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Summary The development and adoption of hybrid seed technology have led to dramatic increases in agricultural productivity. However, it has been a challenge to develop a commercially viable platform for the production of hybrid wheat (Triticum aestivum) seed due to wheat's strong inbreeding habit. Recently, a novel platform for commercial hybrid seed production was described. This hybridization platform utilizes nuclear male sterility to force outcrossing and has been applied to maize and rice. With the recent molecular identification of the wheat male fertility gene Ms1, it is now possible to extend the use of this novel hybridization platform to wheat. In this report, we used the CRISPR/Cas9 system to generate heritable, targeted mutations in Ms1. The introduction of biallelic frameshift mutations into Ms1 resulted in complete male sterility in wheat cultivars Fielder and Gladius, and several of the selected male‐sterile lines were potentially non‐transgenic. Our study demonstrates the utility of the CRISPR/Cas9 system for the rapid generation of male sterility in commercial wheat cultivars. This represents an important step towards capturing heterosis to improve wheat yields, through the production and use of hybrid seed on an industrial scale.

This study focuses on the determination of cavitation rate for samples produced by Direct Metal Laser Sintering (DMLS) of MS1 maraging steel powder without any postprocessing thermal or mechanical treatment (as-built). The test samples, fabricated using an EOSINT M280 machine in a nitrogen atmosphere, were evaluated for their cavitation resistance using the ultrasonic vibration method as proposed by the ASTM G32 standard. Surface characterization to determine the duration of the experiment, as well as during and after the experiment to analyze changes in surface characteristics, was conducted using Scanning Electron Microscopy (SEM). Additionally, chemical analysis was performed using the Energy Dispersive X-ray Spectroscopy (EDS) method before and after the experiment to identify any changes and locations of significant surface damage. The mass loss of the samples over time was measured to calculate the cavitation rate. The goal of this research was to explore the potential applications of laser-sintered MS1 steel powder for manufacturing machine components and parts that are subject to cavitation erosion, due to the numerous benefits of additive manufacturing over traditional conventional technologies.

Reproducible quantification of large biological cohorts is critical for clinical/pharmaceutical proteomics yet remains challenging because most prevalent methods suffer from drastically declined commonly quantified proteins and substantially deteriorated quantitative quality as cohort size expands. MS2-based data-independent acquisition approaches represent tremendous advancements in reproducible protein measurement, but often with limited depth. We developed IonStar, an MS1-based quantitative approach enabling in-depth, high-quality quantification of large cohorts by combining efficient/reproducible experimental procedures with unique data-processing components, such as efficient 3D chromatographic alignment, sensitive and selective direct ion current extraction, and stringent postfeature generation quality control. Compared with several popular label-free methods, IonStar exhibited far lower missing data (0.1%), superior quantitative accuracy/precision [∼5% intragroup coefficient of variation (CV)], the widest protein abundance range, and the highest sensitivity/specificity for identifying protein changes (<5% false altered-protein discovery) in a benchmark sample set (n = 20). We demonstrated the usage of IonStar by a large-scale investigation of traumatic injuries and pharmacological treatments in rat brains (n = 100), quantifying >7,000 unique protein groups (>99.8% without missing data across the 100 samples) with a low false discovery rate (FDR), two or more unique peptides per protein, and high quantitative precision. IonStar represents a reliable and robust solution for precise and reproducible protein measurement in large cohorts.

Widespread deployment of wheat hybrids has been limited by high seed production costs in a commercial environment. This is due to poor outcrossing characteristics of wheat which can, in part, be attributed to poor female organ receptivity to airborne pollen. Here we underpin a detailed characterization of cross-pollination seed set progression in both emasculated wild-type, and a genetically male-sterile wheat line with a comprehensive RNA-Seq and co-expression analysis. We identify three clear phases in the fertilization window: stigma development and onset of receptivity, effective pollination period, and senescence onset and subsequent tissue deterioration. By profiling female-specific marker genes, we determined that physical emasculation of wild-type ‘cv. Chris’ ( Ms1 ) advances female development 2 to 3 days relative to genic male sterility ( ms1d ). Gene networks linked to gibberellic acid homeostasis were found associated with styloid and stigma hair elongation, and increased expression of peroxidases associated with peak receptivity (3 days after floret gaping). Further, changes in the expression of exocyst complex components coincided with the onset of stigma hair senescence. Together the applied clustering and differential expression strategy identified biologically meaningful expression signatures, particularly in stigma hairs, that correlate with the effective pollination period and the observed cross-pollination seed set. Overall, the datasets represent a valuable resource to help identify and/or engineer improved cross-pollination seed set, and ultimately leading to greater insights into this complex biological process and a more cost-effective hybrid seed production system.

Male sterility is an essential trait in hybrid seed production, especially for monoclinous and autogamous food crops. Soybean male-sterile ms1 mutant has been known for more than 50 years and could be instrumental in making hybrid seeds. However, the gene responsible for the male-sterile phenotype has remained unknown. Here, we report the map-based cloning and characterization of the MS1 gene in soybean. MS1 encodes a kinesin protein and localizes to the nucleus, where it is required for the male meiotic cytokinesis after telophase II. We further substantiated that MS1 colocalizes with microtubules and is essential for cell plate formation in soybean male gametogenesis through immunostaining. Both ms1 and CRISPR/Cas9 knockout mutants show complete male sterility but are otherwise phenotypically normal, making them perfect tools for producing hybrid seeds. The identification of MS1 has the practical potential for assembling the sterility system and speeding up hybrid soybean breeding.

