Cadmium (Cd) is a toxic heavy metal that can accumulate in crop plants. We reported previously the engineering of a low cadmium-accumulating line (2B) of rice through overexpression of a truncated OsO3L2 gene. As expression of this transgene was highest in plant roots, amplicon and metatranscriptome sequencing were used to investigate the possibility that its expression affects root associated microbes. Based on amplicon sequencing of bacterial 16S rRNA, but less so from fungal ITS, the OTUs (operational taxonomic units) showed less diversity in soil tightly (rhizoplane) than loosely (rhizosphere) associated with plant roots. Significantly changed OTUs caused by the low-Cd accumulating plant 2B, Cd treatment or both were found, and 10 of the 13 OTUs (77%) that were enriched in Cd treated 2B samples over the wild type counterpart have been previously described as involved in tolerance to Cd or other heavy metals. Metatranscriptome sequencing of rhizosphere microbiome found that bacteria accounted for 70–75% of the microbial RNA. Photosynthesis-antenna proteins and nitrogen metabolism pathways were most active in soil microbes treated with Cd and grown with plant 2B. Correspondingly, the relative abundance of Cyanobacteria was enriched to < 1% of Cd treated rhizosphere bacteria, yet accounted for up to 13% of Cd treated 2B rhizospheric transcripts. These enriched microbes by transgene and Cd are worthy candidates for future application on reducing crop uptake of Cd. •Overexpression of a cadmium-reducing gene reshaped rice root microbiome.•Cadmium treatment contributed to shifts of the rice root microbiome.•Microbial taxa involved in low Cd accumulation of transgenic rice were identified.•Cyanobacteria could play a role in rice low Cd accumulation trait formation.