Leaf senescence impacts crop yield, quality, and post-harvest performance; for example leaf senescence limits the shelf life of cut flowers. Although the senescence process has been extensively studied at the whole-leaf level, its regulation at the single-cell-type level remains largely unknown. Here, we tracked the cell death process during leaf senescence at the single-cell-type resolution through integrated anatomical, multiomics, molecular and genetic analyses in chrysanthemum (Chrysanthemum morifolium Ramat.). These analyses revealed that jasmonic acid (JA) induces spongy mesophyll cell death in the initial stage of leaf senescence, mediated by the transcription factors CmNAC055 and CmNAC087. These transcription factors regulate the acyl coenzyme A oxidase gene CmACX5, encoding a key JA biosynthetic enzyme. This regulatory module is functionally conserved across diverse species, including chrysanthemum, Arabidopsis thaliana, tomato (Solanum lycopersicum), and rice (Oryza sativa). Indeed, attenuating mesophyll cell death with a JA biosynthesis inhibitor delays tomato fruit de-greening and increases rice grain yield by approximately 6%. This study establishes a dynamic landscape of cell fate determination and hormonal regulation during leaf senescence, providing strategies to increase plant production by modulating leaf senescence.

In many flowering plants, the transition from vegetative growth to reproductive development is regulated by seasonal changes in photoperiod. Under inductive photoperiods, leaves produce the florigen FT (FLOWERING LOCUS T), which is transported to the shoot apex to promote flowering. The photoperiod is known to have a major effect on the flowering of chrysanthemum. In the perennial short-day (SD) plant Chrysanthemum seticuspe, the expression of CsFTL3 (FT-like gene) does not increase immediately after shifting from long-day (LD) to SD conditions but gradually accumulates under continuous SD conditions, peaking during inflorescence development. However, the underlying mechanism remains elusive. We show that CsFDL1 (an ortholog of FD) is upregulated, while CsFTL3 is downregulated in leaves during the initial stage of SD inductions. Furthermore, the expression of CsFTL3 is upregulated in the leaves of CsFDL1-knockdown transgenic lines. CsFDL1 is expressed in leaves and forms a complex with CsFTL3 to recognize several TCGA- and ACGT-containing motifs in the CsFTL3 promoter. The CsFTL3-CsFDL1 complex downregulates CsFTL3 expression, thereby preventing its excessive induction by SD signals and inhibiting precocious floral transition. This study reveals that CsFDL1 acts as a key early repressor in the photoperiodic flowering pathway of chrysanthemum leaf, mediating negative feedback regulation by forming a complex with CsFTL3 to achieve precise temporal control of SD-dependent flowering responses.

Genotype-to-phenotype prediction remains a fundamental challenge in current genetic research. In recent years, it has become possible to construct different predictive models based on genomic data. However, in many horticultural crops, it is difficult to accurately verify genomic variations because of the complexity of their genome, making the application of these genome-based methods challenging. Gene expression reflects both genetic regulatory mechanisms and environmental stimuli, offering potential for predicting phenotypes in plants with complex genomes. Thus, in this paper, we tested the possibility for predicting adult plant phenotypes using the gene expression data from seedlings. By applying the transcriptional-based deep learning methods on cut chrysanthemums (Chrysanthemum spp.), which exhibits a complex genetic background characterized by high repetitiveness, heterozygosity, and genome size and is recognized as a segmental allopolyploid, we found that the method is robust and accurate for predicting continuous variables such as leaf vase life, as well as categorical variables such as flower types on the basis of gene expression data. Moreover, the power and performance of transcriptional-based deep learning methods for prediction was validated in rice (Oryza sativa). Our research shows the good performance of phenotype prediction based on gene expression, with potential applications in future gene chip-based breeding practices.

