Flower formation is important for plant development and analyzing early-flowering genes is crucial for accelerating plant reproduction. Exploring the early fruiting genes present in Vitis vinifera (V. vinifera) can provide a molecular theoretical basis for breeding high-quality resistant varieties using the excellent quality of V. vinifera. In this study, the effects of a V. vinifera intraspecific hybrid population on the flowering characteristics of one-year-old seedlings were studied, and samples of flowering and nonflowering plants in the Ecolly × Dornfelder population were screened. High-throughput sequencing was subsequently used to perform transcriptome sequencing and related differential gene expression analysis for each sample. The results revealed that spiral vine trellising showed potential for accelerating flowering in grapevine seedlings, and 93 genes (77 upregulated and 16 downregulated) were differentially expressed between the flowering and the nonflowering groups. GO (Gene Ontology) analysis of the differentially expressed genes revealed that the pathways associated with the flowering and nonflowering groups were enriched mainly in polysaccharide binding, DNA integration, defense reactions, carbohydrate binding, and dioxygenase activity. KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis of the differentially expressed genes revealed pathways that were enriched mainly in protein processing, amino sugar and nucleotide sugar metabolism, glycolysis/gluconeogenesis, pentose and glucuronic acid conversion, and pyruvate metabolism in the endoplasmic reticulum. An analysis of flowering related genes involved in various pathways revealed that the flowering genes of V. vinifera were related mainly to carbohydrate metabolism, secondary metabolite metabolism, amino acid metabolism, and transcription factor regulation.

Although vineyard soils are regarded as having substantial potential for carbon sequestration, the coupled relationship between conventional viticulture, the implementation of vineyard ecological management, and the resulting alterations in soil conditions and sources of soil organic carbon (SOC) remains underexplored. Accordingly, we conducted a five-year vineyard trial in Penglai, Yantai, China to compare different management practices, including traditional viticulture (TV) and extra-simplified eco-viticulture (EV) modes applied to each of Cabernet Sauvignon and Italian Riesling vineyards, with adjacent uncultivated land (CK) as a control. This study investigated the SOC content and carbon stock in soils at various depths within each sample plot, and tracked the dynamics of plant-and microbial-derived compounds using two groups of biomarkers (lignin phenols and amino sugars) along with their relative contributions to SOC. The results showed that long-term TV significantly improved the SOC pool, and based on this, the implementation of EV resulted in a further increase in SOC content and carbon stocks by 7.21-24.05 % and 10.82-30.00 %, respectively. Plant-(31-53 %) and microbial-derived carbon (30-47 %) contributed similarly to SOC in vineyards, but their contents in the EV were generally higher than those in TV. Lignin phenol accumulation was predominantly regulated by plant properties (grapevine basal diameter and grapevine biomass [GB]) and soil abiotic factors (pH, total nitrogen, total phosphorus, total potassium, etc.), while amino sugars were influenced by a smaller set of factors (grapevine root biomass, GB, pH, SOC, and microbial biomass phosphorus). GB and pH were particularly important as co-driver factors for soil carbon sequestration. Furthermore, the effect of viticulture mode on the SOC pool was most pronounced in the 0-10 cm soil depth, gradually decreasing with soil depth. In conclusion, we considered that the EV mode was a recommended practice for sustainable vineyard.

Maintaining high final viable cell counts remains a major challenge in high-cell-density Lacticaseibacillus paracasei fermentation. In this study, L. paracasei ProSci-92 was cultured with glucose alone (G) or a glucose–inulin composite carbon source (G–inulin), and transcriptomic and untargeted intracellular metabolomic analyses were performed at 2, 6, and 12 h. At 12 h, the G–inulin group showed an approximately 5.9-fold higher final viable cell count than the G group under the composite carbon source condition. At 6 h, cross-omics pathway-level associations mainly involved starch and sucrose metabolism, lysine degradation, the two-component system, and ATP-binding cassette (ABC) transporters, indicating the most evident transcriptomic and intracellular metabolomic divergence at the middle fermentation stage. By 12 h, shared pathway-level signals were mainly associated with the phosphotransferase system, galactose metabolism, and the citrate cycle, together with changes in stress- or protein quality control-associated genes and selected intracellular metabolite features, indicating a distinct late-stage molecular profile under G–inulin culture. Overall, G–inulin culture was associated with a higher final viable cell count and stage-dependent transcriptomic and intracellular metabolomic changes during fermentation. These findings provide an omics-based exploratory framework for understanding carbon source-associated molecular responses related to the final viable cell count in L. paracasei ProSci-92.

