Exploring Nitric Oxide as a Regulator in Salt Tolerance: Insights into Photosynthetic Efficiency in Maize.

The growing issue of salinity is a significant threat to global agriculture, affecting diverse regions worldwide. Nitric oxide (NO) serves as an essential signal molecule in regulating photosynthetic performance under physiological and stress conditions. The present study reveals the protective effects of different concentrations (0-300 µM) of sodium nitroprusside (SNP, a donor of NO) on the functions of the main complexes within the photosynthetic apparatus of maize (Zea mays L. Kerala) under salt stress (150 mM NaCl). The data showed that SNP alleviates salt-induced oxidative stress and prevents changes in the fluidity of thylakoid membranes (Laurdan GP) and energy redistribution between the two photosystems (77K chlorophyll fluorescence ratio F735/F685). Chlorophyll fluorescence measurements demonstrated that the foliar spray with SNP under salt stress prevents the decline of photosystem II (PSII) open reaction centers (qP) and improves their efficiency (Φexc), thereby influencing QA- reoxidation. The data also revealed that SNP protects the rate constants for two pathways of QA- reoxidation (k1 and k2) from the changes caused by NaCl treatment alone. Additionally, there is a predominance of QA- interaction with plastoquinone in comparison to the recombination of electrons in QA QB- with the oxygen-evolving complex (OEC). The analysis of flash oxygen evolution showed that SNP treatment prevents a salt-induced 10% increase in PSII centers in the S0 state, i.e., protects the initial S0-S1 state distribution, and the modification of the Mn cluster in the OEC. Moreover, this study demonstrates that SNP-induced defense occurs on both the donor and acceptor sides of the PSII, leading to the protection of overall photosystems performance (PIABS) and efficient electron transfer from the PSII donor side to the reduction of PSI end electron acceptors (PItotal). This study clearly shows that the optimal protection under salt stress occurs at approximately 50-63 nmoles NO/g FW in leaves, corresponding to foliar spray with 50-150 µM SNP.

Abiotic Stress Tolerance in Crop and Medicinal Plants.

Climate change and the increased need for crop production highlight the urgent importance of introducing crops with increased tolerance to adverse environmental conditions [...].

Impact of Salinity on the Energy Transfer between Pigment-Protein Complexes in Photosynthetic Apparatus, Functions of the Oxygen-Evolving Complex and Photochemical Activities of Photosystem II and Photosystem I in Two Paulownia Lines.

The present study shows the effect of salinity on the functions of thylakoid membranes from two hybrid lines of Paulownia: Paulownia tomentosa x fortunei and Paulownia elongate x elongata, grown in a Hoagland solution with two NaCl concentrations (100 and 150 mM) and different exposure times (10 and 25 days). We observed inhibition of the photochemical activities of photosystem I (DCPIH2 → MV) and photosystem II (H2O → BQ) only after the short treatment (10 days) with the higher NaCl concentration. Data also revealed alterations in the energy transfer between pigment-protein complexes (fluorescence emission ratios F735/F685 and F695/F685), the kinetic parameters of the oxygen-evolving reactions (initial S0-S1 state distribution, misses (α), double hits (ß) and blocked centers (SB)). Moreover, the experimental results showed that after prolonged treatment with NaCl Paulownia tomentosa x fortunei adapted to the higher concentration of NaCl (150 mM), while this concentration is lethal for Paulownia elongata x elongata. This study demonstrated the relationship between the salt-induced inhibition of the photochemistry of both photosystems and the salt-induced changes in the energy transfer between the pigment-protein complexes and the alterations in the Mn cluster of the oxygen-evolving complex under salt stress.

Protective Effects of Sodium Nitroprusside on Photosynthetic Performance of Sorghum bicolor L. under Salt Stress.

