Sainfoin (Onobrychis viciifolia) is rich in condensed tannins (CT). CT function includes inhibiting bacterial and fungi activity during the ensiling process. We used polyethylene glycol (PEG) to deactivate tannin activity to find out the effects of CT. The results show that the addition of PEG increased dry-matter loss (8.32% vs. 14.15%, on a dry-matter basis) after 60 d of ensiling, and also increased lactic acid (10.90% vs. 15.90%, on a dry-matter basis) and acetic-acid content (7.32% vs. 13.85%, on a dry-matter basis) after 30 d of ensiling. The PEG-treated group increased its Pediococcus relative abundance (0.37–3.38% vs. 7.82–23.5%,) during the ensiling process, increased its Gibellulopsis relative abundance after 3 d of ensiling (5.96% vs. 19.52%), increased its Vishniacozyma relative abundance after 3 d and 7 d of ensiling (2.36% vs. 17.02%, 3.65% vs. 17.17%), and increased its Aspergillus relative abundance after 7 d, 14 d and 60 d of ensiling (0.28% vs. 1.32%, 0.49% vs. 2.84% and 1.74% vs. 7.56%). However, the PEG-treated group decreased its Alternaria relative abundance during entire ensiling process (14.00–25.21% vs. 3.33–7.49%). These results suggest that condensed tannins inhibit lactic-acid bacteria fermentation though reducing Pediococcus activity, and inhibiting fungi activity depending on different strains.
Condensed tannins (CT) from sainfoin have a high capacity to inhibit proteolysis. A previous study reported that CT from sainfoin can inhibit lactic acid bacteria activity and decrease ammonium-nitrogen (N) content during sainfoin ensiling; however, no study has focused on the metabolome of ensiled sainfoin. The objective of the present study was to investigate the effects of CT [following supplementation of deactivated CT with polyethylene glycol (PEG)] on protease activity, keystone bacteria, and metabolome during sainfoin ensiling. According to the results, PEG amendment increased non-protein N, amino acid, and soluble protein contents significantly (in the 49.08–59.41, 116.01–64.22, and 23.5–41.94% ranges, respectively, p < 0.05) during ensiling, whereas neutral detergent-insoluble protein and acid detergent-insoluble protein were decreased significantly (in the 55.98–64.71 and 36.58–57.55% ranges, respectively, p < 0.05). PEG supplementation increased aminopeptidase and acid protease activity after 3 days of ensiling (p < 0.05) and increased carboxypeptidase activity during the entire ensiling process (p < 0.05). The keystone bacteria changed following PEG addition (Stenotrophomonas, Pantoea, and Cellulosimicrobium in the control vs. Microbacterium, Enterococcus, and Brevundimonas in the PEG-treated group). In total, 510 metabolites were identified after 60 days of sainfoin ensiling, with 33 metabolites annotated in the Kyoto Encyclopedia of Genes and Genomes database. Among the metabolites, phospholipids were the most abundant (72.7% of 33 metabolites). In addition, 10 upregulated and 23 downregulated metabolites were identified in the PEG-treated group when compared with the control group, after 60 days of ensiling (p < 0.05). Pediococcus (correlated with 20 metabolites, R2 > 0.88, p < 0.05) and Lactobacillus (correlated with 16 metabolites, R2 > 0.88, p < 0.05) were the bacteria most correlated with metabolites. The results suggested antagonistic effects between Lactobacillus and Pediococcus during ensiling. The decreased proteolysis during sainfoin ensiling was mainly attributed to the inhibition of protease activity by CT, particularly carboxypeptidase activity. In addition, proteolysis decreased partly due to CT inhibiting Pediococcus activity during ensiling, with Pediococcus being significantly and positively correlated with dopamine after 60 days of ensiling (R2 = 0.8857, p < 0.05).
