Plastoquinone Research Articles

Photo-reducible plastoquinone pools in chloroplasts of Tradescentia plants acclimated to high and low light.

In photosynthetic systems of oxygenic type, plastoquinone (PQ) molecules are reduced by photosystem II (PSII). The turnover of PQ determines the rate of PSII operation. PQ molecules are present in surplus with respect to PSII. In this work, using the pulse amplitude modulation-fluorometry technique, we quantified photo-reducible PQ pools in chloroplasts of two contrasting ecotypes of Tradescantia, acclimated either to low light (~100μmolphotons·m-2 ·s-1 , LL) or to high light (~1000μmolphotons·m-2 ·s-1 , HL). The LL-grown plants are characterized by higher capacity of rapidly reducible PQ pool ([PQ]0 /[PSII]≈8) as compared to HL-grown plants of both species ([PQ]0 /[PSII]≈4). The elevated content of PQ in LL plants favours photosynthetic electron flow at low-solar irradiance.

The ratio of single-turnover to multiple-turnover fluorescence varies predictably with growth rate and cellular chlorophyll in the green alga Dunaliella tertiolecta.

Marine phytoplankton experience a wide range of nutrient and light conditions in nature and respond to these conditions through changes in growth rate, chlorophyll concentration, and other physiological properties. Chlorophyll fluorescence is a non-invasive and efficient tool for characterizing changes in these physiological properties. In particular, the introduction of fast repetition rate fluorometry (FRRf) into studies of phytoplankton physiology has enabled detailed studies of photosynthetic components and kinetics. One property retrieved with an FRRf is the 'single-turnover' maximum fluorescence (FmST) when the primary electron acceptor, Qa, is reduced but the plastoquinone (PQ) pool is oxidized. A second retrieved property is the 'multiple-turnover' fluorescence (FMT) when both Qa and PQ are reduced. Here, variations in FmST and FMT were measured in the green alga Dunaliella tertiolecta grown under nitrate-limited, light-limited, and replete conditions. The ratio of FmST to FMT (ST/MT) showed a consistent relationship with cellular chlorophyll in D. tertiolecta across all growth conditions. However, the ST/MT ratio decreased with growth rate under nitrate-limited conditions but increased with growth rate under light-limited conditions. In addition, cells from light-limited conditions showed a high accumulation of Qb-nonreducing centers, while cells from nitrate-limited conditions showed little to none. We propose that these findings reflect differences in the reduction and oxidation rates of plastoquinone due to the unique impacts of light and nitrate limitation on the stoichiometry of light-harvesting components and downstream electron acceptors.

Spatiotemporal heterogeneity of photosystem II function during acclimation to zinc exposure and mineral nutrition changes in the hyperaccumulator Noccaea caerulescens.

We investigated changes in mineral nutrient uptake and translocation and photosystem II (PSII) functionality, in the hyperaccumulator Noccaea caerulescens after exposure to 800μM Zn in hydroponic culture. Exposure to Zn inhibited the uptake of K, Mn, Cu, Ca, and Mg, while the uptake of Fe and Zn enhanced. Yet, Ca and Mg aboveground tissue concentrations remain unchanged while Cu increased significantly. In the present study, we provide new data on the mechanism of N. caerulescens acclimation to Zn exposure by elucidating the process of photosynthetic acclimation. A spatial heterogeneity in PSII functionality in N. caerulescens leaves exposed to Zn for 3days was detected, while a threshold time of 4days was needed for the activation of Zn detoxification mechanism(s) to decrease Zn toxicity and for the stomatal closure to decrease Zn supply at the severely affected leaf area. After 10-day exposure to Zn, the allocation of absorbed light energy in PSII under low light did not differ compared to control ones, while under high light, the quantum yield of non-regulated energy loss in PSII (ΦNO) was lower than the control, due to an efficient photoprotective mechanism. The chlorophyll fluorescence images of non-photochemical quenching (NPQ) and photochemical quenching (qp) clearly showed spatial and temporal heterogeneity in N. caerulescens exposure to Zn and provided further information on the particular leaf area that was most sensitive to heavy metal stress. We propose the use of chlorophyll fluorescence imaging, and in particular the redox state of the plastoquinone (PQ) pool that was found to display the highest spatiotemporal heterogeneity, as a sensitive bio-indicator to measure the environmental pressure by heavy metals on plants.

