Enhanced Reduction of Ferredoxin in PGR5-Deficient Mutant of Arabidopsis thaliana Stimulated Ferredoxin-Dependent Cyclic Electron Flow around Photosystem I.

The molecular entity responsible for catalyzing ferredoxin (Fd)-dependent cyclic electron flow around photosystem I (Fd-CEF) remains unidentified. To reveal the in vivo molecular mechanism of Fd-CEF, evaluating ferredoxin reduction-oxidation kinetics proves to be a reliable indicator of Fd-CEF activity. Recent research has demonstrated that the expression of Fd-CEF activity is contingent upon the oxidation of plastoquinone. Moreover, chloroplast NAD(P)H dehydrogenase does not catalyze Fd-CEF in Arabidopsis thaliana. In this study, we analyzed the impact of reduced Fd on Fd-CEF activity by comparing wild-type and pgr5-deficient mutants (pgr5hope1). PGR5 has been proposed as the mediator of Fd-CEF, and pgr5hope1 exhibited a comparable CO2 assimilation rate and the same reduction-oxidation level of PQ as the wild type. However, P700 oxidation was suppressed with highly reduced Fd in pgr5hope1, unlike in the wild type. As anticipated, the Fd-CEF activity was enhanced in pgr5hope1 compared to the wild type, and its activity further increased with the oxidation of PQ due to the elevated CO2 assimilation rate. This in vivo research clearly demonstrates that the expression of Fd-CEF activity requires not only reduced Fd but also oxidized PQ. Importantly, PGR5 was found to not catalyze Fd-CEF, challenging previous assumptions about its role in this process.

Evaluating the Oxidation Rate of Reduced Ferredoxin in Arabidopsis thaliana Independent of Photosynthetic Linear Electron Flow: Plausible Activity of Ferredoxin-Dependent Cyclic Electron Flow around Photosystem I.

The activity of ferredoxin (Fd)-dependent cyclic electron flow (Fd-CEF) around photosystem I (PSI) was determined in intact leaves of Arabidopsis thaliana. The oxidation rate of Fd reduced by PSI (vFd) and photosynthetic linear electron flow activity are simultaneously measured under actinic light illumination. The vFd showed a curved response to the photosynthetic linear electron flow activity. In the lower range of photosynthetic linear flow activity with plastoquinone (PQ) in a highly reduced state, vFd clearly showed a linear relationship with photosynthetic linear electron flow activity. On the other hand, vFd increased sharply when photosynthetic linear electron flow activity became saturated with oxidized PQ as the net CO2 assimilation rate increased. That is, under higher photosynthesis conditions, we observed excess vFd resulting in electron flow over photosynthetic linear electron flow. The situation in which excess vFd was observed was consistent with the previous Fd-CEF model. Thus, excess vFd could be attributed to the in vivo activity of Fd-CEF. Furthermore, the excess vFd was also observed in NAD(P)H dehydrogenase-deficient mutants localized in the thylakoid membrane. The physiological significance of the excessive vFd was discussed.

Behavior of Photosystems II and I Is Modulated Depending on N Partitioning to Rubisco in Mature Leaves Acclimated to Low N Levels and Senescent Leaves in Rice.

In mature leaves acclimated to low N levels and in senescent leaves, photosystems II and I (PSII and PSI, respectively) show typical responses to excess light energy. As CO2 assimilation is not transiently suppressed in these situations, the behavior of PSII and PSI is likely caused by endogenous biochemical changes in photosynthesis. In this study, this subject was studied in rice (Oryza sativa L.). Analysis was performed on mature and senescent leaves of control and N-deficient plants. Total leaf-N, Rubisco and chlorophyll (Chl) levels and their ratios were determined as biochemical parameters of photosynthesis. Total leaf-N, Rubisco and Chl levels decreased in the mature leaves of N-deficient plants and senescent leaves. The percentage of Rubisco-N in the total leaf-N decreased in these leaves, whereas that of Chl-N tended to remain almost constant in mature leaves but increased in senescent leaves. Changes in PSII and PSI parameters were best accounted for by the Rubisco-N percentage, strongly suggesting that the behavior of PSII and PSI is modulated depending on changes in N partitioning to Rubisco in mature leaves acclimated to low N levels and in senescent leaves. It is likely that a decrease in N partitioning to Rubisco leads to a decrease in Rubisco capacity relative to other photosynthetic capacities that inevitably generate excess light energy and that the operation of PSII and PSI is modulated in such situations.

