Effects of low temperature and respiratory inhibitors on calcium flux in plant mitochondria

The effects of low temperature on uptake and release of ⁴⁵Ca²⁺ were studied with sound, well-coupled mitochondria extracted at room temperature from avocado (Persea americana Mill, cv Fuerte) fruits. Low Ca²⁺ concentrations (10 micromolar) were employed to simulate physiological conditions. At 25°C, the rate of Ca²⁺ uptake decreased with time, whereas at 5°C the initial rate, though lower, remained linear. As a consequence total uptake at 5°C was substantially greater than at 25°C for periods greater than 5 min. Preincubation of mitochondria at 5°C enhanced subsequent Ca²⁺ uptake at 25°C. Ca²⁺ uptake was inhibited by carbonyl cyanide-m-chlorophenyl hydrazone (CCCP) and by ruthenium red, but neither KCN nor salicylhydroxamic acid separately or together had any major inhibitory effect. Preloaded mitochondria held for 60 min in a Ca-free medium lost little Ca²⁺ at 25°C and none at 5°C, except in the presence of ruthenium red or CCCP.