Cell and plastid membranes play a critical role in plant response to chilling stress. Fall color retention (chilling tolerance) of bermudagrass (Cynodon sp.) is known to vary with cultivar and management practices. A growth chamber study was conducted to characterize the lipid composition of three bermudagrasses in response to chilling stress. The grasses selected were ‘Tahoma 31’ (chilling-sensitive) and ‘Tifway’ (chilling-tolerant) interspecific hybrid bermudagrass (C. dactylon × C. transvaalensis) and ‘Celebration’ common bermudagrass (C. dactylon), which served as an internal standard. Plants were subjected to simulated fall conditions defined as an 8/2 °C (day/night) temperature regime with 10-hour photoperiod and evaluated for chilling response for 42 days before allowing plants to enter an apparent dormancy. Plant leaves were sampled for lipidomics analysis at 0, 14, and 42 days of chilling treatment (DOT) and again after 40 days of recovery from dormancy (during which temperatures were adjusted to mimic average spring conditions for Oklahoma). ‘Tifway’ demonstrated the lowest electrolyte leakage (EL) and visual discoloration at 42 DOT, while ‘Tahoma 31’ had the greatest EL and discoloration on the same date, and ‘Celebration’ was intermediate of the two. Prolonged exposure to chilling stress generally increased digalactosyldiacylglycerol and phosphatidylcholine (PC) content and decreased monogalactosyldiacylglycerol (MGDG) content, with ‘Tahoma 31’ showing the greatest increase in PC and decrease in MGDG. The double bond index, an indicator of fatty acid unsaturation, was greatest in ‘Tifway’ at 42 DOT. Each cultivar increased in fatty acid unsaturation, with Tifway demonstrating the greatest increase in MGDG unsaturation. Multivariate discriminant analysis identified six individual lipid species that contributed most to the cultivar response to chilling. These findings suggest unsaturation level of plastid lipids, particularly MGDG, is important for chilling tolerance and therefore fall color retention of bermudagrass. Furthermore, this study provides evidence that chilling tolerance can be negatively associated with freezing tolerance in bermudagrass.