In this study, the impacts of the foliar application of different sodium nitroprusside (SNP, as a donor of nitric oxide) concentrations (0-300 µM) on two sorghum varieties (Sorghum bicolor L. Albanus and Sorghum bicolor L. Shamal) under salt stress (150 mM NaCl) were investigated. The data revealed that salinity leads to an increase in oxidative stress markers and damage of the membrane integrity, accompanied by a decrease in the chlorophyll content, the open photosystem II (PSII) centers, and the performance indexes (PI ABS and PI total), as well as having an influence on the electron flux reducing photosystem I (PSI) end acceptors (REo/RC). Spraying with SNP alleviated the NaCl toxicity on the photosynthetic functions; the protection was concentration-dependent, and greater in Shamal than in Albanus, i.e., variety specific. Furthermore, the experimental results revealed that the degree of SNP protection under salt stress also depends on the endogenous nitric oxide (NO) amount in leaves, the number of active reaction centers per PSII antenna chlorophylls, the enhanced electron flux reducing end acceptors at the acceptor side of PSI, as well as the stimulation of the cyclic electron transport around PSI. The results showed better protection in both varieties of sorghum for SNP concentrations up to 150 µM, which corresponds to about a 50% increase in the endogenous NO leaf content in comparison to the control plants. Our study provides valuable insight into the molecular mechanisms underlying SNP-induced salt tolerance in sorghum varieties and might be a practical approach to correcting salt intolerance.

Impact of Sodium Nitroprusside on the Photosynthetic Performance of Maize and Sorghum.

Nitric oxide (NO) is an important molecule in regulating plant growth, development and photosynthetic performance. This study investigates the impact of varying concentrations (0-300 µM) of sodium nitroprusside (SNP, a donor of NO) on the functions of the photosynthetic apparatus in sorghum (Sorghum bicolor L. Albanus) and maize (Zea mays L. Kerala) under physiological conditions. Analysis of the chlorophyll fluorescence signals (using PAM and the JIP-test) revealed an increased amount of open PSII reaction centers (qP increased), but it did not affect the number of active reaction centers per PSII antenna chlorophyll (RC/ABS). In addition, the smaller SNP concentrations (up to 150 µM) alleviated the interaction of QA with plastoquine in maize, while at 300 µM it predominates the electron recombination on QAQB-, with the oxidized S2 (or S3) states of oxygen evolving in complex ways in both studied plant species. At the same time, SNP application stimulated the electron flux-reducing end electron acceptors at the PSI acceptor side per reaction center (REo/RC increased up to 26%) and the probability of their reduction (φRo increased up to 20%). An increase in MDA (by about 30%) and H2O2 contents was registered only at the highest SNP concentration (300 µM). At this concentration, SNP differentially affected the amount of P700+ in studied plant species, i.e., it increased (by 10%) in maize but decreased (by 16%) in sorghum. The effects of SNP on the functions of the photosynthetic apparatus were accompanied by an increase in carotenoid content in both studied plants. Additionally, data revealed that SNP-induced changes in the photosynthetic apparatus differed between maize and sorghum, suggesting species specificity for SNP's impact on plants.

Different Sensitivity Levels of the Photosynthetic Apparatus in Zea mays L. and Sorghum bicolor L. under Salt Stress.

The impacts of different NaCl concentrations (0-250 mM) on the photosynthesis of new hybrid lines of maize (Zea mays L. Kerala) and sorghum (Sorghum bicolor L. Shamal) were investigated. Salt-induced changes in the functions of photosynthetic apparatus were assessed using chlorophyll a fluorescence (PAM and OJIP test) and P700 photooxidation. Greater differences between the studied species in response to salinization were observed at 150 mM and 200 mM NaCl. The data revealed the stronger influence of maize in comparison to sorghum on the amount of closed PSII centers (1-qp) and their efficiency (Φexc), as well as on the effective quantum yield of the photochemical energy conversion of PSII (ΦPSII). Changes in the effective antenna size of PSII (ABS/RC), the electron flux per active reaction center (REo/RC) and the electron transport flux further QA (ETo/RC) were also registered. These changes in primary PSII photochemistry influenced the electron transport rate (ETR) and photosynthetic rate (parameter RFd), with the impacts being stronger in maize than sorghum. Moreover, the lowering of the electron transport rate from QA to the PSI end electron acceptors (REo/RC) and the probability of their reduction (φRo) altered the PSI photochemical activity, which influenced photooxidation of P700 and its decay kinetics. The pigment content and stress markers of oxidative damage were also determined. The data revealed a better salt tolerance of sorghum than maize, associated with the structural alterations in the photosynthetic membranes and the stimulation of the cyclic electron flow around PSI at higher NaCl concentrations. The relationships between the decreased pigment content, increased levels of stress markers and different inhibition levels of the function of both photosystems are discussed.

