Reversible changes in structure and function of photosynthetic apparatus of pea (Pisum sativum) leaves under drought stress.

The effects of drought on photosynthesis have been extensively studied, whereas those on thylakoid organization are limited. We observed a significant decline in gas exchange parameters of pea (Pisum sativum) leaves under progressive drought stress. Chl a fluorescence kinetics revealed the reduction of photochemical efficiency of photosystem (PS)II and PSI. The non-photochemical quenching (NPQ) and the levels of PSII subunit PSBS increased. Furthermore, the light-harvesting complexes (LHCs) and some of the PSI and PSII core proteins were disassembled in drought conditions, whereas these complexes were reassociated during recovery. By contrast, the abundance of supercomplexes of PSII-LHCII and PSII dimer were reduced, whereas LHCII monomers increased following the change in the macro-organization of thylakoids. The stacks of thylakoids were loosely arranged in drought-affected plants, which could be attributed to changes in the supercomplexes of thylakoids. Severe drought stress caused a reduction of both LHCI and LHCII and a few reaction center proteins of PSI and PSII, indicating significant disorganization of the photosynthetic machinery. After 7 days of rewatering, plants recovered well, with restored chloroplast thylakoid structure and photosynthetic efficiency. The correlation of structural changes with leaf reactive oxygen species levels indicated that these changes were associated with the production of reactive oxygen species.

Change in the photochemical and structural organization of thylakoids from pea (Pisum sativum) under salt stress.

Salt can induce adverse effects, primarily on the photosynthetic process, ultimately influencing plant productivity. Still, the impact of salt on the photosynthesis process in terms of supercomplexes organization of thylakoid structure and function is not understood in Pea (Pisum sativum). To understand the structure and function in the leaves and thylakoids under salt (NaCl) treatment, we used various biophysical and biochemical techniques like infrared gas analyzer, chlorophyll a fluorescence, circular dichroism, electron microscopy, blue native gels, and western blots. The net photosynthetic rate, transpiration rate, and stomatal conductance were reduced significantly, whereas the water use efficiency was enhanced remarkably under high salt conditions (200 mM NaCl). The photochemical efficiency of both photosystem (PS) I and II was reduced in high salt by inhibiting their donor and acceptor sides. Interestingly the non-photochemical quenching (NPQ) is reduced in high salt; however, the non-regulated energy dissipation (NO) of PSII increased, leading to inactivation of PSII. The obtained results exhibit inhibition of NAD(P)H dehydrogenase (NDH) mediated pathway-dependent cyclic electron transport under salinity caused a decrease in proton motive force of ΔpH and Δψ. Further, the electron micrographs show the disorganization of grana thylakoids under salt stress. Furthermore, the macro-organization and supercomplexes of thylakoids were significantly affected by high salt. Specifically, the mega complexes, PSII-LHCII, PSI-LHCI, and NDH complexes were notably reduced, ultimately altering the electron transport. The reaction center proteins of oxygen-evolving complexes, D1 and D2 proteins were affected to high salt indicating changes in photochemical activities.

Restoration of photosynthetic activity and supercomplexes from severe iron starvation in Chlamydomonas reinhardtii.

The eukaryotic alga Chlamydomonas (C.) reinhardtii is used as a model organism to study photosynthetic efficiency. We studied the organization and protein profile of thylakoid membranes under severe iron (Fe2+) deficiency condition and iron supplement for their restoration. Chlorophyll (Chl) a fluorescence fast OJIP transients were decreased in the severe Fe2+ deficient cells resulting in the reduction of the photochemical efficiency. The circular dichroism (CD) results from Fe2+ deficient thylakoid membranes show a significant change in pigment-pigment and pigment-protein excitonic interactions. The organization of super-complexes was also affected significantly. Furthermore, super-complexes of photosystem (PS) II and PSI, along with its dimers, were severely reduced. The complexes separated using sucrose gradient centrifugation shows that loss of super-complexes and excitonic pigment-pigment interactions were restored in the severely Fe2+ deficient cells upon Fe supplementation for three generations. Additionally, the immunoblots demonstrated that both PSII, PSI core, and their light-harvesting complex antenna proteins were differentially decreased. However, reduced core proteins were aggregated, which in turn proteins were unfold and destabilized the supercomplexes and its function. Interestingly, the aggregated proteins were insoluble after n-Dodecyl ß-D-maltoside solubilization. Further, they were identified in the pellet form. When Fe2+ was added to the severely deficient cells, the photosynthetic activity, pigment-proteins complexes, and proteins were restored to the level of control after 3rd generation.