The mitotic exit network (MEN), regulated by the small GTPase Tem1, plays a crucial role in coordinating cytokinesis and cell cycle progression in eukaryotes. In this study, we identified MoTem1, a functional homolog of Saccharomyces cerevisiae Tem1, in the rice blast fungus Magnaporthe oryzae, and investigated its role in mitotic regulation and pathogenesis. Using targeted mutagenesis, we generated a series of mutant strains: ΔMotem1 (knockout), MoTem1-OE (overexpression), as well as MoTem1-CA (constitutively active) and MoTem1-DN (dominant-negative) variants created via single-nucleotide substitutions. Phenotypic characterization revealed that MoTem1's activity states are critical for fungal growth, development, stress tolerance, and pathogenicity. While ΔMotem1 and MoTem1-CA strains showed reduced virulence, the MoTem1-DN mutant exhibited hypervirulence. Transcriptomic profiling and weighted gene co-expression network analysis (WGCNA) identified chitin synthase MoCHS1 as a downstream gene whose expression is directly or indirectly influenced by MoTem1 activity states. Pharmacological inhibition of chitin synthesis using Polyoxin B in MoTem1-CA showed increased sensitivity, confirming a decreased expression of chitin synthase in the MoTem1-CA strain. Subcellular localization studies revealed GTP-dependent spindle pole body (SPB) targeting, with inactive MoTem1 failing to localize to SPBs, while constitutive MEN activation in MoTem1-CA disrupted spindle position checkpoint (SPOC) controls, resulting in multinucleate hyphae and a range of developmental defects. In conclusion, our work establishes MoTem1 not merely as a cell cycle regulator, but as a global upstream factor that influences nuclear division, cell wall integrity, and broadly reshapes the genomic regulatory network to govern development and pathogenesis in M. oryzae.

Cysteine palmitoylation (S-palmitoylation or S-acylation) is a reversible post-translational modification dynamically controlled by opposing enzymes: palmitoyl acyltransferases (PATs) and depalmitoylases. Despite its established roles in other systems, the mechanistic details of S-acylation in plants, particularly its spatiotemporal regulation during plant-virus interactions, remain poorly understood, largely due to the lack of validated enzyme-substrate pairs. Using the geminivirus tomato yellow leaf curl Chuxiong virus (TYLCCxV) as a model, we show that the viral effector C4 undergoes S-palmitoylation at Cys-4, a modification essential for its plasma membrane anchorage and subsequent mediation of viral pathogenesis. NbPAT4, a palmitoyl acyltransferase from Nicotiana benthamiana, catalyzes C4 S-palmitoylation, promoting its membrane localization, protein stability, and viral infection. Conversely, the C4S substitution (C4C4S) abolishes S-palmitoylation, leading to cytoplasmic redistribution, protein destabilization, and impaired viral pathogenicity. We further identify NbABHD6 as a depalmitoylase that interacts with C4 and catalyzes its S-depalmitoylation, triggering C4 degradation via the 26S proteasome pathway. This study reveals a regulatory axis in plant-geminivirus interactions, identifying NbPAT4 and NbABHD6 as antagonistic enzymes that dynamically regulate the stoichiometry of C4 S-acylation. These findings support a host-pathogen enzymatic tug-of-war model in which competitive S-palmitoylation homeostasis governs viral subcellular trafficking and pathogenicity.

Heat stress (HS) negatively affects cattle welfare, productivity, and sustainability, with marked differences between heat-tolerant breeds such as Nellore and heat-susceptible breeds such as Angus. In this study, an untargeted UHPLC-Orbitrap-MS metabolomic approach was used to investigate breed-specific differences in the blood metabolome associated with HS. Blood samples were collected from the same Nellore and Angus steers during peak HS exposure and after recovery, allowing both inter-breed and within-animal longitudinal comparisons. Multivariate analyses revealed a clear metabolic divergence between breeds during the challenge period, involving metabolites related to amino acid metabolism, lipid remodeling, energy pathways, and steroid metabolism. Longitudinal analyses accounting for repeated measures showed pronounced breed-specific metabolic responses. Nellore cattle displayed extensive and statistically robust metabolic changes across phases, with 114 metabolites remaining significant after false discovery rate (FDR) correction, including amino acids and peptides, nucleotide derivatives, carbohydrates, acyl-carnitines, steroids, and tetrapyrrole-related compounds. In contrast, Angus cattle showed only a limited number of FDR-significant metabolites, indicating a less consistent systemic response to HS. Pathway analysis identified porphyrin, purine, and pyrimidine metabolism as key pathways modulated by HS, particularly in Nellore cattle. Overall, these results suggest that heat tolerance is associated with coordinated metabolic adaptations rather than isolated metabolite changes. Further studies on larger cohorts are needed to validate the functional relevance of these findings.

