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Plants have evolved multiple regulatory mechanisms to cope with natural light fluctuations. The interplay between these mechanisms leads presumably to the resilience of plants in diverse light patterns. We investigated the energy‐dependent nonphotochemical quenching (qE) and cyclic electron transports (CET) in light that oscillated with a 60‐s period with three different amplitudes. The photosystem I (PSI) and photosystem II (PSII) function‐related quantum yields and redox changes of plastocyanin and ferredoxin were measured in Arabidopsis thaliana wild types and mutants with partial defects in qE or CET. The decrease in quantum yield of qE due to the lack of either PsbS‐ or violaxanthin de‐epoxidase was compensated by an increase in the quantum yield of the constitutive nonphotochemical quenching. The mutant lacking NAD(P)H dehydrogenase (NDH)‐like‐dependent CET had a transient significant PSI acceptor side limitation during the light rising phase under high amplitude of light oscillations. The mutant lacking PGR5/PGRL1‐CET restricted electron flows and failed to induce effective photosynthesis control, regardless of oscillation amplitudes. This suggests that PGR5/PGRL1‐CET is important for the regulation of PSI function in various amplitudes of light oscillation, while NDH‐like‐CET acts' as a safety valve under fluctuating light with high amplitude. The results also bespeak interplays among multiple photosynthetic regulatory mechanisms. Summary statement: PGR5/PGRL1‐dependent cyclic electron transports (CET) protect photosystems against light fluctuations of varying amplitudes, while NDH‐like‐dependent CET acts as a safety valve to optimise photosynthesis in high amplitude light fluctuations. Interplays may exist among multiple regulatory mechanisms, including quenching and CET. [ABSTRACT FROM AUTHOR]

We present here the research contributions of Jan Amesz (1934–2001) on deciphering the details of the early physico-chemical steps in oxygenic photosynthesis in plants, algae and cyanobacteria, as well as in anoxygenic photosynthesis in purple, green, and heliobacteria. His research included light absorption and the mechanism of excitation energy transfer, primary photochemistry, and electron transfer steps until the reduction of pyridine nucleotides. Among his many discoveries, we emphasize his 1961 proof, with L. N. M. Duysens, of the "series scheme" of oxygenic photosynthesis, through antagonistic effects of Light I and II on the redox state of cytochrome f. Further, we highlight the following research on oxygenic photosynthesis: the experimental direct proof that plastoquinone and plastocyanin function at their respective places in the Z-scheme. In addition, Amesz's major contributions were in unraveling the mechanism of excitation energy transfer and electron transport steps in anoxygenic photosynthetic bacteria (purple, green and heliobacteria). Before we present his research, focusing on his key discoveries, we provide a glimpse of his personal life. We end this Tribute with reminiscences from three of his former doctoral students (Sigi Neerken; Hjalmar Pernentier, and Frank Kleinherenbrink) and from several scientists (Suleyman Allakhverdiev; Robert Blankenship; Richard Cogdell) including two of the authors (G. Garab and A. Stirbet) of this Tribute. [ABSTRACT FROM AUTHOR]

The kinetics of oxidation/reduction of the primary donor of photosystem 1 (PS1) P700, plastocyanin (PC), and ferredoxin (Fd) in the first leaves of barley seedlings under exposure to high-intensity light (2000 μmol∙quanta∙m–2·s–1, 30 min) and elevated temperature (40°C, 3 h) were studied using differential absorption photometry. Exposure to high-intensity light increased the accumulation of the oxidized form of PS1 reaction center P700+, oxidized PC, and reduced Fd. Reduced Fd was not reoxidized under the same conditions on exposure to red and far-red light. The accumulation of P700+, PC+, and Fd – increased in barley seedlings exposed to elevated temperatures. Also, reoxidation of leaf Fd accelerated under red light. Oxidized Fd did not accumulate under far-red light. It was concluded that photo-independent electron flow through Fd under light stress and alternative electron flows involving the plastoquinone pool under heat stress were activated. [ABSTRACT FROM AUTHOR]

The function of ascorbate peroxidase–related (APX-R) proteins, present in all green photosynthetic eukaryotes, remains unclear. This study focuses on APX-R from Chlamydomonas reinhardtii , namely, ascorbate peroxidase 2 (APX2). We showed that apx2 mutants exhibited a faster oxidation of the photosystem I primary electron donor, P700, upon sudden light increase and a slower re-reduction rate compared to the wild type, pointing to a limitation of plastocyanin. Spectroscopic, proteomic and immunoblot analyses confirmed that the phenotype was a result of lower levels of plastocyanin in the apx2 mutants. The redox state of P700 did not differ between wild type and apx2 mutants when the loss of function in plastocyanin was nutritionally complemented by growing apx2 mutants under copper deficiency. In this case, cytochrome c6 functionally replaces plastocyanin, confirming that lower levels of plastocyanin were the primary defect caused by the absence of APX2. Overall, the results presented here shed light on an unexpected regulation of plastocyanin level under copper-replete conditions, induced by APX2 in Chlamydomonas. [ABSTRACT FROM AUTHOR]

Human immunodeficiency virus (HIV) infection has been a risk to public health due to its long-life treatment. The current medications were limited due to their single therapeutic targets, drug resistance, and adverse side effects. The discovery and study of novel anti-HIV agents are necessary. From our previous study, Canna indica L. leaf extract showed anti-HIV activities with plastocyanin as a potential active compound. In this study, the plastocyanin gene of Canna indica L. was cloned using the cDNA library established, the leaf extract mRNA and expressed in pET28(+); E. coli.DE3(BL21). The 408-bp plastocyanin gene produced 14-kDa recombinant protein plastocyanin (Pc) including a histidine tag (his-tag). Pc was characterized using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS--PAGE) and liquid chromatography coupled with mass spectrometry and further purified using N-terminal 6xHis-tagged fusion binding through column affinity chromatography. As expected, Pc was determined to be approximately 14 kDa on SDS--PAGE. To study the anti-HIV activity of Pc and its primary arrangement, the his-tag was removed by thrombin cleavage. Further, Pc and its product cleaved using thrombin (Pc/T) were evaluated for their cytotoxic and anti-HIV-1 activities using the syncytium reduction and anti-HIV reverse transcriptase assays. The syncytium reduction assay revealed that Pc and Pc/T exhibited anti-HIV activity at EC50 of 38.54 and 128.84 µg/ml; with calculated therapeutic indices of >6.49, active and >1.94, active; respectively. Both Pc and Pc/T also exhibited anti-HIV reverse transcriptase inhibitory activity at IC50 of 6.55 and 4.89 µg/ml, respectively. This study demonstrated that recombinant Pc could be used as an anti-HIV agent. [ABSTRACT FROM AUTHOR]

