Metabolic profiling provides insights into the accumulation patterns of flavonoids and phenolic acids in different parts of Lactuca indica L.

Lactuca indica L. contains high amounts of flavonoids and phenolic acids. However, there is limited information on the composition of these compounds in different parts of the plant. The present study analyzed the secondary metabolite profiles of the stem, leaf, flower, and seed of Lactuca indicaL.cv. Mengzao (LIM) using a widely targeted metabolomic approach. A total of 576 secondary metabolites were identified, including 218, 267, 232, 286, 302, and 308 differentially accumulated metabolites (DAMs) in the stem_vs_leaf, stem_vs_flower, stem_vs_seed, leaf_vs_flower, leaf_vs_seed, and flower_vs_seed comparisons. In particular, considerable differences were detected in the flavonoids and phenolic acids, five flavonoids, five phenolic acids, one triterpenoid and one alkaloid being differentially accumulated in the four parts. Compared to the stem and flower, the leaf and seed had higher total flavonoid content and total phenolic content. Thses findings provide comprehensive insights into utilizing different parts of LIM in developing functional food products.

Chemical profile and quantitative comparison of constituents in different medicinal parts of Lactuca indica during varied harvest periods using UPLC-MS/MS method.

Lactuca indica L. cv. Mengzao (LIM), acknowledged as a pivotal "One Root of Medicine and Food", boasts dual applications in both culinary and medicinal domains. This research delves into the influence of various harvest periods (vegetative, budding, blossom, and fruiting) on distinct medicinal parts (roots, stems, leaves, flowers, and seeds) of LIM, employing plant metabolomics to assess its chemical constituents. A total of 66 chemical constituents were identified in LIM, with 11 chemical components emerging as potential markers for distinguish medicinal parts. Notably, nutritional organs exhibited elevated levels of cichoric acid, rutin and chlorogenic acid. Specifically, leaves during the budding stage displayed the highest chicoric acid content at 11.70 mg·g-1. Conversely, reproductive organs showed heightened concentrations of cichoric acid, rutin and chlorogenic acid, with seeds exhibiting the peak cichoric acid content at 4.53 mg g-1. This study enriches our understanding of LIM by offering novel insights into quality assessment and the comprehensive utilization of its diverse parts.

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

Objective: 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. Methods: 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. Results: 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. Conclusion: 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.

Optimization of ultrasonic-assisted extraction of flavonoids from Lactuca indica L. cv. Mengzao and their antioxidant properties.

In this study, the ultrasound-assisted extraction (UAE) conditions of flavonoids from Lactuca indica L.cv. Mengzao (LIM) leaves was optimized, and the flavonoids content and their antioxidant potential in different parts were analyzed. The optimal extraction parameters to obtain the highest total flavonoids content (TFC) were a a ratio of liquid to solid of 24.76 mL/g, ultrasonic power of 411.43 W, ethanol concentration of 58.86% and an extraction time of 30 min, the average TFC of LIM leaves could reach 48.01 mg/g. For the yield of flavonoids, the UAE method had the best extraction capacity compared with solvent extraction and microwave-assisted extraction (MAE). In general, the TFC in different parts of LIM followed the order flower > leaf > stem > root, the flowering period is the most suitable harvesting period. From ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) quantification, the flower samples showed significantly higher six flavonoids and had the highest radical scavenging capacities compared to other samples. A high positive correlation was observed between the antioxidant activity and TFC, luteolin-7-O-glucoside and rutin were significantly (p < 0.05) correlated with all antioxidant evaluations. This study provides valuable information for the development and utilization of flavonoids in Lactuca indica as ingredients in food, feed and nutritional health products.

Effect of isolated lactic acid bacteria on the quality and bacterial diversity of native grass silage.

