Fe deficiency was imposed by omission of Fe (-Fe), or by inclusion of bicarbonate (supplied as 20 mM NaHCO3) in the nutrient solution in two contrasting peach rootstocks (GF-677; tolerant to Fe deficiency and Cadaman; sensitive to Fe deficiency) for 4 months. In the Fe-deprived leaves and roots, and especially in those treated with bicarbonate, a decrease in Fe concentrations was recorded. Omission of Fe resulted in an increase of the activity of root Fe(III)-chelate reductase (FCR) in both rootstocks, whereas FCR activity decreased in the bicarbonate-treated roots of Cadaman. The results obtained from the FCR assay were confirmed by an agarose-based staining technique used to localize FCR activity. Also, an agar-pH-test revealed that the roots of GF-677 exposed to (-Fe) treatment induced a strong H+ extrusion. In addition, Fe deficiency resulted in reduction of the total chlorophyll (CHL) content. Apart from the (-Fe)-treated leaves of GF-677, Fe deficiency caused a decline in the photosynthetic rate (P(n)) and stomatal conductance (g(s)), without changes of the intercellular CO2 concentration (C(i)), as well as a reduction in the maximum quantum yield of PSII (F(v)/F(m)) and the ratio between variable to initial fluorescence F(v)/F0. The above changes were particularly evident for the bicarbonate-treated leaves of Cadaman. On the other hand, Fe deficiency resulted in an increase of leaf superoxide dismutase (SOD) activity and a depression of catalase (CAT) activity in the leaves and roots, irrespective of the rootstock. Although the non-enzymatic antioxidant activity (FRAP values) was increased in the roots of both rootstocks exposed to -Fe treatment, however, FRAP values were stimulated in the (-Fe)-treated leaves of GF-677 and decreased in the bicarbonate-treated leaves of Cadaman. The H2O2 content was increased in Fe-deprived tissues except for the (-Fe)-treated leaves and roots of GF-677. As a result of Fe deficiency, peroxidase (POD) activity and isoform expression were diminished in the tissues of Cadaman. However, in the tissues of GF-677 subjected to -Fe treatment POD activity was increased whereas an additional POD isoform was detected in the roots suggesting that expression of POD isoforms might be an important attribute linked to the tolerance to Fe deficiency.