Plant height is an important agronomic trait that affects the architecture of rapeseed, one of the most important oil crops worldwide. Reduced height can significantly improve rapeseed lodging resistance, which benefits mechanized harvesting of the crop. The semi-dwarf rapeseed mutant zs-d was obtained from the elite variety Zhongshuang11 (ZS11) by ethyl methylsulfonate mutagenesis. Compared with ZS11, zs-d had purple petioles and dark green leaves, a more compact architecture (due to a smaller branch angle), shorter internodes, and significantly lower plant height. This semi-dwarfism is controlled by a single, partially dominant gene. A comparison of the pigment content of zs-d and ZS11 in leaves revealed that the anthocyanin content of zs-d was dramatically higher; however, its total chlorophyll content was significantly lower. Observation of stem paraffin sections revealed that the number of zs-d cells was significantly higher than that of ZS11 cells in identical area fields of view. Furthermore, the length and area of cortical cells in zs-d were significantly reduced compared with those in ZS11 plants. Determining and analyzing endogenous hormone content showed that while the indole acetic acid content was significantly decreased, the 5-deoxystrigol content of strigolactone synthetic precursors was significantly increased in the zs-d hypocotyls. Integrative analysis of stem and leaf RNA-seq data for zs-d and ZS11 revealed that the mutant phenotype of zs-d may be attributed to phytohormones (especially auxins) and anthocyanins. Overall, this study lays a theoretical foundation for further elucidating the molecular mechanism of semi-dwarfism in zs-d, offering valuable insights for dwarfing in rapeseed breeding.

The absorption of carotenoids, plant pigments with significant health benefits, is promoted when ingested with lipids. However, rapeseed carotenoid contents are relatively low. Our previous study identified that the inactivation of BnaC3.CCD4, a carotenoid cleavage dioxygenase which degrades carotenoids, in rapeseed petals, increased carotenoid accumulation. The present study created loss-of-function single, double, and triple mutants of three other CCD4 copies using a CRISPR/Cas9 system. The results showed that some double and triple mutants exhibited a more than 20-fold increase in seed (3-carotene contents without harming quality or yield. In addition, the carotenoid content of BnaC1.ccd4 was close to those of some triple and double mutants. During seed maturation and leaf senescence, BnaCCD4 mutations delayed carotenoid degradation. The reduced power activities and hydroxyl radical scavenging activities of mutant seed oils indicated increased antioxidation. Enzyme activity analysis of BnaCCD4s in Arabidopsis ccd4 mutant seeds confirmed diverging oxidative cleavage activities on carotenoids.

Alcohol-induced acute gastric mucosal damage (AGMD) remains a significant health concern, driven by oxidative stress and inflammatory responses. Current therapeutic approaches (e.g., acid-suppressive agents, mucosal protectants) are limited by side effects and suboptimal bioavailability, while conventional extraction of herbal medicines (e.g., Honeysuckle-Cassia seeds) yields low bioactive compound solubility. This study explored the potential of Lactobacillus acidophilus and Bacillus subtilis co-fermented Honeysuckle-Cassia seed extracts in mitigating AGMD. In vitro antioxidant assays revealed that mixed bacterial fermentation extract (MBF) exhibited superior scavenging activity against superoxide anion (86.14 %), ABTS (88.11 %), and DPPH (42.96 %) radicals compared to unfermented aqueous extract (AE) and single-strain fermented extract (LAF). In vivo experiments in ethanol-induced AGMD mice showed that MBF significantly restored gastric mucosal redox balance, reducing MDA levels by 60.92 % and enhancing SOD activity and GSH content. Mechanistically, MBF suppressed neutrophil infiltration (MPO ↓61.36 %) and pro-inflammatory cytokines (IL-1β, TNF-α, IL-6), while upregulating tight junction proteins (ZO-1, Claudin-1, Occludin) to protect mucosal integrity. These protective effects were mainly mediated by modulation of the MAPK signaling axis-specifically, inhibiting JNK/p38 and activating ERK-thereby enhancing antioxidant defenses and maintaining tight junction integrity. These findings indicate that probiotic fermentation significantly enhances the therapeutic potential of herbal formulations. Importantly, given its safety and efficacy, MBF may be further developed as a functional food, nutraceutical, or adjunctive dietary therapy to prevent alcohol-related gastric injury, offering a sustainable and food-based preventive strategy.

