Volatile Metabolome Research Articles

Bacterial dynamics and volatile metabolome changes of vacuum-packaged beef with different pH during chilled storage

This study aimed to assess the growth of spoilage bacteria in Brazilian vacuum-packed beef across different pH ranges (5.4–5.8, 5.8–6.1, ≥6.1) stored at temperatures of 0 °C, 4 °C, and 7 °C. Additionally, the research sought to identify predominant spoilage bacteria at the genus level using 16S rDNA gene sequencing and analyze the principal volatile organic compounds (VOCs) produced by this microbiota through HS-SPME/GC–MS. Lactic acid bacteria (LAB) consistently exhibited counts exceeding 6.0 Log CFU/g, regardless of temperature and pH conditions. The bacterial diversity in the meat samples reflected the influence of slaughterhouse environments, with Pseudomonas and Serratia remaining dominant across different cuts and pH levels. Post-storage, variations in pH and temperature modulated the initial bacterial diversity, leading to a reduction in diversity and an increase in LAB such as Lactobacillus, Lactococcus, Leuconostoc, and Carnobacterium. Notably, these changes were observed within pH ranges of 5.4–5.8 and 5.8–6.1, irrespective of beef cuts and storage temperatures. Based on high throughput sequencing and VOCS, correlation analysis revealed a relationship between the growth of specific spoilage microorganisms under vacuum conditions and the presence of VOCs such as alcohols (e.g., 1-propanol, 2-methyl-) and ketones (e.g., 2-nonanone, 2-octanone, 2-heptanone), identifying them as potential indicators of spoilage bacteria growth.

The detection and utilization of volatile metabolomics in Klebsiella pneumoniae by gas chromatography-ion mobility spectrometry

This research aimed to analyze the volatile compounds emitted during the proliferation of Klebsiella pneumoniae (K. pneumoniae) in the laboratory setting using gas chromatography-ion mobility spectrometry (GC-IMS) and to investigate the potential of volatile metabolomics for detecting carbapenemase-producing strains of K. pneumoniae. The volatile metabolomics of K. pneumoniae were comprehensively analyzed using GC-IMS in tryptic soy broth (TSB) as the culture medium. Afterward, the growth stabilization period (T2) served as the primary time point for analysis, with the introduction of imipenem and carbapenemase inhibitors (avibactam sodium or EDTA) during the exponential growth phase (T0) to further investigate alterations in volatile molecules associated with K. pneumoniae. Standard strains were utilized as references, while clinical strains were employed for validation purposes. At T2, a total of 22 volatile organic compounds (VOCs) associated with K. pneumoniae were identified (3 VOCs found in both monomer and dimer forms). Significant differences in VOCs were observed between carbapenemase-negative and carbapenemase-positive strains, both standard and clinical, following the introduction of imipenem. Furthermore, the addition of avibactam sodium led to distinct changes in the VOC content of strains producing class A carbapenemase, while the addition of EDTA resulted in specific alterations in the volatile metabolic profiles of strains producing class B carbapenemase. GC-IMS demonstrated significant promise for analyzing bacterial volatile metabolomics, and its application in evaluating the volatolomics of K. pneumoniae may facilitate the timely detection of carbapenemase-producing strains.

The Effect of Reduced Nitrogen Fertilizer Application on japonica Rice Based on Volatile Metabolomics Analysis.

Nitrogen is critical for rice yield and quality, but its overuse can be detrimental to efficiency and the environment. To identify changes in the quality of rice in response to the reduced application of nitrogen fertilizer, we carried out a comprehensive metabolomics study of SuiJing 18 using volatile metabolomics methods. Our results showed that SuiJing 18 had a total of 358 volatile metabolites, mainly lipids (16.25%), terpenoids (15.41%), heterocyclic compounds (15.13%), and hydrocarbons (13.45%). SuiJing 18 underwent significant changes in response to the reduced application of nitrogen fertilizer. Key sweet volatile compounds such as 4-methyl-benzeneacetaldehyde, hexyl acetate, and 2-methylnaphthalene were present at significantly higher levels when nitrogen fertilizer was applied at a rate of 68 kg of pure nitrogen per hectare, and their flavor characteristics also differed significantly from the compounds resulting from the other two treatments. Focusing on 16 differential volatile metabolites, we further investigated their effects on flavor and quality, thus laying the foundation for a greater understanding of the biomarkers associated with changes in rice quality. This study contributes to a better understanding of the mechanisms underlying changes in rice quality after reduced nitrogen fertilizer application.

