TaGSK3 regulates wheat development and stress adaptation through BR-dependent and BR-independent pathways.

The GSK3/SHAGGY-like kinase plays critical roles in plant development and response to stress, but its specific function remains largely unknown in wheat (Triticum aestivum L.). In this study, we investigated the function of TaGSK3, a GSK3/SHAGGY-like kinase, in wheat development and response to stress. Our findings demonstrated that TaGSK3 mutants had significant effects on wheat seedling development and brassinosteroid (BR) signalling. Quadruple and quintuple mutants showed amplified BR signalling, promoting seedling development, while a sextuple mutant displayed severe developmental defects but still responded to exogenous BR signals, indicating redundancy and non-BR-related functions of TaGSK3. A gain-of-function mutation in TaGSK3-3D disrupted BR signalling, resulting in compact and dwarf plant architecture. Notably, this mutation conferred significant drought and heat stress resistance of wheat, and enhanced heat tolerance independent of BR signalling, unlike knock-down mutants. Further research revealed that this mutation maintains a higher relative water content by regulating stomatal-mediated water loss and maintains a lower ROS level to reduces cell damage, enabling better growth under stress. Our study provides comprehensive insights into the role of TaGSK3 in wheat development, stress response, and BR signal transduction, offering potential for modifying TaGSK3 to improve agronomic traits and enhance stress resistance in wheat.

Identification of sulfhydryl-containing proteins and further evaluation of the selenium-tagged redox homeostasis-regulating proteins.

Chemoproteomic profiling of sulfhydryl-containing proteins has consistently been an attractive research hotspot. However, there remains a dearth of probes that are specifically designed for sulfhydryl-containing proteins, possessing sufficient reactivity, specificity, distinctive isotopic signature, as well as efficient labeling and evaluation capabilities for proteins implicated in the regulation of redox homeostasis. Here, the specific selenium-containing probes (Se-probes) in this work displayed high specificity and reactivity toward cysteine thiols on small molecules, peptides and purified proteins and showed very good competitive effect of proteins labeling in gel-ABPP. We identified more than 6000 candidate proteins. In TOP-ABPP, we investigated the peptide labeled by Se-probes, which revealed a distinct isotopic envelope pattern of selenium in both the primary and secondary mass spectra. This unique pattern can provide compelling evidence for identifying redox regulatory proteins and other target peptides. Furthermore, our examiation of post-translational modification (PTMs) of the cysteine site residues showed that oxidation PTMs was predominantly observed. We anticipate that Se-probes will enable broader and deeper proteome-wide profiling of sulfhydryl-containing proteins, provide an ideal tool for focusing on proteins that regulate redox homeostasis and advance the development of innovative selenium-based pharmaceuticals.

Integration of genome-wide association study, linkage analysis, and population transcriptome analysis to reveal the TaFMO1-5B modulating seminal root growth in bread wheat.

Bread wheat, one of the keystone crops for global food security, is challenged by climate change and resource shortage. The root system plays a vital role in water and nutrient absorption, making it essential for meeting the growing global demand. Here, using an association-mapping population composed of 406 accessions, we identified QTrl.Rs-5B modulating seminal root development with a genome-wide association study and validated its genetic effects with two F5 segregation populations. Transcriptome-wide association study prioritized TaFMO1-5B, a gene encoding the flavin-containing monooxygenases, as the causal gene for QTrl.Rs-5B, whose expression levels correlate negatively with the phenotyping variations among our population. The lines silenced for TaFMO1-5B consistently showed significantly larger seminal roots in different genetic backgrounds. Additionally, the agriculture traits measured in multiple environments showed that QTrl.Rs-5B also affects yield component traits and plant architecture-related traits, and its favorable haplotype modulates these traits toward that of modern cultivars, suggesting the application potential of QTrl.Rs-5B for wheat breeding. Consistently, the frequency of the favorable haplotype of QTrl.Rs-5B increased with habitat expansion and breeding improvement of bread wheat. In conclusion, our findings identified and demonstrated the effects of QTrl.Rs-5B on seminal root development and illustrated that it is a valuable genetic locus for wheat root improvement.