Cucumber (Cucumis sativus L.) is a widely cultivated crop with rich germplasm resources, holding significant nutritional value. It also serves as an important model for studying epidermal cell fate and sex determination. Cucumbers are covered with multicellular and unbranched trichomes, including a specific type called spines found on the surface of the fruit. The presence and density of these fruit spines determine the visual quality of cucumber fruits. However, the key regulatory genes and mechanisms underlying cucumber fruit spine development remain poorly understood. In this study, we identified a WUSCHEL-related homeobox (WOX) family gene CsWOX3, which functioned as a typical transcriptional repressor and played a negative role in fruit spine development. Spatial-temporal expression analysis revealed that CsWOX3 exhibited a relatively high expression level in the cucumber female floral organs, particularly in the fruit exocarp. Knockout of CsWOX3 using CRISPR/Cas9 resulted in a significant 2-to-3-fold increase in the diameter of fruit spines base, while overexpression led to a 17% decrease in the diameter compared to the wild-type. A SQUAMOSA PROMOTER BINDING PROTEIN-LIKE transcription factor CsSPL15 could directly bind and activate the expression of CsWOX3, thereby suppressing the expression of downstream auxin-related genes, such as CsARF18. Additionally, the RING-finger type E3 ubiquitin ligase CsMIEL1-like interacted with the HD domain of CsWOX3, which might result in the ubiquitination and subsequent alteration in protein stability of CsWOX3. Collectively, our study uncovered a WOX transcription factor CsWOX3 and elucidated its expression pattern and biological function. This discovery enhances our comprehension of the molecular mechanism governing cucumber fruit spine morphogenesis.

The multicellular trichomes of cucumber (Cucumis sativus L.) serve as the primary defense barrier against external factors, whose impact extends beyond plant growth and development to include commercial characteristics of fruits. The aphid (Aphis gossypii Glover) is one of prominent pests in cucumber cultivation. However, the relationship between physical properties of trichomes and the aphid resistance at molecular level remains largely unexplored. Here, a spontaneous mutant trichome morphology (tm) was characterized by increased susceptibility towards aphid. Further observations showed the tm exhibited a higher and narrower trichome base, which was significantly distinguishable from that in wild-type (WT). We conducted map-based cloning and identified the candidate, CsTM, encoding a C-lectin receptor-like kinase. The knockout mutant demonstrated the role of CsTM in trichome morphogenesis. The presence of SNP does not regulate the relative expression of CsTM, but diminishes the CsTM abundance of membrane proteins in tm. Interestingly, CsTM was found to interact with CsTIP1;1, which encodes an aquaporin with extensive reports in plant resistance and growth development. The subsequent aphid resistance experiments revealed that both CsTM and CsTIP1;1 regulated the development of trichomes and conferred resistance against aphid by affecting cytoplasmic H2O2 contents. Transcriptome analysis revealed a significant enrichment of genes associated with pathogenesis, calcium binding and cellulose synthase. Overall, our study elucidates an unidentified mechanism that CsTM-CsTIP1;1 alters multicellular trichome morphology and enhances resistance against aphid, thus providing a wholly new perspective for trichome morphogenesis in cucumber.

Leucine-Rich Repeats Extensins (LRX) are a type of cell wall protein that participate in the formation of the plant cell wall and play a crucial role in plant growth and development. However, the study of LRX genes in Cucurbitaceae has not been reported. Here, 40 LRX genes were identified from seven cucurbit species in the Cucurbitaceae database, including cucumber (Cucumis sativus), wax gourd (Benincasa hispida), watermelon melon). These LRX genes were divided into two subfamilies: LRX, controlling vegetative growth, and PEX, controlling reproductive growth. The gene structure, domain, and motif were relatively conserved, indicating that genes within each subfamily have similar functions. The differences in the number of LRX genes among the seven cucurbit species indicate the presence of gene loss or duplication events during evolution. Analysis of cis-regulatory elements showed that these LRX genes may be involved in plant growth and development, phytohormone response, and biotic and abiotic stress responses. In addition, the expression pattern of CsLRX genes in different tissues of cucumber and its expression analysis under salt stress and drought stress were detected by real-time quantitative PCR (qRT-PCR). The results showed that CsLRX genes showed organ-specific expression pattern in cucumber and responded to adversity stress. In summary, the results of the study provide a reference for further understanding the role of these genes in cell wall formation and the growth and development of Cucurbitaceae crops.

