Acid rain is an important phenomena in global climate change and it can strongly influence forest ecology. Many studies have shown the impacts of acid rain on soil respiration, but the results are very different due to the different species and environmental conditions. The persistent acidification of soil caused by acid rain has an important impact on the carbon cycle of the Moso bamboo forest ecosystem. To quantify the effect of acid rain on the soil ecosystem of Moso bamboo forest in subtropical China, the experiment of soil by simulated acid rain in the field was carried out for two years of 2016-2018. Two treatments of the simulated acid rain with pH value of 4.0 (T1) and 2.0 (T2) were set, at the same time taking pH value of 5.8 (CK) as control. The relationship between soil CO₂ emission and soil microbial diversity in the Moso bamboo forest of Tianmu Mountain National Nature Reserve under the simulated acid rain was analyzed, and the relationship between soil respiration, soil microbial and soil physical and chemical properties in subtropical Moso bamboo forest was determined. The results showed that soil respiration rate increased firstly and then decreased after the buffer period under the action of acid rain, and the intensity of the action was T2>T1. The sensitivity of soil respiration to temperature in different treatments from high to low was T2, CK, T1, and increased seasonally with increasing temperature. PCR-DGGE analysis showed that strong acid rain treatment inhibited the growth of soil bacterial community and the order of soil bacteria diversity and richness from high to low was CK, T1, and T2. The weak acid rain had a positive effect on the diversity and abundance of soil fungi, and the opposite was strong acid rain. The effects of acid rain resulted in the decrease or disappearance of Bacillus, Caulobacter, Cystobacteraceae, Bradyrhizobium and Candida, as well as inhibiting soil respiration, while Carnobacterium, Propionibacterium, and Clostridium played greater roles in soil respiration under the simulated acid rain. Soil pH, available potassium, soluble organic carbon and microbial biomass carbon were closely and positively correlated with the soil microbial community structure under simulated acid rain treatment, while alkali nitrogen and available phosphorus were negatively correlated with the soil microbial community structure. In general, simulated acid rain can significantly inhibit soil respiration in Moso bamboo forests, and change soil microbial community structure and diversity. These results provide theoretical basis for further study on the response mechanism of soil ecosystem of Moso bamboo forest to environmental problems.