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The study evaluated the effects of growth stage and ensiling time on silage fermentation characteristics, bacterial communities and their predicted functional potential in Italian ryegrass (Lolium multiflorum Lam.) silage. Italian ryegrass (IR) was harvested at heading (Earing rate >50 %, IR1) and blooming stages (>50% bloom, IR2), respectively. The harvested IR was ensiled in 15-L plastic silos. The experiment had a completely randomized design. Triplicate silos were sampled after 1, 3, 7, 15, 30 and 60 days of fermentation, respectively. The fermentation products were analyzed on each sampling day. The bacterial communities and their functional potential and phenotype on day 3 and 60 were analyzed by high-throughput sequencing technique and PICRUSt2 (Phylogenetic Investigation of Communities by Reconstruction of Unobserved States) method. With the advancing maturity of IR, the neutral detergent fiber …