Plastic greenhouse vegetable cultivation (PGVC) has been widely developed around the world and has resulted in great changes in soil properties and potential contamination by phthalate esters (PAEs). Using high-throughput sequencing, this study investigated the succession and potential factors impacting soil microbial community structures over 20 years of PGVC. The results showed that the pH of soils under PGVC were significantly lower, while the nutrient contents of soils were higher, relative to those of open field soil. The residue concentrations of PAEs in soil under PGVC increased with increasing periods of PGVC. The fungal community diversity, rather than the bacterial community diversity, was significantly reduced in soils under PGVC. However, both the soil bacterial and fungal community structures were changed by long-term PGVC. Among the tested soil physicochemical properties, soil pH and clay were the top two factors affecting the soil bacterial community, while pH and phosphorus (P) mainly affected the soil fungal community structures. No relationship between the changes of microbial communities and PAE residues in soil was observed. This study indicates that the soil acidification and nutrient accumulation under PGVC mainly shifted the changes of soil microbial community structures, which could occur after only 5 years of PGVC.