Shorten spreading duration enhance the quality of summer Meitan Cuiya tea

Meitan Cuiya (MTCY), a representative green tea from Guizhou, China, may exhibit lower quality in summer due to increased bitterness and astringency. Spreading is a common method to enhance tea quality, but its impact on summer MTCY remains unclear. This study combined transcriptomics and volatile metabolomics to investigate the effects of spreading duration on quality of summer fresh tea leaves and MTCY. Results showed that spreading time shortened to 4 h improved the taste of MTCY, due to lower catechins and higher theanine levels. This duration also yielded woody floral scent in MTCY, marked by high levels of trans-Cubebol, linalool, (Z)-linalool oxide. Transcriptomic analysis linked the 4-h spreading to proteasome activities. Aroma formation was related to diterpenoid and flavonoid biosynthesis. Additionally, gibberellins and auxin were associated with quality formation in fresh tea leaves. This research lays a foundation for improving quality of fresh tea leaves and MTCY in summer.

Integrated volatile metabolome and transcriptome analyses provide insights into the warm aroma formation elicited by methyl jasmonate in carrot root.

Carrot is a highly significant vegetable cultivated worldwide and possesses a unique aroma with abundant edible and medicinal values. However, it remains largely unknown whether jasmonic acid could regulate aroma formation in carrot. Here, an integrated analysis of the volatile metabolome and transcriptome of carrot roots exposed to different concentrations of methyl jasmonate (MeJA) was performed. The results revealed 1,227 volatile organic compounds and 972 differential accumulated metabolites, with terpenes representing the largest portion. MeJA treatment evidently increased the relative odor activity values as well as the accumulation of most volatile compounds. In addition, 4,787 differentially expressed genes were identified and subjected to function enrichment analysis, indicating a role of terpene biosynthesis and metabolism in response to MeJA application. A network consisting of 4,680 transcription factor-structural pairs that showed highly significant positive correlations was constructed, which may be utilized as genetic targets for examining terpene accumulation and aroma formation elicited by methyl jasmonate. The results from the present work substantially improved our understanding of MeJA-mediated aroma formation in carrot.

Elevated levels of branched chain fatty acids in low-salt fish sauce by co-fermentation: flavor improvement and metabolism analysis.

Traditional fish sauce products rely on relatively long fermentation time and high salt concentration, resulting in inconsistent quality and health risks. Branched-chain fatty acids (BCFAs) are associated with nutritional benefits and health-care effects, mainly derived from food fermentation. This study aimed to screen BCFAs-producing bacteria with high protease and aminotransferase activity as starter cultures for fish sauce fermentation. The low-salt fish sauce products were obtained by co-fermentation with three chosen strains. Trichloroacetic acid (TCA)-soluble peptides and amino acid nitrogen concentrations were higher in the co-fermentation group (FH group). The organoleptic evaluation showed co-fermentation optimized flavor composition and endured with rich taste. The levels of BCFAs and branched-chain amino acids (BCAAs) significantly increased by co-fermentation. Volatile metabolomics analysis indicated that BCFAs, branched-chain esters, and pyrazines were the key flavor compounds in the co-fermented group. The co-fermentation system with selected strains to ferment low-salt fish sauce has the potential to increase BCFA content and improve flavor and nutrition. © 2024 Society of Chemical Industry.

