Cold-hardiness of olive (Olea europaea L.) cultivars in cold-acclimated and non-acclimated stages: seasonal alteration of antioxidative enzymes and dehydrin-like proteins

Seasonal patterns of antioxidative enzymes and proteins and their relations to cold-hardiness of nine olive (Olea europaea L.) cultivars (Ascolona, Domat, Gemlik, Hojoblanca, Lecquest, Manzanilla, Meski, Samanli and Uslu) are documented in the current study. Fully expanded, uniformly sized leaves from 2-year-old Shoots Of the cultivars were collected from 20-year-old trees in cold-acclimated (CA, in January) and non-acclimated (NA, in July) stages. Leaf samples were exposed to low temperature at 4, -5, -10 and -20 C for 12 It to determine their cold-hardiness (LT50; assessed by electrolyte leakage). Cold-acclimation produced an increase in freezing tolerance of all cultivars (by lowering LT50). Domat and Lecquest were found to have the highest cold-hardiness among the nine cultivars investigated. Ascolona, Gemlik, Hojoblanca had moderate cold-hardiness, while Samanli, Meski, Uslu and Manzanilla were more sensitive. Activities of catalase (CAT: EC 1.11.1.6), ascorbate peroxidase (APX: EC 1.11.1.11) and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase significantly varied depending on the cold-acclimation stage and the cold-hardiness level of the cultivars. Activities of the three antioxidative enzymes and total soluble proteins (TSP) were higher in the CA stage than in the NA stage. Although no accumulation or major polypeptides, except a 23 kDa protein, was detected either in CA samples or NA samples by SDS-PAGE, anti-dehydrin immunoblots revealed that the 43 and 23 kDa polypeptides were detectable during cold-acclimation of olive cultivars, Accumulation of both 43 and 23 kDa dehydrin was significantly higher in the CA stage than in the NA stage in all cultivars. Accumulation of 43 kDa dehydrin was correlated with cold-hardiness of the cultivars, while 23 kDa dehydrin was considered as cultivar-dependent since its accumulation was not parallel to LT50 values of the cultivars. Indeed, the tissues of cvs Domat, Lecquest, Ascolona, Hojoblanca and Gemlik were found to enhance the structural stability Of Cellular membranes in the CA stage by increasing both the activity Of Such enzymes as CAT, APX and NADPII oxidase to activate the antioxidative systems and the expression of 43 kDa dehydrins.