperature: B ⟂ c, s-polarized pump, s-polarized THz detection, temperature dependence 5. B_parallel_c_LCPRCP_pump_sprobe_field: B ∥ c, circularly polarized pump (LCP & RCP), s-polarized THz detection, magnetic field dependence <b>→Theoretical model code (theoretical_model.zip):</b> The Python script depends on packages “numpy” and “matplotlib”. The script generates a plot of the dispersion relations of the theoretical model introduced in the Main Text. More precisely, it plots the real (red) and imaginary (blue) parts of the frequency (ω) as a function of wavenumber (k) as obtained by solving the characteristic equation, equation (6) of the Supplemental Information, with σ_E and σ_Μ given respectively by equations (3) and (2) of the Main Text. All branches of the dispersion relations are plotted simultaneously. All model parameters are adjustable. The included Mathematica notebook (printout also provided in .pdf format) was used to obtain symbolic expressions for the coefficients of powers of ω appearing in the characteristic determinant. These coefficients were copied directly into the Python function detCoeffs(). <b>→ Standalone scripts (not in subfolders):</b> • ChiralPointGroups.m Outputs a table summarizing the 2D matrix representation of σ_Μ in the 11 enantiomorphic point groups. ChiralPointGroups.pdf is a compiled version of chiral point groups table, identical to the output of ChiralPointGroups.m. • THz_current_estimation.m Estimates the photoinduced THz current in tellurium under magnetic field. The script evaluates a phenomenological resonant contribution to the magnetoelectric coupling (with negligible dependence on NIR polarization), leading to excitation of s-polarized, B-antisymmetric mode S_odd at ~0.37 THz. These standalone scripts provide additional physical discussion and calculation detail that are intentionally streamlined or omitted from the published manuscript and its supplementary materials for clarity and space.

Miscanthus is a close relative of saccharum and a potentially valuable genetic resource for improving sugarcane. Differences in flowering time within and between miscanthus and saccharum hinders intra- and interspecific hybridizations. A series of greenhouse experiments were conducted over three years to determine how to synchronize flowering time of saccharum and miscanthus genotypes. We found that day length was an important factor influencing when miscanthus and saccharum flowered. Sugarcane could be induced to flower in a central Illinois greenhouse using supplemental lighting to reduce the rate at which days shortened during the autumn and winter to 1 min d-1, which allowed us to synchronize the flowering of some sugarcane genotypes with Miscanthus genotypes primarily from low latitudes. In a complementary growth chamber experiment, we evaluated 33 miscanthus genotypes, including 28 M. sinensis, 2 M. floridulus, and 3 M. ×giganteus collected from 20.9° S to 44.9° N for response to three day lengths (10 h, 12.5 h, and 15 h). High latitude-adapted M. sinensis flowered mainly under 15 h days, but unexpectedly, short days resulted in short, stocky plants that did not flower; in some cases, flag leaves developed under short days but heading did not occur. In contrast, for M. sinensis and M. floridulus from low latitudes, shorter day lengths typically resulted in earlier flowering, and for some low latitude genotypes, 15 h days resulted in no flowering. However, the highest ratio of reproductive shoots to total number of culms was typically observed for 12.5 h or 15 h days. Latitude of origin was significantly associated with culm length, and the shorter the days, the stronger the relationship. Nearly all entries achieved maximal culm length under the 15 h treatment, but the nearer to the equator an accession originated, the less of a difference in culm length between the short-day treatments and the 15 h day treatment. Under short days, short culms for high-latitude accessions was achieved by different physiological mechanisms for M. sinensis genetic groups from the mainland in comparison to those from Japan; for mainland accessions, the mechanism was reduced internode length, whereas for Japanese accessions the phyllochron under short days was greater than under long days. Thus, for M. sinensis, short days typically hastened floral induction, consistent with the expectations for a facultative short-day plant. However, for high latitude accessions of M. sinensis, days less than 12.5 h also signaled that plants should prepare for winter by producing many short culms with limited elongation and development; moreover, this response was also epistatic to flowering. Thus, to flower M. sinensis that originates from high latitudes synchronously with sugarcane, the former needs day lengths >12.5 h (perhaps as high as 15 h), whereas that the latter needs day lengths <12.5 h.