Leaf shape plays a crucial role in plant growth and development. Among various leaf traits, marginal lobation serves as an ideal morphological marker for breeding programs. However, the genetic mechanism underlying leaf margin lobation in Brassica juncea L. remains unclear. Through RNA sequencing and map-based cloning, we identified an incompletely dominant gene, BjA10.LL, which encodes an HD-ZIP I protein and is responsible for the formation of leaf margin lobation in B. juncea. Sequence analysis of parental alleles revealed no critical variations in the coding region but identified substantial variations in regulatory regions. Heterologous expression of BjA10.LL in Arabidopsis thaliana confirmed its sufficiency to induce lobed leaves. To functionally link the regulatory variations to the phenotype, we analyzed promoter activity and developed a co-dominant molecular marker targeting key indels in a core enhancer. The promoter activity was significantly affected by these sequence variations, and the marker exhibited perfect co-segregation with the lobed-leaf phenotype in an F₂ population, collectively establishing these regulatory polymorphisms as the causal basis for divergent BjA10.LL expression and leaf morphology. These results demonstrate that BjA10.LL positively regulates marginal lobe formation, providing insights into leaf shape regulation in B. juncea and facilitating the genetic improvement of rapeseed.

Advanced genotyping technologies for understanding the genetic intricacies of fungal pathogens have broad applications in crop protection. Here, we introduce a novel genotyping-by-target sequencing (GBTS) chip, a versatile tool designed for comprehensive genetic analysis of fungal populations. This technology overcomes key limitations of traditional molecular marker-based approaches by providing a more efficient, economic, and streamlined solution while bypassing the need for labor-intensive pathogen culturing. We demonstrate its utility by applying it to profile Puccinia striiformis f. sp. tritici (Pst), the causal agent of wheat stripe rust. Our analysis involved 225 infected leaves collected from wheat fields in the northwest oversummering region for Pst in China. We delineated three genetic groups and revealed frequent gene flow, with closer connectivity between Qinghai and Gansu than either province with Ningxia, a pattern consistent with wind trajectory models. These findings illustrate a highly connected regional epidemic system and highlight the value of the GBTS chip for genomic epidemiology. The methodology established here provides a scalable framework for population genetic studies in other fungal pathogens, promising to enhance disease monitoring and management across agricultural systems.

The domestication and selective breeding of horses have profoundly influenced the emergence of adaptive traits and stress resistance mechanisms, shaping modern equine populations. This comprehensive review examines the genomic foundations of these traits, emphasizing recent advancements in high-throughput sequencing technologies and bioinformatics. These tools have elucidated the genetic underpinnings of key characteristics such as endurance, speed, metabolic efficiency, and disease resistance. Importantly, the review identifies and connects gene variants associated with thermoregulation, immune function, and cellular repair mechanisms, shedding light on their synergistic roles in enabling horses to adapt to diverse environmental challenges and physiological stressors. By establishing these causal links, this review enhances the coherence between genomic findings and their implications for equine biology. Furthermore, the integration of genomic insights provides a framework for addressing contemporary challenges in horse management and conservation. Issues such as climate change, disease outbreaks, and the preservation of genetic diversity demand innovative strategies grounded in genomics. By bridging the findings on equine adaptation and stress resistance mechanisms with practical applications in breeding and management, this review highlights the potential of genomics to ensure the sustainability and resilience of equine populations in the face of evolving environmental and societal pressures. This expanded perspective underscores the critical role of genomics in both understanding the evolutionary trajectory of horses and guiding future practices in equine health and conservation.

Plants are continuously exposed to environmental abiotic and biotic stressors that can significantly impact their growth, development, productivity, and lifespan. However, plants have developed exceptionally complex signaling pathways that enable their ability to sense, transduce, and respond to these diverse stress stimuli. Salicylates (SA) and jasmonates (JA) are two key phytohormones that significantly influence plant adaptation to environmental and biotic stressors, pivotal in enhancing stress resilience. The interaction and crosstalk between SA and JA signaling cascades are essential for orchestrating appropriate physiological and biochemical responses to biotic (e.g., pathogen attack, herbivory) and abiotic (e.g., oxidative stress, drought, temperature extremes, UV radiation, salinity, heavy metal toxicity) stresses. Salicylates are primarily recognized for being involved in systemic acquired resistance (SAR) against biotic stressors like pathogens. Conversely, jasmonates are well-documented in their function in defenses aimed at herbivorous insects and in mitigating the outcomes of abiotic conditions such as salinity and drought. However, the crosstalk between SAs and JAs is complex, involving both synergistic and antagonistic interactions that finely tune the natural defensive mechanism of the plant toward both biotic and abiotic stresses. This comprehensive review summarizes the most recent research on how SA and JA biosynthesis, signaling, and interactions govern diverse stress adaptive mechanisms in plants. It covers emerging evidence on the importance of SA-JA crosstalk in regulating physiological, biochemical, and molecular adaptations to combined biotic and abiotic stresses.

