The cultivated strawberry Fragaria x ananassa Duch. is the natural hybrid of F chiloensis (L.) Mill. and F. virginiana Mill. The progenitor species have high genetic diversity compared with the cultivated genotypes; therefore, the use of wild relative in F. chiloensis breeding could provide a good for broadening the available genetic variations of cultivated species. In present study, 13 genotypes selected from strawberry super core collection were tested under Fe (-) and Fe (+) conditions for their response against Fe deficiency conditions in a growth medium (GM) (soil + sand + perlite), potentially able to simulate the actual GM in nature. SPAD-meter readings indicating chlorophyll levels of the leaf, shoot dry matter yield, Fe-efficiency rate, shoot total and active Fe concentrations were determined to evaluate the resistance levels of strawberry genotypes against Fe deficiency. Results of this study indicated that different response for strawberry subspecies and genotypes of the same subspecies grown in GM against Fe deficiency. Symptom for Fe deficiency of genotypes varied between 1-5, SPAD-meter readings 3, shoot dry matter yields and shoot Fe concentrations varied between 6.5-38, 1.02-6.06 g plant(-1) and 41.8-233.1 mg kg(-1) respectively. Iron-efficiencies of genotypes were found between 58-98%. Strawberry subspecies, E virginiana spp. glauca, F. chiloensis ssp. chiloensis and F. chiloensis ssp. pacifica showed Fe-efficiency values of 93 8, 79.5 and 79.1% respectively. We concluded that shoot growth performance, Fe intake from GM, transfer of Fe from roots to shoots, shoot Fe-use efficiency, Fe deficiency symptom levels and SPAD-meter readings indicating chlorophyll levels were significant parameters to evaluate the resistance of strawberry genotypes against Fe deficiency. The most Fe-efficient genotypes belonging to F. virginiana spp.glauca could be used in breeding programs aiming at developing new strawberry genotypes suitable for growing under Fe deficient conditions.