Dataset for "Using Stochastic Block Models for Community Detection". This contains synthetic networks with ground-truth community structure generated using synthetic network generators (specifically, ABCD+o) based on real-world networks and computed clusterings on these real-world networks. Note: * networks.zip contains the synthetic networks

Perennial crops have been the focus of bioenergy research and development for their sustainability benefits associated with high soil carbon (C) and reduced nitrogen (N) requirements. However, perennial crops mature over several years and their sustainability benefits can be negated through land reversion. A photoperiod‐sensitive energy sorghum (Sorghum bicolor) may provide an annual crop alternative more ecologically sustainable than maize (Zea mays) that can more easily integrate into crop rotations than perennials, such as miscanthus (Miscanthus × giganteus). This study presents an ecosystem‐scale comparison of C, N, water and energy fluxes from energy sorghum, maize and miscanthus during a typical growing season in the Midwest United States. Gross primary productivity (GPP) was highest for maize during the peak growing season at 21.83 g C m−2 day−1, followed by energy sorghum (17.04 g C m−2 day−1) and miscanthus (15.57 g C m−2 day−1). Maize also had the highest peak growing season evapotranspiration at 5.39 mm day−1, with energy sorghum and miscanthus at 3.81 and 3.61 mm day−1, respectively. Energy sorghum was the most efficient water user (WUE), while maize and miscanthus were comparatively similar (3.04, 1.75 and 1.89 g C mm−1 H2O, respectively). Maize albedo was lower than energy sorghum and miscanthus (0.19, 0.26 and 0.24, respectively), but energy sorghum had a Bowen ratio closer to maize than miscanthus (0.12, 0.13 and 0.21, respectively). Nitrous oxide (N2O) flux was higher from maize and energy sorghum (8.86 and 12.04 kg N ha−1, respectively) compared with miscanthus (0.51 kg N ha−1), indicative of their different agronomic management. These results are an important first look at how energy sorghum compares to maize and miscanthus grown in the Midwest United States. This quantitative assessment is a critical component for calibrating biogeochemical and ecological models used to forecast bioenergy crop growth, productivity and sustainability.

2′-Fucosyllactose (2′-FL), a human milk oligosaccharide with confirmed benefits for infant health, is a promising infant formula ingredient. Although Escherichia coli, Saccharomyces cerevisiae, Corynebacterium glutamicum, and Bacillus subtilis have been engineered to produce 2′-FL, their titers and productivities need be improved for economic production. Glucose along with lactose have been used as substrates for producing 2′-FL, but accumulation of by-products due to overflow metabolism of glucose hampered efficient production of 2′-FL regardless of a host strain. To circumvent this problem, we used xylose, which is the second most abundant sugar in plant cell wall hydrolysates and is metabolized through oxidative metabolism, for the production of 2′-FL by engineered yeast. Specifically, we modified an engineered S. cerevisiae strain capable of assimilating xylose to produce 2′-FL from a mixture of xylose and lactose. First, a lactose transporter (Lac12) from Kluyveromyces lactis was introduced. Second, a heterologous 2′-FL biosynthetic pathway consisting of enzymes Gmd, WcaG, and WbgL from E. coli was introduced. Third, we adjusted expression levels of the heterologous genes to maximize 2′-FL production. The resulting engineered yeast produced 25.5 g/L of 2′-FL with a volumetric productivity of 0.35 g/L∙h in a fed-batch fermentation with lactose and xylose feeding to mitigate the glucose repression. Interestingly, the major location of produced 2′-FL by the engineered yeast can be changed using different culture media. While 72% of the produced 2′-FL was secreted when a complex medium was used, 82% of the produced 2′-FL remained inside the cells when a minimal medium was used. As yeast extract is already used as food and animal feed ingredients, 2′-FL enriched yeast extract can be produced cost-effectively using the 2′-FL-accumulating yeast cells.

