Glyphosate alone or a tank mixture of glyphosate and 2,4-D is commonly used for broad-spectrum weed control under fallow conditions in Australia. Air temperature or mixing glyphosate with 2,4-D, may influence the efficacy of glyphosate on feather fingergrass (Chloris virgata Sw.), a problematic summer-season weed of Australia. Dose–response studies were conducted with four populations of feather fingergrass under temperature-controlled glasshouse conditions (35/25 °C and 25/15 °C at 12 h/12 h) to assess the level of glyphosate resistance in relation to temperature regimes. Four parameter log-logistic models were used to develop dose–response curves. Based on plant mortality percentage, LD50 (lethal dose for 50% mortality) values of glyphosate at 25/15 °C for populations Ch, SGM2, SGW2, and CP2 were 137, 60, 650, and 1067 g ae ha−1, respectively. However, at 35/25 °C, the corresponding LD50 values were 209, 557, 2108, and 2554 g ae ha−1, respectively. A similar response was observed for the parameter GR50 (dose for 50% growth reduction) values of glyphosate. These results indicate that populations SGW2 and CP2 are highly glyphosate-resistant and in the summer season, it may be very difficult to control these populations due to poor glyphosate efficacy. These results further suggest that the efficacy of glyphosate for feather fingergrass control can be improved if applied during cooler temperatures (25/15 °C) or the spring season compared with warmer temperatures (35/25 °C) or the summer season. In another study, 2,4-D antagonized glyphosate remarkably in the CP2 (glyphosate-resistant) population but only marginally in the Ch (glyphosate-susceptible) population. Thus, it is not advisable to mix 2,4-D with glyphosate for the control of glyphosate-resistant feather fingergrass populations. The results further suggest that the use of this mixture is useful if the feather fingergrass is not glyphosate-resistant; however, the use of the mixture is to be avoided if the population is glyphosate-resistant in order to not exacerbate the potential resistance problem.