Given that lactoferrin (LF) exerts an excellent protection of intestinal homeostasis, the underlying mechanisms, especially epigenetic regulations, are still unknown. This study aimed to investigate the effects of dietary LF epigenetically modulates the oxidative genes by histone modifications to ameliorate ileum inflammation of mice exposed to DON contaminated diet. As expected, we found in the morphology analysis that DON exposure increased ileum crypt depth (CD) and villus width (VW) but reduced villus height (VH) and VH: CD ratio compared to those of the vehicle group. Consistently, the elevated ROS and MDA, along with the decreased ATP, SOD, CAT, GSH, and complex I, III, V were observed in the DON-exposed mice ileum. In contrast, LF markedly ameliorated the impairments of morphological and biochemical indexes. Next, we conducted transcriptome analysis to explore the changed signaling pathways using the ileum RNA of the mice treated with DON or LF. Firstly, the cell cycle pathway genes were significantly downregulated in the DON-exposed mice, and LF improved the cell cycle profile. Again, gene ontology analysis showed that inflammation and oxidative stress were significantly activated by DON exposure, and these were recovered when the DON-exposed mice were supplemented with an LF diet. Consistent with these findings, the signaling pathways of the reduced oxidative phosphorylation and elevated TNFα were also observed to be ameliorated by LF treatment. Importantly, histone modifications, including acetylation, methylation, and lactylation were suggested to be the vital players involved in the DON or LF treatment, in which LF significantly increased the loss of histone modifications on these genes. With a bioinformatics analysis and validation by qRT-PCR, the nuclear receptor NR5A2 was selected as a key master in the ileum of mice stimulated by DON. LF performed the benefit function on the NR5A2-mediated oxidative stress genes Ncoa4 and Prdx3 in the DON-exposed mice. Moreover, a ChIP-qPCR was used to verify that histone marks involving H3K9ac, H3K18ac, H3k27ac, H3K4me1, H3K9la, and H3K18la facilitated the epigenetic regulation of NR5A2-modulated actions. We conclude that dietary LF effectively ameliorated ileum lesions induced by DON in mice by modulating oxidative genes Ncoa4 and Prdx3 through histone modifications.

Heat stress (HS) is a major environmental stress that inhibits plant growth and development. Plants have evolved various mechanisms to cope with heat stress, a key one being the HSF-HSP (Heat stress transcription factor-Heat shock protein) signaling pathway. HSFs can be divided into three classes: A, B, and C. In this study, we report the identification and functional characterization of a specific B2 member LdHSFB2a in Lilium davidii var. unicolor. RT-qPCR (Real-time Quantitative Polymerase Chain Reaction) analyses indicated that LdHSFB2a was highly expressed in HS-exposed leaves. LdHSFB2a was localized in the nucleus, consistent with the characterization of transcription factors. In contrast to other HSFBs, LdHSFB2a did not contain the typical B3 repression domain but exhibited transcriptional repression activity in yeast and plant cells. Transient overexpression and virus-induced gene silencing (VIGS) of LdHSFB2a in lily petals suggested that LdHSFB2a functions positively in lily thermotolerance. Consistent with the implication of LdHSFB2a function in thermotolerance, further analysis revealed that the expression levels of HSFA1, HSFA2, and MBF1c were increased as LdHSFB2a was overexpressed but reduced as LdHSFB2a was silenced. Furthermore, LdHSFB2a bound to the promoters of HSFA3 A, WRKY33, CAT2, and GLOS1. And LdHSFB2a overexpression and silencing enhanced and reduced their expressions, respectively. Therefore, we speculated that LdHSFB2a may be a coactivator that interacts with transcriptional activators to promote thermotolerance in lily by enhancing the expression of heat-responsive genes such as HSFA3 A, WRKY33, CAT2, and GLOS1.

Drought is a common environmental condition that significantly impairs plant growth. In response to drought, plants close their stomata to minimize transpiration and meanwhile activate many stress-responsive genes to mitigate damage. These stress-related mRNA transcripts require the assistance of RNA-binding proteins throughout their metabolic process, culminating in protein synthesis in the cytoplasm. In this study, we identified HLN1 (Hyaluronan 1), an RNA-binding protein with similarity to the animal hyaluronan-binding protein 4 / serpin mRNA binding protein 1 (HABP4/SERBP1), as crucial for plant drought tolerance. The hln1 loss-of-function mutant exhibited higher transpiration rates due to impaired stomatal closure, making it highly susceptible to drought. Drought stress increased HLN1 expression, and the protein underwent liquid–liquid phase separation (LLPS) to form mRNA-ribonucleoprotein (mRNP) condensates in the cytoplasm under osmotic stress. We identified GAD2 as a potential mRNA target of HLN1. GAD2 encodes the predominant glutamate decarboxylase synthesizing γ‐aminobutyric acid (GABA), a non-proteinogenic amino acid that modulates stomatal movement. RIP-qPCR and EMSA showed that HLN1 binds GAD2 mRNA, which promotes HLN1 condensate formation. In hln1 mutants, GAD2 transcripts were less stable, reducing steady-state mRNA levels. As a result, hln1 accumulated less GABA and exhibited impaired stomatal closure under drought. Conversely, HLN1 overexpression stabilized GAD2 mRNA, increased GABA levels, and enhanced drought tolerance in transgenic plants. GAD2 overexpression in hln1 mutants also rescued the drought-sensitive phenotypes. Overall, our study reveals a mechanism whereby HLN1 stabilizes GAD2 mRNA to enhance GABA production and drought tolerance. These findings provide novel strategies for engineering drought-resistant crops.