Particle size and metal species are important to both soil microbial toxicity and phytotoxicity in the soil ecosystem. The effects of CuO and ZnO nanoparticles (NPs) and microparticles (MPs) on soil microbial toxicity, phytotoxicity, and bioaccumulation in two crops (Cucumis sativus and Zea mays) were estimated in a soil microcosm. In the microcosm system, soil was artificially contaminated with 1,000 mg/kg CuO and ZnO NPs and MPs. After 15 days, we compared the microbial toxicity and phytotoxicity by particle size. In addition, C. sativus and Z. mays were cultivated in soils treated with CuO NPs and ZnO NPs, after which the treatment effects on bioaccumulation were evaluated. NPs were more toxic than MPs to microbes and plants in the soil ecosystem. We found that the soil enzyme activity and plant biomass were inhibited to the greatest extent by CuO NPs. However, in a Biolog test, substrate utilization patterns were more dependent upon metal type than particle size. Another finding indicated that the metal NP uptake amounts of plants depend on the plant species. In the comparison between C. sativus and Z. mays, the accumulation of Cu and Zn by C. sativus was noticeably higher. These findings show that metal oxide NPs may negatively impact soil bacteria and plants. In addition, the accumulation patterns of NPs depend on the plant species.