Recent phylogenetic studies have unveiled a novel class of ascorbate peroxidases called "ascorbate peroxidase-related" (APX-R). These enzymes, found in green photosynthetic eukaryotes, lack the amino acids necessary for ascorbate binding. This study focuses on the sole APX-R from Chlamydomonas reinhardtii referred to as ascorbate peroxidase 2 (APX2). We used immunoblotting to locate APX2 within the chloroplasts and in silico analysis to identify key structural motifs, such as the twin-arginine transport (TAT) motif for lumen translocation and the metal-binding MxxM motif. We also successfully expressed recombinant APX2 in Escherichia coli. Our in vitro results showed that the peroxidase activity of APX2 was detected with guaiacol but not with ascorbate as an electron donor. Furthermore, APX2 can bind both copper and heme, as evidenced by spectroscopic, and fluorescence experiments. These findings suggest a potential interaction between APX2 and plastocyanin, the primary copper-containing enzyme within the thylakoid lumen of the chloroplasts. Predictions from structural models and evidence from 1H-NMR experiments suggest a potential interaction between APX2 and plastocyanin, emphasizing the influence of APX2 on the copper-binding abilities of plastocyanin. In summary, our results propose a significant role for APX2 as a regulator in copper transfer to plastocyanin. This study sheds light on the unique properties of APX-R enzymes and their potential contributions to the complex processes of photosynthesis in green algae. [ABSTRACT FROM AUTHOR]

Summary: In natural environments, plants are exposed to rapidly changing light. Maintaining photosynthetic efficiency while avoiding photodamage requires equally rapid regulation of photoprotective mechanisms. We asked what the operation frequency range of regulation is in which plants can efficiently respond to varying light.Chlorophyll fluorescence, P700, plastocyanin, and ferredoxin responses of wild‐types Arabidopsis thaliana were measured in oscillating light of various frequencies. We also investigated the npq1 mutant lacking violaxanthin de‐epoxidase, the npq4 mutant lacking PsbS protein, and the mutants crr2‐2, and pgrl1ab impaired in different pathways of the cyclic electron transport.The fastest was the PsbS‐regulation responding to oscillation periods longer than 10 s. Processes involving violaxanthin de‐epoxidase dampened changes in chlorophyll fluorescence in oscillation periods of 2 min or longer. Knocking out the PGR5/PGRL1 pathway strongly reduced variations of all monitored parameters, probably due to congestion in the electron transport. Incapacitating the NDH‐like pathway only slightly changed the photosynthetic dynamics.Our observations are consistent with the hypothesis that nonphotochemical quenching in slow light oscillations involves violaxanthin de‐epoxidase to produce, presumably, a largely stationary level of zeaxanthin. We interpret the observed dynamics of photosystem I components as being formed in slow light oscillations partially by thylakoid remodeling that modulates the redox rates. [ABSTRACT FROM AUTHOR]

Blue copper proteins are models for illustrating how proteins tune metal properties. Nevertheless, the mechanisms by which the protein controls the metal site remain to be fully elucidated. A hindrance is that the closed shell Cu(I) site is inaccessible to most spectroscopic analyses. Carbon deuterium (C-D) bonds used as vibrational probes afford nonperturbative, selective characterization of the key cysteine and methionine copper ligands in both redox states. The structural integrity of Nostoc plastocyanin was perturbed by disrupting potential hydrogen bonds between loops of the cupredoxin fold via mutagenesis (S9A, N33A, N34A), variably raising the midpoint potential. The C-D vibrations show little change to suggest substantial alteration to the Cu(II) coordination in the oxidized state or in the Cu(I) interaction with the cysteine ligand. They rather indicate, along with visible and NMR spectroscopy, that the methionine ligand distinctly interacts more strongly with the Cu(I) ion, in line with the increases in midpoint potential. Here we show that the protein structure determines the redox properties by restricting the interaction between the methionine ligand and Cu(I) in the reduced state. Blue copper proteins serve as models for understanding how proteins tune metal binding properties, however, the mechanism through which they do so remains unclear. Here, the authors investigate carbon-deuterium bond vibrations in Nostocplastocyanin to study key copper-ligand interactions in different redox states, and reveal specific impacts of perturbation to the cupredoxin scaffold on copper coordination and the redox properties of plastocyanin. [ABSTRACT FROM AUTHOR]

Plastocyanin and cytochrome c6, abundant proteins in photosynthesis, are readouts for cellular copper status in Chlamydomonas and other algae. Their accumulation is controlled by a transcription factor copper response regulator (CRR1). The replacement of copper-containing plastocyanin with heme-containing cytochrome c6 spares copper and permits preferential copper (re)-allocation to cytochrome oxidase. Under copper-replete situations, the quota depends on abundance of various cuproproteins and is tightly regulated, except under zinc-deficiency where acidocalcisomes over-accumulate Cu(I). CRR1 has a transcriptional activation domain, a Zn-dependent DNA binding SBP-domain with a nuclear localization signal, and a C-terminal Cys-rich region that represses the zinc regulon. CRR1 activates >60 genes in Chlamydomonas through GTAC-containing CuREs; transcriptome differences are recapitulated in the proteome. The differentially-expressed genes encode assimilatory copper transporters of the CTR/SLC31 family including a novel soluble molecule, redox enzymes in the tetrapyrrole pathway that promote chlorophyll biosynthesis and photosystem 1 accumulation, and other oxygen-dependent enzymes, which may influence thylakoid membrane lipids, specifically polyunsaturated galactolipids and γ-tocopherol. CRR1 also down-regulates 2 proteins in Chlamydomonas: for plastocyanin, by activation of proteolysis, while for the di‑iron subunit of the cyclase in chlorophyll biosynthesis, through activation of an upstream promoter that generates a poorly-translated 5' extended transcript containing multiple short ORFs that inhibit translation. The functions of many CRR1-target genes are unknown, and the copper protein inventory in Chlamydomonas includes several whose functions are unexplored. The comprehensive picture of cuproproteins and copper homeostasis in this system is well-suited for reverse genetic analyses of these under-investigated components in copper biology.