Objective: The objective of this study was to isolate lactic acid bacteria (LAB) from native grasses and naturally fermented silages, determine their identity, and assess their effects on silage quality and bacterial communities of the native grasses of three steppe types fermented for 60 days. Methods: Among the 58 isolated LAB strains, Limosilactobacillus fermentum (BL1) and Latilactobacillus graminis (BL5) were identified using 16S rRNA sequences. Both strains showed normal growth at 15- 45°C temperature, 3-6.5% NaCl concentration, and pH 4-9. Two isolated LAB strains (labeled L1 and L5) and two commercial additives (Lactiplantibacillus plantarum and Lentilactobacillus buchneri; designated as LP and LB, respectively) were added individually to native grasses of three steppe types (meadow steppe, MS; typical steppe, TS; desert steppe, DS), and measured after 60 d of fermentation. The fresh material (FM) of different steppe types was treated with LAB (1 × 105 colony forming units/g fresh weight) or distilled water (control treatment [CK]). Results: Compared with CK, the LAB treatment showed favorable effects on all three steppe types, i.e., reduced pH and increased water-soluble carbohydrate content, by modulating the microbiota. The lowest pH was found in the L5 treatment of three steppe types, at the same time, the markedly (p < 0.05) elevated acetic acid (AA) concentration was detected in the L1 and LB treatment. The composition of bacterial community in native grass silage shifted from Pantoea agglomerans and Rosenbergiella nectarea to Lentilactobacillus buchneri at the species level. The abundance of Lentilactobacillus buchneri and Lactiplantibacillus plantarum increased significantly in L1, L5, LP, and LB treatments, respectively, compared with CK (p < 0.05). Conclusion: In summary, the addition of LAB led to the shifted of microbiota and modified the quality of silage, and L. fermentum and L. graminis improved the performance of native grass silage.

Alternations of ensiling performance and bacterial community in response to different native grassland in Mongolian Plateau.

AIMS: The aim of this study was to investigate the dynamics of bacterial communities and natural fermentation quality in three steppe types [meadow steppe (MS); typical steppe (TS); and desert steppe (DS)] on the Mongolian Plateau. METHODS AND RESULTS: PacBio single molecule with real-time sequencing technology was applied to provide insights into the dynamics of the physicochemical characteristics and the complex microbiome of native grass after 1, 7, 15, and 30 days of fermentation process. The dry matter, crude protein, and water soluble carbohydrate (WSC) contents of the three groups slowly decreased after 1 day of fermentation process, and the lowest WSC concentration after 30 days of ensiling was detected in the DS group compared to that in the MS and TS groups. There was no significant effect of steppe types on lactic acid and butyric acid content (P > 0.05). The pH was higher in the early stages of fermentation. After 30 days of fermentation, the pH of MS and DS dropped to ∼5.60, while TS was as high as 5.94. At different ensiling days, the pH of TS was significantly higher than that of MS (P < 0.05). The ammonia nitrogen content of MS was significantly higher than TS and DS (P < 0.05). During the whole fermentation process, Leuconostoc mesenteroides and Pseudocitrobacter faecalis were the main species of DS, while Enterobacter roggenkampii and Faecalibacterium prausnitzii dominated the fermentation process in MS and TS, respectively. CONCLUSIONS: The fermentation quality of native grass silage of different steppe types was less satisfactory, with the silage quality ranging from DS, MS, and TS in descending order. The epiphytic bacteria dominating the fermentation process differed between steppe types of silage. Leuconostoc mesenteroides as the main strain of DS had a modulating effect on pH and LA content, while the main strains of MS and TS (Enterobacter roggenkampii and Faecalibacterium prausnitzii) dominated the silage without significant effect on improving fermentation characteristics and nutritional quality.

Optimization of Extraction Process and Dynamic Changes in Triterpenoids of Lactuca indica from Different Medicinal Parts and Growth Periods.

