Search Results (341)

The evolutionarily conserved Notch signalling pathway regulates the fate, proliferation and differentiation of cells in most developing organs, thus affecting their morphogenesis and function. Here, we investigated the role of the Notch2 receptor in the generation and function of epithelial cells of the continuously erupting rodent incisors. We used transgenic Notch1-Cre^ERT2/+;Rosa26^mT/mG and Notch2-Cre^ERT2/+;Rosa26^mT/mG mice to compare the contribution of Notch1- and Notch2-expressing cells and their progeny in the generation of the different epithelial cell populations. Furthermore, we examined if the dental epithelium organisation and enamel structure are affected in early postnatal incisors of Keratin14^Cre/+;Notch2^fl/fl mice using immunofluorescent staining, gene expression analysis, microcomputed tomography and scanning electron microscopy. Our results showed that Notch2 deletion resulted in smaller incisors with disorganised dental epithelium and defective enamel. Delayed eruption was correlated with alterations in the proliferative and differentiation status of epithelial stem cells in the cervical loop area of the incisors. Similar results were obtained with in vitro studies, where inhibition of the Notch signalling by the CB103 blocker recapitulated the in vivo phenotype. In conclusion, this study demonstrates for the first time the importance of Notch2 in epithelial cell fate acquisition, dental epithelium organisation and enamel structure in rodent incisors. Full article

The midgut of Antheraea pernyi plays a critical role in antiviral defense. However, its transcriptional complexity remains poorly understood. Here, a full-length (FL) transcriptome atlas of A. pernyi midgut was developed by integrating PacBio Iso-Seq and RNA-seq techniques. The transcriptome sequences included 1850 novel protein-coding genes, 17,736 novel alternative isoforms, 1664 novel long non-coding RNAs (lncRNAs), and 858 transcription factors (TFs). In addition, 2471 alternative splicing (AS) events and 3070 alternative polyadenylation (APA) sites were identified. Moreover, 3426 and 4796 differentially expressed genes (DEGs) and isoforms were identified after ApNPV infection, respectively, besides the differentially expressed lncRNAs (164), TFs (171), and novel isoforms of ApRelish (1) and ApSOCS2 (4). Enrichment analyses showed that KEGG pathways related to metabolism were suppressed, whereas GO terms related to DNA synthesis and replication were induced. Furthermore, the autophagy and apoptosis pathways were significantly enriched among the upregulated genes. Protein–protein interaction network (PPI) analysis revealed the coordinated downregulation of genes involved in mitochondrial ribosomes, V-type and F-type ATPases, and oxidative phosphorylation, indicating the disruption of host energy metabolism and organelle acidification. Moreover, coordinated upregulation of genes associated with cytoplasmic ribosomes was observed, suggesting that the infection by ApNPV interferes with host translational machinery. These results show that ApNPV infection reprograms energy metabolism, biosynthetic processes, and immune response in A. pernyi midgut. Our study provides a foundation for elucidating the mechanisms of A. pernyi–virus interactions, particularly how the viruses affect host defense strategies. Full article

Huanglongbing (HLB), caused by Candidatus Liberibacter asiaticus (CLas), is the most devastating disease threatening global citrus production. Although no commercial citrus varieties exhibit complete HLB resistance, genotype-specific tolerance variations remain underexplored. This study conducted a comparative transcriptomic profiling of six commercially citrus cultivars in South China, four susceptible cultivars (C. reticulata cv. Tankan, Gongkan, Shatangju, and C. sinensis Osbeck cv. Newhall), and two tolerant cultivars (C. limon cv. Eureka; C. maxima cv Guanxi Yu) to dissect molecular mechanisms underlying HLB responses. Comparative transcriptomic analyses revealed extensive transcriptional reprogramming, with tolerant cultivars exhibiting fewer differentially expressed genes (DEGs) and targeted defense activation compared to susceptible genotypes. The key findings highlighted the genotype-specific regulation of starch metabolism, where β-amylase 3 (BAM3) was uniquely upregulated in tolerant varieties, potentially mitigating starch accumulation. Immune signaling diverged significantly: tolerant cultivars activated pattern-triggered immunity (PTI) via receptor-like kinases (FLS2) and suppressed ROS-associated RBOH genes, while susceptible genotypes showed the hyperactivation of ethylene signaling and oxidative stress pathways. Cell wall remodeling in susceptible cultivars involved upregulated xyloglucan endotransglucosylases (XTH), contrasting with pectin methylesterase induction in tolerant Eureka lemon for structural reinforcement. Phytohormonal dynamics revealed SA-mediated defense and NPR3/4 suppression in Eureka lemon, whereas susceptible cultivars prioritized ethylene/JA pathways. These findings delineate genotype-specific strategies in citrus–CLas interactions, identifying BAM3, FLS2, and cell wall modifiers as critical targets for breeding HLB-resistant cultivars through molecular-assisted selection. This study provides a foundational framework for understanding host–pathogen dynamics and advancing citrus immunity engineering. Full article