Cadmium toxicity in Salvia sclarea L.: An integrative response of element uptake, oxidative stress markers, leaf structure and photosynthesis.

The herbal plant Salvia sclarea L. (clary sage) is classified to cadmium (Cd) accumulators and considered as a potential plant for phytoremediation of heavy metal polluted soil. However, the effect of Cd only treatment on the function of the photosynthetic apparatus of S. sclarea, as well as the mechanisms involved in Cd tolerance have not yet been studied in detail. This study was conducted to examine the integrative responses of S. sclarea plants exposed to a high Cd supply (100 µM) for 3 and 8 days by investigating element nutrient uptake, oxidative stress markers, pigment composition, photosynthetic performance and leaf structure. Measurements of the functional activities of photosystem I (PSI, by P700 photooxidation), photosystem II (PSII, by chlorophyll fluorescence parameters), the oxygen-evolving complex (oxygen evolution by Joliot- and Clark-type electrodes), as well as the leaf pigment and phenolic contents, were used to evaluate the protective mechanisms of the photosynthetic apparatus under Cd stress. Data suggested that the molecular mechanisms included in the photosynthetic tolerance to Cd toxicity involve strongly increased phenolic and anthocyanin contents, as well as an increased non-photochemical quenching and accelerated cyclic electron transport around PSI up to 61%, which protect the function of the photosynthetic apparatus under stress. Furthermore, the tolerance of S. sclarea to Cd stress is also associated with increased accumulation of Fe in leaves by 25%. All the above, clearly suggest that S. sclarea plants employ several different mechanisms to protect the function of the photosynthetic apparatus against Cd stress, which are discussed here.

Different sensitivities of photosystem II in green algae and cyanobacteria to phenylurea and phenol-type herbicides: effect on electron donor side.

The effects of short-term treatment with phenylurea (DCMU, isoproturon) and phenol-type (ioxynil) herbicides on the green alga Chlorella kessleri and the cyanobacterium Synechocystis salina with different organizations of photosystem II (PSII) were investigated using pulse amplitude modulated (PAM) chlorophyll fluorescence and photosynthetic oxygen evolution measured by polarographic oxygen electrodes (Clark-type and Joliot-type). The photosynthetic oxygen evolution showed stronger inhibition than the PSII photochemistry. The effects of the studied herbicides on both algal and cyanobacterial cells decreased in the following order: DCMU>isoproturon>ioxynil. Furthermore, we observed that the number of blocked PSII centers increased significantly after DCMU treatment (204-250 times) and slightly after ioxynil treatment (19-35 times) in comparison with the control cells. This study suggests that the herbicides affect not only the acceptor side but also the donor side of PSII by modifications of the Mn cluster of the oxygen-evolving complex. We propose that one of the reasons for the different PSII inhibitions caused by herbicides is their influence, in different extents, on the kinetic parameters of the oxygen-evolving reactions (the initial S0-S1 state distribution, the number of blocked centers SB, the turnover time of Si states, misses and double hits). The relationship between the herbicide-induced inhibition and the changes in the kinetic parameters is discussed.

The wheat mutant DELLA-encoding gene (Rht-B1c) affects plant photosynthetic responses to cadmium stress.