Tight relationship between two photosystems is robust in rice leaves under various nitrogen conditions.

Leaf nitrogen (N) level affects not only photosynthetic CO2 assimilation, but also two photosystems of the photosynthetic electron transport. The quantum yield of photosystem II [Y(II)] and the non-photochemical yield due to the donor side limitation of photosystem I [Y(ND)], which denotes the fraction of oxidized P700 (P700+) to total P700, oppositely change depending on leaf N level, and the negative correlation between these two parameters has been reported in leaves of plants cultivated at various N levels in growth chambers. Here, we aimed to clarify whether this correlation is maintained after short-term changes in leaf N level, and what parameters are the most responsive to the changes in leaf N level under field conditions. We cultivated rice varieties at two N fertilization levels in paddy fields, treated additional N fertilization to plants grown at low N, and measured parameters of two photosystems of mature leaves. In rice leaves under low N condition, the Y(ND) increased and the photosynthetic linear electron flow was suppressed. In this situation, the accumulation of P700+ can function as excess energy dissipation. After the N addition, both Y(ND) and Y(II) changed, and the negative correlation between them was maintained. We used a newly-developed device to assess the photosystems. This device detected the similar changes in Y(ND) after the N addition, and the negative correlation between Y(ND) and photosynthetic O2 evolution rates was observed in plants under various N conditions. This study has provided strong field evidence that the Y(ND) largely changes depending on leaf N level, and that the Y(II) and Y(ND) are negatively correlated with each other irrespective of leaf N level, varieties and annual variation. The Y(ND) can stably monitor the leaf N status and the linear electron flow under field conditions.

Overexpression of Chloroplast Triosephosphate Isomerase Marginally Improves Photosynthesis at Elevated CO2 Levels in Rice.

We recently suggested that chloroplast triosephosphate isomerase (cpTPI) has moderate control over the rate of CO2 assimilation (A) at elevated CO2 levels via the capacity for triose phosphate utilization (TPU) in rice (Oryza sativa L.) from its antisense-suppression study. In the present study, the effects of cpTPI overexpression on photosynthesis were examined in transgenic rice plants overexpressing the gene encoding cpTPI. The amounts of cpTPI protein in the two lines of transgenic plants were 4.8- and 12.1-folds higher than in wild-type plants, respectively. The magnitude of the increase approximately corresponded to the increase in transcript levels of cpTPI. A at CO2 levels of 100 and 120 Pa increased by 6-9% in the transgenic plants, whereas those at ambient and low CO2 levels were scarcely affected. Similar increases were observed for TPU capacity estimated from the CO2 response curves of A. These results indicate that the overexpression of cpTPI marginally improved photosynthesis at elevated CO2 levels via improvement in TPU capacity in rice. However, biomass production at a CO2 level of 120 Pa did not increase in transgenic plants, suggesting that the improvement in photosynthesis by cpTPI overexpression was not sufficient to improve biomass production in rice.

Order-of-magnitude enhancement in photocurrent generation of Synechocystis sp. PCC 6803 by outer membrane deprivation.

Biophotovoltaics (BPV) generates electricity from reducing equivalent(s) produced by photosynthetic organisms by exploiting a phenomenon called extracellular electron transfer (EET), where reducing equivalent(s) is transferred to external electron acceptors. Although cyanobacteria have been extensively studied for BPV because of their high photosynthetic activity and ease of handling, their low EET activity poses a limitation. Here, we show an order-of-magnitude enhancement in photocurrent generation of the cyanobacterium Synechocystis sp. PCC 6803 by deprivation of the outer membrane, where electrons are suggested to stem from pathway(s) downstream of photosystem I. A marked enhancement of EET activity itself is verified by rapid reduction of exogenous electron acceptor, ferricyanide. The extracellular organic substances, including reducing equivalent(s), produced by this cyanobacterium serve as respiratory substrates for other heterotrophic bacteria. These findings demonstrate that the outer membrane is a barrier that limits EET. Therefore, depriving this membrane is an effective approach to exploit the cyanobacterial reducing equivalent(s).

The ability of P700 oxidation in photosystem I reflects chilling stress tolerance in cucumber.

