A study was carried out to assess the mitigation mechanism of exogenously applied sodium hydrosulfide (NaHS) as a donor of H2S on strawberry seedlings under iron deficiency. The ameliorative effects of NaHS on oxidative damage, ion hemostasis and uptake, and availability of Fe were investigated by spraying solution of 0.2 mM NaHS or 0.2 mM NaHS plus 0.2 mM hypotaurine (HT), a scavenger of H2S to plant leaves. Iron deficiency was created using 0.1 mM FeSO4 instead of 0.1 mM EDTA-Fe in Hoagland's nutrient solution. After a 28-day treatment, strawberry plants exhibited leaf interveinal chlorosis under Fe deficiency, but these apparent symptoms of iron deficiency were overcome by foliar application of NaHS. Exogenously applied NaHS enhanced chlorophyll contents and available iron and Fe accumulation in young leaves, but application of H2S scavenger hypotaurine with NaHS did not change those parameters under Fe deficiency. This clearly shows that NaHS improved iron availability in the strawberry plants. Furthermore, exogenously applied NaHS increased endogenous H2S and iron levels in the roots and leaves. Moreover, NaHS enhanced the levels of zinc (Zn2+), calcium (Ca2+), and magnesium (Mg2+) in both leaves and roots of the strawberry plants grown at Fe deficiency, except for Zn in roots which decreased significantly. This also suggests that NaHS maintains the levels of inorganic ions restricted by Fe deficiency. Fe deficiency increased electrolyte leakage (EL) and the levels of malondialdehyde (MDA) and hydrogen peroxide (H2O2) in plant leaves. Exogenous NaHS reduced the accumulation of H2O2, MDA, and EL and upregulated the activities of key antioxidant enzymes. Overall, NaHS improved Fe uptake and activation by improving endogenous H2S, maintained balance of mineral nutrients and activities of the antioxidant enzymes, and reduced the generation of MDA and H2O2 as well as electrolyte leakage caused by Fe deficiency. So NaHS proved to be effective in ameliorating iron chlorosis caused by iron deficiency.