Mitochondria were extracted from seedling radicles of Pima S-5 and Pima E-14 cottons and the state 3 respiration, state 4 respiration, ADP:O ratio and respiratory control (RC) ratio were measured in vitro over a range of temperatures from 6 to 18C. Mitochondria from E-14 seedlings exhibited a mean state 3 respiration rate of 13.42 μMO₂/min/gm tissue while mitochondria from S-5 seedlings showed a mean state 3 rate of 17.94 μMO₂/min/gm tissue. Mean state 4 respiration exhibited a similar trend with measurements of 73.4 μMO₂/min/gm tissue and 11.73 μMO₂/min/gm tissue for E-14 and S-5. Mitochondria from E-14 seedlings exhibited a mean ADP:O ratio of 3.73 compared to an ADP:O of 3.28 for S-5, across all assay temperatures. Mean respiratory control ratio was 1.79 for E-14 and 1.53 for S-5. These lower respiration rates of E-14 coupled with higher ADP:O ratios and RC ratios support a greater respiratory efficiency at low temperatures of this variety compared to S-5. Additionally, the E-14 mitochondrial membranes exhibited an ability to remain in a fluid state to a lower temperature than Pima S-5 mitochondrial membranes as judged by Arrhenius plots of respiration. Since mitochondrial respiration is considered to be regulated by membrane-bound enzymes, any change in membrane fluidity would conceivably affect mitochondrial enzyme activity and thus alter respiration rates. Changes in respiration rates will be reflected as a break in an Arrhenius plot. The mean break point temperature of state 3 respiration was 10.7C for E-14 and 13.4C for S-5. The mean break point temperature for state 4 respiration was 10.9C for E-14 and 13.6C for S-5. The ability of E-14 to withstand a greater degree of chilling under field conditions can be attributed, in part, to the greater fluidity of seedling mitochondrial membranes at low temperatures and concomitant conservation of respiratory energy through a lower rate of respiration. Assays of mitochondrial respiration and Arrhenius plots of mitochondrial respiration versus temperatures could be used to select cotton lines more tolerant to chilling temperatures.