Stellera chamaejasme L. is a fast-spreading unpalatable poisonous plant that grows in the alpine grasslands of the Qinghai-Tibetan Plateau (QTP). The impacts of unpalatable plant species spread on animal health and plant community have been well studied, but studies into their effects on belowground organisms and processes are rare. We carried out a soil metabarcoding study using Illumina MiSeq sequencing to investigate whether the soil bacteria and fungi communities of Stellera are different to the soil microbiome of neighboring palatable grass Elymus nutans Griseb. Total carbon and nitrogen, the ratio of carbon to nitrogen, ammonium nitrogen, and microbial biomass carbon were all significantly greater in Stellera soil compared to Elymus soil, while no significant differences were observed for gravimetric soil moisture, pH or nitrate nitrogen. There were no significant differences in bacterial and fungal abundance between Stellera and Elymus soil. The bacterial species richness was significantly lower in Stellera soil but no significant difference was observed for fungal species richness. The beta diversity and community composition of bacteria and fungi were markedly different between soils. The presence of bacterial phyla Actinobacteria and Verrucomicrobia, and fungal phyla, Basidiomycota and Glomeromycota, were significantly greater under Stellera soil. This study demonstrated that the spread of undesirable unpalatable plants can potentially disrupt existing plant-soil-microbe associations with potential consequences for grassland soil biodiversity and ecosystem functioning.