The objective of this experiment was to investigate the effects of tartary buckwheat flavonoids (TBF) and 25-hydroxyvitamin D₃ (25-OHD) on fatty liver syndrome (FLS) in laying hens. A total of 450 35-wk-old Lohmann laying hens were selected and randomly divided into five groups, with six replicates per treatment and 15 laying hens in each replicate. The control group was fed a corn-soybean meal basal diet. The FLS group was fed a high- energy–low-protein (HELP) diet, and the other three experimental groups were fed HELP diets supplemented with 60 mg/kg TBF, 69 μg/kg 25-OHD, and 60 mg/kg TBF plus 69 μg/kg 25-OHD, respectively. The experiment lasted 8 weeks. The results demonstrated that feeding laying hens with a HELP diet led to a significant accumulation of fat in their livers, liver enlargement and yellowing, as well as a decline in liver antioxidant capacity and an aggravation of inflammation. TBF alone, 25-OHD alone, and their combination had no effect on the laying performance of laying hens fed with a HELP diet. However, 25-OHD significantly enhanced the albumin content, eggshell strength, and eggshell thickness of eggs (p < 0.05). Compared with the HELP group, TBF, 25-OHD, or their combination reduced serum LDL-C and TG (p < 0.05). The combined treatment further lowered serum NEFA and MDA, enhanced liver SOD activity (p < 0.05), and unlike TBF alone (which reduced hepatic TG) or 25-OHD alone (which decreased liver index), reduced both liver index and hepatic TG (p < 0.05). Liver gene expression analysis showed that combined TBF and 25-OHD significantly inhibited the expression of fat synthesis-related genes (ACC, FAS, GPAT1, ChREBP1, LXRα, SREBP-1C, SREBP-2, FABP) as well as inflammation-related genes (IL-6, TNF-α, NF-κB, TLR4) (p < 0.05). At the phylum level of the cecal microbiota, TBF increased the abundance of Bacteroidota (p < 0.05), and combined TBF and 25-OHD tended to increase the abundance of Firmicutes_D. At the genus level, TBF increased the abundance of Phocaeicola_A (p < 0.05). Furthermore, TBF, 25-OHD, or their combination reduced the abundance of Faecalibacterium (p < 0.05). These findings suggest that combined TBF and 25-OHD mitigates FLS in laying hens potentially through remodeling gut microbiota and maintaining lipid metabolic homeostasis. Full article

Follicular lymphoma (FL) is one of the most common types of non-Hodgkin’s lymphomas. The tumor is characterized by a wide range of clinical manifestations, ranging from indolent forms to early transformation and progression with a poor prognosis. The search for clinically significant genetic changes is essential for personalized risk assessment and treatment selection. The CREBBP gene is frequently mutated in this type of lymphoma, with changes occurring at the level of the earliest tumor precursor cells. However, the prognostic and diagnostic significance of the CREBBP gene mutation status in FL has not been fully established. In this study, we analyzed sequencing data of exons 22–30 of the CREBBP gene in 86 samples from patients with different grades of FL (1–3B), including those in the 3A–3B subgroup without the t(14;18) translocation. We also investigated the prognostic significance of CREBBP gene mutations in relation to the treatment options, namely high-dose chemotherapy with autologous hematopoietic stem cell transplantation (HDCT/auto-HSCT) and conventional chemotherapy programs (CCT). It was found that FL patients with a single missense mutation in the KAT domain of the CREBBP gene experienced an extremely low number of early adverse events related to lymphoma and had better long-term survival rates, regardless of treatment option. In contrast, when comparing patients with FL without a missense mutation in the KAT domain or those with multiple mutations in the CREBBP gene, overall and progression free survival were worse, and early progression and histological transformation were more common. Compared to standard therapy, patients who underwent HDCT/auto-HSCT in the FL 1–3B (14;18)-positive group without a single missense mutation in the KAT domain had better survival rates and lower rates of transformation and early progression. In addition, among patients with FL 3A–3B (14;18)-negative, we found that there were no cases of a missense mutation in the KAT domain of the CREBBP gene. This suggests that a single missense mutation in the CREBBP gene may be a feature that discriminates 14;18-positive FL with a favorable prognosis from a high-risk disease. FL 3A–3B (14;18)-negative may represent a distinct variant with different biology and underlying mechanisms of development compared to classical FL. Full article