The present study aimed to investigate the nitrogen (N) and phosphorus (P) fertilization of continuous addition effects plant biomass, the physiological properties of leaves and the antioxi-dant enzyme activities of alfalfa (Medicago sativa L) in the northern Xinjiang region; including the no fertilization (CK), nitrogen fertilization (N, 120 kg·ha−1), phosphorus fertilization (with low amount of N) (P, 100 kg·ha−1 P and 23.5 kg·ha−1 N) and combined nitrogen and phosphorus fertilization (NP, 120 kg·ha−1 N and 100 kg·ha−1 P) on the K well supplied soil. After three consecutive years of the supply of N and P fertilization, samples were taken at the first flowering of alfalfa (four clippings in the total year) to determine its pigment concentration, stomatal aperture, antioxidant enzyme activity and hay yield. The results showed that NP fertilization promoted growth with a higher number of branches and hay yield of alfalfa, while N or P fertilization alone had a positive effect on the growth of alfalfa. However, P fertilization significantly increased the carotenoid (Car) content at the early flowering stage of alfalfa leaves (during four clippings) (p < 0.05), In addition, NP ferti-lization enhanced stomatal aperture, increased the antioxidant enzyme activity and decreased the oxidized substance at the early flowering stage of alfalfa leaves. The results showed that a N and P balance rather than an absolute amount of either enhanced the growth of alfalfa, and N or P fertili-zation affects physiological traits differently. We propose that NP fertilization increases the nutri-tional characteristics and physiological characteristics, enhancing the adaptive capacity of alfalfa and making it better adapted to external environmental changes.
Nitrogen (N) and phosphorus (P) fertilization significantly affect alfalfa production and chemical composition; however, the effect of combined N and P application on protein fractions and the nonstructural carbohydrate content of alfalfa is not fully understood. This two-year study investigated the effects of N and P fertilization on the protein fractions, nonstructural carbohydrates (NSC), and alfalfa hay yield. Field experiments were carried out using two nitrogen application rates (N60, 60 and N120, 120 kg N ha−1) and four phosphorus application rates (P0, 0; P50, 50; P100, 100; and P150, 150 kg P ha−1), total 8 treatment (N60P0, N60P50, N60P100, N60P150, N120P0, N120P50, N120P100 and N120P150). Alfalfa seeds were sown in the spring of 2019, uniformly managed for alfalfa establishment, and tested in the spring of 2021–2022. Results indicated that P fertilization significantly increased the hay yield (3.07–13.43% ranges), crude protein (6.79–9.54%), non-protein nitrogen of crude protein (fraction A) (4.09–6.40%), and NSC content (11.00–19.40%) of alfalfa under the same treatment of N application (p < 0.05), whereas non-degradable protein (fraction C) decreased significantly (6.85–13.30%, p < 0.05). Moreover, increasing N application resulted in a linear increase the content of non-protein N (NPN) (4.56–14.09%), soluble protein (SOLP) (3.48–9.70%), and neutral detergent-insoluble protein (NDIP) (2.75–5.89%) (p < 0.05), whereas acid detergent-insoluble protein (ADIP) content was significantly decreased (0.56–5.06%, p < 0.05). The regression equations for nitrogen and phosphorus application indicated a quadratic relationship between yield and forage nutritive values. Meanwhile, the comprehensive evaluation scores of NSC, nitrogen distribution, protein fractions, and hay yield by principal component analysis (PCA) revealed that the N120P100 treatment had the highest score. Overall, 120 kg N ha−1 coupled with 100 kg P ha−1 (N120P100) promoted the growth and development of perennial alfalfa, increased soluble nitrogen compounds and total carbohydrate content, and reduced protein degradation, thus improving the alfalfa hay yield and nutritional quality.
Nettle (Urtica cannabina) is an excellent feed resource widely distributed worldwide. Phyllosphere microbes are important as they have living conditions similar to those of the above-ground parts of host plants. Exploring amino acids (AA) and microorganisms can further understand the growth of plants in different seasons. The present study investigated the content of AA and phyllosphere microbes’ structure of nettle plants in different seasons. The results found that AA contents varied significantly with the season, such as alanine, aspartate, cysteine, glutamate, glycine, and methionine contents decreased significantly from spring to winter (p < 0.05), the contents of arginine, histidine, serine, and lysine were highest in summer (p < 0.05). The results suggested that the diversity of bacteria and fungi both increased during winter. During winter, Sphingomonas (relative abundance 25.22–28.45%) and Filobasidum (27.6–41.14%) became dominant. According to the redundancy analysis (RDA) of the correlation between AA and microbes, these two microbes were both the most important factors and showed a negative correlation with AA during winter. Thus, seasons could significantly affect the distribution of phyllosphere microbial communities on the nettle, especially in winter. According to the function prediction(PICRUS2 (KEGG pathway) and FUNGuild) results, the bacteria in the phyllosphere of U. cannabina mainly participated in metabolism. Pathogenic fungi were relatively high in autumn. The present study reveals the influence of seasonal change on the phyllosphere microbial community in U. cannabina.
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