The aim of this study was to determine carbohydrate recovery from hemp for ethanol production and quantify biodiesel from TAG (triacylglycerol) present in hemp. The structural composition of five different hemp varieties (Seward County-SC, York County-YC, Loup County-LC, 19 m96136-19 m, and CBD Hemp-CBD) were analyzed. Concentration of glucan and xylan ranged between 32.63 to 44.52% and 10.62 to 15.48% respectively. The biomass was then pretreated with Liquid hot water followed by disk milling and then hydrolyzed enzymatically to yield monomeric sugars. High glucose (63-85%) and xylose (73-88%) recovery was achieved. Lipids were extracted from hemp using hexane and isopropanol and then transesterified to produce biodiesel. Approximately, 50% of total fatty acids in SC, LC, and CBD hemp were linoleic acid. Palmitic acid was present between 32 to 50% in varieties YC and 19 m. Highest TAG concentration at 25% of total lipids was observed in CBD hemp. The analysis on lipid composition and high sugar recovery demonstrates hemp as a potential bioenergy crop for ethanol and biodiesel coproduction.

A novel process applying high solids loading in chemical-free pretreatment and enzymatic hydrolysis was developed to produce sugars from bioenergy sorghum. Hydrothermal pretreatment with 50% solids loading was performed in a pilot scale continuous reactor followed by disc refining. Sugars were extracted from the enzymatic hydrolysis at 10% to 50% solids content using fed-batch operations. Three surfactants (Tween 80, PEG 4000, and PEG 6000) were evaluated to increase sugar yields. Hydrolysis using 2% PEG 4000 had the highest sugar yields. Glucose concentrations of 105, 130, and 147 g/L were obtained from the reaction at 30%, 40%, and 50% solids content, respectively. The maximum sugar concentration of the hydrolysate, including glucose and xylose, obtained was 232 g/L. Additionally, the glucose recovery (73.14%) was increased compared to that of the batch reaction (52.74%) by using two-stage enzymatic hydrolysis combined with fed-batch operation at 50% w/v solids content.

Mitochondria play a key role in energy production and metabolism, making them a promising target for metabolic engineering and disease treatment. However, despite the known influence of passenger proteins on localization efficiency, only a few protein-localization tags have been characterized for mitochondrial targeting. To address this limitation, we leverage a Variational Autoencoder to design novel mitochondrial targeting sequences. In silico analysis reveals that a high fraction of the generated peptides (90.14%) are functional and possess features important for mitochondrial targeting. We characterize artificial peptides in four eukaryotic organisms and, as a proof-of-concept, demonstrate their utility in increasing 3-hydroxypropionic acid titers through pathway compartmentalization and improving 5-aminolevulinate synthase delivery by 1.62-fold and 4.76-fold, respectively. Moreover, we employ latent space interpolation to shed light on the evolutionary origins of dual-targeting sequences. Overall, our work demonstrates the potential of generative artificial intelligence for both fundamental research and practical applications in mitochondrial biology.

The files in this dataset include the now-public domain full raw text and illustrations for the novel Gentlemen Prefer Blondes (GBP) by Anita Loos, and files comparing the two published versions of the novel in 1925, one in Harper's Bazar magazine and the other in book format by Boni & Liveright. These files comprise the underlying data for the scholarly digital edition of the novel edited by Daniel G. Tracy. The full citation for the publication, including the DOI link for those wishing access the text, is: Loos, Anita. Gentlemen Prefer Blondes. Edited by Daniel G. Tracy, Critical Edition. Windsor & Downs Press, 2025. https://doi.org/10.21900/wd.13