Quinoa (Chenopodium quinoa Willd) is a tetraploid crop that has provided vital subsistence, nutrition, and medicine for Andean indigenous cultures. In recent years, quinoa has gained global importance all over the world. However, variations in fertility have been frequently observed during the flower development of quinoa, severely affecting quinoa production. To comprehend the fundamental causes of fertility variation in quinoa, this research examined hormonal metabolism and gene expression across three ecotypes: normal fertility (F), absent stamens (S1), and abnormal stamens (S3). S1 and S3 presented absent and abnormal stamens, respectively, compared with F. Phytohormone profiling yielded 60 metabolites and revealed the clear separation between different ecotypes at different developmental stages according to principal component analysis (PCA). The results of transcriptomics showed more DEGs (differentially expressed genes) identified between F and S1 ecotypes (8002 and 10,716 for earlier and later stages, respectively) than F vs. S3 (4500 and 9882 for earlier and later stages, respectively) and S1 vs. S3 (4203 and 5052 for earlier and later stages, respectively). Zeatin biosynthesis and hormone signal transduction pathways were enriched among 19 KEGG (Kyoto Encyclopedia of Genes and Genomes) terms, indicating their potential roles in quinoa flower fertility regulation. The correlation-based network presented the associations between selected hormones and genes, possibly regulating fertile ecotypes. Furthermore, we explored the expression of flower development-related genes in three ecotypes using RT-PCR, showing the higher expressions of AP1, AP3, and FLS in sterile ecotypes than fertile ecotypes at both stages. These findings reveal new insights into the hormonal and genetic regulations of floral fertility in quinoa, which may have consequences for developing high-yielding cultivars. Full article

A 14-day study was conducted to evaluate the effect of litter type (dirty litter, DL; fresh litter, FL) and Salmonella Enteritidis SE challenge (no challenge, NC; challenge, SE) on the growth performance and cecal microbial composition of neonate chicks. Day-old chicks (n = 240, Ross 708 male) were allocated to a 2 × 2 factorial design consisting of four treatments: chicks raised on dirty litter (CONDL), chicks raised on fresh litter (CONFL); and chicks raised on litter types similar to CONDL and CONFL but inoculated with 7.46 × 108 CFU SE/mL at d 1 (CONDLSE and CONFLSE). The performance indices measured included body weight (BW), body weight gain (BWG), feed intake (FI), mortality, and feed conversion ratio (FCR). Cecal SE concentration was assessed on d 3 and 14, and ceca were collected from chicks on day 14 for DNA extraction. The Illumina Miseq platform was used for microbiome analysis of the V3–V4 region of the 16S rRNA gene. The interaction of litter type and SE influenced FCR and FI. CONDL recorded the poorest FCR (1.832). FI was highest and similar in CONFLSE, CONDL, and CONDLSE (0.655, 0.692, and 0.677, respectively). Cecal SE concentration was significantly reduced in CONDLSE at d 3 and 14. Alpha diversity was higher (p < 0.05) in the DL compared to that in NC. Beta diversity showed a separation (p < 0.05) between the DL and the FL. Comparative tree analysis revealed 21 differential significant genera, with 14 prevalent in the DL and 7 in the FL, specifically, bacteria genera such as Lactobacillus, Clostridia_vadinBB60_group, Lachnospira, Oscillospiraceae UCG_005, and Marvinbryantia, which play significant roles relating to improved growth performance, metabolic homeostasis within the gut, energy metabolism, and short-chain fatty acid (SCFA) utilization. Our results concluded that litter management regimen differentially alters the microbiome of chicks, which accounts for the improved performance and exclusion of pathogens in the study. Full article