Due to the similarity in the basic coordination behavior of their mono-charged cations, silver biochemistry is known to be linked to that of copper in biological systems. Still, Cu+/2+ is an essential micronutrient in many organisms, while no known biological process requires silver. In human cells, copper regulation and trafficking is strictly controlled by complex systems including many cytosolic copper chaperones, whereas some bacteria exploit the so-called "blue copper" proteins. Therefore, evaluating the controlling factors of the competition between these two metal cations is of enormous interest. By employing the tools of computational chemistry, we aim to delineate the extent to which Ag+ might be able to compete with the endogenous copper in its Type I (T1Cu) proteins, and where and if, alternatively, it is handled uniquely. The effect of the surrounding media (dielectric constant) and the type, number, and composition of amino acid residues are taken into account when modelling the reactions in the present study. The obtained results clearly indicate the susceptibility of the T1Cu proteins to a silver attack due to the favorable composition and geometry of the metal-binding centers, along with the similarity between the Ag+/Cu+-containing structures. Furthermore, by exploring intriguing questions of both metals' coordination chemistry, an important background for understanding the metabolism and biotransformation of silver in organisms is provided. [ABSTRACT FROM AUTHOR]

Recent phylogenetic studies have unveiled a novel class of ascorbate peroxidases called “ascorbate peroxidase-related” (APX-R). These enzymes, found in green photosynthetic eukaryotes, lack the amino acids necessary for ascorbate binding. This study focuses on the sole APX-R from Chlamydomonas reinhardtii referred to as ascorbate peroxidase 2 (APX2). We used immunoblotting to locate APX2 within the chloroplasts and in silico analysis to identify key structural motifs, such as the twin-arginine transport (TAT) motif for lumen translocation and the metal-binding MxxM motif. We also successfully expressed recombinant APX2 in Escherichia coli. Our in vitro results showed that the peroxidase activity of APX2 was detected with guaiacol but not with ascorbate as an electron donor. Furthermore, APX2 can bind both copper and heme, as evidenced by spectroscopic, and fluorescence experiments. These findings suggest a potential interaction between APX2 and plastocyanin, the primary copper-containing enzyme within the thylakoid lumen of the chloroplasts. Predictions from structural models and evidence from 1H-NMR experiments suggest a potential interaction between APX2 and plastocyanin, emphasizing the influence of APX2 on the copper-binding abilities of plastocyanin. In summary, our results propose a significant role for APX2 as a regulator in copper transfer to plastocyanin. This study sheds light on the unique properties of APX-R enzymes and their potential contributions to the complex processes of photosynthesis in green algae.

In this manuscript, a concise library of 2‐Amino‐4‐chlorophenol‐based organosilanes 5(a‐c) was designed and synthesized by employing a multistep strategy and characterized with the help of NMR (1H and 13C) spectroscopic and mass spectrometry techniques. The silica nanoparticles loaded with 5a have been successfully synthesized and confirmed with the help of various techniques. Further, the interaction of probes 5a and its hybrid silica nanoparticles (A‐NPs) have been investigated towards various metal ions and exhibited favorable selectivity towards Cu2+ ions over other common metal ions in methanol with both colorimetric and fluorimetric read‐outs. The binding of probes 5a and A‐NPs with Cu2+ can be monitored by change of color in daylight and also by turn‐off fluorescence response in UV light. The sensing mechanism is ascribed to paramagnetic nature of Cu2+ ion leading to ligand to metal charge transfer and, hence, shows chelation enhanced fluorescence quenching (CHEQ) effect in the vicinity of ligands 5a and A‐NPs. The results demonstrated that the hybrid nanocomposites (A‐NPs) has even lower limit of detection and higher binding affinity as compared to probe 5a. The possible mechanism of 1:1 binding of 5a with Cu2+ could be substantiated by the density functional theory. Thus, this facile and appropriate sensing strategy may serve as a promising tool for selective and sensitive detection of Cu2+ ions. Moreover, the molecular docking studies provide valuable information regarding the interaction of compound 5a with the blue copper protein (plastocyanin). Hence, the current investigation provides a new avenue to design more potent organosilane analogs. [ABSTRACT FROM AUTHOR]

Cytochrome c6 is a redox carrier in the thylakoid lumen of cyanobacteria and some eukaryotic algae. Although the isofunctional plastocyanin is present in land plants and the green alga Chlamydomonas reinhardtii, these organisms also possess a cytochrome c6-like protein designated as cytochrome c6A. Two other cytochrome c6-like groups, c6B and c6C, have been identified in cyanobacteria. In this study, we have identified a novel c6-like cytochrome called PetJ2, which is encoded in the nuclear genome of Cyanophora paradoxa, a member of the glaucophytes - the basal branch of the Archaeplastida. We propose that glaucophyte PetJ2 protein is related to cyanobacterial c6B and c6C cytochromes, and that cryptic green algal and land plant cytochromes c6A evolved froman ancestral archaeplastidial PetJ2 protein. In vitro import experiments with isolated muroplasts revealed that PetJ2 is imported into plastids. Although it harbors a twin-arginine motif in its thylakoid-targeting peptide, which is generally indicative of thylakoid import via the Tat import pathway, our import experiments with isolated muroplasts and the heterologous pea thylakoid import system revealed that PetJ2 uses the Sec pathway instead of the Tat import pathway. [ABSTRACT FROM AUTHOR]

Photosynthetic Control is defined as the control imposed on photosynthetic electron transport by the lumen-pH-sensitive re-oxidation of plastoquinol (PQH2) by cytochrome b6f. Photosynthetic Control leads at higher actinic light intensities to an electron transport chain with a (relatively) reduced photosystem (PS) II and PQ pool and a (relatively) oxidized PS I. Making Light Curves of more than 33 plant species with the recently introduced DUAL-KLAS-NIR (Chl a fluorescence + the redox states of plastocyanin (PC), P700, and ferredoxin (Fd)) the light intensity-dependent induction of Photosynthetic Control was probed and characterized. It was observed that PC became completely oxidized at light intensities ≤ 400 µmol photons m−2 s−1 (at lower light intensities in shade than in sun leaves). The relationship between qP and P700(red) was used to determine the extent of Photosynthetic Control. Instead of measuring the whole Light Curve, it was shown that a single moderate light intensity can be used to characterize the status of a leaf relative to that of other leaves. It was further found that in some shade-acclimated leaves Fd becomes again more oxidized at high light intensities indicating that electron transfer from the PQ pool to P700 cannot keep up with the outflow of electrons on the acceptor side of PS I. It was observed as well that for NPQ-induction a lower light intensity (less acidified lumen) was needed than for the induction of Photosynthetic Control. The measurements were also used to make a comparison between the parameters qP and qL, a comparison suggesting that qP was the more relevant parameter. [ABSTRACT FROM AUTHOR]

