Microbes live throughout the soil profile. Microbial communities in subsurface horizons are impacted by a saltwater–freshwater transition zone formed by seawater intrusion (SWI) in coastal regions. The main purpose of this study is to explore the changes in microbial communities within the soil profile because of SWI. The study characterizes the depth-dependent distributions of bacterial and archaeal communities through high-throughput sequencing of 16S rRNA gene amplicons by collecting surface soil and deep core samples at nine soil depths in Longkou City, China. The results showed that although microbial communities were considerably impacted by SWI in both horizontal and vertical domains, the extent of these effects was variable. The soil depth strongly influenced the microbial communities, and the microbial diversity and community structure were significantly different (p < 0.05) at various depths. Compared with SWI, soil depth was a greater influencing factor for microbial diversity and community structure. Furthermore, soil microbial community structure was closely related to the environmental conditions, among which the most significant environmental factors were soil depth, pH, organic carbon, and total nitrogen.