In recent years, natural wines have become a hot research topic due to their unique flavor complexity and diversity. However, the dynamic changes in constituents during the aging process of natural wines are highly intricate, and the mechanism of microorganisms has not yet been fully and deeply analyzed. This situation has seriously restricted the quality control of natural wines and the optimization of the winemaking process. This paper presents a critical review synthesizing existing knowledge on the aging of natural wines. From the physicochemical point of view, redox and polymerization reactions drive the transformation of phenolic compounds, which in turn trigger the evolution of the wine's color and the softening of the palate. From the microbiological point of view, the activity of microorganisms such as acetic acid bacteria and lactic acid bacteria tends to elevate the content of volatile acids in wines, which produces undesirable flavors, whereas polysaccharides released by yeasts in the process of autolysis help to enhance the complexity of the wine body. Moreover, aging conditions can regulate oxidation rates and microbial communities, thereby achieving a balance between wine stability and sensory characteristics; lees aging can delay oxidative deterioration in wine; it highlights the unique challenges and strategies faced by natural wines due to minimal intervention. This paper synthesizes and critically evaluates the current research on wine aging process, aiming to improve the understanding of the compositional changes during wine aging, and to provide a theoretical basis for microecological regulation and process optimization during natural wine aging.

While microbial community composition during wine fermentation has been extensively characterized, how different vinification strategies shape active microbial metabolic pathways and flavor formation across various cultivation systems remains poorly understood. Moreover, previous studies have largely relied on correlation-based relationships between microbiota and flavor compounds, with limited mechanistic interpretation. To address this gap, we integrated metatranscriptomic (RNA-seq) and metabolomics (GC-MS and UHPLC-Q-TOF/MS) in a factorial design (2 cultivation systems × 2 vinification strategies × 3 biological replicates) based on Cabernet Sauvignon fermentations. Spontaneous fermentation exhibited a higher diversity of active microbes, dominated by Saccharomyces cerevisiae, Kazachstania humilis, Hanseniaspora uvarum, and Leuconostoc pseudomesenteroides. Transcript-level analysis revealed enhanced amino acid and lipid metabolism in indigenous consortia, consistent with elevated concentrations of ethyl acetate, phenethyl alcohol, unsaturated fatty acids, and amino acids. In contrast, controlled fermentation showed higher levels of phenolic compounds, as well as medium- and long-chain fatty acids and their ethyl esters. Multi-omics integration provided mechanistic evidence linking microbial gene expression to flavor formation. Coordinated expression of amino acid transaminases and fatty acid synthases in multispecies consortia was associated with increased production of higher alcohols and fatty acids, whereas elevated alcohol acetyltransferase expression in S. cerevisiae promoted ester biosynthesis in controlled fermentations. These findings establish a functional framework for harnessing indigenous microbiota to modulate natural wine flavor.

Ecological viticulture with minimal intervention is increasingly emphasized for sustainable wine production, but its effects on grape microbial communities and wine flavor metabolism remain insufficiently understood. This study investigated vineyard soil/grape-associated microbiota and wine flavor metabolites under conventional management (CM) and extra-simplified eco-viticulture (ES) across 2023-2025 vintages. Results showed that ES positively influenced soil microbial diversity and grape epidermal bacterial diversity, while reducing epidermal fungal diversity relative to CM. Spontaneous fermentation (SF) enriched organic acids, amino acids, and their derivatives, whereas inoculated fermentation (IF) accumulated glycolysis/TCA cycle intermediates and nucleosides. Volatile metabolomics revealed that ES-S natural wine had higher levels of terpenoids, aldehydes, and heterocyclic compounds with distinct floral/fruity aromas (rOAV>1), while IF wines were dominated by alcohols, esters, and ketones with homogeneous ripe fruit/mushroom notes. Collectively, extra-simplified eco-viticulture enhanced microbial diversity and drove more complex flavor metabolites in natural wines, providing a sustainable strategy for high-quality wine production.

