In this study, we investigated the inhibitory effect of low temperature on biogas fermentation system. Biogas fermentation inoculum was domesticated at 4 C, and subsequently inoculated into pig manure feedstock in a batch biogas fermentation process. A low-temperature biogas fermentation system was maintained at 9 C, and its abiotic factors, bacterial community, and archaeal community were determined. The results showed that (1) the biogas fermentation lasted for 160 days, with a total gas production of 19,150 mL, including N2production of 11,370 mL, CO2production of 3,534 mL, and CH4production of 3,031 mL. (2) The average relative abundance of the primary dominant bacterium operational taxonomic unit (OTU) was 30%, with 100% similarity to Pseudomonas caeni, atypical denitrifying bacterium. The average relative abundance of the secondary dominant bacterium OTU was 20.36%, with 99% similarity to Clostridium cellulovorans, a typical cellulose- and hemicellulose-degrading bacterium. The average relative abundance of the primary dominant methanogenicarchaeon OTU was 7.22%, with 99% similarity to Methanosphaera cuniculi, a typical methylotrophic methanogen, and the secondary dominant methanogenic archaeon OTU had an average relative abundance of 4.15%, with 98% similarity to Methanocorpusculum sinense, a typical hydrogenotrophic methanogen. (3) The primary dominant bacterial genus Pseudomonas exhibited highly significant negative correlations with the dominant biogas-producing microorganisms such as Clostridium, Terrisporobacter, Turicibacter, Methanosphaera, and Methanocorpusculum. We concluded that (1) high concentrations of abiotic factors such as ammonia nitrogen, acetic acid, and propionic acid were important indicators of relatively poor operation of low-temperature biogas fermentation systems, while (2) high numbers of MCGarchaea and Pseudomonas were important biotic factors showing comparatively poor operation of the system.