In order to investigate the contribution of polyamines and related amino acids in the maintenance of zucchini fruit quality during cold storage, two varieties of Cucurbita pepo with different degrees of chilling tolerance were used, Natura (more tolerant) and Sinatra (moresensitive). After harvest, free putrescine levels decreased during storage at 20 °C, whereas in fruit kept at 4 °C this polyamine accumulated in both varieties, but with higher levels in the sensitive variety (Sinatra). This behavior suggests that putrescine is accumulated as a response to low temperature in zucchini fruit by stress-induced chilling injury, and not due to the postharvest storage itself. ADC activity responds quickly to chilling but sharply decreases after 14 days, whereas its expression remains high in both varieties. ODC activity takes over when the cold stress is relatively severe, as this activity was found to be much higher in Sinatra. ODCexpression also correlated with ODC activity. DAO activity increased in Natura fruit, and conversely decreased in Sinatra fruit during storage at 4 °C, whereas the proline content was higher in Natura and lower in Sinatra. Therefore, we suggest that putrescine degradation and proline accumulation contribute to the acquisition of chilling tolerance in zucchini fruit. GABA content decreased in both varieties, with a greater reduction in Natura fruit and less in Sinatra fruit. In addition, GABA transaminase showed a higher activity in Natura fruit than in Sinatra fruit during cold storage, suggesting that GABA catabolism could be involved in the tolerance to postharvest cold storage in zucchini fruit.