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Vicia sativa L., commonly known as the common vetch, is an annual, self-pollinating legume used primarily as fodder both by livestock and wildlife. Additionally, it contributes to environmental balance through nitrogen fixation and the improvement of soil properties. The phenotypic, genetic, and metabolite variability among four advanced lines (BK45, BK29, BK23, BK27) and two commercial varieties (M-6900, BI-65) of V. sativa were evaluated in order to be used for future breeding programs aimed at producing genetically improved varieties. BK45 was the most promising line due to its high genetic polymorphism, but also because it exhibited a significant amount of seed production and high seed quality based on its metabolomics profile. A stepwise multiple regression analysis (MRA) revealed a relationship between SCoT alleles, seed, and biomass yield. Additionally, several statistically significant marker bands linked to metabolites were found using the SCoT marker analysis. Hence, data assessed via MRA may be helpful in marker-assisted breeding programs. Finally, the two commercial varieties can be further exploited in breeding programs due to their high genetic diversity. [ABSTRACT FROM AUTHOR]

Congenital birth defects contribute significantly to preterm birth, stillbirth, perinatal death, infant mortality, and adult disability. As a first step to exploring the mechanisms underlying this major clinical challenge, we analyzed the embryonic phenotypes of lethal strains generated by random mutagenesis. In this study, we report the gross embryonic and perinatal phenotypes of 55 lethal strains randomly picked from a collection of mutants that carry piggyBac (PB) transposon inserts. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses suggested most of the analyzed mutations hit genes involved in heart and nervous development, or in Notch and Wnt signaling. Among them, 12 loci are known to be associated with human diseases. We confirmed 53 strains as embryonic or perinatal lethal, while others were subviable. Gross morphological phenotypes such as body size abnormality (29/55, 52.73%), growth or developmental delay (35/55, 63.64%), brain defects (9/55, 16.36%), vascular/heart development (31/55, 56.36%), and other structural defects (9/55, 16.36%) could be easily observed in the mutants, while three strains showed phenotypes similar to those of human patients. Furthermore, we detected body weight or body composition alterations in the heterozygotes of eight strains. One of them was the TGF-β signaling gene Smad2. The heterozygotes showed increased energy expenditure and a lower fat-to-body weight ratio compared to wild-type mice. This study provided new insights into mammalian embryonic development and will help understand the pathology of congenital birth defects in humans. In addition, it expanded our understanding of the etiology of obesity. [ABSTRACT FROM AUTHOR]

Understanding the genetic basis of salt resistance in crops is crucial for agricultural productivity. This study investigates the phenotypic and genetic basis of salt stress response in rice (Oryza sativa L.), focusing on germination and seedling traits. Under salt stress conditions, significant differences were observed in seed germination and seedling traits between parental LH99 (Indica rice LuHui 99) and SN265 (japonica rice ShenNong 265). Transgressive segregation was evident within the RIL population, indicating complex genetic interactions. Nine QTLs were detected at germination and seedling stages under salt stress, namely qSGE5 and qSGE7 for seed germination energy (SGE); qSGP7 for seed germination percentage (SGP); qSSH7, qSSH9-1, and qSSH9-2 for seeding height (SSH); qSRN6 for root number (SRN); and qSDW6 and qSDW9 for dry weight (SDW). Among them, qSSH9-1 and qSDW9 were localized in the same interval, derived from the salt-resistant parent SN265. PCA revealed distinct trait patterns under salt stress, captured by six PCs explaining 81.12% of the total variance. PC composite scores were used to localize a QTL associated with early salt resistance in rice qESC9, which was located in the same interval as qSSH9-1 and qSDW9, and was subsequently unified under the name qESC9, an important QTL for early-growth salt tolerance in rice. Correlation analysis also confirmed a relationship between alleles of qESC9 and the resistance to salt, underscoring the critical role this locus plays in the determination of overall salt tolerance in rice. Physiological analyses of extreme phenotype lines highlighted the importance of ion exclusion mechanisms in salt-resistant lines, while salt-susceptible lines exhibited elevated oxidative stress and impaired antioxidant defense, contributing to cellular damage. This comprehensive analysis sheds light on the genetic and physiological mechanisms underlying salt stress response in rice, providing valuable insights for breeding programs aimed at enhancing salt resistance in rice. [ABSTRACT FROM AUTHOR]

Tea plant (Camellia sinensis [L.]) is one of the most important crops in China, and tea branch is an important agronomic trait that determines the yield of tea plant. In previous work focused on GWAS that detecting GWAS signals related to plant architecture through whole genome re‐sequencing of ancient tea plants, a gene locus TEA 029928 significantly related to plant type was found. Sequence alignment results showed that this gene belonged to the F‐box family. We named it CsBRC. CsBRC‐GFP fusion proteins were mainly localized in the plasma membrane. By comparing the phenotypes of CsBRC transgenic tobacco and WT tobacco, it was found that the number of branches of transgenic tobacco was significantly higher than that of wild‐type tobacco. Through RNA‐seq analysis, it was found that CsBRC affects the branching development of plants by regulating the expression of genes related to brassinosteroid synthesis pathway in plants. In addition, overexpression of CsBRC in rice could increase tiller number, grain length and width, and 1,000‐grain weight. [ABSTRACT FROM AUTHOR]

Key Message: Innovatively, we consider stomatal detection as rotated object detection and provide an end-to-end, batch, rotated, real-time stomatal density and aperture size intelligent detection and identification system, RotatedeStomataNet. Stomata acts as a pathway for air and water vapor in the course of respiration, transpiration, and other gas metabolism, so the stomata phenotype is important for plant growth and development. Intelligent detection of high-throughput stoma is a key issue. Nevertheless, currently available methods usually suffer from detection errors or cumbersome operations when facing densely and unevenly arranged stomata. The proposed RotatedStomataNet innovatively regards stomata detection as rotated object detection, enabling an end-to-end, real-time, and intelligent phenotype analysis of stomata and apertures. The system is constructed based on the Arabidopsis and maize stomatal data sets acquired destructively, and the maize stomatal data set acquired in a non-destructive way, enabling the one-stop automatic collection of phenotypic, such as the location, density, length, and width of stomata and apertures without step-by-step operations. The accuracy of this system to acquire stomata and apertures has been well demonstrated in monocotyledon and dicotyledon, such as Arabidopsis, soybean, wheat, and maize. The experimental results that the prediction results of the method are consistent with those of manual labeling. The test sets, the system code, and their usage are also given (). [ABSTRACT FROM AUTHOR]

The probiotic potential of Lactobacillus helveticus LH10, derived from vinegar Pei, a brewing mixture, was assessed through genotype and phenotype analyses. The assembled genome was comprised of 1,810,276 bp and predicted a total of 2044 coding sequences (CDSs). Based on the whole genome sequence analysis, two bacteriocin gene clusters were identified, while no pathogenic genes were detected. In in vitro experiments, L. helveticus LH10 exhibited excellent tolerance to simulated gastrointestinal fluid, a positive hydrophobic interaction with xylene, and good auto-aggregation properties. Additionally, this strain demonstrated varying degrees of resistance to five antibiotics, strong antagonistic activity against four tested pathogens, and no hemolytic activity. Therefore, L. helveticus LH10 holds great promise as a potential probiotic candidate deserving further investigation for its beneficial effects on human health. [ABSTRACT FROM AUTHOR]