Low temperature inhibits photosynthesis and negatively affects plant growth. Cucumber (Cucumis sativus L.) is a chilling-sensitive plant, and its greenhouse production requires considerable energy during the winter. Therefore, a useful stress marker for selecting chilling-tolerant cucumber cultivars is desirable. In this study, we evaluated chilling-stress damage in different cucumber cultivars by measuring photosynthetic parameters. The majority of cultivars showed decreases in the quantum yield of photosystem (PS) II [Fv/Fm and Y(II)] and the quantity of active PS I (Pm) after chilling stress. In contrast, Y(ND)-the ratio of the oxidized state of PSI reaction center chlorophyll P700 (P700+)-differed among cultivars and was perfectly inversely correlated with Y(NA)-the ratio of the non-photooxidizable P700. It has been known that P700+ accumulates under stress conditions and protects plants to suppress the generation of reactive oxygen species. In fact, cultivars unable to induce Y(ND) after chilling stress showed growth retardation with reductions in chlorophyll content and leaf area. Therefore, Y(ND) can be a useful marker to evaluate chilling-stress tolerance in cucumber.

Dissection of respiratory and cyclic electron transport in Synechocystis sp. PCC 6803.

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.

Identification of Twelve Different Mineral Deficiencies in Hydroponically Grown Sunflower Plants on the Basis of Short Measurements of the Fluorescence and P700 Oxidation/Reduction Kinetics.

The photosynthetic electron transport chain is mineral rich. Specific mineral deficiencies can modify the electron transport chain specifically. Here, it is shown that on the basis of 2 short Chl fluorescence and P700+ measurements (approx. 1 s each), it is possible to discriminate between 10 out of 12 different mineral deficiencies: B, Ca, Cu, Fe, K, Mg, Mn, Mo, N, P, S, and Zn. B- and Mo-deficient plants require somewhat longer measurements to detect the feedback inhibition they induce. Eight out of twelve deficiencies mainly affect PS I and NIR measurements are, therefore, very important for this analysis. In Cu- and P-deficient plants, electron flow from the plastoquinone pool to PS I, is affected. In the case of Cu-deficiency due to the loss of plastocyanin and in the case of P-deficiency probably due to a fast and strong generation of Photosynthetic Control. For several Ca-, K-, and Zn-deficient plant species, higher levels of reactive oxygen species have been measured in the literature. Here, it is shown that this not only leads to a loss of Pm (maximum P700 redox change) reflecting a lower PS I content, but also to much faster P700+ re-reduction kinetics during the I2-P (~30-200 ms) fluorescence rise phase. The different mineral deficiencies affect the relation between the I2-P and P700+ kinetics in different ways and this is used to discuss the nature of the relationship between these two parameters.

Effects of suppression of chloroplast phosphoglycerate kinase on photosynthesis in rice.

As chloroplast phosphoglycerate kinase (cpPGK) is one of the enzymes which has the highest capacity among the Calvin-Benson cycle enzymes, it has not been regarded as a determinant for photosynthetic capacity. However, it was reported that the rate of CO2 assimilation decreased under high irradiance and normal [CO2] levels in the Arabidopsis cpPGK-knockdown mutant, implying that cpPGK has a control over photosynthetic capacity at a normal [CO2] level. In the present study, the contribution of cpPGK to photosynthetic capacity was evaluated in transgenic rice plants with decreased amounts of cpPGK protein under high irradiance and various [CO2] levels. The gene encoding cpPGK was suppressed using RNA interference techniques. Two lines of transgenic plants, Pi3 and Pi5, in which the amount of cpPGK protein decreased to 21% and 76% of that in wild-type plants, respectively, were obtained. However, there was no substantial difference in the rates of CO2 assimilation between wild-type and transgenic plants. The rates of CO2 assimilation decreased only slightly at elevated [CO2] levels in the transgenic line Pi3 and did not differ between wild-type plants and the transgenic line Pi5, irrespective of [CO2] level. These results clearly indicate that cpPGK does not have a strong control over photosynthetic capacity at various [CO2] levels in rice.

NADPH production in dark stages is critical for cyanobacterial photocurrent generation: a study using mutants deficient in oxidative pentose phosphate pathway.

Live cyanobacteria and algae integrated onto an extracellular electrode can generate a light-induced current (i.e., a photocurrent). Although the photocurrent is expected to be correlated with the redox environment of the photosynthetic cells, the relationship between the photocurrent and the cellular redox state is poorly understood. Here, we investigated the effect of the reduced nicotinamide adenine dinucleotide phosphate [NADP(H)] redox level of cyanobacterial cells (before light exposure) on the photocurrent using several mutants (Δzwf, Δgnd, and ΔglgP) deficient in the oxidative pentose phosphate (OPP) pathway, which is the metabolic pathway that produces NADPH in darkness. The NAD(P)H redox level and photocurrent in the cyanobacterium Synechocystis sp. PCC 6803 were measured noninvasively. Dysfunction of the OPP pathway led to oxidation of the photosynthetic NADPH pool in darkness. In addition, photocurrent induction was retarded and the current density was lower in Δzwf, Δgnd, and ΔglgP than in wild-type cells. Exogenously added glucose compensated the phenotype of ΔglgP and drove the OPP pathway in the mutant, resulting in an increase in the photocurrent. The results indicated that NADPH accumulated by the OPP pathway before illumination is a key factor for the generation of a photocurrent. In addition, measuring the photocurrent can be a non-invasive approach to estimate the cellular redox level related to NADP(H) pool in cyanobacteria.

