This study employed integrated metabolomic and transcriptomic analyses to investigate the impacts of moderate withering (approximately 30% weight loss) on organic acid and flavonoid accumulation in blueberries and the resultant blueberry wines. Moderate withering led to decreases in aliphatic organic acids but increases in phenolic acids and flavonoids in blueberries. The decrease in malate could be linked to the downregulated VcMDH and VcDTC, and the decrease in citrate and 2-oxoglutarate could be a consequence of gamma-aminobutyric acid shunt activation. The increases of phenolic acids were associated with upregulated VcCAD and VcPOD in withered blueberries. Additionally, moderate withering upregulated the expression of VcLAR and VcUFGT, promoting flavonoid accumulation. In blueberry wines, moderate withering decreased total aliphatic organic acid content but increased anthocyanin content, consistent with the findings in blueberries. Overall, this study provided references for applying moderate withering in blueberry winemaking, contributing to modulating acidity and enriching phenolic substances. (c) 2026 The authors. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co., Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

To reduce reliance on chemical fungicides, this study developed microencapsulated Artemisia argyi essential oil (AAEO) in β-cyclodextrin/whey protein microcapsules (AAEO-M) as a natural antifungal preservative for blueberries. AAEO-M exhibited high encapsulation efficiency (94.28%) and loading capacity (20.43%), with porous spherical structure and sustained release properties. It showed strong in vitro antifungal activity against Botrytis cinerea, Alternaria alternata, and Penicillium digitatum. Treatment with 2% AAEO-M significantly maintained fruit marketability above 80.8% after 7 d cold storage plus 12 d shelf life. It also suppressed respiration, reduced membrane damage, delayed protopectin degradation, and preserved bioactive compounds. Furthermore, AAEO-M enhanced defense-related enzymes (PAL, β-1,3-GA, CHI) while inhibiting PPO activity. These findings demonstrate that AAEO-M serves as an effective active packaging material with sustained antifungal action and enhanced disease resistance.

To investigate the impact of geographically distinct strains of the same fungal species on the aroma of fermented tea, three Aspergillus cristatus strains were isolated from Fu brick teas produced in Shaanxi, Hunan, and Sichuan Provinces, China. These strains were subsequently used as inoculants to produce three distinct loose-leaf dark teas, designated SXFT, HNFT, and SCFT, respectively. Sensory analysis demonstrated that SXFT exhibited stronger flowery, sweet, woody, and minty aroma attributes than HNFT and SCFT. GC-MS analysis identified 89 volatile compounds and showed that total volatile levels increased significantly after fermentation, with SXFT showing the highest concentration (666.07 μg/L). Aroma recombination and omission assays revealed several distinct odorants: methyl salicylate, nonanal, linalool, and 6-methyl-5-hepten-2-one in the SXFT; linalool, β-ionone, and 6-methyl-5-hepten-2-one in the SCFT; and linalool, 6-methyl-5-hepten-2-one, and 1-octen-3-ol in the HNFT. These results provide a chemical basis for targeted starter selection in fermented tea production.

The significant anthocyanin loss in blueberry wine can lead to color degradation during aging. This study showed that blending blueberries with ‘Cabernet Sauvignon’ grapes notably enhanced acylated anthocyanins and copigmentation levels, delaying the decrease in the copigment anthocyanin ratio and significantly increasing the polymerized anthocyanin ratio over time. A model blueberry wine system was applied to evaluate the copigmentation effects of primary copigments from grapes on anthocyanins. The thermodynamic parameter analysis and molecular docking showed that epigallocatechin gallate (EGCG) exhibited superior copigmentation effects compared to other compounds. Additionally, the copigmentation effects of EGCG and ferulic acid (FA) together were more pronounced than those of EGCG alone, suggesting a synergistic enhancement of copigmentation. These findings provide valuable insights for the blueberry wine industry, indicating that blending techniques and the use of composite copigments can significantly enhance color stability.