This study constructed a high-throughput method for the acquisition and analysis of three-dimensional phenotypes of cotton, and proposes a method for evaluating the degree of wilting of cotton varieties based on phenotype. The upgraded version of the self-developed data acquisition platform MVS-Pheno V2 was used to continuously collect point cloud data. PointSegAt deep learning network model was used to establish plant stem and leaf segmentation and leaf overlap distinction models, realising the segmentation of cotton plant stems and leaves and the distinction of leaf overlap. In addition, an algorithm called "Active Boundary Segmentation" has been developed, which achieved automatic segmentation of overlapping cotton leaves. Based on point cloud technology, the automation of plant height, leaf count, and wilted leaf area based on voxels was realised, and a set of wilt measurement methods for cotton plants was designed. The results show that the PointSegAt model proposed has good performance in stem and leaf segmentation, with a segmentation accuracy of 0.995 and mean intersection over union of 0.924. In terms of single leaf segmentation, the average accuracy reached 0.95, and the average f1-score reached 0.94. Compared with manual measurements of plant height, leaf count, leaf area, and canopy area, the correlation coefficients were 0.99, 0.96, 0.90, and 0.99, respectively, and the root mean square errors were 0.01, 0.04, 0.19, and 0.02, respectively. Finally, the proposed method was used to perform wilting quantification experiments on two different varieties of cotton plants, and quantitative analysis of drought resistance of different varieties was conducted. • Addressing challenges in plant point cloud data acquisition through device upgrades. • Introducing an organ-level segmentation method for cotton plants. • Automating phenotype calculation and wilting degree measurement. • Developing Cotton MVS software suite. [ABSTRACT FROM AUTHOR]

Abstract Tea plant (Camellia sinensis [L.]) is one of the most important crops in China, and tea branch is an important agronomic trait that determines the yield of tea plant. In previous work focused on GWAS that detecting GWAS signals related to plant architecture through whole genome re‐sequencing of ancient tea plants, a gene locus TEA 029928 significantly related to plant type was found. Sequence alignment results showed that this gene belonged to the F‐box family. We named it CsBRC. CsBRC‐GFP fusion proteins were mainly localized in the plasma membrane. By comparing the phenotypes of CsBRC transgenic tobacco and WT tobacco, it was found that the number of branches of transgenic tobacco was significantly higher than that of wild‐type tobacco. Through RNA‐seq analysis, it was found that CsBRC affects the branching development of plants by regulating the expression of genes related to brassinosteroid synthesis pathway in plants. In addition, overexpression of CsBRC in rice could increase tiller number, grain length and width, and 1,000‐grain weight.

Vicia sativa L., commonly known as the common vetch, is an annual, self-pollinating legume used primarily as fodder both by livestock and wildlife. Additionally, it contributes to environmental balance through nitrogen fixation and the improvement of soil properties. The phenotypic, genetic, and metabolite variability among four advanced lines (BK45, BK29, BK23, BK27) and two commercial varieties (M-6900, BI-65) of V. sativa were evaluated in order to be used for future breeding programs aimed at producing genetically improved varieties. BK45 was the most promising line due to its high genetic polymorphism, but also because it exhibited a significant amount of seed production and high seed quality based on its metabolomics profile. A stepwise multiple regression analysis (MRA) revealed a relationship between SCoT alleles, seed, and biomass yield. Additionally, several statistically significant marker bands linked to metabolites were found using the SCoT marker analysis. Hence, data assessed via MRA may be helpful in marker-assisted breeding programs. Finally, the two commercial varieties can be further exploited in breeding programs due to their high genetic diversity.

Abstract Layered double hydroxides (LDHs) have been widely used in the field of plant engineering, such as DNA/RNA transformation and enhancing plant disease resistance. However, few studies have examined the direct effects of LDHs on plants and their potential utility as nanofertilizers. In this study, the retention capacity of Cu/Fe-layered double hydroxide nanoparticles (CuFe-LDHs) was assessed by comparative experiments on vegetables. The results showed that the retention of CuFe-LDHs in leafy vegetables was high, such as lettuce. Phenotypic analysis revealed that the fresh and dry weights of lettuce leaves were both increased by spraying 10–100 μg/mL CuFe-LDHs. Using the optimal concentration of 10 μg/mL, we conducted further experiments to elucidate the mechanism of CuFe-LDHs promoting lettuce growth. It was found that the application of CuFe-LDHs had a significant effect on growth and induced physiological, transcriptomic, and metabolomic changes, including an increase in the chlorophyll b content, net photosynthetic rate, and intercellular carbon dioxide concentration, as well as modifications in gene expression patterns and metabolite profiles. This work provides compelling evidence that CuFe-LDHs can efficiently adsorb on the surface of lettuce leaves through hydrogen bonding, promote lettuce growth, mitigate the toxicity of heavy metal ions compared to their raw materials at the same concentration and offer a molecular-scale insight into the response of leafy vegetables to CuFe-LDHs. Graphical Abstract

Identification and improvement of salt-tolerant melon (Cucumis melo L.) genotypes is needed to temper salinization threats to growth and physiological development under cultivation conditions. To assess the potential of five inbred lines of melon, a factorial experiment with different salinity treatments was performed. Plants exposed to NaCl showed a different leaf color and phenotypic changes compared to plants in the control group. The inbred line Cm.UT.Kh at 14 ds. m−1 had the highest significant necrosis ratio compared to other inbreeds. LAB and RGB leaf colors showed a significant high coefficient of determination for the regression models selected, which indicated their useful information for assessing salinity effects in melon leaves. Generally, the drawbacks of high-salt treatment in all five inbred lines manifested in decreased total dry and fresh weight of shoots and roots as well as reduced leaf area and plant height compared to the controls. The lowest reduction in leaf leakage was recorded for Cm.UT.Gr and Cm.UT.Jaf. Furthermore, Cm.UT.Gr and Cm.UT.1025 showed the highest membrane stability with increasing salt concentration. The potassium content of leaves revealed positive correlations with high biomass production and membrane stability index, but negative correlations with Na+ leaf content and electrolyte leakage. Principal component analysis (PCA) results for growth parameters, leaf ion content, membrane stability index, and root characteristics could help to indirectly identify salt-tolerant plants. The results of appropriate inbred melon lines implies that selecting the right varieties might lead to higher salinity tolerance in future melon hybridization and breeding programs. [ABSTRACT FROM AUTHOR]

Simple Summary: Ovum pick up and in vitro embryo production (OPU-IVEP) is a critical technique in the dairy industry that is attracting a lot of attention, especially in China's dairy industry. The aim of the technique is to produce as many embryos with excellent genetic material as possible. However, there is a large variation in the ability of donor cows to produce embryos, and this ability is heritable. In this case, genetic selection and animal breeding will be beneficial to improving efficiency. Considering the large differences among different populations, the genetic parameters regarding the Chinese population are currently essential for genetic selection in the future. In this study, we analyzed the phenotypic and genetic characteristics of in vitro embryo production traits in Chinese Holstein cattle. The results of this study may help to understand the breeding potential of in vitro embryo production traits. Ovum pick up and in vitro embryo production (OPU-IVEP) is an essential technique in the dairy industry. The production efficiency of OPU-IVEP is significantly influenced by various factors, and phenotypic and genetic characteristics are highly variable in different populations. The objectives of this study were (1) to reveal the phenotypic characteristics, including population distribution, and impacts of donor age and month on in vitro embryo production and (2) to estimate genetic parameters for five in vitro embryo production traits in Chinese Holstein cattle. A total of 7311 OPU-IVEP records of 867 Holstein heifers from August 2021 to March 2023 were collected in this study. Five in vitro embryo production traits were defined, including the number of cumulus–oocyte complexes (NCOC), the number of cleaved embryos (NCLV), the number of grade I embryos (NGE), and the proportion of NCLV to NCOC (PCLV) and NGE to NCOC (PGE). A univariate repeatability animal model was employed to estimate heritability and repeatability, and a bivariate repeatability animal model was employed to estimate the genetic correlations among five in vitro embryo production traits. It was found that the in vitro embryo production traits were significantly influenced by season, as the NGE and PGE were significantly decreased from June to August. In addition, the production efficiency of OPU-IVEP was also influenced by donor age. On the observed scale, the estimates of heritability were 0.33 for NCOC, 0.24 for NCLV, 0.16 for NGE, 0.06 for PCLV, and 0.10 for PGE, respectively. On the log-transformed scale, the estimates of heritability of NCOC, NCLV, and NGE were 0.34, 0.18, and 0.13. The genetic correlations among NCOC, NCLV, and NGE ranged from 0.61 (NCLV and NGE) to 0.95 (NCOC and NCLV), considering both scales. However, there were low genetic correlations between NCOC and proportion traits (PCLV and PGE) on both the observed scale and the log-transformed scale. In the end, the variation in Chinese Holstein cattle was found to be considerable. The EBV value and average NCOC, NGE, and PGE for the top 10% donors presented extreme differences to those for the bottom 10% donors for NCOC (24.02 versus 2.60), NGE (3.42 versus 0.36), and PGE (30.54% versus 3.46%). Overall, the results of this study reveal that in vitro embryo production traits are heritable with low to high heritability, and the count traits (NCOC, NCLV, and NGE) and proportion traits (PCLV and PGE) reflect different aspects of in vitro embryo production and should be incorporated into genetic selection for improving the embryo production efficiency of dairy cattle. [ABSTRACT FROM AUTHOR]