Higher Reduced State of Fe/S-Signals, with the Suppressed Oxidation of P700, Causes PSI Inactivation in Arabidopsis thaliana.

Environmental stress increases the risk of electron accumulation in photosystem I (PSI) of chloroplasts, which can cause oxygen (O2) reduction to superoxide radicals and decreased photosynthetic ability. We used three Arabidopsis thaliana lines: wild-type (WT) and the mutants pgr5hope1 and paa1-7/pox1. These lines have different reduced states of iron/sulfur (Fe/S) signals, including Fx, FA/FB, and ferredoxin, the electron carriers at the acceptor side of PSI. In the dark, short-pulse light was repetitively illuminated to the intact leaves of the plants to provide electrons to the acceptor side of PSI. WT and pgr5hope1 plants showed full reductions of Fe/S during short-pulse light and PSI inactivation. In contrast, paa1-7/pox1 showed less reduction of Fe/S and its PSI was not inactivated. Under continuous actinic-light illumination, pgr5hope1 showed no P700 oxidation with higher Fe/S reduction due to the loss of photosynthesis control and PSI inactivation. These results indicate that the accumulation of electrons at the acceptor side of PSI may trigger the production of superoxide radicals. P700 oxidation, responsible for the robustness of photosynthetic organisms, participates in reactive oxygen species suppression by oxidizing the acceptor side of PSI.

The difficulty of estimating the electron transport rate at photosystem I.

It is still a controversial issue how the electron transport reaction is carried out around photosystem I (PSI) in the photosynthetic electron transport chain. The measurable component in PSI is the oxidized P700, the reaction center chlorophyll in PSI, as the absorbance changes at 820-830 nm. Previously, the quantum yield at PSI [Y(I)] has been estimated as the existence probability of the photo-oxidizable P700 by applying the saturated-pulse illumination (SP; 10,000-20,000 µmol photons m-2 s-1). The electron transport rate (ETR) at PSI has been estimated from the Y(I) value, which was larger than the reaction rate at PSII, evaluated as the quantum yield of PSII, especially under stress-conditions such as CO2-limited and high light intensity conditions. Therefore, it has been considered that the extra electron flow at PSI was enhanced at the stress condition and played an important role in dealing with the excessive light energy. However, some pieces of evidence were reported that the excessive electron flow at PSI would be ignorable from other aspects. In the present research, we confirmed that the Y(I) value estimated by the SP method could be easily misestimated by the limitation of the electron donation to PSI. Moreover, we estimated the quantitative turnover rate of P700+ by the light-to-dark transition. However, the turnover rate of P700 was much slower than the ETR at PSII. It is still hard to quantitatively estimate the ETR at PSI by the current techniques.

Suppression of chloroplast triose phosphate isomerase evokes inorganic phosphate-limited photosynthesis in rice.

The availability of inorganic phosphate (Pi) for ATP synthesis is thought to limit photosynthesis at elevated [CO2] when Pi regeneration via sucrose or starch synthesis is limited. We report here another mechanism for the occurrence of Pi-limited photosynthesis caused by insufficient capacity of chloroplast triose phosphate isomerase (cpTPI). In cpTPI-antisense transgenic rice (Oryza sativa) plants with 55%-86% reductions in cpTPI content, CO2 sensitivity of the rate of CO2 assimilation (A) decreased and even reversed at elevated [CO2]. The pool sizes of the Calvin-Benson cycle metabolites from pentose phosphates to 3-phosphoglycerate increased at elevated [CO2], whereas those of ATP decreased. These phenomena are similar to the typical symptoms of Pi-limited photosynthesis, suggesting sufficient capacity of cpTPI is necessary to prevent the occurrence of Pi-limited photosynthesis and that cpTPI content moderately affects photosynthetic capacity at elevated [CO2]. As there tended to be slight variations in the amounts of total leaf-N depending on the genotypes, relationships between A and the amounts of cpTPI were examined after these parameters were expressed per unit amount of total leaf-N (A/N and cpTPI/N, respectively). A/N at elevated [CO2] decreased linearly as cpTPI/N decreased before A/N sharply decreased, owing to further decreases in cpTPI/N. Within this linear range, decreases in cpTPI/N by 80% led to decreases up to 27% in A/N at elevated [CO2]. Thus, cpTPI function is crucial for photosynthesis at elevated [CO2].

