We describe a selective and sensitive fluorescence platform for the detection of paraquat (PQ) based on competitive host–guest recognition between phosphate pillar[5]arenes (PWP5) and probe (Safranine T, ST) with using PWP5 functionalized reduced graphene (PWP5-rGO) as the receptor. PQ is a positive charge molecule that is captured by PWP5 via electrostatic interactions. The host-guest interaction between PWP5 and PQ is studied by 1H NMR. Therefore, a selective and sensitive fluorescence sensing of detection PQ is developed. It has a linear response ranges of 0.01–2.0 and 2.0–50.0 μM and a low detection limit of 0.0035 μM (S/N = 3) for PQ. The sensing platform is also used to test PQ in two water samples with satisfying results. It suggests that this approach has potential applications for the determination of PQ.