In this study, the triterpenoids in the leaves of Lactuca indica L.cv. Mengzao (LIM) were extracted via microwave-assisted ethanol extraction, and the optimum extraction conditions for triterpenoids were determined through single-factor experiments and the Box-Behnken method. The effects of three factors (solid-liquid ratio, microwave power and extraction time) on the total triterpenoids content (TTC) were evaluated. The TTC of different parts (roots, stems, leaves and flowers) of LIM in different growth stages was studied, and the scavenging effects of the highest TTC parts on DPPH, ABTS and hydroxyl free radicals were investigated. The results showed that the optimum extraction conditions for microwave-assisted extraction of total triterpenoids from LIM leaves were as follows: solid-liquid ratio of 1:20 g/mL; microwave power of 400 W; and extraction time of 60 min. Under these conditions, the TTC was 29.17 mg/g. Compared with the fresh raw materials, the TTC of the materials increased after freeze drying. The leaves of LIM had the highest TTC, and the flowering stage was the best time. The triterpenoids from the leaves had a strong ability to eliminate DPPH and ABTS free radicals, and the elimination effect of dried leaves was better than that of fresh leaves, while the elimination effect of hydroxyl free radicals was not obvious. The tested method was used to extract total triterpenoids from LIM using a simple process at low cost, which provides a reference for developing intensive processing methods for L. indica.

Effects of Different Types of LAB on Dynamic Fermentation Quality and Microbial Community of Native Grass Silage during Anaerobic Fermentation and Aerobic Exposure.

Silage of native grasses can alleviate seasonal forage supply imbalance in pastures and provide additional sources to meet forage demand. The study aimed to investigate the effects of Lactobacillus plantarum (LP), Lactobacillus buchneri (LB), and Lactobacillus plantarum in combination with Lactobacillus buchneri (PB) on the nutritional quality, fermentation quality, and microbial community of native grass silage at 2, 7, 15, and 60 days after ensiling and at 4 and 8 days after aerobic exposure. The results showed that dry matter content, crude protein content, the number of lactic acid bacteria, and lactic acid and acetic acid content increased and pH and ammonia nitrogen content decreased after lactic acid bacteria (LAB) inoculation compared with the control group (CK). LP had the lowest pH and highest lactic acid content but did not have greater aerobic stability. LB maintained a lower pH level and acetic acid remained at a higher level after aerobic exposure; aerobic bacteria, coliform bacteria, yeast, and molds all decreased in number, which effectively improved aerobic stability. The effect of the compound addition of LAB was in between the two other treatments, having higher crude protein content, lactic acid and acetic acid content, lower pH, and ammonia nitrogen content. At the phylum level, the dominant phylum changed from Proteobacteria to Firmicutes after ensiling, and at the genus level, Lactiplantibacillus and Lentilactobacillus were the dominant genera in both LAB added groups, while Limosilactobacillus was the dominant genus in the CK treatment. In conclusion, the addition of LAB can improve native grass silage quality by changing bacterial community structure. LP is beneficial to improve the fermentation quality in the ensiling stage, LB is beneficial to inhibit silage deterioration in the aerobic exposure stage, and compound LAB addition is more beneficial to be applied in native grass silage.

Effects of Isolated LAB on Chemical Composition, Fermentation Quality and Bacterial Community of Stipa grandis Silage.

This study aimed to screen and identify lactic acid bacteria (LAB) strains from the Stipa grandis and naturally fermented silage, and assess their effects on the silage quality and bacterial community of Stipa grandis after 60 days of the fermentation process. A total of 38 LAB were isolated, and strains ZX301 and YX34 were identified as Lactiplantibacillus plantarum and Pediococcus pentosaceus using 16S rRNA sequences; they can normally grow at 10−30 °C, with a tolerance of pH and NaCl from 3.5 to 8.0 and 3 to 6.5%, respectively. Subsequently, the two isolated LAB and one commercial additive (Lactiplantibacillus plantarum) were added to Stipa grandis for ensiling for 60 days and recorded as the ZX301, YX34, and P treatments. The addition of LAB was added at 1 × 105 colony-forming unit/g of fresh weight, and the same amount of distilled water was sprayed to serve as a control treatment (CK). Compared to the CK treatment, the ZX301 and YX34 treatments exhibited a positive effect on pH reduction. The water-soluble carbohydrate content was significantly (p < 0.05) increased in ZX301, YX34, and P treatments than in CK treatment. At the genus level, the bacterial community in Stipa grandis silage involves a shift from Pantoea to Lactiplantibacillus. Compared to the CK treatment, the ZX301, YX34, and P treatments significantly (p < 0.05) increase the abundance of Pediococcus and Lactiplantibacillus, respectively. Consequently, the results indicated that the addition of LAB reconstructed microbiota and influenced silage quality. The strain ZX301 could improve the ensiling performance in Stipa grandis silage.