Efforts to increase product diversity and typicity in blueberry wine have prompted the investigation of indigenous yeasts as complements to commercial Saccharomyces cerevisiae strains. In this study, indigenous Hanseniaspora thailandica CL-49 and S. cerevisiae CL-70, both demonstrating significant β-D-glucosidase and alcohol acetyltransferase (AAT) activities as well as good ecological adaptability, were isolated from spontaneously fermented blueberry must. Single-strain fermentations revealed complementary aroma-producing capabilities: H. thailandica CL-49 showed the highest AAT activity, resulting in increased acetate esters such as isoamyl acetate and phenethyl acetate, while S. cerevisiae CL-70 displayed higher β-D-glucosidase activity, enhancing terpene compounds such as terpineol. Co-fermentation of CL-70 and CL-49 produced a synergistic enzymatic effect, with β-D-glucosidase and AAT activities 1.98- and 1.35-fold higher, respectively, than the commercial mixed-strain control. This synergistic effect resulted in a 105.3% increase in total ester content, including ethyl butyrate, ethyl hexanoate, and ethyl octanoate. Terpene levels were also increased, intensifying the blueberry wine's floral and fruity characteristics. This study is the first to report the isolation and oenological characterization of H. thailandica from blueberry must, and the findings can support the diversification of blueberry wine sensory profiles.

The cation/proton antiporter (CPA2) family plays key roles in regulating plant ion balance and pH homeostasis, particularly in response to salt stress. In this study, a total of 76 CPA2 members were identified in Gossypium hirsutum, phylogenetically classified into 62 GhCHXs and 14 GhKEAs. Through RNA-Seq data analysis, we found that the expression levels of GhCHX6A and GhKEA1D were significantly upregulated under salt stress. The subcellular localization test showed that GhCHX6A is localized to the vacuolar membrane, while GhKEA1D is localized to the thylakoid membrane. Yeast functional complementation assays revealed that GhCHX6A and GhKEA1D exhibit Na+ and K+ transport activities, respectively. Silencing of these genes via virus-induced gene silencing (VIGS) dampened cotton salt tolerance, manifested as aggravated wilting, increased accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA), decreased chlorophyll content, and disrupted Na+/K+ balance in cotton tissues. These results indicate that GhCHX6A and GhKEA1D enhance salt tolerance in upland cotton by regulating Na+/K+ transport to maintain ion homeostasis and alleviate oxidative damage.

Mango (Mangifera indica L.) is one of the main economic crops in Hainan, China, prized for its distinctive flavor and high nutritional value. It is also rich in health-promoting antioxidants such as vitamin C and flavonoids. Enhanced ultraviolet-B (UV-B) radiation, a growing global environmental concern, alters plant antioxidant systems, with increased flavonoid accumulation as a common adaptive response. However, its effects on mango fruit remain largely unexplored. To investigate the antioxidant responses of mango to enhanced UV-B radiation and identify key responsive flavonoid compounds and regulatory genes, we exposed ‘Tainong 1’ mango fruits growing under natural light to 96 kJ · m-2 · d-1 of UV-B radiation to simulate high UV-B conditions. Treated fruits were smaller in size and had a pulp of a more intense yellow colour. Further, malondialdehyde content in treated fruits was higher during the phase of rapid fruit enlargement. Additionally, treated fruits showed increased sugar-acid ratios, total phenol, total flavonoid, carotenoid, and ascorbic acid contents. Furthermore, they showed significantly enhanced antioxidant activity, as measured by the FRAP, ABTS, and DPPH assays. Extensive targeted metabolomic-analysis identified flavonoids as the largest category of compounds differentially expressed in treated and control groups. Quantitative metabolomics of flavonoids identified Hyperoside, Quercimeritrin, and (-)-Catechin gallate as the key flavonoid metabolites responsive to UV-B treatment. Transcriptome analysis revealed an enrichment of the flavonoid biosynthesis pathway, with most associated differentially expressed genes showing upregulation. Furthermore, qRT-PCR analysis confirmed that the expression of the genes MiCHS7, MiCHI1, MiCHI2, MiFLS, MiF3H2, and MiF3H3 correlated with changes in key flavonoid metabolites. Indeed, correlation analysis indicated that MiCHS7, MiCHI1, MiFLS, and MiF3H3 are potential key genes involved in flavonoid accumulation under UV-B treatment. Thus, our study provides a theoretical basis for breeding for new resilient varieties and developing UV-B-resistant mango cultivation techniques.

