Kentucky bluegrass (Poa pratensis L.; drought resistant) and perennial ryegrass (Lolium perenne L.; drought sensitive) are economically important grass species contrasting in drought stress resistance. This study determined the optimal chlorophyll fluorescence parameters to indicate drought incidence and whether the parameters differentiate intraspecies and interspecies variation in drought stress. For each species, nine cultivars were exposed to well‐watered or drought (water withheld) conditions in growth chambers containing high throughput photosynthetic imagers to track real‐time responses of maximal quantum efficiency of photosystem II (Fv/Fm), quantum yield of photosystem II (ΦII), non‐photochemical quenching (NPQ), energy dependent quenching (qE), and photoinhibition quenching (qI). Soil moisture content and relative water content of leaf tissues were evaluated. Due to drought, Fv/Fm and ΦII decreased for cultivars of both species but earlier for perennial ryegrass compared to Kentucky bluegrass. The NPQ, qI, and qE values exhibited more dynamic and earlier changes due to drought compared to Fv/Fm and ΦII and allowed for early, mid, and late drought responses to be illustrated. Drought induced an increase in NPQ, qI, and qE values, which corresponded to activation of photoprotection mechanisms, and was exhibited earlier for perennial ryegrass compared to Kentucky bluegrass. As the drought treatment progressed a decrease or stabilization of low values of NPQ, qI, and qE were observed, and the lowest values were associated with the most drought sensitive cultivars. These results indicate important stress tolerance protection mechanisms for grass species and will broadly impact basic and applied grass research as a non‐destructive phenotyping tool.This article is protected by copyright. All rights reserved