Natural wines are valued for complex flavors and terroir expression, yet the mechanisms underlying flavor and microbial dynamics during aging remain unclear. This study compared spontaneously fermented and inoculated wines produced under conventional management and extra-simplified eco-viticulture, with emphasis on dynamic changes in flavor metabolites, microbial communities, and biogenic amines during aging. The results showed that Kazachstania humilis, Aureobasidium pullulans, and Oenococcus oeni predominated in spontaneously fermented wines throughout the aging process. In contrast, inoculated wines were initially dominated by Saccharomyces cerevisiae and Oenococcus oeni, followed by Burkholderia spp. Spontaneously fermented wines were characterized by higher concentrations of ethyl acetate, ethyl lactate, phenylethyl acetate, and phenethyl alcohol, whereas inoculated wines were enriched in ethyl caproate, ethyl caprylate, ethyl decanoate, methionol, and isovaleric acid. Moreover, various phenolic acids and their derivatives increased during aging, while certain flavonoids decreased. Biogenic amines, mainly tryptamine and putrescine, exhibited an overall increasing trend. Overall, spontaneously fermented wine from extra-simplified eco-viticulture (ES-S wine) showed more stable and distinctive flavor profiles with superior sensory quality, while spontaneously fermented wine from conventional management (CM-S wine) exhibited inferior quality. These findings provide insights into the maturation and stabilization of natural wines and support the optimization of natural winemaking practices.

Biochar is a promising soil amendment for agricultural fields, yet the potential associations between microbial processes and carbon transformation following biochar addition remain unclear. Using paddy soils as the research system, this study conducted a field experiment to explore the effects of biochar on short-term carbon transformation in paddy soils and the potential underlying mechanisms. The biochar amendment was associated with significantly higher soil respiration (SR) and lower soil organic carbon (SOC) content in paddy soils. Microbiomic analyses indicated that soil dissolved organic nitrogen (DON) was significantly reduced after biochar addition, which in turn was associated with shifts in bacterial functional groups related to soil carbon and nitrogen cycling—enhancing fermentation and aromatic compound degradation activities, while reducing ureolysis and xylanolysis activities. To compete for limited DON resources, the soil bacterial community appeared to exhibit reduced network complexity. Structural equation modelling revealed potential associations suggesting that biochar amendment was linked to shifts in the nitrogen utilization strategies of soil bacteria toward organic fertilizer, and DON was significantly correlated with soil carbon transformation. Notably, DON was also significantly correlated with bacterial aromatic compound degradation function, and together with iron oxide (Fep), these variables constitute a potential coupling mechanism related to soil carbon transformation. Among the candidate taxa, Geothrix fermentans, a known Fe-reducing bacterium, emerged from correlational analyses as a species warranting further investigation for its potential involvement in the proposed N–Fe coupling pattern. This study identifies associative patterns and suggests possible directions for future mechanistic research into the relationship between biochar and soil carbon transformation, and these preliminary correlative insights may help formulate hypotheses for nutrient management in short-term paddy soil systems.

Cold resistance is an important characteristic of sustainable development in the grape industry. The intraspecific recurrent selection in the Vitis vinifera (V. vinifera) method uses high-quality varieties as breeding materials and the substitution and accumulation of minor resistance genes, breeding high-quality grapes with cold resistance. This study was conducted to identify and genetically analyse the cold resistance of a V. vinifera hybrid population (Ecolly × Dunkelfelder), screen for highly resistant and sensitive plant samples, and use high-throughput sequencing to perform transcriptome sequencing and related differential gene expression analysis on each sample. The results revealed that the cold resistance of the hybrid offspring population was characterised by continuous quantitative trait inheritance, with 38 differentially expressed genes (7 upregulated genes and 31 downregulated genes) between the high resistance and high-sensitivity types. Analysis of genes related to various pathways, related to cold resistance, revealed that CYP76F10, Dxs, GERD, NMT, GDE1, glgC, and DHQ-SDH, as well as transcription factor MYB, HB, and MADS family genes, are key candidate genes for V. vinifera cold resistance research. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to investigate the expression characteristics of the six genes that were differentially expressed genes, the results of which were essentially consistent with the results of RNA-seq. Specifically, NMT may enhance cold resistance by enhancing membrane lipid stability. The synergistic expression pattern of CYP76F14 and Dxs suggests its key role in terpene synthesis. By exploring potential genes related to micro effects, a theoretical foundation for further exploration of new high-quality cold-resistant grape varieties has been provided.