Irisquinone is an active bioactive constituent isolated from the dried rhizomes of Iris tectorum, a traditional Chinese medicinal herb. Accumulating evidence has confirmed that irisquinone exhibits prominent antitumor activity against a variety of human malignancies. Nevertheless, its biological function and precise molecular mechanisms underlying anti-osteosarcoma effects remain poorly clarified. In the present study, we systematically explored the inhibitory effect of irisquinone on osteosarcoma progression via a combination of in vitro cell experiments and in vivo xenograft model validation. Network pharmacology and molecular docking were applied as preliminary computational strategies to predict potential therapeutic targets and downstream signaling pathways of irisquinone. Bioinformatics analysis indicated that the PI3K/AKT signaling axis is closely implicated in the anti-osteosarcoma action of irisquinone, with AKT1 predicted as a core potential target. Further in vitro functional experiments verified that irisquinone markedly suppressed the proliferation and migration capacity of osteosarcoma cells. Mechanistically, irisquinone restrained the activation of the PI3K/AKT signaling pathway, modulated the expression of apoptosis-related Bcl-2 family proteins (Bax and Bcl-2), and ultimately triggered osteosarcoma cell apoptosis. In addition, the anti-tumor effect and pathway regulatory activity of irisquinone were further validated in vivo. Collectively, our findings demonstrate that irisquinone exerts its anti-osteosarcoma function, at least in part, through suppressing the PI3K/AKT signaling pathway. This work provides reliable experimental support and a theoretical foundation for the preclinical application of irisquinone and the development of novel therapeutic agents against osteosarcoma.

The directed evolution of biomolecules is an iterative process. Although advancements in language models have expedited protein evolution, effectively evolving RNA remains a challenge. RNA aptamers, selected for their binding properties, provide an ideal system to address this challenge, yet traditional aptamer discovery still relies on labor-intensive, multi-round screening. Here we introduce GRAPE-LM (generator of RNA aptamers powered by activity-guided evolution and language model), a generative artificial intelligence framework designed for the one-round evolution of RNA aptamers. GRAPE-LM integrates a transformer-based conditional autoencoder with nucleic acid language models and is guided by CRISPR-Cas-based aptamer screening data derived from intracellular environments. We validate GRAPE-LM on three disparate targets: the human T cell receptor CD3ε, the receptor-binding domain of the SARS-CoV-2 spike protein and the human oncogenic transcription factor c-Myc (an intracellular disordered protein). GRAPE-LM, informed with only a single round of CRISPR-Cas-based screening, successfully obtains RNA aptamers that outperform those driven from multiple rounds of human selection and optimization.

Crop rotation serves as a valuable agronomic practice for addressing succession barriers in crops, particularly in tobacco growing. The effect of different previous crops on the microbiology of soils planted with tobacco is an area that deserves further study.This study investigated the chemical properties, microbial community composition, and functional genes related to nutrient cycling in tobacco-planted soils with no preceding crop (CK), garlic (T1), or faba bean (T2) as preceding crops. The results indicated that the T1 treatment significantly decreased the contents of soil organic matter (SOM, 11.32%), total phosphorus (TP, 29.41%), total potassium (TK, 3.33%), and available potassium (AK, 46.88%), whereas the T2 treatment notably increased the content of hydrolyzable nitrogen (HN, 34.88%). Furthermore, the T2 treatment significantly enhanced the diversity of soil bacteria and fungi, particularly the bacterial Shannon index (1.49%) and fungal Chao1 (24.11%) and Shannon (7.73%) indices. In terms of microbial composition, compared to the CK, the T2 treatment enriched the relative abundance of beneficial bacterial genera (e.g., Sphingomonas, Methyloceanibacter, Rhizophagus) and reduced the relative abundance of pathogenic fungi (e.g., Fusarium). Additionally, T2 treatment increased the abundance of functional genes associated with nitrogen, phosphorus, and potassium, thereby promoting the cycling of soil nutrients. Overall, faba bean as a preceding crop was more beneficial for subsequent tobacco cultivation than fallow periods or garlic.