Identification of a Novel Mutation Exacerbated the PSI Photoinhibition in pgr5/pgrl1 Mutants; Caution for Overestimation of the Phenotypes in Arabidopsis pgr5-1 Mutant.

PSI photoinhibition is usually avoided through P700 oxidation. Without this protective mechanism, excess light represents a potentially lethal threat to plants. PGR5 is suggested to be a major component of cyclic electron transport around PSI and is important for P700 oxidation in angiosperms. The known Arabidopsis PGR5 deficient mutant, pgr5-1, is incapable of P700 oxidation regulation and has been used in numerous photosynthetic studies. However, here it was revealed that pgr5-1 was a double mutant with exaggerated PSI photoinhibition. pgr5-1 significantly reduced growth compared to the newly isolated PGR5 deficient mutant, pgr5hope1. The introduction of PGR5 into pgr5-1 restored P700 oxidation regulation, but remained a pale-green phenotype, indicating that pgr5-1 had additional mutations. Both pgr5-1 and pgr5hope1 tended to cause PSI photoinhibition by excess light, but pgr5-1 exhibited an enhanced reduction in PSI activity. Introducing AT2G17240, a candidate gene for the second mutation into pgr5-1 restored the pale-green phenotype and partially restored PSI activity. Furthermore, a deficient mutant of PGRL1 complexing with PGR5 significantly reduced PSI activity in the double-deficient mutant with AT2G17240. From these results, we concluded that AT2G17240, named PSI photoprotection 1 (PTP1), played a role in PSI photoprotection, especially in PGR5/PGRL1 deficient mutants.

Physiological Roles of Flavodiiron Proteins and Photorespiration in the Liverwort Marchantia polymorpha.

Against the potential risk in oxygenic photosynthesis, that is, the generation of reactive oxygen species, photosynthetic electron transport needs to be regulated in response to environmental fluctuations. One of the most important regulations is keeping the reaction center chlorophyll (P700) of photosystem I in its oxidized form in excess light conditions. The oxidation of P700 is supported by dissipating excess electrons safely to O2, and we previously found that the molecular mechanism of the alternative electron sink is changed from flavodiiron proteins (FLV) to photorespiration in the evolutionary history from cyanobacteria to plants. However, the overall picture of the regulation of photosynthetic electron transport is still not clear in bryophytes, the evolutionary intermediates. Here, we investigated the physiological roles of FLV and photorespiration for P700 oxidation in the liverwort Marchantia polymorpha by using the mutants deficient in FLV (flv1) at different O2 partial pressures. The effective quantum yield of photosystem II significantly decreased at 2kPa O2 in flv1, indicating that photorespiration functions as the electron sink. Nevertheless, it was clear from the phenotype of flv1 that FLV was dominant for P700 oxidation in M. polymorpha. These data suggested that photorespiration has yet not replaced FLV in functioning for P700 oxidation in the basal land plant probably because of the lower contribution to lumen acidification, compared with FLV, as reflected in the results of electrochromic shift analysis.

Photosynthetic Parameters Show Specific Responses to Essential Mineral Deficiencies.

In response to decreases in the assimilation efficiency of CO2, plants oxidize the reaction center chlorophyll (P700) of photosystem I (PSI) to suppress reactive oxygen species (ROS) production. In hydro-cultured sunflower leaves experiencing essential mineral deficiencies, we analyzed the following parameters that characterize PSI and PSII: (1) the reduction-oxidation states of P700 [Y(I), Y(NA), and Y(ND)]; (2) the relative electron flux in PSII [Y(II)]; (3) the reduction state of the primary electron acceptor in PSII, QA (1 - qL); and (4) the non-photochemical quenching of chlorophyll fluorescence (NPQ). Deficiency treatments for the minerals N, P, Mn, Mg, S, and Zn decreased Y(II) with an increase in the oxidized P700 [Y(ND)], while deficiencies for the minerals K, Fe, Ca, B, and Mo decreased Y(II) without an increase in Y(ND). During the induction of photosynthesis, the above parameters showed specific responses to each mineral. That is, we could diagnose the mineral deficiency and identify which mineral affected the photosynthesis parameters.

