This study investigated the effectiveness of lactic acid bacteria (LAB) starters in 140 tons of signal grass silage. We compared a locally developed starter, a combination of Lactiplantibacillus plantarum KUB-SP1-3 and Pediococcus acidilactici KUB-M6 (KUB-G2) with a commercial starter. We monitored the chemical and microbiological properties of silage at ensiling (day 0) and after 21 days. The results indicated that KUB-G2 produced high-quality silage. There were no significant differences in total viable microbes, lactic acid, or propionic acid contents between the starters. However, the silage produced with KUB-G2 had a lower acetic acid content than that produced with the commercial starter. KUB-G2 also resulted in a narrower microbiome profile dominated by Pediococci whereas the control group displayed a broader range of bacterial taxa with colonization by Lactiplantibacillus and Lentilactobacillus. These findings suggested that KUB-G2, with its distinct microbial profile, is a suitable replacement for imported commercial starters in large-scale signal grass silage production.