Lactobacillus plantarum and propionic acid improve the fermentation quality of high-moisture amaranth silage by altering the microbial community composition.

Objective: The objective of this study was to determine the effect of Lactobacillus plantarum (L. plantarum) and propionic acid (PA) on the microbial community and fermentation performance of high-moisture amaranth silage. Methods: Amaranth silages were rown without addition (AhGCK) as a control and with L. plantarum JYLP-002 (AhGLP) or propionic acid (AhGPA) and then were opened after 60 days of ensiling to determine the microbial community and fermentation quality. Results: Crude protein (CP) content, lactic acid (LA) content, and lactic acid bacteria (LAB) counts were significantly higher in AhGLP and AhGPA compared with those in AhGCK (p < 0.05). In contrast, pH, acetic acid (AA) content, and yeast and aerobic bacteria counts were significantly lower in AhGLP and AhGPA compared with those in AhGCK (p < 0.05). In addition, propionic acid (PA) levels were markedly higher in AhGPA (p < 0.05). In terms of microbial communities, the silage in the additive groups showed an increased relative abundance of Lactiplantibacillus plantarum and Lentilactobacillus buchneri and a reduced relative abundance of Enterobacter cloacae and Clostridium tyrobutyricum. The abundance of Xanthomonas oryzae was significantly increased in AhGPA, but completely inhibited in the silage supplemented with L. plantarum. Spearman's correlation analysis revealed that Lentilactobacillus buchneri and Levilactobacillus brevis were positively associated with LA and negatively associated with pH. Conversely, Clostridium tyrobutyricum and Enterobacter cloacae were negatively associated with LA, but positively associated with pH and AA content. AA content was inversely correlated with Lentilactobacillus buchneri. Functional prediction analysis showed that LAB dominated the three groups of silage and the silages containing additives had improved carbohydrate and amino acid metabolism compared with the control silage; in particular, the AhGLP group had more heterotypic fermentation processes and a richer metabolic pathway. Furthermore, the epiphytic Lactiplantibacillus plantarum and Lentilactobacillus buchneri could inhibit the reproductive activity of undesirable microorganisms to a certain extent, thus slowing the spoilage process of the silage. Conclusion: In conclusion, L. plantarum can improve fermentation characteristics by modulating the microbial community attached to high-moisture amaranth silage and will prove useful for preserving high-moisture silage.

Comparative Analysis of Major Flavonoids among Parts of Lactuca indica during Different Growth Periods.

L. indica L. cv. Mengzao, a medicinal plant of the Ixeris genus, is rich in flavonoids. In order to thoroughly analyze the the distribution and dynamic change of major flavonoids in its various parts from different growth periods, the flavonoids extracted from L. indica L. cv. Mengzao were identified and quantitatively analyzed by ultra-high-performance liquid chromatography mass spectrometer (LC-MS/MS). Results indicated that 15 flavonoids were identified from L. indica L. cv. Mengzao, and rutin, luteolin, luteolin-7-O-glucoside, kaempferol, quercetin, and apigenin are the major flavonoids in L. indica L. cv. Mengzao. In general, the total flavonoids' content in different parts of L. indica L. cv. Mengzao followed the order flowers > leaves > stems > roots. Flowers and leaves are the main harvesting parts of L. indica L. cv. Mengzao, and the flowering period is the most suitable harvesting period. This study provides valuable information for the development and utilization of L. indica L. cv. Mengzao and determined the best part to harvest and the optimal time for harvesting.