Purpose This study aims to further the understanding of support structures and the likely impacts on maraging steel MS1 parts fabricated by selective laser melting (SLM) at 45°, 60° and 75° building angles. Design/methodology/approach Two groups of samples, one group with support structures and the other group without support structures, were designed with the same specifications and printed under the same conditions by SLM at 45°, 60° and 75° building angles. Differences in dimensional accuracy, surface roughness, Vickers microhardness, residual stress and microstructure were compared between groups. Findings The results showed that with support structures, more accurate dimension and slightly higher Vickers microhardness could be obtained. Larger compressive stress dominated and was more uniformly distributed on the supporting surface. Without support structures, the dimension became more precise as the building angle increased and alternating compressive and tensile stress was unevenly distributed on the supporting surface. In addition, the surface roughness of the outer surface decreased with the increase of the built angle, regardless of the support structures. Furthermore, whether the building angle was 45°, 60° or 75°, the observed microstructures revealed that the support structures altered the orientation of the molten pool and the direction of grain growth. Originality/value This paper studies the influence of support structures on the workpieces printed at different building angles. Support structures affect the residual stress distribution, heat dissipation rate and microstructure of the parts, and thus affecting the printing quality. Therefore, it is necessary to balance the support strategy and printing quality to better apply or design the support structures in SLM.

Metal powders for additive manufacturing are expensive, and producing new ones from mined metals has a negative ecological impact. In this work, recycled and reused metal powders from MS1 steel for direct metal laser sintering (DMLS) 3D printing were evaluated in the laboratory. The powders were recycled by melting followed by gas atomizing. Virgin, recycled, and reused metal powders were evaluated using scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), metallography analysis, microhardness measurements, particle size distribution (PSD), shape factor by digital image processing (DIP), and flowability testing. The results showed that the particle distribution was modified after recycling. Kurtosis analysis revealed a reduction from −0.64 for virgin powders to −1.29 for recycled powders. The results demonstrated a positive skewness, indicating that the recycled powder contained a greater proportion of smaller particles. The shape factor was also modified and changed from 1.57 for virgin powders to 1.28 for recycled powders. The microstructure also changed, and austenite was found in the recycled powders. The microhardness of recycled powder decreased by 39% compared to the virgin powder. Recycled powders did not flow, using two different funnels to evaluate their flowability. The flowability of used powder was reduced from 4.3 s to 2.9 s.

Proper sample preparation in proteomic workflows is essential to the success of modern mass spectrometry experiments. Complex workflows often require reagents which are incompatible with MS analysis (e.g., detergents) necessitating a variety of sample cleanup procedures. Efforts to understand and mitigate sample contamination are a continual source of disruption with respect to both time and resources. To improve the ability to rapidly assess sample contamination from a diverse array of sources, I developed a molecular library in Skyline for rapid extraction of contaminant precursor signals using MS1 filtering. This contaminant template library is easily managed and can be modified for a diverse array of mass spectrometry sample preparation workflows. Utilization of this template allows rapid assessment of sample integrity and indicates potential sources of contamination. Graphical Abstract ᅟ

Background Mass spectrometry (MS) uses mass-to-charge ratios of measured particles to decode the identities and quantities of molecules in a sample. Interpretation of raw MS depends upon data processing algorithms that render it human-interpretable. Quantitative MS workflows are complex experimental chains and it is crucial to know the performance and bias of each data processing method as they impact accuracy, coverage, and statistical significance of the result. Creation of the ground truth necessary for quantitatively evaluating MS1-aware algorithms is difficult and tedious task, and better software for creating such datasets would facilitate more extensive evaluation and improvement of MS data processing algorithms. Results We present JS-MS 2.0, a software suite that provides a dependency-free, browser-based, one click, cross-platform solution for creating MS1 ground truth. The software retains the first version’s capacity for loading, viewing, and navigating MS1 data in 2- and 3-D, and adds tools for capturing, editing, saving, and viewing isotopic envelope and extracted isotopic chromatogram features. The software can also be used to view and explore the results of feature finding algorithms. Conclusions JS-MS 2.0 enables faster creation and inspection of MS1 ground truth data. It is publicly available with an MIT license at github.com/optimusmoose/jsms.

Pollen allergy caused by sugi (Japanese cedar, Cryptomeria japonica ) is a serious problem in Japan. One of the measures against pollinosis is the use of male-sterile plants (MSPs; pollen-free plants). In this context, the development of a novel technique for the efficient production of sugi MSPs, which combines marker-assisted selection (MAS) with somatic embryogenesis (SE), was recently reported by our research group. To improve the efficiency of MSP production, in this paper we report improved MAS for male-sterile individuals from embryogenic cells, cotyledonary embryos, and somatic plants of sugi using a newly developed marker in the form of the causative mutation of MS1 itself, selecting individuals with ms1-1 and ms1-2 male-sterile mutations. We also describe simplified methods for extracting DNA from different plant materials and for MAS using LAMP diagnostics. Finally, we show that MAS can be efficiently performed using the one-step indel genotyping (ING) marker developed in this study and using InstaGene for DNA extraction. The combination of SE and 100% accurate marker selection during the embryogenic cell stage enables the mass production of MS1 male-sterile sugi seedlings.