Annotation of untargeted high-resolution full-scan LC-MS metabolomics data remains challenging due to individual metabolites generating multiple LC-MS peaks arising from isotopes, adducts, and fragments. Adduct annotation is a particular challenge, as the same mass difference between peaks can arise from adduct formation, fragmentation, or different biological species. To address this, here we describe a buffer modification workflow (BMW) in which the same sample is run by LC-MS in both liquid chromatography solvent with 14NH3–acetate buffer and in solvent with the buffer modified with 15NH3–formate. Buffer switching results in characteristic mass and signal intensity changes for adduct peaks, facilitating their annotation. This relatively simple and convenient chromatography modification annotated yeast metabolomics data with similar effectiveness to growing the yeast in isotope-labeled media. Application to mouse liver data annotated both known metabolite and known adduct peaks with 95% accuracy. Overall, it identified 26% of ∼27 000 liver LC-MS features as putative metabolites, of which ∼2600 showed HMDB or KEGG database formula match. This workflow is well suited to biological samples that cannot be readily isotope labeled, including plants, mammalian tissues, and tumors.

We apply prospect theory to examining farmers’ economic incentives to divert a share of their land to bioenergy crops (miscanthus and switchgrass in this study). Numerical simulation is conducted for 1,919 rain‐fed U.S. counties to identify the impact of loss aversion on bioenergy crop adoption, and how this impact is influenced by biomass price, discount rate, credit constraint status, and policy instruments. Results show that ignoring farmer’s loss aversion causes overestimation of miscanthus production but underestimation of switchgrass production, particularly when farmers are credit constrained and have a high discount rate. We find that establishment cost subsidy induces more miscanthus production whereas subsidized energy crop insurance induces more switchgrass production. The efficacy of these two policy instruments, measured by biomass production increased by per dollar of government outlay, depends on the magnitude of farmers’ loss aversion and discount rate.

Environmental DNA metabarcoding data for fish communities at 50 sites in the Tennessee River watershed of northern Alabama, United States collected in summer 2018 used in the calculation of an Index of Biotic Integrity for biological monitoring. * New in this V2: In response to peer review at a journal and associated revised statistical analyses, we have added four variables to the file Curtis_etal_IBImetrics.csv and edited the Curtis_etal_readme.txt to explain these variables. The files Curtis_etal_FishDetections.csv and Curtis_etal_FishReadsbySite.csv remain unchanged. - 4 new variables in Curtis_etal_IBImetrics.csv are: fishIBI_noDELT, MaxHab, Stressor, and Distance.

Microbial cell factories have been extensively engineered to produce free fatty acids (FFAs) as key components of crucial nutrients, soaps, industrial chemicals, and fuels. However, our ability to control the composition of microbially synthesized FFAs is still limited, particularly, for producing medium‐chain fatty acids (MCFAs). This is mainly due to the lack of high‐throughput approaches for FFA analysis to engineer enzymes with desirable product specificity. Here we report a mass spectrometry (MS)‐based method for rapid profiling of MCFAs in Saccharomyces cerevisiae by using membrane lipids as a proxy. In particular, matrix‐assisted laser desorption/ionization time‐of‐flight (MALDI‐ToF) MS was used to detect shorter acyl chain phosphatidylcholines from membrane lipids and a higher m/z peak ratio at 730 and 758 was used as an indication for improved MCFA production. This colony‐based method can be performed at a rate of ~2 s per sample, representing a substantial improvement over gas chromatography‐MS (typically >30 min per sample) as the gold standard method for FFA detection. To demonstrate the power of this method, we performed site‐saturation mutagenesis of the yeast fatty acid synthase and identified nine missense mutations that resulted in improved MCFA production relative to the wild‐type strain. Colony‐based MALDI‐ToF MS screening provides an effective approach for engineering microbial fatty acid compositions in a high‐throughput manner.

This project evaluates the quality of retraction indexing metadata in Crossref. We investigated 208 DOIs that were indexed as retracted in Crossref in our April 2023 union list (Schneider et al., 2023), but were no longer indexed as retracted in the July 2024 union list (Salami et al., 2024), despite still being covered in the Crossref database. Therefore, we manually checked the current retraction status of these 208 DOIs on their publishers’ websites to ascertain their actual status.