The key flavor compound formation pathways resulting from indigenous microorganisms during the spontaneous fermentation of wine have not been thoroughly described. In this study, high-throughput metagenomic sequencing and untargeted metabolomics were utilized to investigate the evolution of microbial and metabolite profiles during spontaneous fermentation in industrial-scale wine production and to elucidate the formation mechanisms of key flavor compounds. Metabolome analysis showed that the total amount of esters, fatty acids, organic acids, aldehydes, terpenes, flavonoids, and non-flavonoids increased gradually during fermentation. Enrichment analysis indicated that metabolic pathways related to the synthesis, decomposition, transformation, and utilization of sugars, amino acids, and fatty acids were involved in the formation of key flavor compounds in wine. Metagenomic analysis revealed that Saccharomyces, Hanseniaspora, Zygosaccharomyces, Wickerhamiella, Lactobacillus, and Fructobacillus were the dominant taxa during spontaneous fermentation. They were significantly positively correlated with organic acids, fatty acids, esters, phenols, aldehydes, terpenes, and phenols. In conclusion, this research provides new insights into the metabolic pathways of key flavor compounds formed by indigenous microorganisms during wine fermentation.

The metabolism of the crop rhizosphere affects microflora diversity and nutrient cycling. However, understanding rhizosphere metabolism in suitable crops within arid desert environments and its impact on microflora interactions remains limited. Through metagenomic and non-targeted metabolomic sequencing of rhizosphere soils from one uncultivated land and four vineyards with cropping years of 5, 10, 15 and 20 years, the critical importance of rhizosphere metabolites in maintaining bacterial and fungal diversity was elucidated. The results revealed that Nocardioides, Streptomyces, and Solirubrobacter were the relatively abundant bacterial genera in rhizosphere soils, while Rhizophagus, Glomus, and Pseudogymnoascus were the relatively abundant fungal genera. The composition of rhizosphere metabolic changed significantly during the continuous cropping of grapevines. Dimethylglycine, Formononetin, and Dehydroepiandrosterone were the most important metabolites. Enrichment analysis revealed significant involvement of metabolic pathways such as biosynthesis of amino acids, unsaturated fatty acids, and linoleic acid metabolism. Procrustes analysis highlighted stronger correlations between rhizosphere metabolites and bacterial community compared to those of fungal community. This suggests distinct responses of microflora to crop-released chemical elements across different soil habitats. Co-occurrence network analysis demonstrated complex associations between rhizosphere metabolites and soil microflora, the positive correlations between rhizosphere metabolites and microflora networks predominated over negative correlations. Partial least squares path model indicated that the effect of cropping years on rhizosphere metabolites was greater than that on bacterial microflora diversity. Futhermore, pH, total phosphorus, and alkali-hydrolyzed nitrogen were the key environmental factors affecting rhizosphere metabolites and microbial diversity. These results deepen our valuable insights into the complex biological processes that rhizosphere metabolites influence on microorganisms, and provide strong support for maintaining microbial diversity in farmland soils in arid regions.

The molecular mechanism of arbuscular mycorrhizal fungi (AMF) in reducing cadmium (Cd) accumulation in plants remains unclear. In this respect, the effects of Rhizophagus intraradices (Ri) inoculation under Cd stress on rice growth, the uptake of Cd along with other elements, and the expression of Cd transport genes, including OsNRAMP1/5, were studied using wild-type (WT) and osnramp5 mutant rice. The results showed that Ri inoculation did not affect rice growth. The uptake of Cd of the osnramp5 mutant was much lower than the WT, as 27.6%, 17.5%, and 39.9% of Cd were noted in the grains, shoots, and roots, respectively. For the WT, Cd alone significantly promoted the OsNRAMP5 expression in shoots, but Ri inoculation significantly suppressed OsNRAMP5 expression and significantly reduced its grain and shoot Cd by 44.4% and 62.3%, respectively, compared to the Cd alone treatment. In contrast, for the osnramp5 mutant, Ri inoculation did not influence OsNRAMP5 expression or the grain and shoot Cd. Furthermore, the expression of other Cd transporters (OsIRT1, OsZIP3/7, OsCAX1a) in both varieties were not changed under the treatments. In conclusion, Ri inoculating significantly reduced Cd uptake by rice, with the molecular mechanism by negative regulation of expression of the OsNRAMP5 gene.

To investigate the variation and fractionation of stable isotopes from irrigation water to soil, grapes, and wine, 82H, 818O, and 817O in different samples from 10 regions in China were determined using a water isotope analyser. The values were significantly different among regions according to the chemometric analysis. All isotopes were significantly and positively correlated with irrigation water-soil and grape-wine. A significant water isotopic fractionation effect was observed from the irrigation water to the soil, grapes, and wine. Stable isotope distribution characteristics correlated with longitude, latitude, altitude, temperature, precipitation, station pressure and wind speed. The linear discriminant analysis (LDA), random forest (RF), support vector machine (SVM), and feed-forward neural network (FNN) models 58.33-100 %, 80-100 %, 53.33-100 %, and 73.33-100 % accurate for distinguishing the geographical origins of all samples from training and test data, respectively. These findings provide a theoretical basis for authenticating the geographic origin of Chinese wines using stable isotope analysis.