Plant callus cultures are a sustainable alternative for producing bioactive secondary metabolites, but their low yields limit industrial applications. Carob (Ceratonia siliqua L.) is rich in medicinally valuable compounds, yet conventional cultivation faces challenges. To address this, we use biofortified calcium phosphate nanoparticles, which refer to CaP-NPs that have been enriched with bioactive compounds via green synthesis using Jania rubens extract, thereby enhancing their functional properties as elicitors in carob callus. CaP-NPs were green-synthesized using Jania rubens extract and applied to 7-week-old callus cultures at 0, 25, 50, and 75 mg/L concentrations. At the optimal concentration (50 mg/L), CaP-NPs increased callus fresh weight by 23.9% and dry weight by 35.1%. At 50 mg/L CaP-NPs, phenolic content increased by 95.7%, flavonoids by 34.4%, tannins by 131.8%, and terpenoids by 211.9% compared to controls. Total antioxidant capacity rose by 76.2%, while oxidative stress markers malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) decreased by 34.8% and 14.1%, respectively. Gene expression analysis revealed upregulation of PAL (4-fold), CHI (3.15-fold), FLS (1.16-fold), MVK (8.3-fold), and TA (3.24-fold) at 50 mg/L CaP-NPs. Higher doses (75 mg/L) induced oxidative damage, demonstrating a hormetic threshold. These findings indicate that CaP-NPs effectively enhance secondary metabolite production in carob callus by modulating biosynthetic pathways and redox balance, offering a scalable, eco-friendly approach for pharmaceutical and nutraceutical applications. Full article

Comfrey (Symphytum officinale L.) is a traditional medicinal plant, and its growth period has an important effect on the accumulation of active components. Phenolic acids and flavonoids are the most important active components in comfrey, but their accumulation in comfrey has not been studied. At present, most research on comfrey focuses on its roots. There is still a lack of systematic research on the comparison of active components and biological activities in the aerial parts of comfrey in different growth periods. To explore the influence of the growth period on the active components of comfrey, non-targeted metabolomics and transcriptomics were used to comprehensively analyze the active components of comfrey during the vegetative period, blooming period, and maturity period and compare the dynamic changes in phenolic acid and flavonoid accumulation during different growth periods of comfrey. The results revealed that the vegetative period presented the highest total phenol and flavonoid contents. The predominant secondary metabolites associated with phenolic acids and flavonoids were integral to the phenylpropanoid, flavonoid, and isoflavonoid biosynthetic pathways. Critical structural genes governing these metabolic processes—PAL, C4H, 4CL, CHS, FLS, and DFR—exhibited marked upregulation during the vegetative growth stage. Comprehensive transcriptomic analysis and weighted gene co-expression network analysis were used to construct a co-expression network of structural genes and transcription factors that affected the accumulation of specific metabolites, and the transcription factors related to the synthesis of flavonoids and phenols were predicted. These findings elucidate the temporal regulatory mechanisms governing the growth-phase-dependent accumulation of bioactive constituents in comfrey, advancing the understanding of phytochemical dynamics in medicinal plants. Full article