Rice stands as the most significant crop in China,but it fre-quently encounters diseases that lead to an average yield loss of 10％to 30％[1].Over the past decade,the area affected by rice pests and diseases in China has ranged from 18 to 27 million hectares annually,leading to a loss of 1.3 to 2 million tons of rice[2].Iden-tifying genes that confer broad and durable disease resistance,along with elucidating the molecular mechanisms underlying plant immune activation and broad-spectrum resistance regula-tion,constitutes the fundamental theoretical foundation for breed-ing disease-resistant crops.

The mixed fermentation of non-Saccharomyces yeasts with Saccharomyces cerevisiae (S. cerevisiae) is regarded as a promising strategy to improve the quality of fruit wine. In this study, we investigated the effects of sequential inoculation fermentation involving four non-Saccharomyces yeast species (Tourlaspora delbrueckii (T.d.), Metschnikowia pulcherrima (M.p.), Schizosaccharomyces pombe (S.p.) and Issatchenkia terricola (I.t.)) and an S. cerevisiae strain on the physiochemical properties, volatile compounds and sensory profiling of strawberry wine. The sequential fermentations exhibited a significant reduction in ethanol and organic acids content, while free amino acids content increased by 0.28–3.74 times, particularly in the M.p.+S.c. group. The GC-IMS identified 68 volatile compounds, among which isoamyl acetate, ethyl hexanoate, and isobutanol were found to be the predominant volatiles of strawberry wine. Samples more frequently characterized as “strawberry”, “honey” and “preserved fruit” in Check-all-that-apply analysis were preferred by the panelists (n = 93). The I.t.+S.c. elicited emotional responses of “fascinated” and “surprised” leading to a higher liking rating. Furthermore, the PLSR analysis demonstrated a significant contribution of esters and higher alcohols, such as ethyl 3-methylbutanoate-D, ethyl propanoate, along with isoamyl acetate-D, to the fruity and sweet notes. The present study provided useful information into the mixed inoculation fermentation strategy, which involves combining appropriate non-Saccharomyces yeasts with S. cerevisiae, for developing distinct styles and enhancing the oenological quality of strawberry wine.

Intestinal obstruction, a prevalent and serious condition, necessitates deeper understanding of its genetic architecture. This study rigorously employed two-sample Mendelian randomization to dissect the causal influence of gene expression on intestinal obstruction risk, leveraging comprehensive summary-level data from eQTLGen and FinnGen GWAS. To enhance causal inference, Summary-data-based Mendelian Randomization was integrated, utilizing GTEx eQTL data to specifically assess tissue-relevant gene expression. Our multi-pronged analyses provide compelling genetic evidence supporting causal roles for genetically predicted expression of CHRNB2 and MIAT in intestinal obstruction. Specifically, increased genetically proxied CHRNB2 expression was associated with a protective effect, while higher genetically proxied MIAT expression suggested an elevated susceptibility to intestinal obstruction. Colocalization analysis, alongside HEIDI heterogeneity testing within the SMR framework, further bolstered the robustness of these findings by distinguishing causality from linkage disequilibrium. These convergent results offer novel mechanistic insights into intestinal obstruction pathogenesis, positioning CHRNB2 and MIAT as promising therapeutic targets for both prevention and treatment strategies. Future research is crucial to validate these findings across diverse ancestries and to fully elucidate the intricate biological mechanisms underpinning these gene-disease associations. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement The author(s) received no specific funding for this work. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: Ethical approval for the original data collection was obtained by the eQTLGen Consortium, FinnGen, and GTEx (version 8) from their respective Institutional Review Boards. This study used publicly available summary-level data from these consortia and therefore did not require additional ethical approval, as it is considered secondary research using de-identified data. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All relevant data are within the manuscript and its Supporting Information files.

