Herbicide-resistant weed management is one of the greatest agricultural challenges in crop production. Thus, the quick identification of herbicide-resistant weeds is extremely important for management. This study aimed to evaluate resistance to PSI-inhibiting herbicides (diquat) and physiological response to paraquat application in Sumatran fleabane [Conyza sumatrensis (Retz.) E. Walker; syn.: Erigeron sumatrensis Retz.]. The research was conducted with two C. sumatrensis biotypes, one susceptible and the other with multiple resistance to herbicides from five different modes of action (glyphosate, paraquat, diuron, saflufenacil, and 2,4-D). A dose–response assay was carried out to evaluate herbicide resistance to diquat in the paraquat-resistant C. sumatrensis biotype. The enzymatic activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX), hydrogen peroxide (H₂O₂) content, and chlorophyll a (Chl a) fluorescence were measured in both biotypes after paraquat (400 g ai ha^–1) application. The dose–response assay confirmed resistance of C. sumatrensis to diquat with resistance factor levels of 26-fold and 6-fold for LD₅₀ and GR₅₀ values, respectively, compared with the susceptible biotype. Accumulation of H₂O₂ occurred more rapidly in the paraquat-susceptible biotype than in the resistant one. Paraquat treatment caused an increase in SOD and APX activity in the susceptible biotype, but antioxidant enzyme activities were unaffected by paraquat in the resistant one at 5 h after application (HAA). Chl a fluorescence increased across the first 4 HAA in both resistant and susceptible biotypes. However, at 24 HAA, the resistant biotype showed a decline in fluorescence close to untreated plants, while the susceptible biotype died, confirming resistance to diquat in the paraquat-resistant C. sumatrensis biotype. The paraquat-resistant biotype does not induce antioxidative enzymes, as a possible mechanism of resistance to paraquat, but shows rapid recovery of photosynthesis and continuous growth when subjected to paraquat, while the paraquat-susceptible biotype does not survive.