Low temperature is one of the most significant environmental factors that threaten the survival of subtropical and tropical plant species. By conducting a study, which was arranged in a completely randomized design with three replicates, the relative freezing tolerance (FT) of four Iranian pomegranate cultivars, including ‘Alak Torsh’, ‘Tabestaneh Torsh’, ‘Poost Sefid’, and ‘Poost Syah’, as well as its correlation with some biochemical indices, were investigated. From each cultivar, pieces of one-year-old shoot samples were treated with controlled freezing temperatures (−11, −14, and −17 ℃) to determine lethal temperatures (LT₅₀) based on survival percentage, electrolyte leakage, phenolic leakage, and tetrazolium staining test (TST) methods. Results showed that FT was higher in the second year with a lower minimum temperature and a higher concentration of cryoprotectants. The stronger correlation of electrolyte leakage with survival percentage (r = 0.93***) compared to the other three indices explained that this index could be the most reliable injury index in determining the pomegranate FT to investigate freezing effects. Of all four cultivars, ‘Poost Syah’ was the hardest by presenting a higher FT than ~ −14 ℃ in mid-winter. Accordingly, this pomegranate cultivar seems to be promising to grow in regions with a higher risk of freezing and to be involved in breeding programs to develop novel commercial cultivars.

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