Hexavalent chromium [Cr(VI)] contamination has become an emergent concern in China. Previous field investigations have found that hybrid Napier grass is widely distributed in Cr(VI) contaminated areas. This study investigated the phytoremediation potential and biochemical response of hybrid Napier grass ( Pennisetum americanus L. × Pennisetum purpureum Schumach) grown in soil contaminated with Cr(VI) (0, 20, 40, and 60 mg kg−1) with and without Ethylene diamine tetra acetic acid (EDTA) (4 mM) application. The results indicated that root length, shoot height, dry weight, leaf area, chlorophyll, and photosystem II (PSII) parameters viz.; apparent electron transport rate (ETR), effective quantum yield of PSII (ΦPSⅡ), maximal PSII photochemical efficiency (Fv/Fm), potential activity of PSII (Fv/Fo), photochemical quenching (qP), and non-photochemical quenching (qN) decreased with the increasing Cr(VI) concentration. EDTA application further aggravated reduction of dry biomass and photosystem II. The concentration and the accumulation of Cr in shoot and root, and both the bioaccumulation factor (BAF) and transfer factor (TF) increased with increasing Cr(VI) concentrations and further enhanced with EDTA application. Though the Cr(VI) and Ethylene diamine tetra acetic acid (EDTA) stress reduced tolerance, but, even at highest Cr(VI) concentration, plant could exhibited strong resistance, as evidenced by increase in superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities. Hybrid Napier grass, due to its BAF > 1 and a TF < 1, would be applicable for Cr phytostabilization. Moreover, limiting metal transport to aerial parts of plant would prevent animal’s ingestion, restrict soil mobility, and consequently reduce transmission across the food chain.