Hans Freeman was born in Germany and arrived in Australia with his parents in 1938. A brilliant student at the University of Sydney, he spent a seminal year at the California Institute of Technology before joining the staff at Sydney and initiating research on bioinorganic chemistry, studying metal ion complexes of compounds of biological significance such as amino acids, peptides and proteins. In his use of X-ray crystallography he was a pioneer in Australia, constructing his first crystallographic apparatus and mastering the necessary computing, at first by hand but soon with electronic computers. The culmination of his work with a series of collaborators was the structure of the blue, copper-containing metalloprotein, plastocyanin. Freeman also employed another advanced technique—X-ray spectroscopy and the study of X-ray absorption fine structure. He was a leading figure in Australia and internationally, and played an important role in gaining access for Australian scientists to international facilities such as synchrotron radiation sources at the dawning of the era of 'Big Science'. Hans Charles Freeman was a pioneer in crystallography in Australia, establishing his own lab, undertaking many of the first structural determinations of metal complexes and leading the team that determined the first metallo-protein structure in Australia. He also pioneered the use of X-ray spectroscopic techniques, strongly supporting others to make use of these techniques, activities that culminated in the building of Australian synchrotron facilities in Tsukuba and Clayton. [ABSTRACT FROM AUTHOR]

Cytochrome c6 is a redox carrier in the thylakoid lumen of cyanobacteria and some eukaryotic algae. Although the isofunctional plastocyanin is present in land plants and the green alga Chlamydomonas reinhardtii, these organisms also possess a cytochrome c6-like protein designated as cytochrome c6A. Two other cytochrome c6-like groups, c6B and c6C, have been identified in cyanobacteria. In this study, we have identified a novel c6-like cytochrome called PetJ2, which is encoded in the nuclear genome of Cyanophora paradoxa, a member of the glaucophytes - the basal branch of the Archaeplastida. We propose that glaucophyte PetJ2 protein is related to cyanobacterial c6B and c6C cytochromes, and that cryptic green algal and land plant cytochromes c6A evolved froman ancestral archaeplastidial PetJ2 protein. In vitro import experiments with isolated muroplasts revealed that PetJ2 is imported into plastids. Although it harbors a twin-arginine motif in its thylakoid-targeting peptide, which is generally indicative of thylakoid import via the Tat import pathway, our import experiments with isolated muroplasts and the heterologous pea thylakoid import system revealed that PetJ2 uses the Sec pathway instead of the Tat import pathway. [ABSTRACT FROM AUTHOR]

Cyclic electron transport (CET) is an attractive hypothesis for regulating photosynthetic electron transport and producing the additional ATP in oxygenic phototrophs. The concept of CET has been established in the last decades, and it is proposed to function in the progenitor of oxygenic photosynthesis, cyanobacteria. The in vivo activity of CET is frequently evaluated either from the redox state of the reaction center chlorophyll in photosystem (PS) I, P700, in the absence of PSII activity or by comparing PSI and PSII activities through the P700 redox state and chlorophyll fluorescence, respectively. The evaluation of CET activity, however, is complicated especially in cyanobacteria, where CET shares the intersystem chain, including plastoquinone, cytochrome b6/f complex, plastocyanin, and cytochrome c6, with photosynthetic linear electron transport (LET) and respiratory electron transport (RET). Here we sought to distinguish the in vivo electron transport rates in RET and CET in the cyanobacterium Synechocystis sp. PCC 6803. The reduction rate of oxidized P700 (P700+) decreased to less than 10% when PSII was inhibited, indicating that PSII is the dominant electron source to PSI but P700+ is also reduced by electrons derived from other sources. The oxidative pentose phosphate (OPP) pathway functions as the dominant electron source for RET, which was found to be inhibited by glycolaldehyde (GA). In the condition where the OPP pathway and respiratory terminal oxidases were inhibited by GA and KCN, the P700+ reduction rate was less than 1% of that without any inhibitors. This study indicate that the electron transport to PSI when PSII is inhibited is dominantly derived from the OPP pathway in Synechocystis sp. PCC 6803. [ABSTRACT FROM AUTHOR]

Due to the similarity in the basic coordination behavior of their mono-charged cations, silver biochemistry is known to be linked to that of copper in biological systems. Still, Cu+/2+ is an essential micronutrient in many organisms, while no known biological process requires silver. In human cells, copper regulation and trafficking is strictly controlled by complex systems including many cytosolic copper chaperones, whereas some bacteria exploit the so-called “blue copper” proteins. Therefore, evaluating the controlling factors of the competition between these two metal cations is of enormous interest. By employing the tools of computational chemistry, we aim to delineate the extent to which Ag+ might be able to compete with the endogenous copper in its Type I (T1Cu) proteins, and where and if, alternatively, it is handled uniquely. The effect of the surrounding media (dielectric constant) and the type, number, and composition of amino acid residues are taken into account when modelling the reactions in the present study. The obtained results clearly indicate the susceptibility of the T1Cu proteins to a silver attack due to the favorable composition and geometry of the metal-binding centers, along with the similarity between the Ag+/Cu+-containing structures. Furthermore, by exploring intriguing questions of both metals’ coordination chemistry, an important background for understanding the metabolism and biotransformation of silver in organisms is provided.

Manganese (Mn) is considered as an essential element for plant growth. Mn starvation has been shown to affect photosystem II, the site of the Mn 4 CaO 5 cluster responsible for water oxidation. Less is known on the effect of Mn starvation on photosystem I. Here we studied the effects of Mn deficiency in vivo on redox changes of P700 and plastocyanin (Pc) in the liverwort Marchantia polymorpha using the KLAS-NIR spectrophotometer. Far-red illumination is used to excite preferentially photosystem I, thus facilitating cyclic electron transport. Under Mn starvation, we observed slower oxidation of P700 and a decrease in the Pc signal relative to P700. The lower Pc content under Mn deficiency was confirmed by western blots. Re-reduction kinetics of P700+ and Pc+ were faster in Mn deficient thalli than in the control. The above findings show that the kinetics studied under Mn deficiency not only depend on the number of available reductants but also on how quickly electrons are transferred from stromal donors via the intersystem chain to Pc+ and P700+. We suggest that under Mn deficiency a structural reorganization of the thylakoid membrane takes place favoring the formation of supercomplexes between ferredoxin, cytochrome b 6 f complex, Pc and photosystem I, and thus an enhanced cyclic electron transport. • Mn starvation induces a decrease in plastocyanin content relative to P700. • Mn starvation accelerates electron donation to oxidized plastocyanin and toP700+. • Mn starvation seems to favor cyclic electron transport around photosystem I. • Enhanced cyclic flow may be an acclimation strategy under micronutrient deprivation. [ABSTRACT FROM AUTHOR]

