Jing Huang, Qiang Zhang, Fang Wei Yu, Ming Qing Liu, Huai Kang Jing, Shen Yun Wang, Hao Yu Wang, Jia Qi Li, Lu Zheng, Hua Shao, Xiao Lin He, Ren Fang Shen, Xiao Fang Zhu
Cadmium (Cd) pollution in paddy soils threatens global food safety. Identifying molecular gateways controlling grain Cd accumulation is essential for breeding low-Cd rice. A field-based screen of a large-scale CRISPR/Cas9 mutant library identified OsGONST3, a Golgi nucleotide sugar transporter family member localized to the plasma membrane. After 1 μM Cd treatment for 7 d, OsGONST3 transcript abundance increased approximately 3.4-fold in root and 5.7-fold in shoot, accompanied by increased OsGONST3-GFP protein accumulation. Heterologous OsGONST3 expression increased yeast Cd content by approximately 23% relative to the empty-vector control. Under hydroponic exposure to 1 μM Cd for 7 d, two independent osgonst3 mutants contained 22-25% less Cd in root, approximately 13% less in shoot, and 16-21% less in xylem sap than Nipponbare. Conversely, OsGONST3-overexpressing lines contained 40-42%, 38-40%, and 28-37% more Cd in root, shoot, and xylem sap, respectively. Under tested field condition, OsGONST3 disruption reduced grain Cd by approximately 25-26% without a significant yield penalty and was associated with secondary transcriptional adjustments in Cd-homeostasis genes. These findings identify OsGONST3 as a plasma membrane-localized Cd influx-associated protein and a promising target for breeding low-Cd rice.