The E3 ubiquitin ligase CBL-B is a master regulator of effector T cells through its downstream regulation of T-cell receptor and co-stimulatory stimulation, but it has also been shown to play a negative feedback role in NK cells. CBL-B depletion enhances the production of IFNγ and the cytotoxicity of activated NK cells. Similar to the effect of depleting CD8 T cells in mice, depletion of NK cells abolishes the tumor growth delay observed in Cbl-b-/- mice. With emerging data supporting the potential role of exogenous NK cells in anti-tumor immunity, inhibition of CBL-B represents a promising approach to enhance both adaptive and innate immunity for cancer immunotherapy. Using one of our small molecules, allosteric CBL-B inhibitors, HOT-A, we demonstrated that CBL-B inhibition enhanced NK activation, IFNγ production and granzyme B secretion in response to the various stimulation conditions, including cytokines (such as IL-15 and IL12 + IL-18) and stimulatory receptors (such as NKp30 and NKG2D + CD244). NK cell proliferation was also enhanced in the presence of HOT-A. Single cell analysis further suggested that HOT-A increased NK polyfunctional cytokine expression during IL-12 + IL-18 activation. In a coculture system of NK cells and K562 target cells, HOT-A dose-dependently promoted primary human NK cell activation, cytokine production and cytotoxicity in the K562 cells. In the in vivo CT-26 tumor model, granzyme B+ NK cells in the TIL were significantly increased in the HOT-A treated tumor bearing mice and NK function gene signature was also increased by Nanostring analysis, suggesting an enhanced NK function in the tumor microenvironment. Taken together, the pre-clinical data presented here demonstrate that our CBL-B inhibitor enhances NK cell function in vitro and in vivo, suggesting that its enhancement of anti-tumor immunity may be driven through both effector T cell and NK cell-driven biology. Citation Format: Yilin Qi, Jun Kuai, Yingzhi Bi, David Greco, Samira Jaeger, Ken Carson, Hadi Danaee, Timothy Reilly, Geraldine Harriman, Fang Wang. Inhibition of the E3 ligase CBL-B enhances the effector function and proliferation of natural killer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr LB337.

Pre-harvest sprouting (PHS), the germination of seeds on the plant prior to harvest, poses significant challenges to agriculture. It not only reduces seed and grain yield, but also impairs the commodity quality of the fruit, ultimately affecting the success of the subsequent crop cycle. A deeper understanding of PHS is essential for guiding future breeding strategies, mitigating its impact on seed production rates and the commercial quality of fruits. PHS is a complex phenomenon influenced by genetic, physiological, and environmental factors. Many of these factors exert their influence on PHS through the intricate regulation of plant hormones responsible for seed germination. While numerous genes related to PHS have been identified in food crops, the study of PHS in vegetable crops is still in its early stages. This review delves into the regulatory elements, functional genes, and recent research developments related to PHS in vegetable crops. Meanwhile, this paper presents a novel understanding of PHS, aiming to serve as a reference for the study of this trait in vegetable crops.

该文探讨了在创新性实验教学狮子山昆虫种类调查课程中,以兴趣点吸引学生参与,以学生为主体、教师为主导的原则开展实验内容的设计与操作,注重对实验过程的管理,完善以过程为导向的评价体系.通过教学改革,激发了学生的创新思维和创新意识,学生的科研实践能力和科学素养得到显著提升.

开展学科技能竞赛可以提高学生专业认知度,提升综合素质及激发创新思维.该文详细介绍了2019年华中农业大学成功举办第一届种子解剖大赛的组织实施过程,通过赛后总结分析了技能竞赛折射出的问题,并提出有效建议,旨在让技能竞赛真正成为促进创新人才培养的有效实践教学途径.

The underlying mechanisms of thymocyte development and lineage determination remain incompletely understood, and the emerging evidences demonstrated that RNA binding proteins (RBPs) are deeply involved in governing T cell fate in thymus. Serine/arginine-rich splicing factor 1 (SRSF1), as a classical splicing factor, is a pivotal RBP for gene expression in various biological processes. Our recent study demonstrated that SRSF1 plays essential roles in the development of late thymocytes by modulating the T cell regulatory gene networks post-transcriptionally, which are critical in response to type I interferon signaling for supporting thymocyte maturation. Here, we report SRSF1 also contributes to the determination of the CD8+ T cell fate. By specific ablation of SRSF1 in CD4+CD8+ double positive (DP) thymocytes, we found that SRSF1 deficiency impaired the maturation of late thymocytes and diminished the output of both CD4+ and CD8+ single positive T cells. Interestingly, the ratio of mature CD4+ to CD8+ cells was notably altered and more severe defects were exhibited in CD8+ lineage than those in CD4+ lineage, reflecting the specific function of SRSF1 in CD8+ T cell fate decision. Mechanistically, SRSF1-deficient cells downregulate their expression of Runx3, which is a crucial transcriptional regulator in sustaining CD8+ single positive (SP) thymocyte development and lineage choice. Moreover, forced expression of Runx3 partially rectified the defects in SRSF1-deficient CD8+ thymocyte maturation. Thus, our data uncovered the previous unknown role of SRSF1 in establishment of CD8+ cell identity.

