Map-based cloning of a new total loss-of-function allele of OsHMA3 causes high cadmium accumulation in rice grain

Rice (Oryza sativa) is a major dietary source of the toxic metal cadmium (Cd). Reducing Cd transfer from soil to the rice grain is important for food safety. Rice cultivars vary widely in their Cd accumulation, but the genetic basis for this variation is not fully understood. Based on field and pot experiments comparing 26 rice cultivars, we identified a cultivar with high Cd accumulation in grain (BG367, coded as W4) and a cultivar with low grain Cd accumulation (Huajingxian 74, coded as W0). W4 showed a higher Cd translocation from roots to shoots than W0. Using chromosome single segment substitution lines derived from the two cultivars, we mapped a quantitative trait locus for Cd accumulation in grain to a 400 kb region in chromosome 7. Using yeast expression assays and transgenic complementation, we identified OsHMA3 as the causal gene at this locus. Compared with OsHMA3W0, OsHMA3W4 has a deletion of 14 amino acids predicted to be in the ATP binding domain. OsHMA3W4 showed a complete loss of transport activity for Cd in yeast assays. Taking our findings together, we have identified a new allele of OsHMA3 with a total loss-of-function, resulting in greatly elevated Cd translocation to rice shoots and grain.