Soybean frogeye leaf spot (FLS) disease has been reported globally and is caused by the fungus Cercospora sojina, which affects the growth, seed yield, and quality of soybean. Among the 15 physiological microspecies of C. sojina soybean in China, Race 7 is one of the main pathogenic microspecies. A few genes are involved in resistance to FLS, and they cannot meet the need to design molecular breeding methods for disease resistance. In this study, a soybean recombinant inbred line (RIL3613) population and a germplasm resource (GP) population were planted at two sites, Acheng (AC) and Xiangyang (XY). Phenotypic data on the percentage of leaf area diseased (PLAD) in soybean leaves were obtained via image recognition technology after the inoculation of seven physiological species and full onset at the R3 stage. Quantitative trait loci (QTLs) and quantitative trait nucleotides (QTNs) were mapped via linkage analysis and genome-wide association studies (GWASs), respectively. The resistance genes of FLS were subsequently predicted in the linkage disequilibrium region of the collocated QTN. We identified 114 QTLs and 18 QTNs in the RIL3613 and GP populations, respectively. A total of 14 QTN loci were colocalized in the two populations, six of which presented high phenotypic contributions. Through haplotype–phenotype association analysis and expression quantification, three genes (Glyma.06G300100, Glyma.06G300600, and Glyma.13G172300) located near molecular markers AX-90524088 and AX-90437152 (QTNs) are associated with FLS Chinese Race 7, identifying them as potential candidate resistance genes. These results provide a theoretical basis for the genetic mining of soybean antigray spot No. 7 physiological species. These findings also provide a theoretical basis for understanding the genetic mechanism underlying FLS resistance in soybeans. Full article

Flavonoids are an important type of bioactive substance contained in jujubes. Flavonol synthase (FLS) is a key enzyme for the synthesis of flavonoids such as flavonols and anthocyanins. To study the biological functions of FLS in jujubes, we cloned the ZjFLS gene; analyzed its physicochemical properties and evolutionary relationships; and then conducted an expression characteristic analysis, subcellular localization, prokaryotic expression and heterologous overexpression in Arabidopsis thaliana. The results showed that the length of ZjFLS is 951 bp, and it encodes 316 amino acids. A sequence analysis revealed that ZjFLS exhibited a high degree of conservation in evolution. The results of a qRT-PCR analysis indicated that the ZjFLS gene could be expressed in different tissues of jujube: the expression level was the highest in the leaves, followed by the flowers, and the lowest was in the fruits. Within these expression levels, it was higher in young leaves than in mature leaves and higher in the white-ripe-stage fruits than in the semi-red-stage fruits. Subcellular localization indicated that the ZjFLS gene was located in the nucleus, cytoplasmic matrix, and cytoplasmic membrane. Our research findings show that the ZjFLS protein can be induced and obtained in the prokaryotic expression system and successfully purified. It mainly exists in the form of inclusion bodies and has a relatively low content in the soluble supernatant. The total flavonoid content of Arabidopsis thaliana strains with a heterologous overexpression of the ZjFLS gene was significantly higher than that of the wild type, confirming that the ZjFLS gene can promote the biosynthesis of flavonoid substances. Full article

Adult neurogenesis in the hippocampal dentate gyrus (DG) is not only essential for learning and pattern separation, but it is also involved in emotional regulation. This process is vulnerable to local and peripheral inflammation, which is partly mediated by microglia in the DG. As Crohn’s disease (CD) is associated with neuropsychiatric comorbidity, including depression and cognitive impairment, a reduction in adult hippocampal neurogenesis by chronic gut-derived inflammation has been hypothesized. Here, we present the first study that examined the influence of chronic ileocolitis on microglia in the DG and on adult hippocampal neurogenesis in a transgenic mouse model of CD, which is generated by a constitutive knockout of caspase 8 in intestinal epithelial cells (IECs, Casp8^ΔIEC mice). Structural and transcriptional analyses revealed that microglial cell proliferation and density in the DG as well as the expression of genes associated with their homeostasis and activation in the forebrain were maintained in 14- and 24-week-old Casp8^ΔIEC mice compared to Casp8^fl controls. Furthermore, different stages of adult hippocampal neurogenesis, including progenitor cell proliferation, maturation, and apoptosis of newly generated cells, were predominantly unaffected by chronic ileocolitis, except a potential minor phenotypic shift in maturating cells in 24-week-old mice. Together, we demonstrate largely preserved adult hippocampal neurogenesis, lacking signs of local inflammatory microglial activation despite chronic inflammation of the gut. Full article