Тhe sensitivity to cadmium (Cd) stress of two near-isogenic wheat lines with differences at the Rht-B1 locus, Rht-B1a (tall wild type, encoding DELLA proteins) and Rht-B1c (dwarf mutant, encoding modified DELLA proteins), was investigated. The effects of 100 µM CdCl2 on plant growth, pigment content and functional activity of the photosynthetic apparatus of wheat seedlings grown on a nutrient solution were evaluated through a combination of PAM chlorophyll fluorescence, oxygen evolution, oxidation-reduction kinetics of P700 and 77 K fluorescence. The results showed that the wheat mutant (Rht-B1c) was more tolerant to Cd stress compared to the wild type (Rht-B1a), as evidenced by the lower reductions in plant growth and pigment content, lower inhibition of photosystem I (PSI) and photosystem II (PSII) photochemistry and of the oxygen evolution measured with Clark-type and Joliot-type electrodes. Furthermore, the enhanced Cd tolerance was accompanied by increased Cd accumulation within mutant plant tissues. The molecular mechanisms through which the Rht-B1c mutation improves plant tolerance to Cd stress involve structural alterations in the mutant photosynthetic membranes leading to better protection of the Mn cluster of oxygen-evolving complex and increased capacity for PSI cyclic electron transport, protecting photochemical activity of the photosynthetic apparatus under stress. This study suggests a role for the Rht-B1c-encoded DELLA proteins in protective mechanisms and tolerance of the photosynthetic apparatus in wheat plants exposed to heavy metals stress.

Damage and protection of the photosynthetic apparatus from UV-B radiation. II. Effect of quercetin at different pH.

The effect of the exogenously added quercetin against the UV-B inhibition of the photosystem II (PSII) functions in isolated pea thylakoid membranes suspended at different pH of the medium (6.5, 7.6 and 8.4) was investigated. The data revealed that the interaction of this flavonoid with the membranes depends on the pH and influences the initial S0-S1 state distribution of PSII in the dark, the energy transfer between pigment-protein complexes of the photosynthetic apparatus and the membrane fluidity. Quercetin also displays a different UV-protective effect depending on its location in the membranes, as the effect is more pronounced at pH 8.4 when it is located at the membrane surface. The results suggest that quercetin induces structural changes in thylakoid membranes, one of the possible reasons for its protection of the photosynthetic apparatus.

Action and target sites of nitric oxide in chloroplasts.

Nitric oxide (NO) is an important signalling molecule in plants under physiological and stress conditions. Here we review the influence of NO on chloroplasts which can be directly induced by interaction with the photosynthetic apparatus by influencing photophosphorylation, electron transport activity and oxido-reduction state of the Mn clusters of the oxygen-evolving complex or by changes in gene expression. The influence of NO-induced changes in the photosynthetic apparatus on its functions and sensitivity to stress factors are discussed.

Effects of exogenous 24-epibrassinolide on the photosynthetic membranes under non-stress conditions.

In the present work the effects of exogenous 24-epibrassinolide (EBR) on functional and structural characteristics of the thylakoid membranes under non-stress conditions were evaluated 48 h after spraying of pea plants with different concentrations of EBR (0.01, 0.1 and 1.0 mg.L(-1)). The results show that the application of 0.1 mg.L(-1) EBR has the most pronounced effect on the studied characteristics of the photosynthetic membranes. The observed changes in 540 nm light scattering and in the calorimetric transitions suggest alterations in the structural organization of the thylakoid membranes after EBR treatment, which in turn influence the kinetics of oxygen evolution, accelerate the electron transport rate, increase the effective quantum yield of photosystem II and the photochemical quenching. The EBR-induced changes in the photosynthetic membranes are most probably involved in the stress tolerance of plants.

Damage and protection of the photosynthetic apparatus from UV-B radiation. I. Effect of ascorbate.

In this work, the effect of the exogenously added ascorbate (Asc) against the UV-B inhibition of the photosystem II (PSII) functions in isolated pea thylakoid membranes was studied. The results reveal that Asc decreases the UV-B induced damage of the donor and the acceptor side of PSII during short treatment up to 60 min. The exogenous Asc exhibits a different UV-protective effect on PSII centers in grana and stroma lamellae, as the effect is more pronounced on the PSIIß centers in comparison to PSIIα centers. Data also suggest that one of the possible protective roles of the Asc in photosynthetic membranes is the modification of the oxygen-evolving complex by influence on the initial S(0)-S(1) state distribution in the dark.

Effect of partial or complete elimination of light-harvesting complexes on the surface electric properties and the functions of cyanobacterial photosynthetic membranes.