In vineyard mulching research, using biodegradable liquid mulch represents a novel and environmentally conscious approach to mulching. In comparison, grapevine branch return has been identified as the most effective mulching method. The effects of in-row mulching with two materials, biodegradable liquid film (BLF) and grapevine branches (GBM), on soil properties and microbial communities in the vineyard were assessed using a one-way horizontal block test with tillage as a control. The results indicated that the application of mulching resulted in a reduction in soil bulk weight; an increase in soil moisture; an enhancement in soil organic matter; and a notable elevation in soil nutrients content compared to the control treatment. Both mulching techniques increased the abundance and diversity of soil microorganisms, strongly correlated with soil physicochemical properties. The correlation analysis demonstrated that total organic carbon (TOC); total nitrogen (TN); total potassium (TK); nitrate nitrogen (NN); and available phosphorus (AP) had the most significant impact on shaping the microbial community, exhibiting a positive correlation with microbial diversity. Additionally, soil nutrients were identified to exert a more pronounced influence on the composition of the bacterial community.

With the development of people’s diets and working patterns, obesity is an increasingly serious health threat faced globally. Grape pomace is an important by-product generated during the wine production process which is rich in polyphenols. Polyphenols show promising potential in anti-inflammatory, antioxidant, and metabolic regulatory applications. Nevertheless, the effects of grape pomace polyphenols on obesity alleviation and their underlying mechanisms require further investigation. The results of this study indicate that grape pomace polyphenols exhibit a preventive effect against obesity caused by a high-fat diet (HFD), ameliorated gut microbiota dysbiosis, and improved gut short chain fatty acid (SCFA) levels. The present study employed comprehensive bioinformatics approaches to characterize gut microbial profiles in each experimental group, including: alpha and beta diversity analyses, phylum- and genus-level relative abundance analyses, Linear Discriminant Analysis Effect Size (LEfSe), and Pearson correlation analysis between gut microbiota and short chain fatty acids. Both grape seeds extract (GSE) and grape peel extract (GPE) reduced the elevated F/B ratio caused by HFD, raised the abundance of probiotics such as Lachnospiraceae_NK4A136_group, Bifidobacterium, and Blautia, and mitigated the increase of pathogenic bacteria Fusobacteria and Eschericha-Shigella caused by HFD. Moreover, Lactobacillus, Faecalibaculum, Clostridium-sensu-stricto-1, Bifidobacterium, Blautia, Alistipes, and Dubosiella may be regulated by GSE and GPE to produce SCFAs, alleviating obesity and metabolic disorders. In conclusion, our results suggest that GSE and GPE show remarkable efficacy in ameliorating obesity and modulating gut microbiota in mice, providing evidence to support the utilization of grape pomace as a metabolic regulator.

Despite previous reports that autotrophic microorganisms have capable of absorbing atmospheric CO2 and increasing soil organic carbon content, their specific pathways involved in carbon fixation have remained elusive. This study aimed to evaluate the differences of eight known carbon fixation pathways involving soil autotrophic microorganisms in vineyard soils with different planting years, and reveal the effects of soil physicochemical properties on the composition of carbon fixation microorganisms. Thus, we performed metagenomic sequencing on one uncultivated soil and four vineyard soils of different planting years. The results showed that autotrophic microorganisms harboring genes of eight konwn pathways related to carbon fixation were identified at each sampling site. The predominant phyla of autotrophic microorganisms were Actinomycetota, Pseudomonadota, and Acidobacteriota, respectively. The rTCA cycle was the most prominent carbon fixation pathway in this study. The relative abundance of genes related to rTCA cycle were increased by 11 %, 7 %, 4 %, and 8 % in the 5year-old (C5), the 10-year-old (C10), the 15-year-old (C15), and the 20-year-old (C20) vineyard soils, respectively, compared to that in soil of uncultivated land (UL). The abundance of enzyme encoding genes involved in carbon fixation pathways varied significantly among soil samples, and the variation trend was consistent with the abundances of genes related to carbon fixation pathway, indicating their significant involvement in regulating carbon fixation. Moreover, environmental factors significantly impacted to the composition of autotrophic microbial, in particular, pH was primarily factor impacted on the composition of autotrophic microbial involved in carbon fixation. This study clarified the effects of vineyard planting years on the composition of soil autotrophic microbial and their carbon fixation pathway, which provides basic data for understanding the function of soil autotrophic microbial in orchards.