在开放式教学改革的基础上,对大学物理实验教学内容和模式进一步深化革新.在教学模式上,构建虚实结合的BOPPPS教学模式,实现自主化、泛在化的实验课程学习过程;在教学内容上,重构"有趣、有度、有用、有味"的模块化实验教学内容体系,建立了"多元化+信息化+过程化"的教学评价体系.实践表明:虚实结合的大学物理实验BOPPPS教学模式更好地调动了学生学习的主动性与创造性,实现了价值塑造、能力培养、知识传授的育人目标,体现了农业院校大学物理实验的特色.

芥菜型油菜细胞质雄性不育(CMS)系WJS01A是一种稳定不育系,其败育彻底,不受环境条件的影响.该研究从形态、细胞特征、遗传和分子生物学等方面对WJS01A进行鉴定,以揭示其败育机制,为该不育系在油菜育种中的应用提供理论基础.结果 表明:(1)不育系WJS01A的花序结构与正常芥菜型油菜差异不大,但在花蕾饱满度、花朵张开度及花瓣长度和宽度方面略低于正常的芥菜型油菜;它的雌蕊发育正常,但花药、花丝缩短,致使雄蕊高度显著低于柱头,花药白化无花粉产生.(2)将wJS01A衍生的甘蓝型油菜背景不育系WNJ01A以及Polima(Pol)、Ogura (Ogu)和Kosena (Kos)不育系分别与其恢复系或保持系测交,结果显示来源于WJS01A的不育类型与Pol、Ogu和Kos等材料的恢保关系明显不同,仅Hui01可以恢复WNJ01A的育性.(3)不育系WJS01A属于无花粉囊型不育,败育时期为花药原基到孢原细胞时期.(4)线粒体不育基因多重PCR可以明显区分WJS01A、WNJ01A与Pol、Ogu、Kos,但是目前的引物组合不能区分WJS01A与正常的芥菜型油菜.(5)线粒体基因组的限制性片段长度多态性(RFLP)分析表明,在所检测的8个探针/酶组合中均可以将不育系WJS01A与其他4种细胞质雄性不育系区分开,说明WJS01A是一种显著不同于Pol、Ogu和Kos等的细胞质雄性不育类型.WJS01A的利用可以丰富和拓宽当前油菜杂种优势利用的遗传基础,为缓解当前油菜杂种优势利用中不育胞质单一性问题提供新的种质.

Intestinal epithelial barrier dysfunction plays an important role in the development of multiple organ failure during systemic inflammation. The opening of the epithelial barrier leads to increased leukocyte infiltration into the lamina propria, the event therefore promotes microbial translocation and augments intestinal inflammation that contribute to the severity of trauma-associated complications. The aim of the study was to explore the roles of protein kinase 6 (PTK6) and focal adhesion kinase (FAK) in the intestinal epithelial barrier dysfunction following the treatment of TNFα/IFNγ. Epithelial barrier function was analyzed by measuring transcellular electrical resistance (TER) in cultured mouse intestinal epithelial cells. Our data revealed that stimulation of the epithelial cells with TNFα/IFNγ caused a decrease in transcellular electric resistance coupled with decrease expression of cell junctional proteins. The responses were prevented by the inhibitions of either PTK6 or FAK. Western blot analysis showed that elevated levels of active PTK6 and decreased detection of the tight junction protein claudin-3 upon TNFα stimulation. In addition, the downregulation of PTK6 in cells with siRNA attenuates TNFα/IFNγ induced FAK activation via diminished detection of FAK phosphorylation at Y397. Transfection of PTK6-deficient cells with active PTK6 rescued the FAK activation resulting in epithelial barrier dysfunction in response to TNFα/IFNγ. The results suggest that PTK6 mediated FAK activation plays a critical role in gut epithelial barrier dysfunction during inflammatory injury.