Artificial intelligence decision making tools in food metabolomics: Data fusion unravels synergies within the hazelnut (Corylus avellana L.) metabolome and improves quality prediction

This study investigates the metabolome of high-quality hazelnuts (Corylus avellana L.) by applying untargeted and targeted metabolome profiling techniques to predict industrial quality. Utilizing comprehensive two-dimensional gas chromatography and liquid chromatography coupled with high-resolution mass spectrometry, the research characterizes the non-volatile (primary and specialized metabolites) and volatile metabolomes. Data fusion techniques, including low-level (LLDF) and mid-level (MLDF), are applied to enhance classification performance. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) reveal that geographical origin and postharvest practices significantly impact the specialized metabolome, while storage conditions and duration influence the volatilome. The study demonstrates that MLDF approaches, particularly supervised MLDF, outperform single-fraction analyses in predictive accuracy. Key findings include the identification of metabolites patterns causally correlated to hazelnut’s quality attributes, of them aldehydes, alcohols, terpenes, and phenolic compounds as most informative. The integration of multiple analytical platforms and data fusion methods shows promise in refining quality assessments and optimizing storage and processing conditions for the food industry.

Functional characterization of sesquiterpene synthase in Mongolian medicine Syringa oblata in heartwood formation

Lilac (Syringa oblata) is a well-known horticultural plant, and its aromatic heartwood is widely utilized in Traditional Mongolian Medicine for treating angina. However, limited research on the dynamic changes and mechanisms of aromatic substance formation during heartwood development hinders the analysis and utilization of its medicinal components. In this study, volatile metabolome analysis revealed that sesquiterpenes are the primary metabolites responsible for the aroma in heartwood, with cadinane and eremophilane types being the most prevalent. Among the identified sesquiterpene synthases, SoSTPS1-5 exhibited significantly increased expression in heartwood formation and was selected for further investigation. Molecular docking simulations predicted multiple amino acid binding sites and confirmed its ability to catalyze the formation of eremophilane, copaene, cadinane, germacrane, and elemane-type sesquiterpenes from FPP (farnesyl pyrophosphate). Co-expression and promoter analysis suggested a transcriptional regulatory network primarily involving WRKY transcription factors. Additionally, aiotic and biotic stress inducers, such as Ag+, Fusarium oxysporum, and especially MeJA, were found to activate the expression of SoSTPS1-5 and promote sesquiterpene accumulation. This study provides insights into the basis of medicinal substance formation and the potential mechanisms of sesquiterpene accumulation in lilac heartwood, laying a foundation for future research on the biosynthesis and utilization of its medicinal components.

Characterization of the flavor profile of UHT milk during shelf-life via volatile metabolomics fingerprinting combined with chemometrics

Ultra-high temperature (UHT) milk is popular among consumers. However, its flavor and texture change in its shelf life. Flavor is highly determinative for the success of dairy products and for consumers’ willingness to buy. It is important to milk producers to ensure the optimal flavor of their products in the shelf life. In order to be able to control and predict the flavor quality of UHT milk during the shelf life, this study compared the variations in sensory quality, volatile aroma release and backbone flavor factors and developed a discriminant model to assess flavor quality based on flavouromics data of five competing milk sample during storage. Using partial least squares discriminant analysis (PLS-DA) with Electronic-nose (E-nose) data excellent classification sensitivity and specificity were achieved compared to models based on gas chromatography-mass spectrometry (GC–MS) data. The PLS-DA model using E-nose data exhibited a 100% correct classification rate for the storage period, and a 92% correct rate based on the eight variable importance in the projection (VIP) elements screened for volatile components from different groups. The discriminative model developed herein based on E-nose combined with chemometrics demonstrated advantages such as speed, efficiency, and environmental friendliness. This method shows promise as a precise tool for analyzing aroma changes in UHT milk during its shelf life, and provide support for controlling the flavor substances and milk product development.

Volatile metabolomics and transcriptomics analyses provide insights into the mechanism of volatile changes during fruit development of 'Ehime 38' (Citrus reticulata) and its bud mutant.

