Micronutrient homeostasis and chloroplast iron protein expression is largely maintained in a chloroplast copper transporter mutant.

PAAI is a P-Type ATPase that functions to import copper (Cu) into the chloroplast. Arabidopsis thaliana (L.) Heynh. paa1 mutants have lowered plastocyanin levels, resulting in a decreased photosynthetic electron transport rate. In nature, iron (Fe) and Cu homeostasis are often linked and it can be envisioned that paa1 acclimates its photosynthetic machinery by adjusting expression of its chloroplast Fe-proteome, but outside of Cu homeostasis paa1 has not been studied. Here, we characterise paa1 ultrastructure and accumulation of electron transport chain proteins in a paa1 allelic series. Furthermore, using hydroponic growth conditions, we characterised metal homeostasis in paa1 with an emphasis on the effects of Fe deficiency. Surprisingly, the paa1 mutation does not affect chloroplast ultrastructure or the accumulation of other photosynthetic electron transport chain proteins, despite the strong decrease in electron transport rate. The regulation of Fe-related photosynthetic electron transport proteins in response to Fe status was maintained in paa1, suggesting that regulation of the chloroplast Fe proteins ignores operational signals from photosynthetic output. The characterisation of paa1 has revealed new insight into the regulation of expression of the photosynthetic electron transport chain proteins and chloroplast metal homeostasis and can help to develop new strategies for the detection of shoot Fe deficiency.

Regulation of Iron Homeostasis and Use in Chloroplasts.

Iron (Fe) is essential for life because of its role in protein cofactors. Photosynthesis, in particular photosynthetic electron transport, has a very high demand for Fe cofactors. Fe is commonly limiting in the environment, and therefore photosynthetic organisms must acclimate to Fe availability and avoid stress associated with Fe deficiency. In plants, adjustment of metabolism, of Fe utilization, and gene expression, is especially important in the chloroplasts during Fe limitation. In this review, we discuss Fe use, Fe transport, and mechanisms of acclimation to Fe limitation in photosynthetic lineages with a focus on the photosynthetic electron transport chain. We compare Fe homeostasis in Cyanobacteria, the evolutionary ancestors of chloroplasts, with Fe homeostasis in green algae and in land plants in order to provide a deeper understanding of how chloroplasts and photosynthesis may cope with Fe limitation.

A Program for Iron Economy during Deficiency Targets Specific Fe Proteins.

Iron (Fe) is an essential element for plants, utilized in nearly every cellular process. Because the adjustment of uptake under Fe limitation cannot satisfy all demands, plants need to acclimate their physiology and biochemistry, especially in their chloroplasts, which have a high demand for Fe. To investigate if a program exists for the utilization of Fe under deficiency, we analyzed how hydroponically grown Arabidopsis (Arabidopsis thaliana) adjusts its physiology and Fe protein composition in vegetative photosynthetic tissue during Fe deficiency. Fe deficiency first affected photosynthetic electron transport with concomitant reductions in carbon assimilation and biomass production when effects on respiration were not yet significant. Photosynthetic electron transport function and protein levels of Fe-dependent enzymes were fully recovered upon Fe resupply, indicating that the Fe depletion stress did not cause irreversible secondary damage. At the protein level, ferredoxin, the cytochrome-b6f complex, and Fe-containing enzymes of the plastid sulfur assimilation pathway were major targets of Fe deficiency, whereas other Fe-dependent functions were relatively less affected. In coordination, SufA and SufB, two proteins of the plastid Fe-sulfur cofactor assembly pathway, were also diminished early by Fe depletion. Iron depletion reduced mRNA levels for the majority of the affected proteins, indicating that loss of enzyme was not just due to lack of Fe cofactors. SufB and ferredoxin were early targets of transcript down-regulation. The data reveal a hierarchy for Fe utilization in photosynthetic tissue and indicate that a program is in place to acclimate to impending Fe deficiency.