Here, we describe a novel mycovirus, tentatively designated as "Botryosphaeria dothidea mitovirus 3" (BdMV3), isolated from Botryosphaeria dothidea strain FJ, which causes pear ring rot disease in Fujian Province, China. The complete genome nucleotide sequence of BdMV3 is 2538 nt in length and contains a single 2070-nt open reading frame (ORF) encoding a putative RNA-dependent RNA polymerase (RdRp) of 689 amino acids (aa) using the fungal mitochondrial genetic code. BLASTp analysis revealed that the RdRp of BdMV3 shares 28.91%–69.36% sequence identity (query sequence coverage more than 90%) with those of members of the genus Mitovirus, with the highest sequence identity of 69.36% and 68.79% to the corresponding RdRp aa sequences of Rhizoctonia solani mitovirus 10 and Macrophomina phaseolina mitovirus 4, respectively. Phylogenetic analysis based on RdRp aa sequences indicated that BdMV3 is a new member of the genus Mitovirus in the family Mitoviridae.

The kinase PDK1 is a crucial regulator for immune cell development by connecting PI3K to downstream AKT signaling. However, the roles of PDK1 in CD4 + T cell differentiation, especially in T follicular helper (Tfh) cell, remain obscure. Here we reported PDK1 intrinsically promotes the Tfh cell differentiation and germinal center responses upon acute infection by using conditional knockout mice. PDK1 deficiency in T cells caused severe defects in both early differentiation and late maintenance of Tfh cells. The expression of key Tfh regulators was remarkably downregulated in PDK1-deficient Tfh cells, including Tcf7 , Bcl6 , Icos , and Cxcr5 . Mechanistically, ablation of PDK1 led to impaired phosphorylation of AKT and defective activation of mTORC1, resulting in substantially reduced expression of Hif1α and p-STAT3. Meanwhile, decreased p-AKT also suppresses mTORC2-associated GSK3β activity in PDK1-deficient Tfh cells. These integrated effects contributed to the dramatical reduced expression of TCF1 and ultimately impaired the Tfh cell differentiation.

Here we show that FTO as an N6-methyladenosine (m6A) RNA demethylase is degraded by selective autophagy, which is impaired by low-level arsenic exposure to promote tumorigenesis. We found that in arsenic-associated human skin lesions, FTO is upregulated, while m6A RNA methylation is downregulated. In keratinocytes, chronic relevant low-level arsenic exposure upregulated FTO, downregulated m6A RNA methylation, and induced malignant transformation and tumorigenesis. FTO deletion inhibited arsenic-induced tumorigenesis. Moreover, in mice, epidermis-specific FTO deletion prevented skin tumorigenesis induced by arsenic and UVB irradiation. Targeting FTO genetically or pharmacologically inhibits the tumorigenicity of arsenic-transformed tumor cells. We identified NEDD4L as the m6A-modified gene target of FTO. Finally, arsenic stabilizes FTO protein through inhibiting p62-mediated selective autophagy. FTO upregulation can in turn inhibit autophagy, leading to a positive feedback loop to maintain FTO accumulation. Our study reveals FTO-mediated dysregulation of mRNA m6A methylation as an epitranscriptomic mechanism to promote arsenic tumorigenicity.

Invariant natural killer T (iNKT) cells are highly conserved innate-like T lymphocytes that originate from CD4+CD8+ double-positive (DP) thymocytes. Here, we report that serine/arginine splicing factor 1 (SRSF1) intrinsically regulates iNKT cell development by directly targeting Myb and balancing the abundance of short and long isoforms. Conditional ablation of SRSF1 in DP cells led to a substantially diminished iNKT cell pool due to defects in proliferation, survival, and TCRα rearrangement. The transition from stage 0 to stage 1 of iNKT cells was substantially blocked, and the iNKT2 subset was notably diminished in SRSF1-deficient mice. SRSF1 deficiency resulted in aberrant expression of a series of regulators that are tightly correlated with iNKT cell development and iNKT2 differentiation, including Myb, PLZF, Gata3, ICOS, and CD5. In particular, we found that SRSF1 directly binds and regulates pre-mRNA alternative splicing of Myb and that the expression of the short isoform of Myb is substantially reduced in SRSF1-deficient DP and iNKT cells. Strikingly, ectopic expression of the Myb short isoform partially rectified the defects caused by ablation of SRSF1. Furthermore, we confirmed that the SRSF1-deficient mice exhibited resistance to acute liver injury upon α-GalCer and Con A induction. Our findings thus uncovered a previously unknown role of SRSF1 as an essential post-transcriptional regulator in iNKT cell development and functional differentiation, providing new clinical insights into iNKT-correlated disease.

运用磁学理论和仿真软件,从漆包线和通水导线两种方案,分析讨论了在体积较大磁感应强度较高的磁场设计需求下,螺线管的尺寸、重量和散热问题.为工程实践提供参考,设计理念、思路和方案也将为相关设备的设计制造提供借鉴.