Leaf senescence is a critical process in plants and has a direct impact on many important agronomic traits. Despite decades of research on senescence-altered mutants via forward genetics and functional assessment of senescence-associated genes (SAGs) via reverse genetics, the senescence signal and the molecular mechanism that perceives and transduces the signal remain elusive. Here, using dark-induced senescence (DIS) of Arabidopsis leaf as the experimental system, we show that exogenous copper induces the senescence syndrome and transcriptomic changes in lightgrown plants parallel to those in DIS. By profiling the transcriptomes and tracking the subcellular copper distribution, we found that reciprocal regulation of plastocyanin, the thylakoid lumen mobile electron carrier in the Z scheme of photosynthetic electron transport, and SAG14 and plantacyanin (PCY), a pair of interacting small blue copper proteins located on the endomembrane, is a common thread in different leaf senescence scenarios, including DIS. Genetic and molecular experiments confirmed that the PCYSAG14 module is necessary and sufficient for promoting DIS. We also found that the PCY-SAG14 module is repressed by a conserved microRNA, miR408, which in turn is repressed by phytochrome interacting factor 3/4/5 (PIF3/4/5), the key trio of transcription factors promoting DIS. Together, these findings indicate that intracellular copper redistribution mediated by PCYSAG14 has a regulatory role in DIS. Further deciphering the copper homeostasis mechanism and its interaction with other senescenceregulating pathways should provide insights into our understanding of the fundamental question of how plants age. [ABSTRACT FROM AUTHOR]

Interrogative proteome analyses are used to identify and quantify the expression of proteins involved in heat tolerance and to identify associated physiological processes in heat-stressed plants. The objectives of the study were to identify and quantify the expression of proteins involved in heat tolerance and to identify associated physiological processes in chickpea (Cicer arietinum L.) heat-tolerant (Acc#7) and sensitive genotype (Acc#8) from a field study. Proteomic and gene ontological analyses showed an upregulation in proteins related to protein synthesis, intracellular traffic, defence and transport in the heat-tolerant genotype compared to the susceptible one at the warmer site. Results from KEGG analyses indicate the involvement of probable sucrose-phosphate synthase (EC 2.4.1.14) and sucrose-phosphate phosphatase (EC 3.1.3.24) proteins, that were upregulated in the heat-tolerant genotype at the warmer site, in the starch and sucrose pathway. The presence of these differentially regulated proteins including HSP70, ribulose bisphosphate carboxylase/oxygenase activase, plastocyanin and protoporphyrinogen oxidase suggests their potential role in heat tolerance, at flowering growth stage, in field-grown chickpea. This observation supports unaltered physiological and biochemical performance of the heat-tolerant genotypes (Acc#7) relative to the susceptible genotype (Acc#8) in related studies (Makonya et al. 2019). Characterisation of the candidate proteins identified in the current study as well as their specific roles in the tolerance to heat stress in chickpea are integral to further crop improvement initiatives. Increased frequency and duration of high temperatures continues to limit crop development. An analysis of chickpea proteome to help uncover proteins and enzymes contributing to its heat tolerance and associated physiological processes is essential. Proteins related to protein synthesis, intracellular traffic, transport and defence were upregulated in heat-tolerant genotypes consistent with their previously unaltered physiological and biochemical performance. Identified candidate proteins and their specific roles in the tolerance to heat stress in chickpea are integral to further crop improvement initiatives. [ABSTRACT FROM AUTHOR]

Plants develop a series of physiological, biochemical, and molecular responses to survive in drought stress. Many drought-related genes with various functions which regulated by ABA-dependent and independent pathways have been identified in wheat. However, studies on the effect of biostimulants on drought-related genes are limited and unclear. As a biostimulant, humic acid (HA) has enormous roles on plant growth, yield, and protection of resistance in various abiotic stresses, but its relation to drought-related genes is unknown. Here, we aimed to determine the relationships between drought stress generated by − 6 bar and − 8 bar PEG6000 and ABA and HA treatments on total oxidant status (TOS) and total antioxidant status (TAS) and expression of miRNA target genes (AP2, GRF, LAC, CSD1/CSD2, and Plastocyanin) on tolerant (Aksel 2000 and Kırkpınar 79) and susceptible (Atlı 2002 and Kırmızı Kılçık) bread wheat (Triticum aestivum L.) genotypes. According to the results, the expression of the genes varied depending on genotype and tissue and it was determined whether the genes used ABA-dependent or ABA-independent mechanisms to provide tolerance to stress in our promoter analysis. Also, the irregularities of the biochemical changes and expression of genes that were caused by PEG6000 could be improved by HA considering the positive effects on genes. [ABSTRACT FROM AUTHOR]

During photosynthesis, electrons are transferred between the cytochrome b 6 f complex and photosystem I. This is carried out by the protein plastocyanin in plant chloroplasts, or by either plastocyanin or cytochrome c 6 in many cyanobacteria and eukaryotic algal species. There are three further cytochrome c 6 homologs: cytochrome c 6A in plants and green algae, and cytochromes c 6B and c 6C in cyanobacteria. The function of these proteins is unknown. Here, we present a comprehensive analysis of the evolutionary relationship between the members of the cytochrome c 6 family in photosynthetic organisms. Our phylogenetic analyses show that cytochromes c 6B and c 6C are likely to be orthologs that arose from a duplication of cytochrome c 6, but that there is no evidence for separate origins for cytochromes c 6B and c 6C. We therefore propose renaming cytochrome c 6C as cytochrome c 6B. We show that cytochrome c 6A is likely to have arisen from cytochrome c 6B rather than by an independent duplication of cytochrome c 6, and present evidence for an independent origin of a protein with some of the features of cytochrome c 6A in peridinin dinoflagellates. We conclude with a new comprehensive model of the evolution of the cytochrome c 6 family which is an integral part of understanding the function of the enigmatic cytochrome c 6 homologs. [ABSTRACT FROM AUTHOR]

