Plant phyllosphere bacteria are vital for plant health and productivity and are affected by both abiotic and biotic factors. In this study, we surveyed the structure of the phyllosphere bacterial community associated with alfalfa. For two varieties of alfalfa, forty-eight samples of phyllosphere communities were collected at two locations over four seasons in 2020. Proteobacteria and actinobacteria were associated with the dominating phylum in the bacterial communities of the alfalfa phyllosphere. Sphingomonas was the most abundant genus-level bacteria, followed by Methylobacterium, Burkholderia-Caballeronia-Paraburkholderia, and Pseudomonas. Sampling time had a greater affect than site and variety on alfalfa surface microorganisms. The variation in phyllosphere bacterial community assembly was mostly explained by the season–site interaction (43%), season–variety interaction (35%), and season (28%). Variety, site–variety interaction, and season–site–variety interactions did not have a meaningful effect on phyllosphere bacterial diversity and community structure. The bacterial community in the phyllosphere of alfalfa showed seasonal changes over time. The environmental factors that contributed most to the phyllosphere bacterial community of alfalfa were temperature and sunshine duration, which were significantly positively correlated with most of the dominant bacterial genera in the alfalfa phyllosphere.

Broussonetia papyrifera L. (paper mulberry) is an alternative woody plant, which can used to replace part of the protein feed for ruminants. Ensiling is an effective way to preserve fresh pasture and to solve the problem of stable storage and feed conversion of paper mulberry in the rapid growth period. However, low dry matter (DM), water-soluble carbohydrate, and lactic acid bacteria (LAB) reduce the quality of paper mulberry silage. This study assesses the influence of wilting time (0 h and 3.5 h; lighting: 3.43 × 10⁴ Lux) and three additives (Enterococcus durans, CL; cellulase, CE; and formic acid, FA) on the fermentation quality, aerobic stability, and bacterial community of whole plant B. papyrifera silage. The whole plant B. papyrifera sample was mowed and wilted for 0 h and 3.5 h, and then had CL, CE, or FA added, followed by 60 days of ensiling. The results show all silage samples had high fermentation quality with pH below 4.2, ammonia-nitrogen below 100 g/kg DM, and no detectable butyric acid. The additives protected the DM and the crude protein from protease activity (p < 0.05), and CL was the most effective among them. Furthermore, wilting time influenced the silage’s bacterial communities, but overall, CL treatment had the greatest impact on bacterial communities. Wilting time and formic acid treatment significantly improved aerobic stability (p < 0.05). Enterococcus was positively correlated with lactic acid (LA), while negatively correlated with LA and Weissella (p < 0.001). Enterococcus was identified as the main driver of the whole plant paper mulberry ensiling process in the present study. In conclusion, compared to other additives, LAB is the most effective and economical to improve the fermentation quality and reduce the protein degradation of whole plant paper mulberry silage. Our findings provide a theoretical basis to improve the quality and production of paper mulberry silage.

In this study, the influence of two lactic acid bacteria (LAB) strains [Lactobacillus rhamnosus BDy (LR-BDy) and Lactobacillus buchneri TSy (LB-TSy)] selected from Southwest China on the fermentation characteristics and in vitro gas production of Broussonetia papyrifera (paper mulberry) silage were experimentally explored. The experimental groups were a control group (C), an LB-TSy treatment (LB), an LR-BDy treatment (LR), and an LR-BDy + LB-TSy hybrid group (LR × LB). After the LAB were added, the pH value of paper mulberry silage significantly declined (p < 0.05), and the crude protein content was effectively preserved (p < 0.05). However, no significant changes were found in the levels of neutral detergent fiber, acid detergent fiber, and crude ash (p > 0.05). The lactic acid content in paper mulberry silage was evidently increased (p < 0.05). The in vitro gas production in the LR at 36, 48, and 72 h were markedly higher than that in the other treatments (p < 0.05). Owing to the addition of LAB, the microbial diversity in paper mulberry silage was reduced, while the relative bacterial abundance of Lactobacillus was enhanced. Hence, the addition of LAB selected from the warm and humid region in Southwest China can improve the quality of paper mulberry silage and elevate its feeding value in this region.

Italian ryegrass is a high-quality forage grass, and a full understanding of the changes in its microbiome and metabolome during aerobic exposure can prolong its aerobic stability and improve its utilization value. Italian ryegrass silage was prepared with deionized water (CK), Lactobacillus rhamnosus BDy3-10 (LR), Lactobacillus buchneri TSy1-3 (LB), and a mixture of these two lactic acid bacteria (M). The silage was maintained at ambient temperature for 60 days followed by aerobic exposure. The results show that the Italian ryegrass silage in the LB and M groups exhibited aerobic stability for up to 19 days. A total of 1881 chemicals were identified in Italian ryegrass silage. These metabolites are associated with bacterial communities, especially Lactobacillus. The addition of lactic acid bacteria resulted in a common differential metabolic pathway compared to CK: “phenylpropanoid biosynthesis”. “Flavone and flavonol biosynthesis” was the significant differential metabolic pathway between LB and LR. Inoculation with LB significantly increased the concentrations of lactic acid, acetic acid, vitexin, and luteolin. In conclusion, lactic acid bacteria (LAB) additives affect the microbial community and metabolites of silage. The application of LB inoculants is a feasible way to obtain well-fermented Italian ryegrass silage and improve aerobic stability, even at higher moisture content levels.