Regulation of the stem­like properties of estrogen receptor­positive breast cancer cells through NR2E3/NR2C2 signaling.

Cancer stem cells (CSCs) are major drivers of metastasis, drug resistance and recurrence in numerous cancers. However, critical factors that can modulate CSC stemness have not been clearly identified. Nuclear receptor subfamily 2 group E member 3 (nr2e3) expression has been previously reported to be positively associated with drug sensitivity and favorable clinical outcomes in patients with estrogen receptor (ER)+ breast cancer. This suggests that nr2e3 expression may be inversely associated with CSC stemness in this type of tumor cells. The present study aimed to investigate the regulatory roles of NR2E3 in the stem-like properties of ER+ breast cancer cells and to identify the underlying mechanisms. Bioinformatics analysis was performed using the data derived from the Cancer Genome Atlas database. Nr2e3-specific shRNA and nuclear receptor subfamily 2 group C member 2 (nr2c2) overexpressed plasmids were constructed to silence and enhance the expression of nr2e3 and nr2c2, respectively. Transwell and wound healing experiments were conducted to evaluate the migration and invasion ability of MCF7 cells, while colony formation tests were used to evaluate the clonality. Flow cytometry was used to detect the percentage of CD44+CD24-/low cells. Reverse transcription-quantitative PCR and western blotting were performed to detect expression at the mRNA and protein levels. The results showed that compared with normal breast tissues and MCF10A cells, the expression of nr2e3 was increased in ER+ breast tumor tissues and cell lines. Nr2e3 silencing promoted the migration, invasion and colony-forming ability of the ER+ MCF7 cells. It also increased the expression of epithelial-mesenchymal transition markers and stem cell-related transcription factors, in addition to the percentage of CD44+CD24-/low cells. The expression of nr2e3 and nr2c2 was found to be positively correlated. Nr2e3 knockdown decreased the mRNA and protein expression levels of nr2c2, whereas nr2c2 overexpression reversed the elevated CD44+CD24-/low cell ratio and the increased migratory activity caused by nr2e3 silencing. The results of the present study suggest that NR2E3 may serve an important role in modulating the stem-like properties of ER+ breast cancer cells, where NR2E3/NR2C2 signaling may be a therapeutic target in ER+ breast cancer.

Detection of Cysteine Sulfenic Acid on E. coli Proteins with a Biotin-Benzoboroxole Probe.

S-sulfenylation of cysteine residues on proteins can effectively change protein structures and accordingly regulate their functions in vivo. Investigation of S-sulfenylation in different biological environments is thus vital for a systematic understanding of cellular redox regulation. In this work, a functional probe, biotin-benzoboroxole (Bio-ben), was designed for the detection of cysteine sulfenic acid (Cys-SOH). The performance of Bio-ben was characterized by small-molecule sulfenic acid, protein models, and proteome tests via mass spectra and western blotting. The results showed that Bio-ben was validated for cysteine sulfenic acid on proteins with good capture efficiency even at low concentrations. Compared with commonly used probes such as dimedone, the current probe has significantly shortened labeling time and exhibited comparable sensitivity. The proposed method provides a new approach for exploring S-sulfenylation in the oxidative modification of proteins and is helpful for related biological and clinical applications.

Quantitative Chemoproteomic Profiling of Targets of Au(I) Complexes by Competitive Activity-Based Protein Profiling.

Owing to the encouraging pharmacological action and acceptable toxicity profile, Au(I) complexes have attracted growing interest in the application of disease treatment. In order to investigate their potential target proteins and related bioinformation, herein, we screened four Au(I) complexes and explored the binding proteins utilizing a competitive activity-based protein profiling (ABPP) strategy, including identification experiments and reactivity classification experiments, which offers a simple and robust method to identify the target proteins of Au(I) complexes. We quantified the target proteins of the four Au(I) complexes and found that most of proteins were associated with cancer. In addition, the newly Au(I)-binding proteins and biological gold-protein interaction pathways were exhibited. Furthermore, we estimated the correlation between target proteins of Au(I) complexes and various cancers, which will promote the development of the gold anticancer drugs.