Layered double hydroxides (LDHs) have been widely used in the field of plant engineering, such as DNA/RNA transformation and enhancing plant disease resistance. However, few studies have examined the direct effects of LDHs on plants and their potential utility as nanofertilizers. In this study, the retention capacity of Cu/Fe-layered double hydroxide nanoparticles (CuFe-LDHs) was assessed by comparative experiments on vegetables. The results showed that the retention of CuFe-LDHs in leafy vegetables was high, such as lettuce. Phenotypic analysis revealed that the fresh and dry weights of lettuce leaves were both increased by spraying 10–100 μg/mL CuFe-LDHs. Using the optimal concentration of 10 μg/mL, we conducted further experiments to elucidate the mechanism of CuFe-LDHs promoting lettuce growth. It was found that the application of CuFe-LDHs had a significant effect on growth and induced physiological, transcriptomic, and metabolomic changes, including an increase in the chlorophyll b content, net photosynthetic rate, and intercellular carbon dioxide concentration, as well as modifications in gene expression patterns and metabolite profiles. This work provides compelling evidence that CuFe-LDHs can efficiently adsorb on the surface of lettuce leaves through hydrogen bonding, promote lettuce growth, mitigate the toxicity of heavy metal ions compared to their raw materials at the same concentration and offer a molecular-scale insight into the response of leafy vegetables to CuFe-LDHs. [ABSTRACT FROM AUTHOR]

Cotton (Gossypium hirsutum L.) seed morphological structure has a significant impact on the germination, growth and quality formation. However, the wide variation of cotton seed morphology makes it difficult to achieve quantitative analysis using traditional phenotype acquisition methods. In recent years, the application of micro-CT technology has made it possible to analyze the threedimensional morphological structure of seeds, and has shown technical advantages in accurate identification of seed phenotypes. In this study, we reconstructed the seed morphological structure based on micro-CT technology, deep neural network Unet-3D model, and threshold segmentation methods, extracted 11 basics phenotypes traits, and constructed three new phenotype traits of seed coat specific surface area, seed coat thickness ratio and seed density ratio, using 102 cotton germplasm resources with clear year characteristics. Our results show that there is a significant positive correlation (P< 0.001) between the cotton seed size and that of the seed kernel and seed coat volume, with correlation coefficients ranging from 0.51 to 0.92, while the cavity volume has a lower correlation with other phenotype indicators (r<0.37, P< 0.001). Comparison of changes in Chinese self-bred varieties showed that seed volume, seed surface area, seed coat volume, cavity volume and seed coat thickness increased by 11.39%, 10.10%, 18.67%, 115.76% and 7.95%, respectively, while seed kernel volume, seed kernel surface area and seed fullness decreased by 7.01%, 0.72% and 16.25%. Combining with the results of cluster analysis, during the hundred-year cultivation history of cotton in China, it showed that the specific surface area of seed structure decreased by 1.27%, the relative thickness of seed coat increased by 8.70%, and the compactness of seed structure increased by 50.17%. Furthermore, the new indicators developed based on micro-CT technology can fully consider the three-dimensional morphological structure and cross-sectional characteristics among the indicators and reflect technical advantages. In this study, we constructed a microscopic phenotype research system for cotton seeds, revealing the morphological changes of cotton seeds with the year in China and providing a theoretical basis for the quantitative analysis and evaluation of seed morphology. [ABSTRACT FROM AUTHOR]

BackgroundWith 9730 protein-coding genes and a nearly complete gene knockout strain collection, Neurospora crassa is a major model organism for filamentous fungi. Despite this abundance of information, the phenotypes of these gene knockout mutants have not been categorized to determine whether there are broad correlations between phenotype and any genetic features.ResultsHere, we analyze data for 10 different growth or developmental phenotypes that have been obtained for 1168 N. crassa knockout mutants. Of these mutants, 265 (23%) are in the normal range, while 903 (77%) possess at least one mutant phenotype. With the exception of unclassified functions, the distribution of functional categories for genes in the mutant dataset mirrors that of the N. crassa genome. In contrast, most genes do not possess a yeast ortholog, suggesting that our analysis will reveal functions that are not conserved in Saccharomyces cerevisiae. To leverage the phenotypic data to identify pathways, we used weighted Partitioning Around Medoids (PAM) approach with 40 clusters. We found that genes encoding metabolic, transmembrane and protein phosphorylation-related genes are concentrated in subsets of clusters. Results from K-Means clustering of transcriptomic datasets showed that most phenotypic clusters contain multiple expression profiles, suggesting that co-expression is not generally observed for genes with shared phenotypes. Analysis of yeast orthologs of genes that co-clustered in MAPK signaling cascades revealed potential networks of interacting proteins in N. crassa.ConclusionsOur results demonstrate that clustering analysis of phenotypes is a promising tool for generating new hypotheses regarding involvement of genes in cellular pathways in N. crassa. Furthermore, information about gene clusters identified in N. crassa should be applicable to other filamentous fungi, including saprobes and pathogens.

Photosynthetic organisms must cope with environmental challenges, like those imposed by the succession of days and nights or by sudden changes in light intensities, that trigger global changes in gene expression and metabolism. The photosynthesis machinery is particularly susceptible to environmental changes and adaptation to them often involves redox-sensing proteins that are the targets of reactive oxygen species generated by photosynthesis activity. Here we show that EngA, an essential GTPase and ribosome-assembly protein involved in ribosome biogenesis in bacteria and chloroplasts, also plays a role in acclimatization to environmentally relevant stress in Synechococcus elongatus PCC7942 and that PipX, a promiscuous regulatory protein that binds to EngA, appears to fine-tune EngA activity. During growth in cold or high light conditions, the EngA levels rise, with a concomitant increase of the EngA/PipX ratio. However, a sudden increase in light intensity turns EngA into a growth inhibitor, a response involving residue Cys122 of EngA, which is part of the GD1-G4 motif NKCES of EngA proteins, with the cysteine conserved just in the cyanobacteria-chloroplast lineage. This work expands the repertoire of ribosome-related factors transmitting redox signals in photosynthetic organisms and provides additional insights into the complexity of the regulatory interactions mediated by EngA and PipX. [ABSTRACT FROM AUTHOR]

Marine natural products represent a unique source for clinically relevant drugs due to their vast molecular and mechanistic diversity. ZJ-101 is a structurally simplified analog of the marine natural product superstolide A, isolated from the New Caledonian sea sponge Neosiphonia Superstes. The mechanistic activity of the superstolides has until recently remained a mystery. Here, we have identified potent antiproliferative and antiadhesive effects of ZJ-101 on cancer cell lines. Furthermore, through dose–response transcriptomics, we found unique dysregulation of the endomembrane system by ZJ-101 including a selective inhibition of O-glycosylation via lectin and glycomics analysis. We applied this mechanism to a triple-negative breast cancer spheroid model and identified a potential for the reversal of 3D-induced chemoresistance, suggesting a potential for ZJ-101 as a synergistic therapeutic agent. [ABSTRACT FROM AUTHOR]

The probiotic potential of Lactobacillus helveticus LH10, derived from vinegar Pei, a brewing mixture, was assessed through genotype and phenotype analyses. The assembled genome was comprised of 1,810,276 bp and predicted a total of 2044 coding sequences (CDSs). Based on the whole genome sequence analysis, two bacteriocin gene clusters were identified, while no pathogenic genes were detected. In in vitro experiments, L. helveticus LH10 exhibited excellent tolerance to simulated gastrointestinal fluid, a positive hydrophobic interaction with xylene, and good auto-aggregation properties. Additionally, this strain demonstrated varying degrees of resistance to five antibiotics, strong antagonistic activity against four tested pathogens, and no hemolytic activity. Therefore, L. helveticus LH10 holds great promise as a potential probiotic candidate deserving further investigation for its beneficial effects on human health.