In cyanobacteria and most green algae of the eukaryotic green lineage, the copper-protein plastocyanin (Pc) alternatively replaces the heme-protein cytochrome c6 (Cc6) as the soluble electron carrier from cytochrome f (Cf) to photosystem I (PSI). The functional and structural equivalence of 'green' Pc and Cc6 has been well established, representing an example of convergent evolution of two unrelated proteins. However, plants only produce Pc, despite having evolved from green algae. On the other hand, Cc6 is the only soluble donor available in most species of the red lineage of photosynthetic organisms, which includes, among others, red algae and diatoms. Interestingly, Pc genes have been identified in oceanic diatoms, probably acquired by horizontal gene transfer from green algae. However, the mechanisms that regulate the expression of a functional Pc in diatoms are still unclear. In the green eukaryotic lineage, the transfer of electrons from Cf to PSI has been characterized in depth. The conclusion is that in the green lineage, this process involves strong electrostatic interactions between partners, which ensure a high affinity and an efficient electron transfer (ET) at the cost of limiting the turnover of the process. In the red lineage, recent kinetic and structural modeling data suggest a different strategy, based on weaker electrostatic interactions between partners, with lower affinity and less efficient ET, but favoring instead the protein exchange and the turnover of the process. Finally, in diatoms the interaction of the acquired green-type Pc with both Cf and PSI may not yet be optimized. [ABSTRACT FROM AUTHOR]

Inorganic elements, although required only in trace amounts, permit life and primary productivity because of their functions in catalysis. Every organism has a minimal requirement of each metal based on the intracellular abundance of proteins that use inorganic cofactors, but elemental sparing mechanisms can reduce this quota. A well-studied copper-sparing mechanism that operates in microalgae faced with copper deficiency is the replacement of the abundant copper protein plastocyanin with a heme-containing substitute, cytochrome (Cyt) c6. This switch, which is dependent on a copper-sensing transcription factor, copper response regulator 1 (CRR1), dramatically reduces the copper quota. We show here that in a situation of marginal copper availability, copper is preferentially allocated from plastocyanin, whose function is dispensable, to other more critical copper-dependent enzymes like Cyt oxidase and a ferroxidase. In the absence of an extracellular source, copper allocation to Cyt oxidase includes CRR1-dependent proteolysis of plastocyanin and quantitative recycling of the copper cofactor from plastocyanin to Cyt oxidase. Transcriptome profiling identifies a gene encoding a Zn-metalloprotease, as a candidate effecting copper recycling. One reason for the retention of genes encoding both plastocyanin and Cyt c6 in algal and cyanobacterial genomes might be because plastocyanin provides a competitive advantage in copper-depleted environments as a ready source of copper.

We identified a Cu-accumulating structure with a dynamic role in intracellular Cu homeostasis. During Zn limitation, Chlamydomonas reinhardtii hyperaccumulates Cu, a process dependent on the nutritional Cu sensor CRR1, but it is functionally Cu deficient. Visualization of intracellular Cu revealed major Cu accumulation sites coincident with electron-dense structures that stained positive for low pH and polyphosphate, suggesting that they are lysosome-related organelles. Nano-secondary ion MS showed colocalization of Ca and Cu, and X-ray absorption spectroscopy was consistent with Cu(+) accumulation in an ordered structure. Zn resupply restored Cu homeostasis concomitant with reduced abundance of these structures. Cu isotope labeling demonstrated that sequestered Cu(+) became bioavailable for the synthesis of plastocyanin, and transcriptome profiling indicated that mobilized Cu became visible to CRR1. Cu trafficking to intracellular accumulation sites may be a strategy for preventing protein mismetallation during Zn deficiency and enabling efficient cuproprotein metallation or remetallation upon Zn resupply.

Trichoderma biopriming enhances rice growth in drought-stressed soils by triggering various plant metabolic pathways related to antioxidative defense, secondary metabolites, and hormonal upregulation. In the present study, transcriptomic analysis of rice cultivar IR64 bioprimed with Trichoderma harzianum under drought stress was carried out in comparison with drought-stressed samples using next-generation sequencing techniques. Out of the 2,506 significant (p < 0.05) differentially expressed genes (DEGs), 337 (15%) were exclusively expressed in drought-stressed plants, 382 (15%) were expressed in T. harzianum -treated drought-stressed plants, and 1,787 (70%) were commonly expressed. Furthermore, comparative analysis of upregulated and downregulated genes under stressed conditions showed that 1,053 genes (42%) were upregulated and 733 genes (29%) were downregulated in T. harzianum -treated drought-stressed rice plants. The genes exclusively expressed in T. harzianum- treated drought-stressed plants were mostly photosynthetic and antioxidative such as plastocyanin, small chain of Rubisco, PSI subunit Q, PSII subunit PSBY, osmoproteins, proline-rich protein, aquaporins, stress-enhanced proteins, and chaperonins. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis states that the most enriched pathways were metabolic (38%) followed by pathways involved in the synthesis of secondary metabolites (25%), carbon metabolism (6%), phenyl propanoid (7%), and glutathione metabolism (3%). Some of the genes were selected for validation using real-time PCR which showed consistent expression as RNA-Seq data. Furthermore, to establish host– T. harzianum interaction, transcriptome analysis of Trichoderma was also carried out. The Gene Ontology (GO) analysis of T. harzianum transcriptome suggested that the annotated genes are functionally related to carbohydrate binding module, glycoside hydrolase, GMC oxidoreductase, and trehalase and were mainly upregulated, playing an important role in establishing the mycelia colonization of rice roots and its growth. Overall, it can be concluded that T. harzianum biopriming delays drought stress in rice cultivars by a multitude of molecular programming. [ABSTRACT FROM AUTHOR]

We have investigated if the heterologous expression of a functional green alga plastocyanin in the diatom Phaeodactylum tricornutum can improve photosynthetic activity and cell growth. Previous in vitro assays showed that a single‐mutant of the plastocyanin from the green algae Chlamydomonas reinhardtii is effective in reducing P. tricornutum photosystem I. In this study, in vivo assays with P. tricornutum strains expressing this plastocyanin indicate that even the relatively low intracellular concentrations of holo‐plastocyanin detected (≈4 μM) are enough to promote an increased growth (up to 60%) under iron‐deficient conditions as compared with the WT strain, measured as higher cell densities, content in pigments and active photosystem I, global photosynthetic rates per cell, and even cell volume. In addition, the presence of plastocyanin as an additional photosynthetic electron carrier seems to decrease the over‐reduction of the plastoquinone pool. Consequently, it promotes an improvement in the maximum quantum yield of both photosystem II and I, together with a decrease in the acceptor side photoinhibition of photosystem II—also associated to a reduced oxidative stress—a decrease in the peroxidation of membrane lipids in the choroplast, and a lower degree of limitation on the donor side of photosystem I. Thus the heterologous plastocyanin appears to act as a functional electron carrier, alternative to the native cytochrome c6, under iron‐limiting conditions. [ABSTRACT FROM AUTHOR]