Cholestatic liver injury (CLI) is a severe liver disorder caused by impaired bile flow, for which effective therapeutic options remain limited. Although the aryl hydrocarbon receptor (AHR) has been implicated in protection against CLI, the mechanisms underlying its dysregulation and functional role during cholestasis remain incompletely understood. Here, we investigated the role of AHR in cholestatic liver injury and the mechanisms contributing to impaired AHR activation using multi-omics approaches. Using mouse models of CLI induced by a 0.1

Ethylene is a key gaseous phytohormone that critically regulates the senescence of postharvest cut chrysanthemum, where early-stage accumulation can rapidly accelerate quality deterioration. Accurate detection and timely regulation of ethylene are therefore essential for effective preservation. Here, we report an intelligent ethylene degradation system that integrates a high-performance tin dioxide (SnO2) gas senser with a thermally activated titanium dioxide (TiO2) catalytic degradation layer, both driven by a shared laser-induced graphene heater. Building on our previous self-heating sensing platform, the system operates in an intermittent sensing mode and triggers thermal degradation only when ethylene concentration exceeds a predefined threshold. The SnO2 sensor exhibits high sensitivity across a wide range (0.05-100 ppm), an ultralow detection limit of 1.956 ppb, and strong resistance to humidity and long-term drift. Guided by sensor feedback, the TiO2 layer achieves rapid ethylene degradation, removing over 66.89% within 5 min for concentrations from 0.5 to 30 ppm, reaching equilibrium significantly faster than conventional photocatalytic systems. Applied to postharvest cut chrysanthemums, sensor-guided regulation effectively suppresses ethylene accumulation while reducing energy consumption by more than 92% compared to continuous operation. This work demonstrates a closed-loop, sensorenabled strategy for intelligent ethylene control with potential applications in postharvest storage and other ethylene-sensitive environments.

IntroductionIntrahepatic cholestasis of pregnancy (ICP) is a cholestatic liver disorder associated with substantial fetal morbidity, including preterm birth, fetal distress, and even intrauterine demise. Although prior studies have documented structural and transcriptional alterations in the placenta during ICP, the mechanistic underpinnings linking maternal cholestasis to adverse fetal outcomes remain incompletely elucidated.MethodsIn this study, a murine model of ICP was established by feeding pregnant C57BL/6 mice a 0.1% DDC (3,5-dicarboxylic acid-1,4-dihydrocollidine) diet from E0.5 to E18.5. We assessed fetal growth and employed multi-omics approaches, including placental transcriptome sequencing, maternal gut microbiome profiling, and serum/placental metabolome analysis.ResultsPlacental transcriptome sequencing revealed that ICP significantly downregulated the expression of antioxidant-related genes including Mgst1, Gstt1, Ggt1, Gpx8, Gstk1, and GSTA4 leading to reduced total antioxidant capacity in placental tissue and elevated levels of malondialdehyde (MDA), a marker of lipid peroxidation. Furthermore, ICP disrupted the maternal gut microbiota, resulting in decreased production of antioxidant microbial metabolites such as valeric acid and erythritol. This deficiency further aggravated oxidative damage in the placenta.DiscussionCollectively, our findings uncover a novel gut microbiota-placenta axis driven by cholestasis, which contributes to fetal IUGR. The maternal cholestasis induces gut dysbiosis, which diminishes the production of valeric acid and erythritol. The deficiency of these metabolites, coupled with a direct suppression of the placental Nrf2/Keap1 antioxidant signaling pathway by cholestasis, leads to placental oxidative stress. This oxidative damage impairs placental function, ultimately resulting in fetal growth restriction. Disrupting this pathogenic cycle may offer a promising therapeutic strategy for preventing or treating ICP-related reproductive disorders.

Soil salinization poses a major threat to global agricultural productivity and plant biodiversity. The phytohormone abscisic acid (ABA) is central to plant adaptation to abiotic stress; however, the mechanisms by which ABA coordinates posttranslational modifications of signaling proteins with epigenetic regulation remain poorly understood. Here, we show that salt stress-induced ABA accumulation up-regulates Heat Shock Factor 4 (CmHSFA4), a gene that is known to enhance chrysanthemum salt tolerance. The ABA responsive transcription factor ABRE binding factor 1 (CmABF1) binds to the CmHSFA4 promoter to activate its expression and also recruits the chromatin remodeler BRAHMA (CmBRM) to repress transcription by limiting H3 lysine-4 trimethylation (H3K4me3) deposition. We further demonstrate that the ABA-activated sucrose non-fermenting-1-related protein kinase 2.2 (CmSnRK2.2) phosphorylates and stabilizes CmABF1, while concurrently phosphorylating and promoting CmBRM degradation under salt stress. This dual regulation enhances H3K4me3 enrichment at the CmHSFA4 promoter, thereby inducing its transcription and conferring salt tolerance. Together, our findings reveal an ABA-SnRK2.2-ABF1/BRM signaling module that integrates phosphorylation-dependent protein stabilization and degradation with histone methylation dynamics to fine-tune salt stress-responsive gene expression in chrysanthemum.