Soybean is one of the most important crops globally,and its production must be significantly increased to meet increasing demand.Elucidating the genetic regulatory networks underlying soybean organ develop-ment is essential for breeding elite and resilient varieties to ensure increased soybean production under climate change.An integrated transcriptomic atlas that leverages multiple types of transcriptomics data can facilitate the characterization of temporal-spatial expression patterns of most organ development-related genes and thereby help us to understand organ developmental processes.Here,we constructed a comprehensive,integrated transcriptomic atlas for soybeans,integrating bulk RNA sequencing(RNA-seq)datasets from 314 samples across the soybean life cycle,along with single-nucleus RNA-seq and spatially enhanced resolution omics sequencing datasets from five organs:root,nodule,shoot apex,leaf,and stem.Investigating genes related to organ specificity,blade development,and nodule formation,we demonstrate that the atlas has robust power for exploring key genes involved in organ formation.In addition,we developed a user-friendly panoramic database for the transcriptomic atlas,enabling easy ac-cess and queries,which will serve as a valuable resource to significantly advance future soybean functional studies.

Instant Dark Tea (IDT) is processed by fermenting instant green tea (IGT) with Aspergillus niger. To explore the development of aroma characteristics and volatile compounds (VCs) during liquid-state fermentation (LSF), the IDTs were collected at 0, 1, 3, 5 and 7 days. Quantitative descriptive analysis (QDA) revealed that the sample fermented for 3 days (D3) exhibited high levels of sweet and fruity aroma attributes. A total of 85 VCs, including terpenes, higher alcohols, and esters, were identified using gas chromatography-mass spectrometry (GC-MS). The D3 sample contained the most individual VCs (82) and the highest total volatiles content (15.65 mg/L). Gas chromatography-olfactometry (GC-O) analysis verified that D3 sample contained several key odor-active compounds, including linalool, geraniol, phenylethyl alcohol, 1-octen-3-ol, phenylacetaldehyde, acetophenone and alpha-terpineol, that were closely associated with the flowery, sweet and fruity aroma attributes. This study may help to guide industrial production of high-aroma IDTs by LSF in the future.

The trend towards reducing chemical fungicides in the food industry has increased interest in antimicrobial essential oils for food preservation. This study investigates the chemical composition of Artemisia argyi essential oil (AAEO) and its efficacy in inhibiting Botrytis cinerea, Alternaria alternata, and Penicillium digitatum, along with potential antifungal mechanisms involved. GC-MS analysis identified that the main components of AAEO were eucalyptol (19.19 %), L(-)-borneol (6.38 %), camphor (6.17 %), (+)-thujone (5.74 %), terpinen-4-ol (5.55 %), and p-cymene (5.31 %). AAEO demonstrated significant antifungal activity against all three fungi, especially through non-contact exposure. Exposure to AAEO increased AKP, nucleic acid and soluble proteins release, reflecting cell wall and membrane integrity alterations. AAEO also reduced biofilm formation, cell viability, and respiratory activity. SEM images revealed cellular fractures and deformations in the fungal cell walls. The fumigation with AAEO during storage resulted in a significant reduction of both the decay rate (by 24.7 %) and pathogen infestation (by 61.2 %) in blueberries on the 13th day of storage. A 50.8 % reduction in Alternaria alternata incidence was observed in inoculated blueberries by day 4 post-AAEO treatment. Therefore, AAEO is a potent antifungal agent against B. cinerea, A. alternata, and P. digitatum, positioning it a promising option for food preservation.

