Hydrilla verticillata (L.f.) Royle has a great ability to accumulate large amounts of arsenic (As). We studied the influence of phosphorus (P), nitrogen (N), pH, and arsenite (As(III)) on As transformation and fate in the H. verticillata - medium system via orthogonal experimental design. The results showed highest plant growth was under intermediate As(III) in the medium, with Chlorophyll a and Chlorophyll b contents in plant diminishing after 96 h treatment. Exposure to high N, high As(III), intermediate P, and low pH in the medium, the highest total arsenic uptake by plants were 169.1 ± 5.5 mg g^-1 dry weight, with As(III) as the predominant speciation (49.1 ± 4.8% to 88.5 ± 0.2%) in plants. Meanwhile, trace As (mainly arsenate (As(V))) was adsorbed on the surface of H. verticillata, and the adsorption amounts of As(V) increased with increasing As(III) concentrations in the medium. The dominant As species was As(V) in the medium although plant was supplied with As(III), and highest As(III) oxidation proportion in the medium would occur when low N and pH associated with high P and As(III). Collectively, As(III) uptake and transformation by H. verticillata cannot be overlooked in the biogeochemical cycling of As in aquatic environment.