Potassium （K^＋） is one of the essential macronutrients for plant growth and development. K^＋ uptake from environment and K^＋ translocation in plants are conducted by K^＋ channels and transporters. In this study, we demonstrated that KT/HAK/KUP transporter KUP7 plays crucial roles in K^＋ uptake and translocation in Arabidopsis root. The kup7 mutant exhibited a sensitive phenotype on Iow-K^＋ medium, whose leaves showed chlorosis symptoms compared with wild-type plants. Loss of function of KUP7 led to a reduction of K^＋ uptake rate and K^＋ content in xylem sap under W-deficient conditions. Thus, the K^＋ content in kup7 shoot was significantly reduced under Iow-K^＋ conditions. Localization analysis revealed that KUP7 was predominantly targeted to the plasma membrane. The complementation assay in yeast suggested that KUP7 could mediate K^＋ transport. In addition, phosphorylation on S80, S719, and S721 was important for KUP7 activity. KUP7 was ubiquitously expressed in many organs/tissues, and showed a higher expression level inArabidopsis root. Together, our data demonstrated that KUP7 is crucial for K^＋ uptake inArabidopsis root and might be also involved in K^＋ transport into xylem sap, affecting K^＋ translocation from root toward shoot, especially under K^＋-Iimited conditions.