Black spot disease, caused by the fungus Alternaria alternata, is a global plant pathogen that lacks sustainable control measures and poses a serious threat to multiple economically important crops. One approach to enhancing disease resistance in susceptible plants is grafting them onto disease-resistant rootstocks, yet the mechanisms by which roots enhance disease resistance in shoots remain largely unknown. Here, using chrysanthemum-Artemisia vulgaris grafts, we identified that a raffinose synthase-encoding gene, CmRS6, is essential in the susceptible chrysanthemum scion for graft-transmitted resistance from the disease-resistant A. vulgaris rootstock. Exogenous raffinose treatments enhanced A. alternata resistance in chrysanthemum, tomato, cabbage, and apple, highlighting its broad defensive role. The CmERF1B transcription factor activated CmRS6 expression and raffinose accumulation, whereas CmJAZ1-like repressed CmERF1B via direct interaction. While we found evidence for rootstock-to-scion transport of raffinose, long-distance jasmonate (JA) transport from A. vulgaris rootstocks to chrysanthemum scions was the primary mechanism for graft-transmitted resistance, and A. alternata infection further promoted JA transport. Collectively, our study demonstrates that rootstock-to-scion JA transport mediates graft-transmitted A. alternata resistance by upregulating scion raffinose biosynthesis, thereby offering new strategies for the sustainable control of A. alternata in crops.

As the primary fungal pathogen of apple (Malus domestica) blue mold, Penicillium expansum severely affects the post-harvest quality of apples. Protein-protein interactions play a crucial role in plant resistance to biotic stress. Here, we re-analyse the previously obtained proteomic data from apples infected with Penicillium expansum and identify numerous differentially expressed proteins that may significantly contribute to their defence mechanisms. The MdACBP protein is expressed during the defence response of apples against P. expansum infection and exhibits a high binding affinity for acyl-CoA and phospholipids, suggesting a crucial role in the apple defence mechanism. In this study, through bioinformatics analysis, subcellular localization, yeast two-hybrid assays, and interaction site prediction, we identified MdACBP as a member of the ACBP1 family, localised in the cytoplasm. During P. expansum infection, MdACBP interacts with A0A0A2IS21 (Pe25) with a confidence score of 0.8783. In conclusion, our findings suggest that MdACBP and Pe25 act together to defend against P. expansum infection in apples, providing a theoretical basis for the prevention of P. expansum induced blue mold in apples.

Melatonin is synthesized in multiple tissues and organs of pigs, and existing studies have shown the presence of the melatonin-synthesizing enzyme ASMT protein. However, the genomic information for the ASMT gene has been lacking. The aim of this study was to locate the genomic information of the ASMT gene in pigs using comparative genomics analysis and then obtain the coding region information through molecular cloning. First, using the NCBI Genome Data Viewer, we found that in most animals, the AKAP17A gene is often located next to the ASMT gene, with both genes arranged in the same direction. Similarly, the P2RY8 gene is commonly adjacent to the ASMTL gene, also in the same orientation. We also discovered that the ASMTL gene is frequently adjacent to the ASMT gene and arranged in the opposite direction. Using the “three-point localization” principle, we inferred the position of the ASMT gene based on the coordinates of AKAP17A and ASMTL in pigs. Our results revealed that on the pig X chromosome, a gene called LOC110258194 is located next to the AKAP17A and ASMTL genes, and its arrangement aligns with the ASMT gene in other species. Additionally, Ensembl contains a gene, ENSSSCG00000032659, at the same position, with completely overlapping exons, though it is not annotated as ASMT. Further analysis using the TreeFam tool from EMBL-EBI and the CDD tool from NCBI revealed that LOC110258194 and ENSSSCG00000032659 do not contain the typical Maf domain of ASMTL and, thus, should not be annotated as ASMTL, but rather as the ASMT gene. Using a slow-down PCR method for high-GC content genes, we successfully cloned the full CDS region of the pig ASMT gene and identified a new transcript missing Exon 6 and Exon 7. This transcript was submitted to NCBI and assigned the GenBank accession number MW847601. Our results represent the first successful localization of the ASMT gene in pigs, the first cloning of the ASMT gene’s coding region, and the first discovery of a new transcript of the pig ASMT gene.

