Dormancy removal by cold stratification increases glutathione and S-nitrosoglutathione content in apple seeds

Abstract

S-nitrosoglutathione (GSNO), an integral metabolite of nitric oxide (NO) biochemistry is reduced by S-nitrosoglutathione reductase (GSNOR) (EC 1.2.1.46), leading to formation of glutathione in oxidised form (GSSG), further reduced to GSH by glutathione reductase (GR). GSH as a vital antioxidant has a significant role for seed quality and during seed germination. Since early 50th of 20th century it is known that deep dormancy of apple (Malus domestica Borkh.) embryos is removed by 90 days of cold stratification. Our previous studies demonstrated that similar effect is observed after short term (3 h) exposition of isolated embryos to nitric oxide (NO) donors. The aim of our work was to verify the differences in GSNO level and GSNOR activity in embryonic axes isolated after initiation of germination (24 h of imbibition) from dormant embryos (the control) and from 90 days cold stratified seeds. Our data indicated that seed dormancy breakage is accompanied by increased GSNO content and the decrease of GSNOR activity. The abundance of GSNOR protein is similar in both non-dormant and dormant embryonic axes during first hours of water uptake, while GSNOR transcript level increases in non-dormant tissue. Furthermore, in non-dormant embryonic axes we noticed a higher glutathione pool, mostly in its reduced form. These results are linked to the increase of cytosolic GR transcript level and increased enzyme activity in embryonic axes isolated from stratified seeds.