Plaque Characteristics in Young Adults With Symptomatic Intracranial Atherosclerotic Stenosis: A Preliminary Study.

PURPOSE: To determine how intracranial vascular wall and atherosclerosis plaque characteristics differ between young and old adults with sICAS. METHODS: Eighty-four consecutive patients with sICAS who underwent high-resolution magnetic resonance imaging (HRMRI) from December 2017 to July 2020 were retrospectively collected. These participants were divided into young adult group (18-50 years, n = 28) and old adult group (>50 years, n = 56). Reviewers were blinded to any clinical information and HRMRI scans were analyzed for qualitative and quantitative indicators of vascular walls and plaque at the maximal lumen narrowing site using the independent-sample t-test, Mann-Whitney U-test, chi-square test or Fisher exact test, and logistic regression analysis. RESULTS: Young patients with sICAS had significantly smaller maximum wall thickness (1.45 ± 0.38 vs.1.75 ± 0.51 mm2, P = 0.003), higher prevalence of positive remodeling (53.57 vs. 21.43%, P = 0.003), and lower prevalence of diabetes mellitus (14.29 vs. 35.71%, P = 0.04) than old patients. Plaque burden and other plaque features were comparable between young and old patients. CONCLUSION: Young patients with sICAS have smaller maximum wall thickness and greater ability to reconstruct, and are more likely to show positive remodeling, which may lead to some atherosclerotic lesions being missed. Young patients with evidence of vessel narrowing should be carefully examined for presence of high-risk atherosclerotic plaque.

Dual-Channel Imaging of Amyloid-ß Plaques and Peroxynitrite To Illuminate Their Correlations in Alzheimer's Disease Using a Unimolecular Two-Photon Fluorescent Probe.

Alzheimer's disease (AD) involves multiple pathological factors that mutually cooperate and closely contact to form interaction networks for jointly promoting the AD progression. Therefore, the comonitoring of different factors is particularly valuable for elucidating their level dynamics and complex interactions. However, such significant investigations remain a major challenge due to the lack of unimolecular fluorescent probes capable of simultaneous and discriminative visualization of multiple targets. To address this concern, as proof of principle, we rationally designed a unimolecular fluorescent probe to discriminate and simultaneously profile amyloid-ß (Aß) plaques and peroxynitrite (ONOO-), which are both the pronounced AD pathological factors. Herein, a novel ONOO- reaction trigger was installed onto an Aß plaque binding fluorophore to generate a dual functional fluorescent probe, displaying completely separate spectral responses to Aß plaques and ONOO- with high selectivity and sensitivity. With this probe, for the first time, we comonitored the distribution and variation of Aß plaques and ONOO- through two independent fluorescence channels, demonstrating their close apposition and tight correlation during AD course in live cell and mouse models through two-photon imaging mode. Notably, Aß aggregates induce the neuronal ONOO- generation, which conversely facilitates Aß aggregation. The two critical events, ONOO- stress and Aß aggregation, mutually amplify each other through positive feedback mechanisms and jointly promote the AD onset and progression. Furthermore, by coimaging of the level dynamics of Aß plaques and ONOO-, we found that the cerebral ONOO- is a potential biomarker, which emerges earlier than Aß plaques in transgenic mouse models. Overall, the dual-channel responsive performance renders this probe as a powerful imaging tool to decipher Aß plaque-ONOO- interactions, which will facilitate AD-associated molecular pathogenesis elucidation and multitarget drug discovery.

Selenium-oxidizing Agrobacterium sp. T3F4 steadily colonizes in soil promoting selenium uptake by pak choi (Brassica campestris).