Photosynthetic organisms must cope with environmental challenges, like those imposed by the succession of days and nights or by sudden changes in light intensities, that trigger global changes in gene expression and metabolism. The photosynthesis machinery is particularly susceptible to environmental changes and adaptation to them often involves redox-sensing proteins that are the targets of reactive oxygen species generated by photosynthesis activity. Here we show that EngA, an essential GTPase and ribosome-assembly protein involved in ribosome biogenesis in bacteria and chloroplasts, also plays a role in acclimatization to environmentally relevant stress in Synechococcus elongatus PCC7942 and that PipX, a promiscuous regulatory protein that binds to EngA, appears to fine-tune EngA activity. During growth in cold or high light conditions, the EngA levels rise, with a concomitant increase of the EngA/PipX ratio. However, a sudden increase in light intensity turns EngA into a growth inhibitor, a response involving residue Cys122 of EngA, which is part of the GD1-G4 motif NKCES of EngA proteins, with the cysteine conserved just in the cyanobacteria-chloroplast lineage. This work expands the repertoire of ribosome-related factors transmitting redox signals in photosynthetic organisms and provides additional insights into the complexity of the regulatory interactions mediated by EngA and PipX.

Using high-throughput sequencing technology the changes in bacterial community structure, function and phenotype during the fermentation of medium-temperature Daqu were analyzed. The results showed that the bacterial community of medium-temperature Daqu was mainly composed of Welchiella, Staphylococcus, Escherichia, Lactobacillus, etc. Based on OTU level, the samples of medium-temperature Daqu could be clustered into three stages including early stage (0 d and 3 d), middle stage (9 d and 13 d) and late stage (30 d). Staphylococcus was the main bacteria in the early fermentation stage, weissella was the main dominant bacteria in the middle stage, and muribaculum was the main bacteria in the late fermentation stage. The functional abundance of genes such as amino acid transport and metabolism, carbohydrate transport and metabolism from bacteria inhabiting medium- temperature Daqu was high. The bacterial community in the early and late fermentation stages had higher abundance in gene function such as cell wall / membrane / envelope biogenesis, posttranslational modification, protein turnover, chaperones protein than in the middle fermentation stage. Most microorganisms were facultative anaerobic bacteria in the early stage of fermentation while the aerobic bacteria and anaerobic bacteria dominated in the middle and late stage, respectively. [ABSTRACT FROM AUTHOR]

Lactic acid bacteria are generally regarded as alternatives to antibiotics in livestock and poultry farming, especially Lactobacillus strains, which are safe and have probiotic potential. Although Lactobacillus salivarius has long been proposed to be a probiotic, the understanding of the roles of this species is still in its infancy. Here, a strain of L. salivarius CGMCC20700 isolated from the intestinal mucosa of Yunnan black-bone chicken broilers was investigated in the context of its safety and probiotic characteristics by whole-genome sequencing in parallel with phenotypic analysis. Whole-genome sequencing results showed that L. salivarius CGMCC20700 has a single scaffold of 1,737,577 bp with an average guanine-to-cytosine (GC) ratio of 33.51% and 1,757 protein-coding genes. The annotation of Clusters of Orthologous Groups (COG) classified the predicted proteins from the assembled genome as possessing cellular, metabolic, and information-related functions. Sequences related to risk assessment, such as antibiotic resistance and virulence genes, were identified, and the strain was further confirmed as safe according to the results of antibiotic resistance, hemolytic, and acute oral toxicology tests. Two gene clusters of antibacterial compounds and broad-spectrum antimicrobial activity were identified using genome mining tools and antibacterial spectrum tests. Stress resistance genes, active stressor removal genes, and adhesion related genes that were identified and examined with various phenotypic assays (such as stress tolerance tests in acids and bile salts and auto aggregation and hydrophobicity assays). The strain showed a high survival rate in the presence of bile salts and under acidic conditions and exhibited significant auto aggregation capacity and hydrophobicity. Overall, L. salivarius CGMCC20700 demonstrated excellent safety and probiotic potential at both the genomic and physiological levels and can be considered an appropriate candidate probiotic for livestock and poultry farming. [ABSTRACT FROM AUTHOR]

Flax is a dual-purpose crop that is important for oil and fiber production. The growth habit is one of the crucial targets of selection during flax domestication. Wild hybridization between cultivated flax and wild flax can produce superior germplasms for flax breeding and facilitate the study of the genetic mechanism underlying agronomically important traits. In this study, we used pale flax, Linum grandiflorum, and L. perenne to pollinate Longya-10. Only pale flax interspecific hybrids were obtained, and the trait analysis of the F1 and F2 generations showed that the traits analyzed in this study exhibited disparate genetic characteristics. In the F1 generation, only one trait, i.e., the number of capsules per plant (140) showed significant heterosis, while the characteristics of other traits were closely associated with those of the parents or a decline in hybrid phenotypes. The traits of the F2 generation were widely separated, and the variation coefficient ranged from 9.96% to 146.15%. The quantitative trait locus underlying growth habit was preliminarily found to be situated on chromosome 2 through Bulked-segregant analysis sequencing. Then linkage mapping analysis was performed to fine-map GH2.1 to a 23.5-kb interval containing 4 genes. Among them, L.us.o.m.scaffold22.109 and L.us.o.m.scaffold22.112 contained nonsynonymous SNPs with Dindex=1. Combined with the qRT-PCR results, the two genes might be possible candidate genes for GH2.1. This study will contribute to the development of important germplasms for flax breeding, which would facilitate the elucidation of the genetic mechanisms regulating the growth habit and development of an ideal architecture for the flax plant. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance (AMR) is a growing global health threat. This study investigated antibiotic resistance in E. coli isolates from municipal wastewater (86 isolates) and clinical urinary tract infection (UTI) cases (34 isolates) in a Grenadian community, using data from January 2022 to October 2023. Antibiogram data, assessed per WHO guidelines for Critically Important antimicrobials (CIA), showed the highest resistance levels in both clinical and wastewater samples for ampicillin, followed by amoxicillin/clavulanic acid and nalidixic acid, all classified as Critically Important. Similar resistance was observed for sulfamethoxazole-trimethoprim (highly important) in both groups, with nitrofurantoin showing resistance in the important category. According to the WHO AWaRe classification, ampicillin (ACCESS group) had the highest resistance, while nitrofurantoin had the lowest across all samples. The WATCH group antibiotics, cefuroxime and cefoxitin, showed comparable resistance levels, whereas aztreonam from the RESERVE group (tested only in wastewater) was 100% sensitive. Multiple Antibiotic Resistance (MAR) index analysis revealed that 7% of wastewater and 38.2% of clinical samples had MAR values over 0.2, indicating prior antibiotic exposure in clinical isolates. These parallel patterns in wastewater and clinical samples highlight wastewater monitoring as a valuable tool for AMR surveillance, supporting antibiotic stewardship through ongoing environmental and clinical assessment.

With advances in underlying technologies such as complex multicellular systems, synthetic materials, and bioengineering techniques, we can now generate in vitro miniaturized human tissues that recapitulate the organotypic features of normal or diseased tissues. Importantly, these 3D culture models have increasingly provided experimental access to diverse and complex tissues architectures and their morphogenic assembly in vitro. This review presents an analytical toolbox for biological researchers using 3D modeling technologies through which they can find a collation of currently available methods to phenotypically assess their 3D models in their normal state as well as their response to therapeutic or pathological agents., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The unicellular red alga Cyanidioschyzon merolae is a eukaryotic photosynthetic model organism used for basic and applied cell biology studies. Its nuclear genome can be modified by homologous recombination with exogenously introduced DNA. The comparison of mutants with isogenic strains is critical for reliable genetic analyses; however, this has been impossible thus far. Strain construction and genetic complementation studies take a long time and give rise to unstable phenotypes. In this study, we modified the pMKT vector system described in Takemura et al. (2018) by adding an upstream sequence of the target gene, after the URA5.3T marker, to create a double sandwich of repeat elements at the target gene genomic loci after transformation. 5-fluoroorotic acid selection-based DNA excision yields two mutants simultaneously: an epitope-tagged mutant with only the URA5.3 marker eliminated; and a knockout mutant with both URA5.3T and the target gene's open reading frame eliminated. This system can provide multiple sets of isogenic controls in parallel, and further reveal the essentialness of a gene. Here, we introduce the multi-functional mutant design system Gene Tagging and Knock Out (GTKO), and demonstrate its utility by creating GTKO mutants of the CmBTB1 and CmBTB2 genes in C. merolae., (© The Author(s) 2025. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site–for further information please contact journals.permissions@oup.com.)