BACKGROUND:N6-methyladenosine (m6A) is a prevalent and conserved RNA modification in eukaryotes. While its roles in the 3' untranslated regions (3' UTR) are well-studied, its role in the 5' UTR and its relationship with histone modifications remain underexplored. RESULTS:We demonstrate that m6A methylation in the 5' UTR of mRNA triggers a downstream shift in H3K4me3 modification. This regulatory mechanism is conserved in Arabidopsis, rice, and chrysanthemum. The observed shift in H3K4me3 is genetically controlled by m6A modifiers and influences gene expression. MTA, the m6A methylase, preferentially binds to phosphorylated serine 5 (Ser5P)-CTD of RNA Pol II during transcription, leading to the displacement of ATX1, the H3K4me3 methylase. This dynamic binding of MTA and ATX1 to RNA Pol II ultimately results in the shift of H3K4me3 modification. Genetic evidence demonstrates that m6A in the 5' UTR controls H3K4me3 shift, thereby affecting SEDOHEPTULOSE-BISPHOSPHATASE expression and leaf senescence. CONCLUSIONS:Our study provides new insights into the roles of m6A modification and its crosstalk with histone modification in 5' UTRs, shedding light on the mechanism of m6A-mediated gene expression regulation.

Leaf senescence in plants is a coordinated cell death process that impacts the post-harvest performance horticultural plants. While the mechanisms of leaf senescence have been extensively studied, the translational control of this process remains largely unexplored. In this study, we demonstrated that leaf senescence chrysanthemum is controlled by mRNA m6A methylation-mediated translational regulation. We found that methylation is decreased during leaf senescence. Inhibiting the expression of the m6A eraser CmALKBH10B leads to increased m6A modifications and delayed leaf senescence, whereas overexpression of CmALKBH10B results reduced m6A levels and accelerated senescence. The majority of genes differentially modified by m6A exhibited no significant changes at the expression level, implying that translational regulation may contribute to senescence. Genetic and molecular evidences indicated that m6A modification impacts the translation efficiency of CmEIN2 gene, thereby influencing leaf senescence. Our findings provide crucial insights into the translational regulatory mechanism of leaf senescence, adding another layer of complexity to the comprehensive regulatory networks governing this process.

The role of ethylene as an initial signaling molecule in waterlogging stress is well-established. However, the complex molecular mechanisms underlying ethylene biosynthesis and its functional significance in chrysanthemums under waterlogging conditions have remained unclear. In this study, we observed an increase in the expression of 1-aminocyclopropane-1-carboxylate synthase 6 (CmACS6), which encodes a key enzyme responsible for ethylene biosynthesis, in response to waterlogging. This elevation increases ethylene production, induces leaf chlorosis, and enhances the chrysanthemum's sensitivity to waterlogging stress. Moreover, our analysis of upstream regulators revealed that the expression of CmACS6, in response to waterlogging, is directly upregulated by CmHRE2-like (Hypoxia Responsive ERF-like, CmHRE2L), an ethylene response factor. Notably, CmHRE2-L binds directly to the GCC-like motif in the promoter region of CmACS6. Genetic validation assays demonstrated that CmHRE2L was induced by waterlogging and contributed to ethylene production, consequently reducing waterlogging tolerance in a partially CmACS6-dependent manner. This study identified the regulatory module involving CmHRE2L and CmACS6, which governs ethylene biosynthesis in response to waterlogging stress.