Influence of the modification of the cyanobacterial light-harvesting complex [i.e. phycobilisomes (PBS)] on the surface electric properties and the functions of photosynthetic membranes was investigated. We used four PBS mutant strains of Synechocystis sp. PCC6803 as follows: PAL (PBS-less), CK (phycocyanin-less), BE (PSII-PBS-less) and PSI-less/apcE(-) (PSI-less with detached PBS). Modifications of the PBS content lead to changes in the cell morphology and surface electric properties of the thylakoid membranes as well as in their functions, such as photosynthetic oxygen-evolving activity, P700 kinetics and energy transfer between the pigment-protein complexes. Data reveal that the complete elimination of PBS in the PAL mutant causes a slight decrease in the electric dipole moments of the thylakoid membranes, whereas significant perturbations of the surface charges were registered in the membranes without assembled PBS-PSII macrocomplex (BE mutant) or PSI complex (PSI-less mutant). These observations correlate with the detected alterations in the membrane structural organization. Using a polarographic oxygen rate electrode, we showed that the ratio of the fast to the slow oxygen-evolving PSII centers depends on the partial or complete elimination of light-harvesting complexes, as the slow operating PSII centers dominate in the PBS-less mutant and in the mutant with detached PBS.

LHCII organization and thylakoid lipids affect the sensitivity of the photosynthetic apparatus to high-light treatment.

Pulse-amplitude-modulated (PAM) chlorophyll fluorescence and photosynthetic oxygen evolution were used to investigate the role of the different amount and organization of light-harvesting complexes of photosystem II (LHCII) in four pea species on the susceptibility of the photosynthetic apparatus to high-light treatment. In this work we analyzed the thylakoid membrane lipid composition of the studied pea plants. A relationship between the structural organization of LHCII proteins, the amount of the main lipid classes and the sensitivity of the photosynthetic apparatus to high-light treatment was found. The results reveal that the photosynthetic apparatus, enriched in oligomeric forms of LHCII concomitant with decreased amount of anionic lipids and increased content of the monogalactosyldiacylglycerol (MGDG), is less sensitive to high light. Our data also suggest that the degree of LHCII oligomerization, as well as the lipid composition do not influence the degree of recovery of the PSII photochemistry after excess light exposure.

Identification of thylakoid membrane thermal transitions in Synechocystis sp. PCC6803 photosynthetic mutants.

We used differential scanning calorimetry (DSC) as a technique capable of identifying photosynthetic complexes on the basis of their calorimetric transitions. Annotation of thermal transitions was carried out with thylakoid membranes isolated from various photosynthetic mutants of Synechocystis sp. PCC6803. The thylakoid membranes exhibited seven major DSC bands between 40 and 85°C. The heat sorption curves were analyzed both by mathematical deconvolution of the overall endotherms and by a subsequent annealing procedure. The successive annealing procedure proved to be more reliable technique than mathematical deconvolution in assigning thermal transitions. The main DSC band, around 47°C, resulting from the high enthalpy change that corresponds to non-interacting complex of PSII, was assigned using the PSI-less/apcE(-) mutant cells. Another band around 68-70°C relates to the denaturation of PSII surrounded by other proteins of the photosynthetic complexes in wild type and PSI-less/apcE(-) cells. A further major transition found at 82-84°C corresponds to the PSI core complex of wild type and PSII-deficient BE cells. Other transition bands between 50-67 and 65-75°C are believed to relate to ATP synthase and cytochrome b(6)f, respectively. These thermal transitions were obtained with thylakoids isolated from PSI(-)/PSII(-) mutant cells. Some minor bands determined at 59 and 83-84°C correspond to an unknown complex and NADH dehydrogenase, respectively. These annotations were done by PSI-less/apcE(-) and PSI(-)/PSII(-) mutants.

Photoelectron transport ability of chloroplast thylakoid membranes treated with NO donor SNP: changes in flash oxygen evolution and chlorophyll fluorescence.

The nitric oxide (NO) donor sodium nitroprusside (SNP) is frequently used in plant science in vivo. The present in vitro study reveals its effects on the photosynthetic oxygen evolution and the chlorophyll fluorescence directly on isolated pea thylakoid membranes. It was found that even at very low amounts of SNP (chlorophyll/SNP molar ratio∼67:1), the SNP-donated NO stimulates with more than 50% the overall photosystem II electron transport rate and diminishes the evolution of molecular oxygen. It was also found that the target site for SNP-donated NO is the donor side of photosystem II. Compared with other NO-donors used in plant science, SNP seems to be the only one exhibiting stimulation of electron transport through photosystem II.

