“Alto2” is a new biomass sorghum variety, which has the characteristics of fast growth, high growth, and strong cadmium (Cd) resistance, so it has the application prospect of soil remediation plants. In order to reveal the Cd resistance mechanism of this plant and pave the way for genetic breeding and cultivation of efficient remediation plants in the future, in this research, through the determination of Cd content in various tissues of sorghum under Cd stress and the physicochemical response combined with RNA-Seq analysis, the mechanism of Cd resistance of “Alto2” was initially revealed. The results show biomass sorghum “Alto2” was mainly connected with aboveground and underground parts through the MAPK signaling pathway and plant hormone signaling pathway, and transmit stress signal in response to Cd stress. Chelase and metal-binding proteins may be the functional genes mainly responsible for Cd enrichment and transport and regulated by stress signals. However, the expression of aboveground transporters was not significant. This may be because Cd in biomass sorghum is mainly concentrated in the underground part and is enriched by the chelation of secondary metabolites from plant roots by the cell wall leading to inhibition of aboveground transporter expression. The results of this study indicate that the biomass sorghum “Alto2” on Cd has high resistance, but the lack of the aboveground enrichment of transportability requires further research to improve the Cd transportability of this plant.