Selenium (Se) deficiency in soil is linked to its low content in edible crops, resulting in adverse impacts on the health of 15% of the global population. The crop mainly absorbs oxidized selenate and selenite from soil, then converts them into organic Se. However, the role of Se-oxidizing bacteria in soil Se oxidation, Se bioavailability and Se absorption into plants remains unclear. The strain Agrobacterium sp. T3F4, isolated from seleniferous soil, was able to oxidize elemental Se into selenite under pure culture conditions. The green fluorescent protein (gfp)-gene-marked strain (T3F4-GFP) and elemental Se or selenite (5 mg·kg-1) were added to pak choi (Brassica campestris ssp. chinensis) pot cultures. Observation of the fluorescence and viable counting indicated that GFP-expressing bacterial cells steadily colonized the soil in the pots and the leaves of the pak choi, reaching up to 6.6 × 106 and 2.0 × 105 CFU g-1 at 21 days post cultivation, respectively. Moreover, the total Se content (mostly organic Se) was significantly increased in the pak choi under T3F4 inoculated pot culture, with elemental Se(0) being oxidized into Se(IV), and soil Se(IV) being dissolved before being absorbed by the crop. After strain T3F4 was inoculated, no significant differences in microbial diversity were observed in the soils and roots, whereas the abundance of Rhizobium spp. was significantly increased. To our knowledge, this is the first time that Se-oxidizing Agrobacterium sp. T3F4 has been found to steadily colonize soil and plant tissues, and that its addition to soil increases the absorption of Se in plants. This study provides a potential strategy for Se biofortification.

Fluorescence switch of gold nanoclusters stabilized with bovine serum albumin for efficient and sensitive detection of cysteine and copper ion in mice with Alzheimer's disease.

The near-infrared fluorescence of gold nanoclusters stabilized with bovine serum albumin (BSA -AuNCs) centered at 675 nm could be enhanced by cysteine and then effectively quenched by copper ion (Cu2+), therefore, cysteine and copper ion could be detected in sequence. At "on" state, fluorescence enhancement of BSA-AuNCs is generated due to the reaction between cysteine and BSA-AuNCs, via filling the surface defect of gold nanoclusters, while Cu2+ can further oxidize the reductive sulfydryl of cysteine and interact with amino acids presented in the BSA chain, inducing gold nanoclusters to aggregate, thus causing "off" state with fluorescence quenching. Fluorescence switch of BSA-AuNCs can be used for cysteine and Cu2+ detection in mice brain with Alzheimer's disease (AD) in vitro, with fast response, high chemical stability and sensitivity. Besides, it was able to image the endogenous Cu2+ in liver and heart of AD mice in situ. The results are promising, especially in the framework of early diagnosis of Alzheimer's disease.

Covalent organic frameworks-based paper solid phase microextraction combined with paper spray mass spectrometry for highly enhanced analysis of tetrabromobisphenol A.

How to further improve the sensitivity of paper spray mass spectrometry (PS-MS) is very important for its application in low content substance detection. Covalent organic frameworks (COFs) are a kind of novel crystalline materials that exhibit tremendous potential for multifarious applications. Nevertheless, the applications of COFs in PS-MS are still unexplored. Herein, for the first time, a method that used COFs nanoparticles as a coated matrix of PS-MS coupled with solid phase microextraction (SPME) was developed, and as a proof of concept, it was applied for highly sensitive analysis of pollutant tetrabromobisphenol A (TBBPA) in environmental samples. In order to prepare the COFs-coated papers, a variety of factors, such as starch type and amount, loaded COFs amount and spray solvent, have been explored in detail. Compared with the naked paper substrates, a lower limit of quantitation of 1 nmol L-1 with 50-fold enhancement was achieved using COFs-based paper composites, which indicated that papers decorated with COFs might be a promising candidate for paper spray with high sensitivity in the analysis of environmental samples.

Fibrisoma montanum sp. nov., isolated from soil of Mountain Danxia, China.