The goal of this study was to identify the pathogenic gene variants in female patients with severe X-linked hypohidrotic ectodermal dysplasia (XLHED). Whole-exome sequencing (WES) and Sanger sequencing were used to screen for the pathogenic gene variants. The harmfulness of these variations was predicted by bioinformatics. Then, skewed X-chromosome inactivation (XCI) was measured by PCR analysis of the CAG repeat region in the human androgen receptor (AR) gene in peripheral blood cells. Two novel Ectodysplasin-A (EDA) heterozygous variants (c.588_606del19bp and c.837G>A) and one heterozygous variant (c.1045G>A, rs132630317) were identified in the three female XLHED patients. The bioinformatics analysis showed that these variants might be pathogenic. The tertiary structure analysis showed that these variants could cause structural damage to EDA proteins. Analysis of the skewed X-chromosome inactivation revealed that extreme skewed X-chromosome inactivation was found in patient #35 (98:2), whereas it was comparatively moderate in patients #347 and #204 (21:79 and 30:70). Our results broaden the variation spectrum of EDA and the phenotype spectrum of XLHED, which could help with clinical diagnosis, treatment, and genetic counseling. [ABSTRACT FROM AUTHOR]

Autism Spectrum Disorder (ASD) is a developmental condition characterized by social and communication differences. Recent research suggests ASD affects 1-in-44 children in the United States. ASD is diagnosed more commonly in males, though it is unclear whether this diagnostic disparity is a result of a biological predisposition or limitations in diagnostic tools, or both. One hypothesis centers on the ‘female protective effect,’ which is the theory that females are biologically more resistant to the autism phenotype than males. In this examination, phenotypic data were acquired and combined from four leading research institutions and subjected to multivariate linear discriminant analysis. A linear discriminant model was trained on the training set and then deployed on the test set to predict group membership. Multivariate analyses of variance were performed to confirm the significance of the overall analysis, and individual analyses of variance were performed to confirm the significance of each of the resulting linear discriminant axes. Two discriminant dimensions were identified between the groups: a dimension separating groups by the diagnosis of ASD (LD1: 87% of variance explained); and a dimension reflective of a diagnosis-by-sex interaction (LD2: 11% of variance explained). The strongest discriminant coefficients for the first discriminant axis divided the sample in domains with known differences between ASD and comparison groups, such as social difficulties and restricted repetitive behavior. The discriminant coefficients for the second discriminant axis reveal a more nuanced disparity between boys with ASD and girls with ASD, including executive functioning and high-order behavioral domains as the dominant discriminators. These results indicate that phenotypic differences between males and females with and without ASD are identifiable using parent report measures, which could be utilized to provide additional specificity to the diagnosis of ASD in female patients, potentially leading to more targeted clinical strategies and therapeutic interventions. The study helps to isolate a phenotypic basis for future empirical work on the female protective effect using neuroimaging, EEG, and genomic methodologies.

Accurate identification and monitoring of sunflower capitula are crucial for field phenotypic analysis, cultivation management, phenological monitoring, and yield prediction. Manual observation, however, faces significant challenges due to the complexity of field environments and the morphological diversity of sunflower capitula. Unmanned Aerial Vehicles (UAVs) have emerged as an ideal platform for monitoring sunflower capitula due to their low cost and high spatiotemporal resolution. This study introduces Sunflower-YOLO, an enhanced model based on YOLOv7-tiny, designed for detecting sunflower capitula in UAV remote sensing images. The model effectively identifies sunflower capitula and distinguishes between three specific states: open, half-open, and bud. Sunflower-YOLO incorporates several key improvements: the SiLU activation function replaces the original LeakyReLU, enhancing the model's nonlinear expression capability; a shallow high-resolution feature map and an additional detection head for small targets are introduced during the feature fusion stage to improve the detection performance of small capitula; and the integration of deformable convolution and the SimAM attention mechanism enhances the ELAN structure in the backbone, creating a new DeformAtt-ELAN structure that improves the model's ability to capture morphological variations and reduces noise interference. Experimental results demonstrate that Sunflower-YOLO achieves precision, recall, and mAP@0.5 of 92.3 %, 89.7 %, and 93 %, respectively, marking improvements of 4.2 %, 4.2 %, and 3.7 % over the original YOLOv7-tiny model. The average precision (AP) for the three growth states is 98.7 %, 93.4 %, and 87 %, with AP for the half-open and bud states improving by 6.5 % and 4.7 %, respectively. The model's FLOPs is 17.7 G, its size is 13.8MB, and it achieves an FPS of 188.52. Compared to current mainstream state-of-the-art (SOTA) models for object detection, Sunflower-YOLO achieves the highest mAP@0.5 in detecting multiple types of sunflower capitula. The constructed capitulum density map offers a practical view for observing sunflower growth status. This study highlights the immense potential of combining UAV remote sensing technology with YOLO object detection algorithms in monitoring sunflower capitula and their growth processes, providing an innovative and effective approach for precision agriculture practices. [Display omitted] • Sunflower-YOLO model accurately detects three growth stages of sunflower capitula in UAV images. • SiLU, small target detection head, and DeformAtt-ELAN structure enhance model performance. • Generated sunflower capitula density maps for phenotypic analysis and field management. [ABSTRACT FROM AUTHOR]

Aims: Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) is a life‐threatening, inherited, nonhypertensive arteriole disease of the brain. Therapeutic strategy for CARASIL is limited because its pathogenesis is not clear. We previously reported the first family with CARASIL in China, which involves a high‐temperature requirement serine protease gene mutation (HtrA1L364P). Based on this previous study, we constructed a CARASIL mouse model (Mut‐hHtrA1L364P mouse, hereinafter referred to as Mut). This paper aimed to systematically study the behavior, pathology, and molecular biology of Mut mice and explore the pathogenesis and possible therapeutic strategies of CARASIL. Methods: Food maze and water maze experiments were used in the behavioral studies. Pathological studies were carried out by arteriole labeling staining and electron microscopy. The mRNA and protein expression levels of the key factors of TGF‐β/Smad signaling pathway (TGF‐β, Smad2, Smad3, and Smad4) in the brain of the model mice were detected by immunohistochemistry, real‐time quantitative polymerase chain reaction (RT‐PCR), and Western blot assay. Results: The food maze and water maze experiment data showed significant differences between the Mut and wild‐type (WT) mice in the first time to find food, the time to contact the escape table for the first time, and the number of times to travel in the escape table quadrant (p < 0.001). The results of vascular labeling staining showed that some small arteries in the brain of Mut mice lost normal structure. The results of electron microscopy showed that the cell morphologies in the cortex and hippocampus of Mut mice were abnormal; the number of synapses was reduced; the walls of capillaries, venules, and arterioles thickened; lumen stenosis and other abnormal phenomenon occurred; and lipofuscin deposition and autophagosomes were found in the hippocampus. Immunohistochemistry, RT‐PCR, and Western Blot results showed that the mRNA and protein expression levels of TGF‐β, Smad2, and Smad3 in the brain of Mut mice increased to different degrees. Conclusions: The most significant innovation of this study is the first study on the pathogenesis of CARASIL disease using model animals. The Mut mice can well simulate the pathogenesis of CARASIL in behavioral and pathological aspects. The TGF‐β/Smad signaling pathway, which is involved in the pathogenesis of CARASIL, is abnormally upregulated in the brain of Mut mice. [ABSTRACT FROM AUTHOR]

