Biochar management has been proposed as a promising strategy to mitigate climate change. However, the long-term effects of biochar amendment on soil greenhouse gas (GHG) production and microbial community in forest ecosystems under projected warming remain highly uncertain. In this study, we conducted a 49-day incubation experiment to investigate the impact of biochar application on soil physico-chemical properties, GHG production rates, and microbial community at three temperature levels using a temperate forest soil amended with spruce biochar four years ago. Our results showed that temperature exerted a positive effect on soil CO2, CH4 and N2O production, leading to an increase in total global warming potential by 169% and 87% as temperature rose from 5 to 15 °C and from 15 to 25 °C, respectively, and thus a positive feedback to warming. Moreover, warming was found to reduce soil microbial biomass significantly, but at the same time promote the selection of an activated microbial community towards some phyla, e.g. Acidobacteria and Actinobacteria. We observed that biochar amendment reduced soil CH4 consumption and N2O production in the absence of litter by 106% and 94%, respectively, but did not affect soil CO2 production. While biochar had no significant influence of total global warming potential of forest soil, it could promote climate change mitigation by increasing the total soil carbon content by 26% in the presence of litter. In addition, biochar application was shown to enhance soil available phosphorus and dissolved organic carbon concentrations, as well as soil microbial biomass under a warmer environment. Our findings highlighted the potential of spruce biochar as a soil amendment in improving soil fertility and carbon sequestration in temperate forest over the long term, without creating any adverse climatic impacts associated with soil GHG production.