Probing functional and optical cross-sections of PSII in leaves during state transitions using fast repetition rate light induced fluorescence transients.

Plants adjust the relative sizes of PSII and PSI antennae in response to the spectral composition of weak light favouring either photosystem by processes known as state transitions (ST), attributed to a discrete antenna migration involving phosphorylation of light-harvesting chlorophyll-protein complexes in PSII. Here for the first time we monitored the extent and dynamics of ST in leaves from estimates of optical absorption cross-section (relative PSII antenna size; aPSII). These estimates were obtained from in situ measurements of functional absorption cross-section (σPSII) and maximum photochemical efficiency of PSII (φPSII); i.e. aPSII = σPSII/φPSII (Kolber et al. 1998) and other parameters from a light induced fluorescence transient (LIFT) device (Osmond et al. 2017). The fast repetition rate (FRR) QA flash protocol of this instrument monitors chlorophyll fluorescence yields with reduced QA irrespective of the redox state of plastoquinone (PQ), as well as during strong ~1 s white light pulses that fully reduce the PQ pool. Fitting this transient with the FRR model monitors kinetics of PSII → PQ, PQ → PSI, and the redox state of the PQ pool in the 'PQ pool control loop' that underpins ST, with a time resolution of a few seconds. All LIFT/FRR criteria confirmed the absence of ST in antenna mutant chlorina-f2 of barley and asLhcb2-12 of Arabidopsis, as well as STN7 kinase mutants stn7 and stn7/8. In contrast, wild-type barley and Arabidopsis genotypes Col, npq1, npq4, OEpsbs, pgr5 bkg and pgr5, showed normal ST. However, the extent of ST (and by implication the size of the phosphorylated LHCII pool participating in ST) deduced from changes in a'PSII and other parameters with reduced QA range up to 35%. Estimates from strong WL pulses in the same assay were only ~10%. The larger estimates of ST from the QA flash are discussed in the context of contemporary dynamic structural models of ST involving formation and participation of PSII and PSI megacomplexes in an 'energetically connected lake' of phosphorylated LHCII trimers (Grieco et al. 2015). Despite the absence of ST, asLhcb2-12 displays normal wild-type modulation of electron transport rate (ETR) and the PQ pool during ST assays, reflecting compensatory changes in antenna LHCIIs in this genotype. Impaired LHCII phosphorylation in stn7 and stn7/8 accelerates ETR from PSII →PQ, over-reducing the PQ pool and abolishing the yield difference between the QA flash and WL pulse, with implications for photochemical and thermal phases of the O-J-I-P transient.

Chlorophyll Fluorescence Imaging Analysis for Elucidating the Mechanism of Photosystem II Acclimation to Cadmium Exposure in the Hyperaccumulating Plant Noccaea caerulescens.

We provide new data on the mechanism of Noccaea caerulescens acclimation to Cd exposure by elucidating the process of photosystem II (PSII) acclimation by chlorophyll fluorescence imaging analysis. Seeds from the metallophyte N. caerulescens were grown in hydroponic culture for 12 weeks before exposure to 40 and 120 μM Cd for 3 and 4 days. At the beginning of exposure to 40 μM Cd, we observed a spatial leaf heterogeneity of decreased PSII photochemistry, that later recovered completely. This acclimation was achieved possibly through the reduced plastoquinone (PQ) pool signaling. Exposure to 120 μM Cd under the growth light did not affect PSII photochemistry, while under high light due to a photoprotective mechanism (regulated heat dissipation for protection) that down-regulated PSII quantum yield, the quantum yield of non-regulated energy loss in PSII (ΦNO) decreased even more than control values. Thus, N. caerulescens plants exposed to 120 μM Cd for 4 days exhibited lower reactive oxygen species (ROS) production as singlet oxygen (1O2). The response of N. caerulescens to Cd exposure fits the ‘Threshold for Tolerance Model’, with a lag time of 4 d and a threshold concentration of 40 μM Cd required for the induction of the acclimation mechanism.