Volatile compounds are important determinants affecting fruit flavor. Previous study has identified a bud mutant of 'Ehime 38' (Citrus reticulata) with different volatile profile. However, the volatile changes between WT and MT during fruit development and underlying mechanism remain elusive. In this study, a total of 35 volatile compounds were identified in the pulps of WT and MT at five developmental stages. Both varieties accumulated similar and the highest levels of volatiles at stage S1, and showed a downward trend as the fruit develops. However, the total volatile contents in the pulps of MT were 1.4-2.5 folds higher than those in WT at stages S2-S5, which was mainly due to the increase in the content of d-limonene. Transcriptomic and RT-qPCR analysis revealed that most genes in MEP pathway were positively correlated with the volatile contents, of which DXS1 might mainly contribute to the elevated volatiles accumulation in MT by increasing the flux into the MEP pathway. Moreover, temporal expression analysis indicated that these MEP pathway genes functioned at different developmental stages. This study provided comprehensive volatile metabolomics and transcriptomics characterizations of a citrus mutant during fruit development, which is valuable for fruit flavor improvement in citrus.

Dynamical changes of volatile metabolites and identification of core fungi associated with aroma formation in Fu Brick tea during the fungal fermentation

Fu Brick Tea (FBT), a distinguished dark tea, derives its unique sensory identity from a specialized fungal fermentation process. However, the effect of core functional fungi on sensory attributes during FBT fungal fermentation still needs to be systematically explored. Employing a synergistic blend of electronic sensory evaluation, microbiomics, and volatile metabolomics, this study delves into the dynamic evolution of fungi and volatile metabolites (VMs) during fungal fermentation. Utilizing fungal sequencing and Gas Chromatography-Mass Spectrometry (GC-MS) techniques, we unveiled 234 OTUs, with g_Eurotium emerging as a dominant fungus. Additionally, we identified 288 contributing VMs, 35 of which exhibited notable fluctuations during fermentation. These associated fungi influenced crucial metabolic pathways, enhancing the synthesis of sweet-scented components like acetophenone, while repressing woody and fecal aromas, exemplified by nerolidol, skatole, and indole. This study reveals the evolution of fungi and VMs during FBT fungal fermentation, and suggests 4 possible pathways of substance synthesis in which fungi may be involved, delivering valuable insights into the nuanced processes governing the quality formation of FBT.

Fecal microbiota and volatile metabolome pattern alterations precede late-onset meningitis in preterm neonates.

The fecal microbiota and metabolome are hypothesized to be altered before late-onset neonatal meningitis (LOM), in analogy to late-onset sepsis (LOS). The present study aimed to identify fecal microbiota composition and volatile metabolomics preceding LOM. Cases and gestational age-matched controls were selected from a prospective, longitudinal preterm cohort study (born <30 weeks' gestation) at nine neonatal intensive care units. The microbial composition (16S rRNA sequencing) and volatile metabolome (gas chromatography-ion mobility spectrometry (GC-IMS) and GC-time-of-flight-mass spectrometry (GC-TOF-MS)), were analyzed in fecal samples 1-10 days pre-LOM. Of 1397 included infants, 21 were diagnosed with LOM (1.5%), and 19 with concomitant LOS (90%). Random Forest classification and MaAsLin2 analysis found similar microbiota features contribute to the discrimination of fecal pre-LOM samples versus controls. A Random Forest model based on six microbiota features accurately predicts LOM 1-3 days before diagnosis with an area under the curve (AUC) of 0.88 (n=147). Pattern recognition analysis by GC-IMS revealed an AUC of 0.70-0.76 (P<0.05) in the three days pre-LOM (n=92). No single discriminative metabolites were identified by GC-TOF-MS (n=66). Infants with LOM could be accurately discriminated from controls based on preclinical microbiota composition, while alterations in the volatile metabolome were moderately associated with preclinical LOM.

Aroma formation in single- and double-petal jasmines (Jasminum sambac) during flowering via volatile metabolome and transcriptome

Jasminum sambac (L.) Aiton exists in single-petal (SP), double-petal (DP), and multi-petal (MP) phenotypes, which have a sweet floral scent and release aroma when the flowers open. A total of 103 volatiles were discovered from SP and DP jasmines. 16 volatiles were identified as their key odorants. Caryophyllene, geraniol, linalool, α-farnesene, β-ocimene, methyl anthranilate, methyl salicylate, benzeneacetaldehyde and hexanal may form the overall aroma of those two jasmines during flowering. High-resolution transcriptional profiles of those two jasmines at four flowering stages were generated from our recently reported chromosome-level genome assembly, from which we identified 35 differentially expressed genes (DEGs) involved in terpenoid metabolic pathways and 10 DEGs involved in the phenylpropane/benzene metabolic pathway, which may shape the characteristic aroma of those two jasmines at the molecular level. Through co-expression network analysis of key odorants, we found that bZIP8, bHLH6, MYB4, and AP2/ERF3 may contribute to the regulation of aroma volatiles in those two jasmines. These results offer an important theoretical foundation for studying the regulation of aroma formation for SP and DP jasmines.

