‘Rainbow Red’ kiwifruit have been reported to gradually ripen during low temperature storage and on the vine in the absence of detectable ethylene. This study was conducted to compare the expression of ripening-related genes during storage at different temperatures and on the vine. Fruit at 5°C and 10°C ripened faster to eating quality within four weeks accompanied with increased expression of ripening-related genes: AcACO3,AcXET2, AcEXP1, AcPG, AcPMEi, AcSUS, AcβAMY1, AcβAMY2, AcGA2ox2, AcNAC3, AcNAC4, and AcMADS2. Fruit at 15°C required a longer period of eight weeks to attain eating quality in concurrence with delayed accumulation of the ripening-related genes. Fruit at 22°C ripened at the slowest rate and did not attain eating quality even after eight weeks, with very minimal accumulation of ripening-related genes. On-vine ripening occurred slowly at the early stages when the average field temperature was ~20°C, but the rate increased as the temperature dropped to ≤15°C accompanied by increased expression of ripening-related genes. These results indicate that both ripening on-vine and during low temperature storage are modulated by low temperature independent of ethylene.