This study chose meadow steppe, typical steppe, desert steppe, three types of steppes, a total of 13 sampling sites in northern temperate steppe of China, and determined phospholipid fatty acids (PLFAs) composition of soil microbes. Nonmetric multidimensional scaling method was used to analyze the soil microbial community composition. The results showed that microbial community structure significantly differed among three types of steppes. Then, the soil microbial community composition data with 10 kinds of environmental factors were combined to conduct redundancy analysis. 79.87% of the variation of soil microbial community composition was explained. Soil moisture content, soil total carbon, soil total nitrogen could affect the soil microbial community composition of meadow steppe. Soil microbial community composition of typical steppe was closely related to light carbon and nitrogen component of soil. Mean average temperature and soil pH value significantly influenced the soil microbial community composition of desert steppe.