Iron stabilizes thylakoid protein–pigment complexes in Indian mustard during Cd-phytoremediation as revealed by BN-SDS-PAGE and ESI-MS/MS

Two-dimensional BN-SDS-PAGE, ESI-MS/MS and electron microscopy (EM) were used to study the role of iron (Fe) under cadmium (Cd) stress in retention of thylakoidal multiprotein complexes (MPCs) and chloroplast ultrastructure of Indian mustard, a moderate hyperaccumulator plant. Mustard was grown hydroponically with or without iron for 17 days and then exposed to CdCl2 for 3 days. Fe deficiency led to an increase in oxidative stress and damage to chloroplast/thylakoids accompanied by a decrease in chlorophyll content; exposure of plants to Cd further enhanced the oxidative stress and Cd accumulation (more in −Fe plants). However, the presence of iron aided plants in the suppression of oxidative stress and retention of chloroplasts and chlorophylls under Cd stress. Proteomic analyses by 2D BN-SDS-PAGE and mass spectrometry showed that Fe deficiency considerably decreased the amount of LHCII trimer, ATPase-F1 portion, cyt b6/f and RuBisCO. No or less reduction, was observed for PSI(RCI+LHCI), the PSII-core monomer, and the PSII subcomplex, while an increase in the LHCII monomer was noted. Under iron deficiency, Cd proved to be very deleterious to MPCs, except for the PSII subcomplex, the LHCII monomer and free proteins which were increased. Iron proved to be very protective in retaining almost all the complexes. MPCs showed greater susceptibility to Cd than Fe deficiency, mainly at the level of RuBisCO and cyt b6/f; an increase in the amount of the PSII subcomplex, LHCII monomer and free proteins indicates differences in the mechanisms affected by Fe deficiency and Cd stress when compared to Fe-fed plants. This study furthers our understanding of the sites actually damaged in MPCs under Fe deficiency and Cd stress. A role emerges for iron in the protection of MPCs and, hence, of the chloroplast. The present study also indicates the importance of iron for efficient phytoextraction/phytoremediation.