Leaf senescence impacts the quality, productivity and post-harvest performance of crops and ornamental plants. However, the underlying molecular mechanisms governing this process remain poorly understood. In this study, we identified CmbHLH63 as a 'brake signal' of leaf senescence in chrysanthemum and elucidated its critical regulatory role within the CmbHLH1L-CmbHLH63-CmNLP6/7L complex. CmbHLH63 promotes leaf senescence, while it is downregulated during this process. Notably, CmbHLH63 lacks intrinsic transcriptional regulatory activity; however, the expression of a majority of genes was repressed in plants overexpressing CmbHLH63. Further investigation revealed that this repression is achieved through recruiting the transcriptional repressor CmbHLH1L. We further found that CmNLP6/7L, a transcriptional activator, competes with CmbHLH1L for the interaction with CmbHLH63, leading to the release of their inhibition on downstream gene CmNLP6/7L. We found that the accumulation of CmbHLH1L is increased in the cytoplasm and decreased in the nucleus during leaf senescence. Mass spectrometry assays and subcellular localization studies revealed that CmbHLH1L undergoes lysine acetylation at position 140, which prevents its nuclear accumulation, thereby elevating CmNLP6/7L expression during leaf senescence.

Terpenoids represent one of the most varied groups of secondary metabolites and are essential in numerous biological functions in plants, especially in response to biotic stress. However, the molecular mechanisms underlying the biosynthesis of sesquiterpenes in response to aphid infestation and the impact of sesquiterpenes on aphid feeding behavior remain unclear. Here, we focus on CmTPS1 as a starting point to analyze its catalytic activity in the synthesis of sesquiterpenes. Then, the transcriptional regulatory mechanisms of sesquiterpenes and their role in the defense of chrysanthemum against aphids were explored. After aphid feeding, the methyl jasmonate content in chrysanthemum increased, and the CmMYBML1 expression was upregulated. CmMYBML1 directly binds to the CmTGA1 promoter and activates its expression, and CmTGA1 directly binds to the promoter of the chrysanthemum sesquiterpene synthase gene CmTPS1 and promotes its expression. The cascade regulates the synthesis of various sesquiterpenes, including β-Caryophyllene, β-Copaene, (E)-β-Farnesene, Germacrene D, and Aromadendrene, ultimately enhancing the chrysanthemum's defense against aphids. This study is the first to show that the CmMYBML1-CmTGA1-CmTPS1 module regulates sesquiterpene biosynthesis in response to aphid feeding. Our findings provide new insights into a novel regulatory network by which plants modulate the synthesis of sesquiterpenes to defend against herbivores in chrysanthemum.

Type 2 diabetes mellitus (T2DM) poses a significant global health challenge. Genome-wide Association Studies have linked T2DM to genetic variants in the melatonin receptor 1a (MTNR1A) and 1b (MTNR1B) genes, which encode the MT1 and MT2 receptors, respectively. Our results found that the rs2119882 MT1 mutation was associated with higher blood glucose levels and increased body mass index (BMI) in humans. Metabolomic analysis showed elevated levels of palmitic acid (a saturated fatty acid) and reduced levels of oleic acid (an unsaturated fatty acid) in individuals with this mutation. In contrast, the rs10830963 MT2 mutation did not show the significant differences in blood glucose level or BMI compared to normal control individuals. Inhibition of MTNR1A and MTNR1B expression led to lower GLUT-4 mRNA and insulin receptor protein levels in human liver cells, resulting in decreased glycogen synthesis and metabolic disruptions. We used CRISPR/Cas9 to create MTNR1A and MTNR1B knockout (KO) mice, which also exhibited reduced GLUT-4 and INSR mRNA levels, decreased glucose tolerance, and increased insulin resistance. These mice also developed obesity, liver lipid deposition, increased abdominal white adipose tissue, and lower androgen levels. Metabolomic and proteomic analyses of the KO mice revealed increased triglycerides and phospholipids, and decreased unsaturated fatty acids. Proteomic studies showed reduced levels of insulin receptor tyrosine kinase, lipid droplet-associated hydrolase, and glucose-6-phosphate dehydrogenase, disrupting fatty acid metabolism and increasing liver lipid deposition. Additionally, a high-fat diet challenge in MTNR1A and MTNR1B KO male mice accelerate the INSR protein expression suppression, hepatic triglyceride accumulation, blood glucose elevation and weight gain. Finally, we generated AANAT over-expressing sheep, which showed improved glucose tolerance and higher insulin levels after glucose injection compared to WT sheep. These findings underscore the importance of melatonin and its receptors in glucose and lipid metabolism, suggesting their deficiencies may contribute to T2DM.