We extended the fragment molecular orbital (FMO) method interfaced with density functional theory (DFT) into spin unrestricted formalism (UDFT) and developed energy gradients for the ground state and single point excited state energies based on time-dependent DFT. The accuracy of FMO is evaluated in comparison to the full calculations without fragmentation. Electronic excitations in solvated organic radicals and in the blue copper protein, plastocyanin (PDB code: 1BXV), are reported. The contributions of solvent molecules to the electronic excitations are analyzed in terms of the fragment polarization and quantum effects such as interfragment charge transfer. [ABSTRACT FROM AUTHOR]

SUMMARY: The plastoquinone (PQ) pool mediates electron flow and regulates photoacclimation in plants. Here we report the action spectrum of the redox state of the PQ pool in Arabidopsis thaliana, showing that 470–500, 560 or 650–660 nm light favors Photosystem II (PSII) and reduces the PQ pool, whereas 420–440, 520 or 690 nm light favors Photosystem I (PSI) and oxidizes PQ. These data were used to construct a model predicting the redox state of PQ from the spectrum of any polychromatic light source. Moderate reduction of the PQ pool induced transition to light state 2, whereas state 1 required highly oxidized PQ. In low‐intensity PSI light, PQ was more oxidized than in darkness and became gradually reduced with light intensity, while weak PSII light strongly reduced PQ. Natural sunlight was found to favor PSI, which enables plants to use the redox state of the PQ pool as a measure of light intensity. Significance Statement: The redox state of the chloroplast plastoquinone (PQ) pool regulates state transitions and gene expression in response to illumination conditions. We show how wavelengths and intensity of photosynthetically active light determine the redox state of the PQ pool in such a way that plants can measure the intensity of sunlight using the PQ pool as a sensor. [ABSTRACT FROM AUTHOR]

In photosynthetic electron transport, large multiprotein complexes are connected by small diffusible electron carriers, the mobility of which is challenged by macromolecular crowding. For thylakoid membranes of higher plants, a long-standing question has been which of the two mobile electron carriers, plastoquinone or plastocyanin, mediates electron transport from stacked grana thylakoids where photosystem II (PSII) is localized to distant unstacked regions of the thylakoids that harbor PSI. Here, we confirm that plastocyanin is the long-range electron carrier by employing mutants with different grana diameters. Furthermore, our results explain why higher plants have a narrow range of grana diameters since a larger diffusion distance for plastocyanin would jeopardize the efficiency of electron transport. In the light of recent findings that the lumen of thylakoids, which forms the diffusion space of plastocyanin, undergoes dynamic swelling/shrinkage, this study demonstrates that plastocyanin diffusion is a crucial regulatory element of plant photosynthetic electron transport. [ABSTRACT FROM AUTHOR]

In photosynthesis research, non-invasive in vivo spectroscopic analyses have been used as a practical tool for studying photosynthetic electron transport. Klas-NIR spectrophotometer has been recently developed by Klughammer and Schreiber (Photosynth Res 128:195–214, 2016) for in vivo measurements of redox changes of P700, plastocyanin (Pcy) and ferredoxin (Fd). Here we show examples using the Klas-NIR spectrophotometer for the evaluation of the redox states and quantities of these components in plant leaves and cyanobacterial suspensions. The redox poise under light of the electron transport components is different in leaves from higher plants compared with cyanobacteria. During a short illumination with an actinic light, P700, Pcy, and Fd are kept reduced in barley leaves but are oxidized in cyanobacteria. During far-red light illumination, P700 and Pcy are mostly oxidized in the leaves but are partially kept reduced in cyanobacteria. In the cyanobacterium, Thermosynechococcus elongatus, which has no Pcy but uses cytochrome c6 (cyt c6) as the electron donor to photosystem I, a cyt c6 signal was detected in vivo. To show the potential of Klas-NIR spectrophotometer for studying different developmental stages of a leaf, we performed measurements on fully mature and early senescing barley leaves. Pcy content in leaves decreased during senescence at an early stage. The Pcy loss was quantitatively analyzed using Klas-NIR spectrophotometer, giving absolute ratios of Pcy to PSI of 2.5 and 1.6 in younger and older leaves, respectively. For quantification of the signals in vivo, in vitro data (Sétif et al. in Photosynth Res142:307–319, 2019) obtained with Klas-NIR spectrophotometer were used. [ABSTRACT FROM AUTHOR]

A kinetic-LED-array-spectrophotometer (Klas) was recently developed for measuring in vivo redox changes of P700, plastocyanin (PCy), and ferredoxin (Fd) in the near-infrared (NIR). This spectrophotometer is used in the present work for in vitro light-induced measurements with various combinations of photosystem I (PSI) from tobacco and two different cyanobacteria, spinach plastocyanin, cyanobacterial cytochrome c6 (cyt. c6), and Fd. It is shown that cyt. c6 oxidation contributes to the NIR absorption changes. The reduction of (FAFB), the terminal electron acceptor of PSI, was also observed and the shape of the (FAFB) NIR difference spectrum is similar to that of Fd. The NIR difference spectra of the electron-transfer cofactors were compared between different organisms and to those previously measured in vivo, whereas the relative absorption coefficients of all cofactors were determined by using single PSI turnover conditions. Thus, the (840 nm minus 965 nm) extinction coefficients of the light-induced species (oxidized minus reduced for PC and cyt. c6, reduced minus oxidized for (FAFB), and Fd) were determined with values of 0.207 ± 0.004, – 0.033 ± 0.006, – 0.036 ± 0.008, and – 0.021 ± 0.005 for PCy, cyt. c6, (FAFB) (single reduction), and Fd, respectively, by taking a reference value of + 1 for P700+. The fact that the NIR P700 coefficient is larger than that of PCy and much larger than that of other contributing species, combined with the observed variability in the NIR P700 spectral shape, emphasizes that deconvolution of NIR signals into different components requires a very precise determination of the P700 spectrum. [ABSTRACT FROM AUTHOR]

The accuracy of core excitation energies and core electron binding energies computed within a Δself-consistent-field framework is assessed. The variational collapse of the core excited state is prevented by maintaining a singly occupied core orbital using an overlap criterion called the maximum overlap method. When applied to a wide range of small organic molecules, the resulting core excitation energies are not systematically underestimated as observed in time-dependent density functional theory and agree well with experiment. The accuracy of this approach for core excited states is illustrated by the calculation of the pre-edge features in x-ray absorption spectra of plastocyanin, which shows that accurate results can be achieved with Δself-consistent-field calculations when used in conjunction with uncontracted basis functions. [ABSTRACT FROM AUTHOR]

