Dataset Information

Functional Analysis of Sugars in Modulating Bacterial Communities and Metabolomics Profiles of Medicago sativa Silage.

ABSTRACT: This study explored the effects of four sugar source treatments, including no sugar (CON), fructose (FRU), pectin (PEC), and starch (STA), on the microbiota and metabolome of alfalfa (Medicago sativa) silage. The bacterial community was determined via 16S rRNA gene sequencing. The metabolome was analyzed using ultra high-performance liquid chromatography tandem time-of-flight mass spectrometry (UHPLC/TOF-MS). After 60 days of fermentation, the pH values in FRU and PEC were lower than those in STA and CON. FRU had a greater lactic acid concentration compared to STA and CON. Weissella (47.44%) and Lactobacillus (42.13%) were the dominant species in all four groups. The abundance of Pediococcus was lower, and the abundance of Leuconostoc, Pantoea, and Microbacterium was higher, in FRU compared to CON. The abundance of norank_f__Bacteroidales_S24-7_group was higher, and the abundance of Turicibacter was lower, in both FRU and PEC than in CON. Leuconostoc was negatively correlated with the pH value, and Pediococcus was positively correlated with the pH value. No microbiomes were detected as discriminative features between STA and CON. The addition of FRU and PEC presented more peptides, such as Leu-Val-Thr, Leu-Phe, Ile-Pro-Ile, Val-Trp, and Ile-Leu-Leu but a lower abundance of metabolites for triterpene glycosides including sanchinoside B1, medicagenic acid, betavulgaroside IV, and prosapogenin compared to CON. The addition of PEC presented more phenyllactic acid compared to CON. Our study demonstrated that the addition of pectin and fructose improved the quality of alfalfa silage mainly by promoting Leuconostoc, Pantoea, and Microbacterium, and inhibiting Pediococcus in FRU, and promoting norank_f__Bacteroidales_S24-7_group and inhibiting Turicibacter in both FRU and PEC; this was due to altered metabolic profiles resulting from antifungal activity and decreased triterpene glycoside accumulation. This study improves our understanding of ensiling mechanisms related to the contributions of sugar.

SUBMITTER: Wang B

PROVIDER: S-EPMC7232540 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Functional Analysis of Sugars in Modulating Bacterial Communities and Metabolomics Profiles of <i>Medicago sativa</i> Silage.

Wang Bing B Gao Run R Wu Zhe Z Yu Zhu Z

Frontiers in microbiology 20200505

This study explored the effects of four sugar source treatments, including no sugar (CON), fructose (FRU), pectin (PEC), and starch (STA), on the microbiota and metabolome of alfalfa (<i>Medicago sativa</i>) silage. The bacterial community was determined via 16S rRNA gene sequencing. The metabolome was analyzed using ultra high-performance liquid chromatography tandem time-of-flight mass spectrometry (UHPLC/TOF-MS). After 60 days of fermentation, the pH values in FRU and PEC were lower than thos ...[more]

PMID: 32477276

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Project description:BACKGROUND: Plant-associated bacterial communities caught the attention of several investigators which study the relationships between plants and soil and the potential application of selected bacterial species in crop improvement and protection. Medicago sativa L. is a legume crop of high economic importance as forage in temperate areas and one of the most popular model plants for investigations on the symbiosis with nitrogen fixing rhizobia (mainly belonging to the alphaproteobacterial species Sinorhizobium meliloti). However, despite its importance, no studies have been carried out looking at the total bacterial community associated with the plant. In this work we explored for the first time the total bacterial community associated with M. sativa plants grown in mesocosms conditions, looking at a wide taxonomic spectrum, from the class to the single species (S. meliloti) level. RESULTS: Results, obtained by using Terminal-Restriction Fragment Length Polymorphism (T-RFLP) analysis, quantitative PCR and sequencing of 16 S rRNA gene libraries, showed a high taxonomic diversity as well as a dominance by members of the class Alphaproteobacteria in plant tissues. Within Alphaproteobacteria the families Sphingomonadaceae and Methylobacteriaceae were abundant inside plant tissues, while soil Alphaproteobacteria were represented by the families of Hyphomicrobiaceae, Methylocystaceae, Bradyirhizobiaceae and Caulobacteraceae. At the single species level, we were able to detect the presence of S. meliloti populations in aerial tissues, nodules and soil. An analysis of population diversity on nodules and soil showed a relatively low sharing of haplotypes (30-40%) between the two environments and between replicate mesocosms, suggesting drift as main force shaping S. meliloti population at least in this system. CONCLUSIONS: In this work we shed some light on the bacterial communities associated with M. sativa plants, showing that Alphaproteobacteria may constitute an important part of biodiversity in this system, which includes also the well known symbiont S. meliloti. Interestingly, this last species was also found in plant aerial part, by applying cultivation-independent protocols, and a genetic diversity analysis suggested that population structure could be strongly influenced by random drift.