Proper nutrition in late pregnancy is crucial for postpartum recovery and piglet growth. Improper management can lead to various physiological stresses in sows, resulting in reduced reproductive performance. This study aimed to assess the potential benefits of short-term supplementation with tryptophan on late-pregnant sows, fed from day 99 to day 114 of gestation. We divided 21 Landrace hybrid sows into three groups. The control group was fed a basal diet, while the experimental groups were fed basal diets supplemented with 0.1% and 0.2% tryptophan, respectively. Results indicated that feeding 0.1-0.2% tryptophan had no significant effect on sows' glucose and lipid metabolism. However, compared to the control group, tryptophan supplementation significantly increased reproductive hormone (E2) levels in sows, with no significant impact on immune function and oxidative status. Furthermore, sows showed significant improvements in reproductive per-formance after tryptophan supplementation, particularly in weaning survival rate, and average litter weight at birth,. Further investigation revealed that tryptophan supplementation significantly increased melatonin levels in sows. Given melatonin's beneficial effects on mammalian reproductive activity, it is inferred that these effects are partly mediated by tryptophan derived melatonin. In conclusion, supplementation with tryptophan in late pregnancy positively influences reproductive activity in sows and the growth of newborn piglets. Due to the limitation of sample size, these effects are only preliminary assessments. In the future, we will expand the sample size and further explore its mechanism.

Pest image datasets play a crucial role in improving the accuracy of pest recognition models, enabling automated monitoring and precise pest control, and advancing intelligent agriculture. However, existing agricultural pest datasets have limitations in terms of species diversity and the number of samples, and only the appearance of the pest is considered while ignoring professional agricultural knowledge. In this paper, a large-scale dataset, termed AP162, is proposed for the recognition of agricultural pests, which consists of 162 pest categories and 194,700 images. AP162 takes into account both the visual appearance of agricultural pests and their harmful characteristics for crops. Different subset divisions are adopted to explore the appearance features and potential damage to pests from two perspectives. The agriculture-based perspective classifies pests according to crop damage types to support targeted pest control, while the vision-based perspective relies on morphological similarities. Extensive experiments have been conducted on the AP162 dataset utilizing recent deep learning methodologies, thereby establishing it as a new benchmark. This paper proposes a weighted fusion strategy that agricultural domain knowledge is integrated with vision-based fine-grained feature, achieving a recognition accuracy of 89.2% on AP162. It is publicly available at https://github.com/SCNYDX-KL/AP162.

Excessive soil salinity not only hampers plant growth and development but can also lead to plant death. Previously, we found that heat-shock factor A4 (CmHSFA4) enhances the tolerance of chrysanthemum (Chrysanthemum morifolium) to salt. However, the underlying molecular mechanism remains unclear. In this study, we identified a candidate MYB transcription factor, CmMYB121, which responded to salt stress. We observed that the CmMYB121 transcription is suppressed by CmHSFA4. Moreover, overexpression of CmMYB121 exacerbated chrysanthemum sensitivity to salt stress. CmHSFA4 directly bound to the promoter of CmMYB121 at the heat-shock element. Protein-protein interaction assays identified an interaction between CmHSFA4 and CmMYBS3, a transcriptional repressor, and recruited the corepressor TOPLESS (CmTPL) to inhibit CmMYB121 transcription by impairing the H3 and H4 histone acetylation levels of CmMYB121. Our study demonstrated that a CmHSFA4-CmMYBS3-CmTPL complex modulates CmMYB121 expression, consequently regulating the tolerance of chrysanthemum to salt. The findings shed light on the responses of plants to salt stress.