Land‐use change has significant influences on soil respiration (Rs) in terrestrial ecosystems. Soil microbes play critical roles in soil carbon cycling. Nevertheless, the specific mechanism of how changes in soil microbial properties are linked to the variation of Rs rate during land‐use change still remains poorly understood, especially in the Loess Plateau, China. Here, the characteristics of Rs rate and soil microbial community following the land‐use change from farmland to plantation/grassland were analyzed via an automated soil CO2 flux system and high‐throughput 16S rDNA gene sequencing. The afforestation altered soil microbial diversity and community composition, which was mainly explained by soil pH, temperature, organic matter, nitrate, alkali‐hydrolyzed nitrogen, and available phosphorus. The biomarkers of Bacteroidetes, Firmicutes, and Thaumarchaeota were found in farmland soil at the phylum level. The afforestation also significantly decreased Rs rate, which was closely related to Shannon's index, Simpson's index, and some microbial taxa, such as Bacteroidetes, Firmicutes, Nitrospirae, and Acidobacteria. Bacteroidetes, and Firmicutes were particularly expected to be important drivers of high Rs rate in farmland soil. Moreover, the microbial interaction was probably also an important factor affecting Rs rate. Our results indicate that the response of Rs to land‐use change depends on soil microbial community being regulated by soil physicochemical properties in the Loess Plateau, China.