摘要Auxin is unique among plant hormones in that its function requires polarized transport across plant cells. A chemiosmotic model was proposed to explain how polar auxin transport is derived by the H+ gradient across the plasma membrane (PM) established by PM H+-adenosine triphosphatases (ATPases). However, a classical genetic approach by mutations in PM H+-ATPase members did not result in the ablation of polar auxin distribution, possibly due to functional redundancy in this gene family. To confirm the crucial role of PM H+-ATPases in the polar auxin transport model, we employed a chemical genetic approach. Through a chemical screen, we identified protonstatin-1 (PS-1), a selective small-molecule inhibitor of PM H+-ATPase activity that inhibits auxin transport. Assays with transgenic plants and yeast strains showed that the activity of PM H+-ATPases affects auxin uptake as well as acropetal and basipetal polar auxin transport. We propose that PS-1 can be used as a tool to interrogate the function of PM H+-ATPases. Our results support the chem-iosmotic model in which PM H+-ATPase itself plays a fundamental role in polar auxin transport.