Elucidating the host-pathogen interactions is critical for uncovering the mechanisms controlling Mycobacterium tuberculosis (Mtb) infection. Using dual RNA-seq with fluorescent Mtb, we simultaneously profiled macrophage and bacterial transcriptomes to resolve dynamic intracellular responses. Macrophages containing dead Mtb exhibited strong immune activation, including enhanced antigen presentation and lysosomal function, whereas macrophages harboring live Mtb showed persistent NF-κB signaling and metabolic reprogramming. Mtb counteracted host defenses through upregulation of DNA repair genes and manipulation of extracellular matrix signaling via SPP1 and integrins, alongside tryptophan catabolism and lipid binding pathways supporting adaptation. Cross-species correlation analysis revealed coordinated transcriptional programs, notably a strong inverse association between Mtb aromatic compound catabolism and host receptor tyrosine kinase signaling. Additional correlations linked bacterial metabolism with host lipid transport and steroid biosynthesis. Together, these results provide a high resolution view of macrophage and Mtb transcriptional interplay defining bacterial persistence versus immune clearance.

Amyotrophic lateral sclerosis (ALS) is a fatal neurological disorder characterized by progressive degeneration of nerve cells in the spinal cord and brain. We generated and characterized a human induced pluripotent stem cell (iPSC) line from skin fibroblasts of a patient with ALS due to SOD1 Mutation. The pluripotency of these iPSCs was verified by the expression of several pluripotency markers at both RNA and protein levels, as well as their capability to differentiate into all three germ layers.

The increasing production and consumption of tea drinks has led to the generation of large amounts of discarded extracted tea residues. As a result, researchers have attempted to extract tea water-insoluble protein (TP) from discarded tea residues to produce food emulsifiers. Thus, in this study, high-internal-phase Pickering emulsions (HIPPEs) stabilized by TP were developed and characterized. First, the effects of salt ions on the emulsifying properties of TP were examined using interfacial tension and hydrophobicity. Fourier transform infrared spectroscopy was used to determine the suitable range of salt ions in the processing stage. Then, the particle size distribution, microstructure, rheological properties, and stability of the emulsions were systematically investigated by controlling the oil phase volume, particle concentration of TP, and emulsification method. The results showed that TP was effectively adsorbed on the oil-water interface and formed a stable particle layer, which means that TP-stable high-internal-phase Pickering emulsions (TPHIPPEs) has been successfully prepared. Further analysis showed that TPHIPPEs exhibited good stability and gelation properties. The pH range was 7-9, and the salt ion concentration was <0.5 M. Additionally, TPHIPPEs exhibited excellent temperature tolerance and antioxidant ability. Finally, the application development results revealed that the loading and retention rates of β-carotene in TPHIPPEs were significantly higher than those of the control group of camellia oil, and that TPHIPPEs exhibited good resistance to UV light and thermal degradation. This study provides new insights into the high-value utilization of tea residue resources.

Degradation of soil quality, imbalanced microbial community , and reduced crop yields due to continuous cropping are prevalent issues in tobacco agriculture. Rotation is an effective strategy to alleviate these problems. Therefore, this study used flue-cured tobacco monoculture (CK) as a control and set up three treatments: faba bean (YCCD), barley (YCDM), and garlic (YCDS). Various culture media, including NA, LB, PCA, R2A, and modified Gao’s I media, along with Illumina MiSeq highthroughput sequencing, were employed to investigate the impact of these crop rotation patterns on the soil cultivable bacterial community. The results showed that the Ace, Chao1, and Shannon indices were significantly higher in the YCDM treatment (P<0.05). Crop rotation significantly increased the relative abundance of beneficial bacteria, such as Pandoraea, Sporosarcina, and Serratia, as well as the relative abundance of nitrogen, phosphorus, and sulfur cycling genes. Soil pH, organic matter, total phosphorus, available nitrogen, total potassium, and available potassium content were identified as key factors influencing bacterial community structure. In conclusion, different rotation patterns regulate the bacterial community involved in nitrogen, phosphorus, and sulfur cycling by affecting soil pH and nutrient content. The enrichment of cultivable bacterial communities helps mitigate soil degradation and ultimately enhances crop yields.