Additional information on the physiological performances for different cassava genotypes would support better decision-making about desirable genetic resources for water-limited conditions. The objective of this study was to evaluate the physiological expression and yield of eight different cassava genotypes grown under a dry environment during high storage root accumulation. The eight cassava genotypes, i.e., Kasetsart 50, Huay Bong 80, Rayong 5, Rayong 7, Rayong 9, Rayong 11, Rayong 90, and CMR38-125-77 were evaluated under rain-fed upland conditions at Khon Kaen University, Thailand, during 2018 to 2020. A randomized complete block design (RCBD) with three replications was used. Soil moisture contents, chlorophyll fluorescence (Fv/Fm and Fv′/Fm′), net photosynthesis (Pn), stomatal conductance, water use efficiency (WUE), relative water content (RWC) for leaf, leaf area index (LAI), specific leaf area (SLA), starch content, crop dry weight, and starch yield were observed at 180, 270, and 360 days after planting (DAP), and weather data during the experimental period were also recorded. The results from both 2018/2019 and 2019/2020 indicated that Pn was positively and significantly correlated with stomatal conductance and Fv/Fm during the high storage root accumulation stage (270 and 360 DAP) with soil moisture content lower than field capacity. CMR38-125-77 had satisfactory performances in Pn, RWC, Fv/Fm, Fv′/Fm′, stomatal conductance, LAI, SLA, WUE, biomass, starch content, and starch yield at a last growth stage with soil moisture content lower than permanent wilting point. Significant association between crop dry weight and WUE at 360 DAP was recorded, and CMR38-125-77 and Kasetsart 50 were classified as favorable genotypes with high WUE and biomass.