Photosynthetic Linear Electron Flow Drives CO2 Assimilation in Maize Leaves.

Photosynthetic organisms commonly develop the strategy to keep the reaction center chlorophyll of photosystem I, P700, oxidized for preventing the generation of reactive oxygen species in excess light conditions. In photosynthesis of C4 plants, CO2 concentration is kept at higher levels around ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) by the cooperation of the mesophyll and bundle sheath cells, which enables them to assimilate CO2 at higher rates to survive under drought stress. However, the regulatory mechanism of photosynthetic electron transport for P700 oxidation is still poorly understood in C4 plants. Here, we assessed gas exchange, chlorophyll fluorescence, electrochromic shift, and near infrared absorbance in intact leaves of maize (a NADP-malic enzyme C4 subtype species) in comparison with mustard, a C3 plant. Instead of the alternative electron sink due to photorespiration in the C3 plant, photosynthetic linear electron flow was strongly suppressed between photosystems I and II, dependent on the difference of proton concentration across the thylakoid membrane (ΔpH) in response to the suppression of CO2 assimilation in maize. Linear relationships among CO2 assimilation rate, linear electron flow, P700 oxidation, ΔpH, and the oxidation rate of ferredoxin suggested that the increase of ΔpH for P700 oxidation was caused by the regulation of proton conductance of chloroplast ATP synthase but not by promoting cyclic electron flow. At the scale of intact leaves, the ratio of PSI to PSII was estimated almost 1:1 in both C3 and C4 plants. Overall, the photosynthetic electron transport was regulated for P700 oxidation in maize through the same strategies as in C3 plants only except for the capacity of photorespiration despite the structural and metabolic differences in photosynthesis between C3 and C4 plants.

Quantification of NAD(P)H in cyanobacterial cells by a phenol extraction method.

In photosynthetic organisms, it is recognized that the intracellular redox ratio of NADPH is regulated within an appropriate range for the cooperative function of a wide variety of physiological processes. However, despite its importance, there is large variability in the values of the NADPH fraction [NADPH/(NADPH + NADP+)] quantitatively estimated to date. In the present study, the light response of the NADPH fraction was investigated by applying a novel NADP(H) extraction method using phenol / chloroform / isoamyl alcohol (PCI) in the cyanobacterium Synechocystis sp. PCC 6803. The light response of NADP(H) observed using PCI extraction was qualitatively consistent with the NAD(P)H fluorescence time course measured in vivo. Moreover, the results obtained by PCI extraction and the fluorescence-based methods were also consistent in a mutant lacking the ability to oxidize NAD(P)H in the respiratory chain, and exhibiting a unique NADPH light response. These observations indicate that the PCI extraction method allowed quantitative determination of NADP(H) redox. Notably, the PCI extraction method showed that not all NADP(H) was oxidized or reduced by light-dark transition. Specifically, the fraction of NADPH was 42% in the dark-adapted cell, and saturated at 68% in light conditions.

Photochemistry of Photosystems II and I in Rice Plants Grown under Different N Levels at Normal and High Temperature.

Although N levels affect leaf photosynthetic capacity, the effects of N levels on the photochemistry of photosystems II and I (PSII and PSI, respectively) are not well-understood. In the present study, we examined this aspect in rice (Oryza sativa L. 'Hitomebore') plants grown under three different N levels at normal or high temperatures that can occur during rice culture and do not severely suppress photosynthesis. At both growth temperatures, the quantum efficiency of PSII [Y(II)] and the fraction of the primary quinone electron acceptor in its oxidized state were positively correlated with the amount of total leaf-N, whereas the quantum yields of non-photochemical quenching and donor-side limitation of PSI [Y(ND)] were negatively correlated with the amount of total leaf-N. These changes in PSII and PSI parameters were strongly correlated with each other. Growth temperatures scarcely affected these relationships. These results suggest that the photochemistry of PSII and PSI is coordinately regulated primarily depending on the amount of total leaf-N. When excess light energy occurs in low N-acclimated plants, oxidation of the reaction center chlorophyll of PSI is thought to be stimulated to protect PSI from excess light energy. It is also suggested that PSII and PSI normally operate at high temperature used in the present study. In addition, as the relationships between Y(II) and Y(ND) were found to be almost identical to those observed in osmotically stressed rice plants, common regulation is thought to be operative when excess light energy occurs due to different causes.