The purpose of this research was to investigate and identify PAX9 gene variants in four Chinese families with non-syndromic tooth agenesis. We identified pathogenic gene variants by whole-exome sequencing (WES) and Sanger sequencing and then studied the effects of these variants on function by bioinformatics analysis and in vitro experiments. Four novel PAX9 heterozygous variants were identified: two missense variants (c.191G > T (p.G64V) and c.350T > G (p.V117G)) and two frameshift variants (c.352delC (p.S119Pfs*2) and c.648_649insC(p.Y217Lfs*100)). The bioinformatics analysis showed that these variants might be pathogenic. The tertiary structure analysis showed that these four variants could cause structural damage to PAX9 proteins. In vitro functional studies demonstrated that (1) the p.Y217Lfs*100 variant greatly affects mRNA stability, thereby affecting endogenous expression; (2) the p. S119Pfs* 2 variant impairs the subcellular localization of the nuclear expression of the wild-type PAX9 protein; and (3) the four variants (p.G64V, p.V117G, p.S119Pfs*2, and p.Y217Lfs*100) all significantly affect the downstream transcriptional activity of the BMP4 gene. In addition, we summarized and analyzed tooth missing positions caused by PAX9 variants and found that the maxillary second molar (84.11%) and mandibular second molar (84.11%) were the most affected tooth positions by summarizing and analyzing the PAX9-related non-syndromic tooth agenesis positions. Our results broaden the variant spectrum of the PAX9 gene related to non-syndromic tooth agenesis and provide useful information for future genetic counseling. [ABSTRACT FROM AUTHOR]

Wheat (Triticum aestivum L.) is the among the first three agroeconomic important crop plants, with a high carbohydrate and protein content in grains. In this study, 80 cultivars, from the VGRB germplasm collection, were phenotypes in field experiments. Morphological traits were observed on the harvest spike prior to threshing. Awn roughness will be assessed visually and by sliding one's finger along the central part of the awn in the direction from top to bottom. Flowering dates will be retrieved from digitized records of VGRB propagation cycles. Accessions were evaluated in 2020/2021 field trials under a randomized block experimental design. Among the measured phenotypic records there are traits as germination rate, flowering time, disease resistance and yield. Results indicate that VRGB germplasm contains a wide variety of genotypes that have a high agronomical value. This germplasm could be used in future breeding programs. [ABSTRACT FROM AUTHOR]

Under steady-state conditions, conventional CD4+ T lymphocytes are classically divided into naïve (CD44lo CD62Lhi) and memory (CD44hi CD62Llo) cell compartments. While the latter population is presumed to comprise a mixture of distinct subpopulations of explicit foreign antigen (Ag)-specific "authentic" memory and foreign Ag-independent memory-phenotype (MP) cells, phenotypic markers differentially expressed in these two cell types have yet to be identified. Moreover, while MP cells themselves have been previously described as heterogeneous, it is unknown whether they consist of distinct subsets defined by marker expression. In this study, we demonstrate using combined single-cell RNA sequencing and flow cytometric approaches that self-driven MP CD4+ T lymphocytes are divided into CD127hi Sca1lo, CD127hi Sca1hi, CD127lo Sca1hi, and CD127lo Sca1lo subpopulations that are Bcl2lo, while foreign Ag-specific memory cells are CD127hi Sca1hi Bcl2hi. We further show that among the four MP subsets, CD127hi Sca1hi lymphocytes represent the most mature and cell division-experienced subpopulation derived from peripheral naïve precursors. Finally, we provide evidence arguing that this MP subpopulation exerts the highest responsiveness to Th1-differentiating cytokines and can induce colitis. Together, our findings define MP CD4+ T lymphocytes as a unique, self-driven population consisting of distinct subsets that differ from conventional foreign Ag-specific memory cells in marker expression and establish functional relevance for the mature subset of CD127hi Sca1hi MP cells. [ABSTRACT FROM AUTHOR]

Purpose: To compare the clinical characteristics of pediatric urolithiasis patients with positive and negative molecular diagnoses. Methods: The clinical characteristics corresponding to pediatric urolithiasis patients that had undergone exome sequencing at our hospital between January 2016 and May 2021 were collected. Genetic analysis results were used to separate patients into positive and negative molecular diagnosis groups. Multivariate logistic regression analyses adjusted for visiting age, sex, ethnicity, province, and body mass index were used to compare differences in medical history, diagnostic imaging findings, and renal function between individuals with and without molecular diagnoses. Results: In total, 194 patients with pediatric urolithiasis of unknown etiology underwent exome sequencing and were included in the present study, of whom 63 obtained urolithiasis-related molecular diagnoses. Relative to cases without a molecular diagnosis, those with a positive molecular diagnosis were more likely to be associated with a positive family history (OR 2.84, 95% CI 1.29–6.29, p = 0.008), consanguineous parents (OR 24.7, 95% CI 1.34–454, p = 0.002), early onset (OR 1.26, 95% CI 1.09–1.45, p < 0.001), nephrocalcinosis (OR 10.6, 95% CI 3.06–36.6, p < 0.001), cast stone (OR 18.9, 95% CI 4.40–81.1, p < 0.001), multiple stones (OR 13.9, 95% CI 6.39–30.2, p < 0.001), bilateral stones (OR 7.04, 95% CI 3.47–14.2, p < 0.001), a lower estimated glomerular filtration rate (OR 1.17, 95% CI 1.07–1.28, p < 0.001), and chronic kidney disease (OR 26.9, 95% CI 1.42–526, p < 0.001). Conclusion: A positive family history, consanguineous parents, early onset, nephrocalcinosis, severe stone burden, and impaired renal function are signals of concern that are suggestive of inherited urolithiasis. [ABSTRACT FROM AUTHOR]

Abstract Background With 9730 protein-coding genes and a nearly complete gene knockout strain collection, Neurospora crassa is a major model organism for filamentous fungi. Despite this abundance of information, the phenotypes of these gene knockout mutants have not been categorized to determine whether there are broad correlations between phenotype and any genetic features. Results Here, we analyze data for 10 different growth or developmental phenotypes that have been obtained for 1168 N. crassa knockout mutants. Of these mutants, 265 (23%) are in the normal range, while 903 (77%) possess at least one mutant phenotype. With the exception of unclassified functions, the distribution of functional categories for genes in the mutant dataset mirrors that of the N. crassa genome. In contrast, most genes do not possess a yeast ortholog, suggesting that our analysis will reveal functions that are not conserved in Saccharomyces cerevisiae. To leverage the phenotypic data to identify pathways, we used weighted Partitioning Around Medoids (PAM) approach with 40 clusters. We found that genes encoding metabolic, transmembrane and protein phosphorylation-related genes are concentrated in subsets of clusters. Results from K-Means clustering of transcriptomic datasets showed that most phenotypic clusters contain multiple expression profiles, suggesting that co-expression is not generally observed for genes with shared phenotypes. Analysis of yeast orthologs of genes that co-clustered in MAPK signaling cascades revealed potential networks of interacting proteins in N. crassa. Conclusions Our results demonstrate that clustering analysis of phenotypes is a promising tool for generating new hypotheses regarding involvement of genes in cellular pathways in N. crassa. Furthermore, information about gene clusters identified in N. crassa should be applicable to other filamentous fungi, including saprobes and pathogens.

Under steady-state conditions, conventional CD4+ T lymphocytes are classically divided into naïve (CD44lo CD62Lhi) and memory (CD44hi CD62Llo) cell compartments. While the latter population is presumed to comprise a mixture of distinct subpopulations of explicit foreign antigen (Ag)-specific “authentic” memory and foreign Ag-independent memory-phenotype (MP) cells, phenotypic markers differentially expressed in these two cell types have yet to be identified. Moreover, while MP cells themselves have been previously described as heterogeneous, it is unknown whether they consist of distinct subsets defined by marker expression. In this study, we demonstrate using combined single-cell RNA sequencing and flow cytometric approaches that self-driven MP CD4+ T lymphocytes are divided into CD127hi Sca1lo, CD127hi Sca1hi, CD127lo Sca1hi, and CD127lo Sca1lo subpopulations that are Bcl2lo, while foreign Ag-specific memory cells are CD127hi Sca1hi Bcl2hi. We further show that among the four MP subsets, CD127hi Sca1hi lymphocytes represent the most mature and cell division-experienced subpopulation derived from peripheral naïve precursors. Finally, we provide evidence arguing that this MP subpopulation exerts the highest responsiveness to Th1-differentiating cytokines and can induce colitis. Together, our findings define MP CD4+ T lymphocytes as a unique, self-driven population consisting of distinct subsets that differ from conventional foreign Ag-specific memory cells in marker expression and establish functional relevance for the mature subset of CD127hi Sca1hi MP cells.

