A range of agricultural practices influence soil microbial communities, such as tillage and organic C inputs, however such effects are largely unknown at the initial stage of soil formation. Using an eight-year field experiment established on exposed parent material (PM) of a Mollisol, our objectives were to: (1) to determine the effects of field management and soil depth on soil microbial community structure; (2) to elucidate shifts in microbial community structure in relation to PM, compared to an arable Mollisol (MO) without organic amendment; and (3) to identify the controlling factors of such changes in microbial community structure. The treatments included two no-tilled soils supporting perennial crops, and four tilled soils under the same cropping system, with or without chemical fertilization and crop residue amendment. Principal component (PC) analysis of phospholipid fatty acid (PLFA) profiles demonstrated that microbial community structures were affected by tillage and/or organic and inorganic inputs via PC1 and by land use and/or soil depth via PC2. All the field treatments were separated by PM into two groups via PC1, the tilled and the no-tilled soils, with the tilled soils more developed towards MO. The tilled soils were separated with respect to MO via PC1 associated with the differences in mineral fertilization and the quality of organic amendments, with the soils without organic amendment being more similar to MO. The separations via PC1 were principally driven by bacteria and associated with soil pH and soil C, N and P. The separations via PC2 were driven by fungi, actinomycetes and Gram (−) bacteria, and associated with soil bulk density. The separations via both PC1 and PC2 were associated with soil aggregate stability and exchangeable K, indicating the effects of weathering and soil aggregation. The results suggest that in spite of the importance of mineral fertilization and organic amendments, tillage and land-use type play a significant role in determining the nature of the development of associated soil microbial community structures at the initial stages of soil formation. [ABSTRACT FROM AUTHOR]