Cardamine violifolia belongs to the Brassicaceae family and is a selenium (Se) hyperaccumulator found in Enshi, China. In this study, C. violifolia was found to accumulate mercury (Hg) in its roots and aboveground parts at concentrations up to 6000 μg/g. In the seedling and mature stages, the bioaccumulation factors (BAF_S) of Hg reached 1.8–223, while the translocation factor (TF) for Hg reached 1.5. We observed a significant positive correlation between THg concentrations in plant tissues and those in the soil (r² = 0.71–0.84). Synchrotron radiation X-ray fluorescence with focused X-ray (μ-SRXRF) showed that Hg was translocated from the roots to shoots through the vascular bundle and was transported through the leaf veins in leaves. Transmission electron microscopy showed that root cells were more tolerant to Hg than leaf cells. These findings provide insights into the mechanisms of Hg hyperaccumulation in C. violifolia. Overall, we demonstrated that C. violifolia is a promising Hg hyperaccumulator that may be used for phytoremediating Hg-contaminated farmlands.