ABSTRACT Clostridioides difficile is a Gram-positive, spore-forming anaerobic bacteria that is one of the leading causes of antibiotic-associated diarrhea. The cell wall protein 66 gene (cwp66) encodes a cell wall protein, which is the second major cell surface antigen of C. difficile. Although immunological approaches, such as antibodies and purified recombinant proteins, have been implemented to study the role of Cwp66 in cell adhesion, no deletion mutant of the cwp66 gene has yet been characterized. We constructed a cwp66 gene deletion mutant using Clustered Regularly Interspaced Short Palindromic Repeats Cpf1 (CRISPR-Cpf1) system. The phenotypic and transcriptomic changes of the Δcwp66 mutant compared with the wild-type (WT) strain were studied. The deletion of the cwp66 gene led to the decrease of cell adhesive capacity, cell motility, and stresses tolerance (to Triton X-100, acidic environment, and oxidative stress). Interestingly, the Δcwp66 mutant is more sensitive than the WT strain to clindamycin, ampicillin, and erythromycin but more resistant than the latter to vancomycin and metronidazole. Moreover, mannitol utilization capability in the Δcwp66 mutant was lost. Comparative transcriptomic analyses indicated that (i) 22.90-fold upregulation of cwpV gene and unable to express gpr gene were prominent in the Δcwp66 mutant; (ii) the cwp66 gene was involved in vancomycin resistance of C. difficile by influencing the expression of d-Alanine-d-Alanine ligase; and (iii) the mannose/fructose/sorbose IIC and IID components were upregulated in Δcwp66 mutant. The present work deepens our understanding of the contribution of the cwp66 gene to cell adhesion, stress tolerance, antibiotic resistance, and mannitol transportation of C. difficile. IMPORTANCE The cell wall protein 66 gene (cwp66) encodes a cell wall protein, which is the second major cell surface antigen of C. difficile. Although immunological approaches, such as antibodies and purified recombinant proteins, have been implemented to study the role of Cwp66 in cell adhesion, no deletion mutant of the cwp66 gene has yet been characterized. The current study provides direct evidence that the cwp66 gene serves as a major adhesion in C. difficile, and also suggested that deletion of the cwp66 gene led to the decrease of cell adhesive capacity, cell motility, and stresses tolerance (to Triton X-100, acidic environment, and oxidative stress). Interestingly, the antibiotic resistance and carbon source utilization profiles of the Δcwp66 mutant were significantly changed. These phenotypes were detrimental to the survival and pathogenesis of C. difficile in the human gut and may shed light on preventing C. difficile infection.

[Objective] This study investigated the functions of CsMADS08 and its downstream regulatory genes to provide basis for cultivating new cucumber varieties. [Method] Cucumber CsMADS08 gene was introduced to Arabisopis agl8 homozygous mutant by Agrobacterium-mediated method. The CsMADS08 transgenic resistant plants were obtained through screening. The phenotypic changes of the Arabidopsis agl8 mutant plants, transgenic plants and wild-ype plants were compared. The structure of cauline leaves of different plants was observed by microstructures. The expressions of CsMADS08 and downstream regulatory genes SHP1, SHP2, ALC and IND in roots, stems, rosette leaves, stem leaves, flowers and fruits were analyzed by real-time fluorescence quantitative PCR. [Result]There were no significant differences in root length, plant height, number and shape of rosette leaves between ag18 mutant plants, CsMADS08 transgenic plants and wildtype Arabidopsis. Compared with wildtype plants, the bolting and flowering time of 2-8 mutant pants was delayed. There was no remarkable change in the expression of CsMADS08 gene in flowers of the agl8 mutant plants. The bolting and flowering time of CsMADS08 transgenic plants was 4 -- 6 days earlier than that of the agl8 mutant plants and the expression of CsMADS08 gene in the flower of transgenic plants was increased. There were significant differences in number of branches, fruits, number and shape of stem leaves between three types of plants. The CsMADS08 transgenic plants had small and dense leaves, an approximate circular shape, as well as long and full fruit pods without premature cracking. Microscopc observation of stem leaves showed that, the leaf cells of CsMADS08 transgenic plants enlarged and arranged orderly compared with wildtype plants. The vascuar bundle cells increased, and the leaves became thicker. Realtime fluorescent quantitative PCR results showed that CsMADS08 gene expressed in all tissues of the transgenic plants with the highest expression in stem leaves. In comparison with agl8 mutant plants, SHP1, SHP2,ALC and IND genes were up-regulated in flowers and down-regulated in other tissues of the CsMADS08 transgenic plants with the largest decrease in stem leaves. [Conclusion] CsMADS08 gene had the functions of promoting lowering, increasing lateral branches,and regulating leaf morphology and fruit development. [ABSTRACT FROM AUTHOR]

Cyanobacteria, phototrophic organisms performing oxygenic photosynthesis, must adapt their metabolic processes to important environmental challenges, like those imposed by the succession of days and nights. Not surprisingly, certain regulatory proteins are found exclusively in this phylum. One of these unique proteins, PipX, provides a mechanistic link between signals of carbon/nitrogen and of energy, transduced by the signaling protein PII, and the control of gene expression by the global nitrogen regulator NtcA. PII, required for cell survival unless PipX is inactivated or downregulated, functions by protein–protein interactions with transcriptional regulators, transporters, and enzymes. PipX also functions by protein–protein interactions, and previous studies suggested the existence of additional interacting partners or included it into a relatively robust six-node synteny network with proteins apparently unrelated to the nitrogen regulation system. To investigate additional functions of PipX while providing a proof of concept for the recently developed cyanobacterial linkage network, here we analyzed the physical and regulatory interactions between PipX and an intriguing component of the PipX synteny network, the essential ribosome assembly GTPase EngA. The results provide additional insights into the functions of cyanobacterial EngA and of PipX, showing that PipX interacts with the GD1 domain of EngA in a guanosine diphosphate-dependent manner and interferes with EngA functions in Synechococcus elongatus at a low temperature, an environmentally relevant context. Therefore, this work expands the PipX interaction network and establishes a possible connection between nitrogen regulation and the translation machinery. We discuss a regulatory model integrating previous information on PII–PipX with the results presented in this work. [ABSTRACT FROM AUTHOR]

Cyanobacteria, phototrophic organisms performing oxygenic photosynthesis, must adapt their metabolic processes to important environmental challenges, like those imposed by the succession of days and nights. Not surprisingly, certain regulatory proteins are found exclusively in this phylum. One of these unique proteins, PipX, provides a mechanistic link between signals of carbon/nitrogen and of energy, transduced by the signaling protein PII, and the control of gene expression by the global nitrogen regulator NtcA. PII, required for cell survival unless PipX is inactivated or downregulated, functions by protein–protein interactions with transcriptional regulators, transporters, and enzymes. PipX also functions by protein–protein interactions, and previous studies suggested the existence of additional interacting partners or included it into a relatively robust six-node synteny network with proteins apparently unrelated to the nitrogen regulation system. To investigate additional functions of PipX while providing a proof of concept for the recently developed cyanobacterial linkage network, here we analyzed the physical and regulatory interactions between PipX and an intriguing component of the PipX synteny network, the essential ribosome assembly GTPase EngA. The results provide additional insights into the functions of cyanobacterial EngA and of PipX, showing that PipX interacts with the GD1 domain of EngA in a guanosine diphosphate-dependent manner and interferes with EngA functions in Synechococcus elongatus at a low temperature, an environmentally relevant context. Therefore, this work expands the PipX interaction network and establishes a possible connection between nitrogen regulation and the translation machinery. We discuss a regulatory model integrating previous information on PII–PipX with the results presented in this work.