Strong-flavor Baijiu, a type of Chinese liquor, is produced through anaerobic solid-state fermentation in a sealed mud pit. Ethyl caproate, the characteristic flavor compound of strong-flavor Baijiu, is influenced by caproic acidproducing bacteria in the pit mud. To better understand the formation of caproic acid, this study investigated the microbial composition and physicochemical parameters of pit mud from different layers (top, middle, and bottom) in Hubei and Sichuan provinces, China. The results revealed that Caproiciproducens plays a key role in caproic acid production by using lactic acid as a substrate, with its abundance increasing with the depth of the pit mud. A strain Caproiciproducens sp. R1, isolated from the pit mud, was shown to produce caproic acid from lactic acid within an initial pH range of 5.5-9.0 and lactic acid concentrations of 1 %-5 % (m/v). In addition, inoculation of strain R1 into Huangshui (a lactic acid-rich liquid from Baijiu production) resulted in 40.72 mM caproic acid production. This study demonstrates that Caproiciproducens plays a crucial role in caproic acid production from lactic acid during the fermentation process of strong-flavor Baijiu.

The regional characteristics of wine are shaped by the synergistic effects of vineyard climate conditions, soil microbial microorganisms, soil properties, and grape must microorganisms; however, their role in shaping regional wine quality is still poorly understood. In this study, soil, grape must, and fermentation samples were collected from Cabernet Sauvignon vineyards in five regions of China. High-throughput sequencing technology was used to analyze the microbiota, and Headspace-Solid Phase Microextraction-Gas Chromatography-Mass Spectrometry (HS-SPME-GC-MS) was used to determine the wine metabolite profile. The results showed that the wine metabolite profiles from different vineyards were significantly different and could be distinguished by their volatile compounds, with each vineyard possessing unique characteristic metabolites. The geographical origin of vineyards significantly influenced the microbial diversity of both soil and winery environments. Although the microbiota changed during fermentation, regional microbial signatures were preserved at the end of fermentation. The random forest model indicated that fungal diversity and weather are key predictors influencing wine regionality, with fungal diversity in grape must having the greatest impact. Partial least squares path modeling further revealed that fungal diversity in grape must had the most significant impact on wine metabolite profiles, followed by weather and then soil fungal diversity. In contrast, soil properties and soil bacterial diversity had weaker effects on these profiles and were significantly influenced by the weather. Overall, this study provides a novel perspective for understanding the mechanisms underlying wine regionality and clarifies the key role of microorganisms, particularly fungal communities, in shaping wine regionality.

Nine-carbon aldehydes and their relative alcohols (C9 aromas) are the main aroma compounds of cucumber (Cucumis sativus L.) fruits and provide a unique cucumber-like note. However, the key regulators of C9 aroma accumulation in cucumber fruit are poorly characterized. Based on C9 aroma dynamic analysis and transcriptome analysis during fruit development of two different cucumber inbred lines, Q16 and Q24, Lipoxygenase09 (CsLOX09) was identified as a candidate gene for C9 aroma accumulation. Additionally, Q24 with higher CsLOX09 expression accumulated more C9 aromas than Q16. To verify the function of CsLOX09, Cslox09 homozygote knockout lines were created. C9 aroma content decreased by 80.79% to 99.16% in these mutants compared to the wild type. To further explore the reasons for the difference in CsLOX09 expression between Q16 and Q24 fruits, a co-expression network was constructed by integrating the C9 aroma-associated metabolism and transcriptomic data. Eighteen candidate transcription factors were highly correlated with the expression of CsLOX09. DNA binding with One Finger 1.8 (CsDof1.8) was confirmed to bind directly to the A/TAAAG motif of the CsLOX09 promoter through dual-luciferase, yeast one-hybrid, chromatin immunoprecipitation-qPCR and electrophoretic mobility shift assays. Furthermore, C9 aroma content and CsLOX09 expression were significantly increased in the CsDof1.8 overexpression lines. Overall, these data elucidate the metabolic regulation of C9 aromas in cucumber and provide a foundation for facilitating the regulation of flavor in cucumber breeding.