Effect of phosphatidylglycerol depletion on the surface electric properties and the fluorescence emission of thylakoid membranes.

To explore the possible effect of phosphatidylglycerol (PG) on the surface electric properties and chlorophyll fluorescence characteristics we used electric light scattering technique and 77K chlorophyll fluorescence of thylakoid membranes from a cyanobacterium, Synechocystis PCC6803 (wild type) and its pgsA mutant defective in PG synthesis. We found a strong decrease in the permanent and induced electric dipole moments of the mutant thylakoids, following long-term PG depletion parallel with a decrease of the emission peak from PSI and an increase of the emission peak from PSII. Partial recovery of the electric state of thylakoid membranes was observed at re-addition of PG to the mutant cells depleted of PG for 21 days. This change in the electric dipole moments is probably due to a decrease in PG content and progressive structural alterations in the macroorganization of the photosynthetic complexes induced by PG deprivation. Our results suggest that the depletion of a lipid, which carries a negative charge, despite its small contribution to the overall lipid content, significantly perturbs the surface charge of the membranes. These changes are related with the chlorophyll fluorescence emission ratios of two photosystems and may partly explain our earlier results concerning the PG requirement for the function and assembly of photosystems I and II reaction centers.

Sensitivity of the photosynthetic apparatus to UV-A radiation: role of light-harvesting complex II-photosystem II supercomplex organization.

In this work we study the effect of UV-A radiation on the function of the photosynthetic apparatus in thylakoid membranes with different organization of the light-harvesting complex II-photosystem II (LHCII-PSII) supercomplex. Leaves and isolated thylakoid membranes from a number of previously characterized pea species with different LHCII size and organization were subjected to UV-A treatment. A relationship was found between the molecular organization of the LHCII (ratio of the oligomeric to monomeric forms of LHCII) and UV-A-induced changes both in the energy transfer from PSII to PSI and between the chlorophyll-protein complexes within the LHCII-PSII supercomplex. Dependence on the organization of the LHCII was also found with regard to the degree of inhibition of the photosynthetic oxygen evolution. The susceptibility of energy transfer and oxygen evolution to UV-A radiation decreased with increasing LHCII oligomerization when the UV-A treatment was performed on isolated thylakoid membranes, in contrast to the effect observed in thylakoid membranes isolated from pre-irradiated pea leaves. The data suggest that UV-A radiation leads mainly to damage of the PSIIalpha centers. Comparison of membranes with different organization of their LHCII-PSII supercomplex shows that the oligomeric forms of LHCII play a key role for sensitivity to UV-A radiation of the photosynthetic apparatus.

Relationship between the organization of the PS II super complex and the functions of the photosynthetic apparatus.

The chlorophyll fluorescence and the photosynthetic oxygen evolution (flash-induced oxygen yield patterns and oxygen bursts under continuous irradiation) were investigated in the thylakoid membranes with different stoichiometry and organization of the chlorophyll-protein complexes. Data show that the alteration in the organization of the photosystem II (PS II) super complex, i.e. the amount and the organization of the light-harvesting chlorophyll a/b protein complex (LHCII), which strongly modifies the electric properties of the membranes, influences both the energy redistribution between the two photosystems and the oxygen production reaction. The decrease of surface electric parameters (charge density and dipole moments), associated with increased degree of LHCII oligomerization, correlates with the strong reduction of the energy transfer from PS II to PSI. In the studied pea thylakoid membranes (wild types Borec, Auralia and their mutants Coeruleovireus 2/16, Costata2/133, Chlorotica XV/1422) with enhanced degree of oligomerization of LHCII was observed: (i) an increase of the S(0) populations of PS II in darkness; (ii) an increase of the misses; (iii) an alteration of the decay kinetics of the oxygen bursts under continuous irradiation. There is a strict correlation between the degree of LHCII oligomerization in the investigated pea mutants and the ratio of functionally active PS II alpha to PS II beta centers, while in thylakoid membranes without oligomeric structure of LHCII (Chlorina f2 barley mutant) the PS II alpha centers are not registered.