Silage fermentation characteristics and microbial diversity of alfalfa (Medicago sativa L.) in response to exogenous microbiota from temperate grasses

The objective of this study was to explore the microbiological factors that cause the difference in silage fermentation characteristics between grass and legume. Specifically, the effects of epiphytic microbiota from alfalfa, oat and Italian ryegrass on ensiling characteristics and microbial community of alfalfa were assessed. By γ-ray irradiation sterilization and microbiota transplantation technology, the sterile alfalfa was inoculated as follows: (i) aseptic water (STAL); (ii) epiphytic bacteria from alfalfa (ALAL); (iii) epiphytic bacteria from oat (ALOT); (iv) epiphytic bacteria from Italian ryegrass (ALIR). Alfalfa at the initial flowering stage was ensiled in laboratory-scale silos for 1, 3, 7, 14, 30 and 60 days. Compared with ALAL and ALIR, higher lactic acid contents and ratio of lactic acid to acetic acid, and lower acetic acid, propionic acid, ethanol and ammonia nitrogen contents were observed in ALOT after 60 days of fermentation. In each treated group, Lactobacillus was the most dominant genus after 60 days of ensiling. Relatively higher abundance of Weissella , Hafnia-Obesumbacterium , Enterobacteriaceae or hetero-fermentative Lactobacillus was found in ALAL and ALIR after 60 days. Co-occurrence network analysis proved Pediococcus and Lactococcus were pivotal in deciding the fermentation pattern of alfalfa silage. According to the 16S rRNA gene-predicted functional profiles, the metabolism of amino acids was inhibited by the epiphytic microbiota from oat. Overall, ALOT showed a homo-fermentative process, whereas ALAL and ALIR exhibited a hetero-fermentative pattern. Furthermore, the exogenous microorganisms inhibiting the metabolism of amino acids can be a good potential source to improve the silage quality of legume forage.