A Gram-stain-negative, non-motile, rod-shaped, aerobic bacterium, designated HYT19T, was isolated from soil of Mountain Danxia in southern China. It showed the highest similarity of 16S rRNA gene sequence (97.0%) and formed a monophyletic clade with Fibrisoma limi BUZ 3T. Strain HYT19T grew at 16-37 °C (optimum 28-30 °C) and at pH 6-7. The draft genome size of strain HYT19T was 7.8 Mb with a DNA G+C content of 54.0 mol%. The digital DDH and average nucleotide identity values between strain HYT19T and F. limi BUZ 3T were 28.8% and 85.1%, respectively. MK-7 was the sole respiratory quinone. The major polar lipids were phosphatidylethanolamine, unidentified aminophospholipid, two unidentified aminolipids, unidentified phospholipid and unidentified lipid. The strain contained C16:1ω5c, iso-C15:0, summed feature 3 (C16:1ω6c and/or C16:1ω7c), C16:0, iso-C17:0 3-OH and anteiso-C15:0 as the major fatty acids. On the basis of phylogenetic, genomic, phenotypic and chemotaxonomic analysis, we propose a new species Fibrisoma montanum sp. nov. of genus Fibrisoma. The type strain is HYT19T (= CCTCC AB 2018342T = JCM 33105T).

Sphingomonas gilva sp. nov., isolated from mountain soil.

A Gram-stain-negative, strictly aerobic, motile, yellow, rod-shaped bacterium, designated ZDH117T, was isolated from soilsampled atthe Danxialandformin Guangdong Province, PR China. The 16S rRNA gene sequence of strain ZDH117T had highest similarityvalues to Sphingomonas adhaesivaDSM 7418T (97.5 %), SphingomonasdesiccabilisCP1DT (97.3 %) and Sphingomonas ginsenosidimutans KACC 14949T (97.2 %). However, phylogenetic analyses based on 16S rRNA gene sequences demonstrated that strain ZDH117T clustered with Sphingomonas zeicaulis 541T (96.17 %) and Sphingomonas sanxanigenens DSM 19645T (95.95 %). The genomic average nucleotide identity values of ZDH117T with S. adhaesiva DSM 7418T, S. desiccabilis CP1DTand S. ginsenosidimutans KACC TT were 75.1, 75.2 and 75.0 %, respectively. The G+C content of the genomic DNA was 67.6 mol%. Strain ZDH117T was characterized to have ubiquinone-10 as the predominant respiratory quinone, sym-homospermidine as the major polyamine and summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c), C14 : 0-2OH, C16 : 0 and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) as the major cellular fatty acids (>5 % of total). The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, sphingoglycolipid, an unidentified phospholipid and three unidentified lipids. On the basis of its phenotypic, chemotaxonomic and phylogenetic characteristics, strain ZDH117T represents a novel species of the genus Sphingomonas, for which the name Sphingomonas gilva sp. nov. is proposed. The type strain is ZDH117T (=KCTC 62894T=CCTCCAB 2018262T).

A fast, highly sensitive and selective assay of iodide ions with single-stranded DNA-templated copper nanoparticles as a fluorescent probe for its application in Kunming mice samples.

The development of fast, sensitive, selective and flexible methods for the detection of iodide is highly demanded and is of great significance. In this work, single-stranded DNA-templated copper nanoparticles (ssDNA-CuNPs) generated by sodium ascorbate reduction of Cu2+ along the single-stranded DNA of poly-T were utilized as a fluorescent probe for the determination of iodide ions (I-). The detection scheme is based on the instant quenching of the fluorescence of ssDNA-CuNPs by iodide ions. I- can be quantified in the concentration range from 0.050 to 40 µM and from 40 to 80 µM, and the limit of detection is as low as 15 nM. This method provides a simple and convenient strategy for the biochemical assay of I-, which is also helpful for early diagnosis of related diseases. The establishment of a low cost and fast detection method would be particularly important in developing countries where medical supplies are lacking.