Biochemical Responses in Populus tremula: Defending against Sucking and Leaf-Chewing Insect Herbivores.

The main biochemical traits were estimated in poplar leaves under biotic attack (aphids and spongy moth infestation). Changes in the abundance of bioactive compounds in genetically uniform individuals of European aspen (Populus tremula), such as proline, polyphenolic compounds, chlorophylls a and b, and volatile compounds, were determined between leaves damaged by sucking insects (aphid-Chaitophorus nassonowi) and chewing insects (spongy moth-Lymantria dispar) compared to uninfected leaves. Among the nine analyzed phenolic compounds, only catechin and procyanidin showed significant differences between the control leaves and leaves affected by spongy moths or aphids. GC-TOF-MS volatile metabolome analysis showed the clear separation of the control versus aphids-infested and moth-infested leaves. In total, the compounds that proved to have the highest explanatory power for aphid-infested leaves were 3-hexenal and 5-methyl-2-furanone, and for moth-infested leaves, trans-α-farnesene and 4-cyanocyclohexane. The aphid-infested leaves contained around half the amount of chlorophylls and twice the amount of proline compared to uninfected leaves, and these results evidenced that aphids influence plant physiology more than chewing insects.

GC-MS with Headspace Extraction for Non-Invasive Diagnostics of IBD Dynamics in a Model of DSS-Induced Colitis in Rats.

Inflammatory bowel diseases are extremely common throughout the world. However, in most cases, it is asymptomatic at the initial stage. Therefore, it is important to develop non-invasive diagnostic methods that allow identification of the IBD risks in a timely manner. It is well known that gastrointestinal microbiota secrete volatile compounds (VOCs) and their composition may change in IBD. We propose a non-invasive method to identify the dynamics of IBD development in the acute and remission stage at the level of VOCs in model of dextran sulfate sodium (DSS) with chemically induced colitis measured by headspace GC/MS (HS GC/MS). Methods: VOCs profile was identified using a headspace GC/MS (HS GC/MS). GC/MS data were processed using MetaboAnalyst 5.0 and GraphPad Prism 8.0.1 software. The disease activity index (DAI) and histological method were used to assess intestinal inflammation. The peak of intestinal inflammation activity was reached on day 7, according to the disease activity index. Histological examination data showed changes in the intestine due to different stages of inflammation. As the acute inflammation stage was reached, the metabolomic profile also underwent changes, especially at the short-fatty acids level. A higher relative amounts of acetic acid (p value < 0.025) and lower relative amounts of propanoic acid (p value < 0.0005), butanoic acid (p value < 0.005) and phenol 4-methyl- (p value = 0.053) were observed in DSS7 group on day 7 compared to the control group. In remission stage, disease activity indexes decreased, and the histological picture also improved. But metabolome changes continued despite the withdrawal of the DSS examination. A lower relative amounts of propanoic acid (p value < 0.025), butanoic acid (p value < 0.0005), pentanoic acid (p value < 0.0005), and a significant de-crease of hexanoic acid (p value < 0.0005) relative amounts were observed in the DSS14 group compared to the control group on day 14. A model of DSS-induced colitis in rats was successfully implemented for metabolomic assessment of different stages of inflammation. We demonstrated that the ratios of volatile compounds change in response to DSS before the appearance of standard signs of inflammation, determined by DAI and histological examination. Changes in the volatile metabolome persisted even after visual intestine repair and it confirms the high sensitivity of the microbiota to the damaging effects of DSS. The use of HS GC/MS may be an important addition to existing methods for assessing inflammation at early stages.