This work evaluates the consistency and reliability of the title flag, i.e., retraction labeling that appears in the title of retracted publications, using 925 sampled retracted publications indexed in the Crossref only (Lee & Schneider, 2023), that are indexed in three other sources, Retraction Watch, Scopus, and Web of Science as of April 2023. We presume the retraction status of an item based on its title flag. For example, the flag "removal notice" is a retraction notice, and "retracted article" is a retracted paper. We compared the item's likely retraction status from the flag with the item's actual retraction status from the publisher's website.

Plant bioengineering is a time-consuming and labor-intensive process with no guarantee of achieving desired traits. Here, we present a fast, automated, scalable, high-throughput pipeline for plant bioengineering (FAST-PB) in maize (Zea mays) and Nicotiana benthamiana. FAST-PB enables genome editing and product characterization by integrating automated biofoundry engineering of callus and protoplast cells with single-cell matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). We first demonstrated that FAST-PB could streamline Golden Gate cloning, with the capacity to construct 96 vectors in parallel. Using FAST-PB in protoplasts, we found that PEG2050 increased transfection efficiency by over 45%. For proof-of-concept, we established a reporter-gene-free method for CRISPR editing and phenotyping via mutation of high chlorophyll fluorescence 136. We show that diverse lipids were enhanced up to 6-fold using CRISPR activation of lipid controlling genes. In callus cells, an automated transformation platform was employed to regenerate plants with enhanced lipid traits through introducing multigene cassettes. Lastly, FAST-PB enabled high-throughput single-cell lipid profiling by integrating MALDI-MS with the biofoundry, protoplast, and callus cells, differentiating engineered and unengineered cells using single-cell lipidomics. These innovations massively increase the throughput of synthetic biology, genome editing, and metabolic engineering and change what is possible using single-cell metabolomics in plants.

The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids. However, its broader application is hampered by the lack of efficient genetic tools to enable sophisticated metabolic manipulations. We recently constructed an episomal plasmid based on the autonomously replicating sequence (ARS) from Saccharomyces cerevisiae (ScARS) in I. orientalis and developed a CRISPR/Cas9 system for multiplexed gene deletions. Here we report three additional genetic tools including: (1) identification of a 0.8 kb centromere-like (CEN-L) sequence from the I. orientalis genome by using bioinformatics and functional screening; (2) discovery and characterization of a set of constitutive promoters and terminators under different culture conditions by using RNA-Seq analysis and a fluorescent reporter; and (3) development of a rapid and efficient in vivo DNA assembly method in I. orientalis, which exhibited ~100% fidelity when assembling a 7 kb-plasmid from seven DNA fragments ranging from 0.7 kb to 1.7 kb. As proof of concept, we used these genetic tools to rapidly construct a functional xylose utilization pathway in I. orientalis.

Sugar alcohols are commonly used as low-calorie sweeteners and can serve as potential building blocks for bio-based chemicals. Previous work has shown that the oleaginous yeast Rhodosporidium toruloides IFO0880 can natively produce arabitol from xylose at relatively high titers, suggesting that it may be a useful host for sugar alcohol production. In this work, we explored whether R. toruloides can produce additional sugar alcohols. Rhodosporidium toruloides is able to produce galactitol from galactose. During growth in nitrogen-rich medium, R. toruloides produced 3.2 ± 0.6 g/L, and 8.4 ± 0.8 g/L galactitol from 20 to 40 g/L galactose, respectively. In addition, R. toruloides was able to produce galactitol from galactose at reduced titers during growth in nitrogen-poor medium, which also induces lipid production. These results suggest that R. toruloides can potentially be used for the co-production of lipids and galactitol from galactose. We further characterized the mechanism for galactitol production, including identifying and biochemically characterizing the critical aldose reductase. Intracellular metabolite analysis was also performed to further understand galactose metabolism. Rhodosporidium toruloides has traditionally been used for the production of lipids and lipid-based chemicals. Our work demonstrates that R. toruloides can also produce galactitol, which can be used to produce polymers with applications in medicine and as a precursor for anti-cancer drugs. Collectively, our results further establish that R. toruloides can produce multiple value-added chemicals from a wide range of sugars.