It has been experimentally shown that the folding of apoplastocyanin (apoPC) accompanies a very large enthalpic loss [N. Baden et al., J. Chem. Phys. 127, 175103 (2007)]. This implies that an even larger entropic gain occurs in stabilizing the folded structure to overcome the enthalpic loss. Here, we calculate the water-entropy gain upon the folding of apoPC using the angle-dependent integral equation theory combined with the multipolar water model and the recently developed morphometric approach. It is demonstrated that the calculated value is in quantitatively good accord with the value estimated from the experimental data by accounting for the conformational-entropy loss. According to a prevailing view, the water adjacent to a hydrophobic group is unstable especially in terms of the rotational entropy and the folding is driven primarily by the release of such unfavorable water to the bulk through the burial of nonpolar side chains. We show, however, that the resultant entropic gain is too small to elucidate the experimental result. The great entropic gain observed is ascribed to the reduction in the restriction for the translational motion of water molecules in the whole system. [ABSTRACT FROM AUTHOR]

We report applications of analytical formalisms and molecular dynamics (MD) simulations to the calculation of redox entropy of plastocyanin metalloprotein in aqueous solution. The goal of our analysis is to establish critical components of the theory required to describe polar solvation at the mesoscopic scale. The analytical techniques include a microscopic formalism based on structure factors of the solvent dipolar orientations and density and continuum dielectric theories. The microscopic theory employs the atomistic structure of the protein with force-field atomic charges and solvent structure factors obtained from separate MD simulations of the homogeneous solvent. The MD simulations provide linear response solvation free energies and reorganization energies of electron transfer in the temperature range of 280–310 K. We found that continuum models universally underestimate solvation entropies, and a more favorable agreement is reported between the microscopic calculations and MD simulations. The analysis of simulations also suggests that difficulties of extending standard formalisms to protein solvation are related to the inhomogeneous structure of the solvation shell at the protein-water interface combining islands of highly structured water around ionized residues along with partial dewetting of hydrophobic patches. Quantitative theories of electrostatic protein hydration need to incorporate realistic density profile of water at the protein-water interface. [ABSTRACT FROM AUTHOR]

Plastocyanin functions as an electron carrier in the photosynthetic electron transport chain, located at the thylakoid membrane. In several species, endogenous plastocyanin levels are correlated with the photosynthetic electron transport rate. Overexpression of plastocyanin genes in Arabidopsis thaliana increases plant size, but this phenomenon has not been observed in crop species. Here, we investigated the effects of heterologous expression of a gene encoding a plastocyanin isoform from Arabidopsis, AtPETE2 , in the oil seed crop Camelina sativa under standard growth conditions and under salt stress. AtPETE2 heterologous expression enhanced photosynthetic activity in Camelina, accelerating plant development and improving seed yield under standard growth conditions. Additionally, CsPETE2 from Camelina was induced by salt stress and AtPETE2 expression lines had larger primary roots and more lateral roots than the wild type. AtPETE2 expression lines also had larger seeds and higher total seed yield under long-term salt stress compared with non-transgenic Camelina. Our results demonstrate that increased plastocyanin levels in Camelina can enhance photosynthesis and productivity, as well as tolerance to osmotic and salt stresses. Heterologous expression of plastocyanin may be a useful strategy to mitigate crop stress in saline soils. • Heterologous expression of Arabidopsis plastocyanin gene PETE2 in Camelina sativa optimizes photosynthetic activity. • Optimized photosynthetic activity leads to accelerated plant development and improved seed yield. • Increased plastocyanin level in Camelina sativa improves tolerance to salt stress. [ABSTRACT FROM AUTHOR]

Extensive tests have been performed to benchmark and to compare with second-order perturbation theory based on a complete active space self-consistent field reference function (CASPT2), the recently developed n-electron valence state perturbation theory at second order (NEVPT2). Test calculations included the group fifteen diatomic molecules X[sub 2] (X=N, P, As, and Sb) and the [sup 4]S/[sup 2]D and [sup 4]S/[sup 2]P splittings for the corresponding atoms, the [sup 1]A[sub 1]–[sup 3]B[sub 1] splittings for CH[sub 2] and SiH[sub 2], and the absorption spectra of pyrrole and of Cu(Imidazole)[sub 2](SH)(SH[sub 2])[sup +], which is a model for plastocyanin. Comparisons with full configuration-interaction calculations and experimental data show that the accuracy of NEVPT2 is in most cases even better than CASPT2. Where intruder states hamper the CASPT2 calculations, NEVPT2 performs significantly better. Care is needed in the choice of active orbitals, for example in the calculation of the [sup 4]S/[sup 2]D and [sup 4]S/[sup 2]P splittings for the group fifteen atoms. This is due to the different treatment of orbitals belonging to the inactive or active spaces, making the NEVPT2 not invariant for the choice of active space, even in cases where the multiconfiguration self-consistent field energy is invariant. © 2004 American Institute of Physics. [ABSTRACT FROM AUTHOR]

In the face of rising salinity along coastal regions and in irrigated areas, molecular breeding of tolerant crops and reforestation of exposed areas using tolerant woody species is a two-way strategy. Thus, identification of tolerant plants and of existing tolerance mechanisms are of immense value. In the present study, three Eucalyptus ecotypes with potentially differential salt sensitivity were compared. Soil-grown Eucalyptus plants were exposed to 80 and 170 mM NaCl for 30 days. Besides analysing salt effects on ionic/osmotic balance, and hydrolytic enzymes, plants were compared for dynamics of light-induced redox changes in photosynthetic electron transport chain (pETC) components, namely plastocyanin (PC), photosystem I (PSI) and ferredoxin (Fd), parallel to traditional chlorophyll a fluorescence-based PSII-related parameters. Deconvoluted signals for PC and Fd from PSI allowed identification of PC and PSI as the prime salinity-sensitive components of pETC in tested Eucalyptus species. Eucalyptus loxophleba portrayed efficient K+-Na+ balance (60–90% increased K+) along with a more dynamic range of redox changes for pETC components in old leaves. Young leaves in Eucalyptus loxophleba showed robust endomembrane homeostasis, as underlined by an increased response of hydrolytic enzymes at lower salt concentration (~1.7–2.6-fold increase). Findings are discussed in context of salinity dose dependence among different Eucalyptus species.