Lactobacillus paracasei ZFM54 alters the metabolomic profiles of yogurt and the co-fermented yogurt improves the gut microecology of human adults

Gut microbiota imbalance could lead to various diseases, making it important to optimize the structure of the gut flora in adults. Lactobacillus paracasei ZFM54 is a bacteriocin- and folic acid-producing Lactobacillus strain. Herein, L. paracasei ZFM54 was used as the potentially probiotic bacterium to ferment milk together with a yogurt starter. We optimized the fermentation conditions, and the obtained yogurts were then subjected to volatile and nonvolatile metabolome analysis, showing that L. paracasei ZFM54 can not only improve the acidity, water holding capacity and live lactic acid bacteria counts, but also improve many volatile acid contents and increase some beneficial nonvolatile metabolites, such as N-ethyl glycine and l-lysine, endowing the yogurt with more flavor and better function. The regulatory effects of the co-fermented yogurt on the intestinal microecology of volunteers were investigated by 16S rRNA sequencing and short-chain fatty acid (SCFA) analysis after consuming the yogurt for a 2-wk period, showing a better effect to increase the relative abundance of beneficial bacteria such as Ruminococcus and Alistipes, decrease harmful bacteria (Escherichia-Shigella and Enterobacter), and enhance the production of SCFA (acetate, propionate, and butyric acid) compared with the control yogurt. We found that L. paracasei ZFM54 can significantly improve the health benefits of yogurt, laying the foundation for its commercial application in improving gut microbiota.

A veterinary cerumenomic assay for bovine laminitis identification.

Bovine laminitis disorder results in animal welfare and economic concerns in dairy and beef farms worldwide. However, the affected metabolic pathways, pathophysiologic characteristics, and inflammatory mechanisms remain unclear, hampering the development of new diagnostics. Using cerumen (earwax) as a source of volatile metabolites (cerumenomic) that carry valuable biological information has interesting implications for veterinary medicine. Nonetheless, up to now, no applications of veterinary cerumenomic assays have been made to identify bovine laminitis. This work aims to develop a veterinary cerumenomic assay for bovine laminitis identification that is non-invasive, robust, accurate, and sensitive to detecting the metabolic disturbances in bovine volatile metabolome. Twenty earwax samples (10 from healthy/control calves and 10 from laminitis calves) were collected from Nellore cattle, followed by Headspace/Gas Chromatography-Mass Spectrometry (HS/GC-MS) analysis and biomarker selection in two multivariate approaches: semiquantitative (intensity data) and semiqualitative (binary data). Following the analysis, cerumen volatile metabolites were indicated as candidate biomarkers for identifying bovine laminitis by monitoring their intensity or occurrence. In the semiquantitative strategy, the p-cresol presented the highest diagnostic figures of merit (area under the curve: 0.845, sensitivity: 0.700, and specificity: 0.900). Regarding the binary approach, a panel combining eight variables/volatiles, with formamide being the most prominent one, showed an area under the curve, sensitivity, and specificity of 0.97, 0.81, and 0.90, respectively. In summary, this work describes the first veterinary cerumenomic assay for bovine laminitis that indicates new metabolites altered during the inflammatory condition, paving the way for developing laminitis early diagnosis by monitoring the cerumen metabolites.

A comparative analysis of morphology, microstructure, and volatile metabolomics of leaves at varied developmental stages in Ainaxiang (Blumea balsamifera (Linn.) DC.).