The taproot of radish (Raphanus sativus L.) is an important sink organ; it is morphologically diverse and contains large amounts of secondary metabolites. Sucrose metabolism is believed to be important in the development of sink organs. We measured the amounts of glucose, fructose, and sucrose in the roots of sixty three radish accessions and analyzed the association between the sugar content and the root phenotype. Fructose content correlated with the root color and length characteristics, glucose was the most abundant sugar in the roots, and the sucrose content was very low, compared to that of the hexoses in most of the accessions. Expression analysis of the genes involved in sucrose metabolism, transportation, starch synthesis, and cell wall synthesis was performed through RNA sequencing. The genes encoding sucrose synthases (SUSY) and the enzymes involved in the synthesis of cellulose were highly expressed, indicating that SUSY is involved in cell wall synthesis in radish roots. The positive correlation coefficient (R) between the sucrose content and the expression of cell wall invertase and sugar transporter proteins suggest that hexose accumulation could occur through the apoplastic pathway in radish roots. A positive R score was also obtained when comparing the expression of genes encoding SUSY and fructokinase (FK), suggesting that the fructose produced by SUSY is mostly phosphorylated by FK. In addition, we concluded that sucrose was the most metabolized sugar in radish roots.

Efficient and accurate methods of analysis are needed for the huge amount of biological data that have accumulated in various research fields, including genomics, phenomics, and genetics. Artificial intelligence (AI)-based analysis is one promising method to manipulate biological data. To this end, various algorithms have been developed and applied in fields such as disease diagnosis, species classification, and object prediction. In the field of phenomics, classification of accessions and variants is important for basic science and industrial applications. To construct AI-based classification models, three types of phenotypic image data were generated from 156 Brassica rapa core collections, and classification analyses were carried out using four different convolutional neural network architectures. The results of lateral view data showed higher accuracy compared with top view data. Furthermore, the relatively low accuracy of ResNet50 architecture suggested that definition and estimation of similarity index of phenotypic data were required before the selection of deep learning architectures.

RR-TZF gene plays an important role in plant growth, development and stress responses. OsTZF9 gene is a member of rice RR-TZF gene family. In order to analyze the biological function of OsTZF9 gene, a plant overexpression vector pCXUN-TZF9 driven by Ubiqutin promoter was constructed, and 6 clones of transgenic regenerated plants were obtained by using the agrobacterium-mediated method. Further analysis of the phenotype of the progenies of the transgenic plants showed that plant height, tiller number, effective panicle number, panicle length and 1000-grain weight of the OsTZPP-overexpressed plants were decreased compared with those of the wild-type Nipponbare; compared with the transgenic negative isolated plants, heading date of the O$TZF9-overexpressed plants was delayed. [ABSTRACT FROM AUTHOR]

The taproot of radish (Raphanus sativus L.) is an important sink organ; it is morphologically diverse and contains large amounts of secondary metabolites. Sucrose metabolism is believed to be important in the development of sink organs. We measured the amounts of glucose, fructose, and sucrose in the roots of sixty three radish accessions and analyzed the association between the sugar content and the root phenotype. Fructose content correlated with the root color and length characteristics, glucose was the most abundant sugar in the roots, and the sucrose content was very low, compared to that of the hexoses in most of the accessions. Expression analysis of the genes involved in sucrose metabolism, transportation, starch synthesis, and cell wall synthesis was performed through RNA sequencing. The genes encoding sucrose synthases (SUSY) and the enzymes involved in the synthesis of cellulose were highly expressed, indicating that SUSY is involved in cell wall synthesis in radish roots. The positive correlation coefficient (R) between the sucrose content and the expression of cell wall invertase and sugar transporter proteins suggest that hexose accumulation could occur through the apoplastic pathway in radish roots. A positive R score was also obtained when comparing the expression of genes encoding SUSY and fructokinase (FK), suggesting that the fructose produced by SUSY is mostly phosphorylated by FK. In addition, we concluded that sucrose was the most metabolized sugar in radish roots. [ABSTRACT FROM AUTHOR]

Efficient and accurate methods of analysis are needed for the huge amount of biological data that have accumulated in various research fields, including genomics, phenomics, and genetics. Artificial intelligence (AI)-based analysis is one promising method to manipulate biological data. To this end, various algorithms have been developed and applied in fields such as disease diagnosis, species classification, and object prediction. In the field of phenomics, classification of accessions and variants is important for basic science and industrial applications. To construct AI-based classification models, three types of phenotypic image data were generated from 156 Brassica rapa core collections, and classification analyses were carried out using four different convolutional neural network architectures. The results of lateral view data showed higher accuracy compared with top view data. Furthermore, the relatively low accuracy of ResNet50 architecture suggested that definition and estimation of similarity index of phenotypic data were required before the selection of deep learning architectures. [ABSTRACT FROM AUTHOR]

This article discusses the use of droplet-based technology to study the potential phenotypic effect of microplastics (MPs) on antimicrobial resistance (AMR). Microplastics and nanoplastics provide a micro-environment called the "plastisphere" for bacteria to form biofilms, which exacerbates the threat of AMR. The study used droplets containing Escherichia coli and cefotaxime antibiotic, with or without polystyrene microspheres. The results showed that the presence of microspheres in the droplets may enhance the resistance of E. coli against cefotaxime, possibly due to the observed tendency for clumping (indicating biofilm formation) of E. coli when microspheres are present. The authors suggest that further experiments with different antibiotics and types/sizes of microplastics are needed to better understand the potential of microplastics to enhance AMR. [Extracted from the article]

The phylogenetic structure of bacteria is not always consistent with the traditional classification scheme based on the phenotypic properties of bacteria. That is one of the problems of modern bacterial taxonomy. In addition, traditional methods to identify bacteria using phenotypic properties have a number of disadvantages. In recent decades, significant progress was achieved in the investigation of microbial world using molecular methods for fast identification. The aim of the study was to clarify the species status of four strains of bacteria isolated from black lichens of the cliffs of Galindez Island in the Antarctic on the basis of phenotypic and genetic analysis. Methods. Morphological and cultural properties of bacteria were studied according to generally accepted microbiological methods. Physiological and biochemical ones were investigated using test systems API Coryne and API 20E (bioMerieux SA, France), according to the manufacturer's instructions. Phylogenetic analysis was performed based on the nucleotide sequences of the 16S rRNA gene. To identify closely related species, a comparative analysis of the nucleotide sequences of 16S rRNA genes was performed using the BLAST software package. The phylogenetic position was determined by constructing trees (dendrograms) to show the position of the studied strains among closely related and typical species (programs ClustalX 2.1, Mega 6.06). The tree was constructed applying ClustalX 2.1 by comparing the nearest neighbors with bootstrap analysis (bootstrap NJ tree) using 1000 bootstrap tests (1000 alternative trees). Then the phylogenetic tree was edited by the program Mega v. 6.00. Results. Based on the results of comparative, phylogenetic and phenotypic analysis, the studied Antarctic strains 180n1, 181n2, 188n2, 190n2 were identified as Pseudomonas fluorescens, Microbacterium foliorum, Sporosarcina aquimarina and Rothia sp., respectively. The coefficient of similarity of 16S rRNA genes of strain 180n1 with such a closely related species from the database P. fluorescens NBRC 14160 was 99.5%; 181n2 with M. foliorum P 333/02 - 99.4%; 188n2 with S. aquimarina SW28 - 99.7%. These strains form common clusters with closely related species on phylogenetic dendrograms. The strain 190n2 can be considered as Rothia sp., since has the remote position from closely related strains in the cluster Rothia and a low percentage of similarity (97.3%) with the species Rothia endophytica YIM 67072. These strains belong to the phyla: Firmicutes, Actinobacteria, Proteobacteria. Conclusions. Phylogenetic and phenotypic analyzes allowed determining the taxonomic position of isolated aerobic chemoorganotrophic microbial strains of the Antarctic. Nucleotide sequences of 16S rRNA genes are deposited in the International GenBank database under numbers HG518622, HG518623, HG518625, HG518626. [ABSTRACT FROM AUTHOR]