Background/Objectives: Antibiotics have a significant impact on the gut microbiota, and we hypothesized that human milk oligosaccharides may alleviate antibiotic-induced gut microbiota dysbiosis. Methods: Six groups of eight mice were administered drinking water with or without ampicillin for one week. We then introduced the human milk oligosaccharide 2′-fucosyllactose (2′FL), either alone or in combination with difucosyl-lactose (DFL), for two weeks after the termination of ampicillin treatment. Results: Ampicillin reduced microbiota diversity and the abundance of specific bacteria. One week after the termination of ampicillin treatment, the 2′FL + DFL mixture counteracted the ampicillin-induced reduction in diversity, although this effect was not sustained. Over the subsequent two weeks, the 2′FL + DFL mixture had a significant impact on the relative abundances of Lactobacillus spp. and Bacteroides spp. Ampicillin also reduced caecal propionate levels, downregulated the gene Gzmb for Granzyme B, and upregulated the gene Reg3a for Regenerating islet-derived protein 3 alpha, all of which were counteracted by the 2′FL + DFL mixture. Ampicillin had a minor impact on ileal cytokine levels. The 2′FL + DFL mixture showed a cytokine effect indicating reduced adaptive and innate inflammation. Ampicillin reduced water intake and growth in the mice. The oligosaccharides did not affect water intake, but the 2′FL + DFL mixture slightly reduced body weight. Conclusions: The 2′FL + DFL mixture appears to hold potential for counteracting some of the side effects of ampicillin treatment. Full article

Drought ranks among the key abiotic stresses that limit the growth and yield of grapevines (Vitis vinifera L.). Flavonols, a class of antioxidants commonly found in grapevines, play a crucial role in combating drought stress. In this study, we characterized the function and regulatory mechanism of the grapevine VviMYC4 in mediating flavonol biosynthesis in response to drought stress. VviMYC4 encodes a protein of 468 amino acids with conserved bHLH-MYC_N and bHLH domains. Phylogenetic analysis confirmed its homology with the grapevine VviMYC2 and similarity in function. The expression of VviMYC4 in ‘Cabernet Sauvignon’ grapevine seedling leaves increased initially and then decreased during prolonged drought stress. The homologous and heterologous transformation of VviMYC4 in grape suspension cells, Arabidopsis plants, tobacco leaves, and grapevine leaves demonstrated its ability to positively regulate flavonol biosynthesis and accumulation by promoting the expression of flavonol-related genes, thereby enhancing the drought tolerance of transgenic plants. Furthermore, VviMYC4 could bind to specific E-box sites on the promoters of VviF3H and VviFLS to improve their activities. This study highlights VviMYC4 as a pivotal positive regulator of drought tolerance in grapevines and proposes that VviMYC4 enhances the antioxidant and reactive oxygen species (ROS) scavenging abilities of grapevines in challenging environments and improves their stress resilience by mediating flavonol biosynthesis. Our findings offer crucial candidate genes and valuable insights for the molecular breeding of grapevine drought resistance. Full article

Iris dichotoma Pall., renowned for its high ornamental value, is frequently cultivated in flowerbeds and courtyards, endowing garden landscapes with unique allure. Dark-hued flowers are widely regarded as more aesthetically appealing. This study utilized the petals of two distinct Iris dichotoma Pall. phenotypes as research materials to investigate the underlying mechanism of flower color formation. The purple-flowered Iris dichotoma Pall. was designated as Group P, and the white-flowered one as Group W. A comprehensive integrative analysis of the transcriptome and metabolome of the two petal types was carried out. Metabolomic analysis revealed that the contents of several anthocyanin derivatives, including delphinidin, petunidin, malvidin, peonidin, and procyanidin, were significantly higher in purple petals compared to white petals, with delphinidin exhibiting the highest content. The transcriptomic analysis detected 6731 differentially expressed genes (DEGs) between the white and purple petal types. Specifically, 3596 genes showed higher expression levels in purple petals, while 3135 genes exhibited lower expression levels in purple petals compared to white petals. Ten phenylalanine ammonia-lyase (PAL) genes, two chalcone synthase (CHS) genes, one anthocyanidin reductase (ANR) gene, one 4-coumarate-CoA ligase (4CL) gene, one dihydroflavonol 4-reductase (DFR) gene, one flavanone 3′-hydroxylase (F3′H) gene, and one flavonol synthase (FLS) gene were identified; they all had purple petals displaying higher expression levels than white petals. This research uncovers the potential formation mechanism of anthocyanins in the two Iris dichotoma Pall. types, thereby furnishing a theoretical foundation for floral breeding endeavors. Full article