Ainaxiang (Blumea balsamifera (Linn.) DC.) is cultivated for the extraction of (-)-borneol and other pharmaceutical raw materials due to its abundant volatile oil. However, there is limited knowledge regarding the structural basis and composition of volatile oil accumulation in fresh B. balsamifera leaves. To address this problem, we compare the fresh leaves' morphology, microstructure, and volatile metabonomic at different development stages, orderly defined from the recently unfolded young stage (S1) to the senescent stage (S4). Distinct differences were observed in the macro-appearance and microstructure at each stage, particularly in the B. balsamifera glandular trichomes (BbGTs) distribution. This specialized structure may be responsible for the accumulation of volatile matter. 213 metabolites were identified through metabolomic analysis, which exhibited spatiotemporal accumulation patterns among different stages. Notably, (-)-borneol was enriched at S1, while 10 key odor metabolites associated with the characteristic balsamic, borneol, fresh, and camphor aromas of B. balsamifera were enriched in S1 and S2. Ultra-microstructural examination revealed the involvement of chloroplasts, mitochondria, endoplasmic reticulum, and vacuoles in the synthesizing, transporting, and storing essential oils. These findings confirm that BbGTs serve as the secretory structures in B. balsamifera, with the population and morphology of BbGTs potentially serving as biomarkers for (-)-borneol accumulation. Overall, young B. balsamifera leaves with dense BbGTs represent a rich (-)-borneol source, while mesophyll cells contribute to volatile oil accumulation. These findings reveal the essential oil accumulation characteristics in B. balsamifera, providing a foundation for further understanding.

Volatile metabolomics reveals the characteristics of the unique flavor substances in oats

Oats is a cereal well known for its high nutritional value and unique flavor. This study investigated the metabolomics data from oats, wheat, and barley using broadly targeted GC–MS metabonomic techniques. A total of 437 volatile organic compounds (VOCs) were identified, of which 414 were shared metabolites, with three metabolites unique to oats. Three hundred and seven differentially accumulated metabolites (DAMs) were screened from all the comparison groups, of which 27 metabolites were shared by oats and barley, and 121 shared by oats and wheat. Terpenoids and esters were the key metabolites determining the differences in flavor. A KEGG analysis indicated that the alpha-linolenic acid and phenylalanine pathways were the most significant metabolic pathways. The 42 DAMs found may be the main substances leading to the flavor differences between the different varieties. Overall, this study reveals the main reasons for the unique flavor of oats through metabolomic evidence.

Microbiomics and volatile metabolomics-based investigation of changes in quality and flavor of oat (Avena sativa L.) silage at different stages.

This study aimed to analyze the fermentation quality, microbial community, and volatile metabolites of oat silage harvested at two different stages, while examining the correlation between microorganisms and volatile metabolites. Oats were harvested at two growth stages (pre-heading [PRH] and post-heading [POH] stages), followed by 90 days of natural fermentation, with 6 replicates per treatment. Pre- and post-silage samples were randomly selected for nutrient composition, fermentation parameters, microbial population, and high-throughput sequencing analysis. Volatile metabolomics analysis was also performed on samples after 90 days of fermentation to detect differences in flavor quality after silage. The effect of growth stage on the nutrient content of oats was significant, with pre-heading oats having higher crude protein and post-heading oats having higher water soluble carbohydrates content (p < 0.05). Following a 90-day fermentation period, the pH and ammonia nitrogen/total nitrogen levels in the PRH-90 (silage from pre-heading oats after 90 days of fermentation) group demonstrated a significant decrease (p < 0.05), whereas the lactic acid content was notably higher compared to the POH-90 (silage from post-heading oats after 90 days of fermentation) group (p <0.05). Lactiplantibacillus dominated in the PRH-90 group and Enterococcus dominated in the POH-90 group, with abundances of (> 86%) and (> 87%), respectively. The differential volatile metabolites of the two treatment groups were dominated by esters and terpenoids, and the differences in flavor were mainly concentrated in sweet, green, and fruity odors. The results of Kyoto encyclopedia of genes and genomes pathway enrichment analysis demonstrated three major metabolic pathways: phenylpropanoid biosynthesis, phenylalanine metabolism, and biosynthesis of secondary metabolites. Specific microorganisms were significantly correlated with flavor indicators and flavor metabolites. Lactiplantibacillus was significantly positively correlated with flavor substances indicating sweet and fruity flavors, contributing to good flavor, while Enterococcus was significantly and positively correlated with flavor substances indicating bad flavors. In summary, growth stage had significant effects on nutritional components, fermentation parameters and flavor quality of oats, with the fermentation process dominated by Lactiplantibacillus leading to good flavor, while the fermentation process dominated by Enterococcus led to the development of poor flavor.