Project description:BackgroundHeterosis, the superior performance of hybrids relative to parents, has clear agricultural value, but its genetic control is unknown. Our objective was to test the hypotheses that hybrids expressing heterosis for biomass yield would show more gene expression levels that were different from midparental values and outside the range of parental values than hybrids that do not exhibit heterosis.ResultsWe tested these hypotheses in three Medicago sativa (alfalfa) genotypes and their three hybrids, two of which expressed heterosis for biomass yield and a third that did not, using Affymetrix M. truncatula GeneChip arrays. Alfalfa hybridized to approximately 47% of the M. truncatula probe sets. Probe set signal intensities were analyzed using MicroArray Suite v.5.0 (MAS) and robust multi-array average (RMA) algorithms. Based on MAS analysis, the two heterotic hybrids performed similarly, with about 27% of genes showing differential expression among the parents and their hybrid compared to 12.5% for the non-heterotic hybrid. At a false discovery rate of 0.15, 4.7% of differentially expressed genes in hybrids (approximately 300 genes) showed nonadditive expression compared to only 0.5% (16 genes) in the non-heterotic hybrid. Of the nonadditively expressed genes, approximately 50% showed expression levels that fell outside the parental range in heterotic hybrids, but only one of 16 showed a similar profile in the non-heterotic hybrid. Genes whose expression differed in the parents were three times more likely to show nonadditive expression than genes whose parental transcript levels were equal.ConclusionThe higher proportions of probe sets with expression level that differed from the parental midparent value and that were more extreme than either parental value in the heterotic hybrids compared to a non-heterotic hybrid were also found using RMA. We conclude that nonadditive expression of transcript levels may contribute to heterosis for biomass yield in alfalfa.

Project description:Alfalfa (Medicago sativa L.) is a kind of roughage frequently utilized as an animal feed but challenging to be ensiled due to its low water-soluble carbohydrate (WSC), high water content, and elevated buffering capacity, thus requiring the application of lactic acid bacteria (LAB) to improve its fermentation. This study employed high-throughput metagenomic sequence technology to reveal the effects of homofermentative LAB, Lactobacillus plantarum (Lp), or Pediococcus pentosaceus (Pp), and heterofermentative LAB, L. buchneri (Lb), or their combinations (LbLp or LbPp) (applied at 1.0 × 109 colony forming units (cfu) per kilogram of alfalfa biomass fresh material) on the fermentation, microbial community, and functional profiles of alfalfa silage after 7, 14, 30, and 60 ensiling days. The results indicated a reduction (P < 0.05) in glucose and pH and higher (P < 0.05) beneficial organic acid contents, xylose, crude protein, ammonia nitrogen, and aerobic stability in Lb-, LbPp-, and LbLp-inoculated alfalfa silages after 30 and 60 d. Also, higher (P < 0.05) WSC contents were recorded in LbLp-inoculated alfalfa silages after 30 d (10.84 g/kg dry matter [DM]) and 60 d (10.92 g/kg DM). Besides, LbLp-inoculated alfalfa silages recorded higher (P < 0.05) LAB count (9.92 log10 cfu/g) after 60 d. Furthermore, a positive correlation was found between the combined LAB inoculants in LbLp-inoculated alfalfa silages and dominant LAB genera, Lactobacillus and Pediococcus, with fermentation properties after 30 and 60 d. In addition, the 16S rRNA gene-predicted functional analyses further showed that the L. buchneri PC-C1 and L. plantarum YC1-1-4B combination improved carbohydrate metabolism and facilitated further degradation of polysaccharides in alfalfa after 60 d of ensiling. These findings reveal the significant performance of L. buchneri and L. plantarum in combination with dominant LAB species in suppressing the growth of Clostridia, molds, and yeasts and improving the fermentation characteristics and functional carbohydrate metabolism of alfalfa after 60 d ensiling, thus suggesting the need for further studies to uncover the diverse performance of the LAB combination and their consortium with other natural and artificial inoculants in various kinds of silages.

Dataset Information

Functional Analysis of Sugars in Modulating Bacterial Communities and Metabolomics Profiles of Medicago sativa Silage.

Publications

Functional Analysis of Sugars in Modulating Bacterial Communities and Metabolomics Profiles of <i>Medicago sativa</i> Silage.

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