Self-Adaptable Quinone-Quinol Exchange Mechanism of Photosystem II.

The step of plastoquinone (PQ) reduction to plastoquinol (PQH2) can regulate the photoreaction rate of photosystem II (PSII). To experimentally unravel the PQ-PQH2 exchange mechanism of PSII, we investigate the reaction kinetics of plant PSII membranes and the subunits-trimmed PSII core complexes with various PQ analogues and directly probe the reductions of PQ and other quinones by 257 nm resonance Raman scattering. Two phases of quinone concentration effect on the reaction rate originate from the quinone-quinol exchange mechanism. The results indicate that high concentrations of quinone, more than one movable quinone molecule per PSII reaction center, could trigger quinone-quinol exchange adapting to the unidirectional route: quinones enter through channel I and/or III, and quinols leave through channel II. A weak quinone binding site near QB probably plays a crucial role in pushing quinone-quinol exchange forward in the unidirectional route. Our work provides experimental proofs demonstrating a self-adaptable quinone-quinol exchange mechanism of PSII.

Oxidation of the plastoquinone pool in chloroplast thylakoid membranes by superoxide anion radicals.

The plastoquinone (PQ)-pool in chloroplast thylakoid membranes is a key electron carrier in the photosynthetic electron transport chain (PETC), and its redox state plays an essential role in the control of plant metabolism. Oxygen reduction in thylakoid membranes producessuperoxide anion radicals ( ), which may react with the PQ-pool. Here, using isolated thylakoids, we show for the first time the oxidation of the PQ-pool by . The xanthine-xanthine oxidase system was used to supply externally to the thylakoid membrane and the redox state of the PQ-pool was monitored by tracking chlorophyll a fluorescence. We propose that,invivo,the reaction of produced in Photosystem I with reduced PQ (plastohydroquinone) creates hydrogen peroxide, which serves as a messenger that signals the redox state of the PETC.

Time-resolved step-scan FTIR difference spectroscopy for the study of photosystem I with different benzoquinones incorporated into the A1 binding site

Time-resolved step-scan FTIR difference spectroscopy has been used to study photosystem I (PSI) with plastoquinone-9 (PQ) and two other benzoquinones (2,6-dimethyl-1,4-benzoquinone and 2,3,5,6-tetrachloro-1,4-benzoquinone) incorporated into the A1 binding site. By subtracting a (P700+A1− − P700A1) FTIR difference spectrum for PSI with the native phylloquinone (PhQ) incorporated from corresponding spectra for PSI with different benzoquinones (BQs) incorporated, FTIR double difference spectra are produced, that display bands associated with vibrational modes of the quinones, without interference from features associated with protein vibrational modes.Molecular models for BQs involved in asymmetric hydrogen bonding were constructed and used in vibrational mode frequency calculations. The calculated data were used to aid in the interpretation and assignment of bands in the experimental spectra. We show that the calculations capture the general trends found in the experimental spectra.By comparing four different FTIR double difference spectra we are able to verify unambiguously bands associated with phyllosemiquinone in PSI at 1495 and 1415 cm−1. We also resolve a previously unrecognized band of phyllosemiquinone at 1476 cm−1 that calculations suggest is due in part to a C4−⃛O stretching mode.For PSI with PQ incorporated, calculations and experiment taken together indicate that the C1−⃛O and C4−⃛O vibrational modes of the semiquinone give rise to bands at 1487 and 1444 cm−1, respectively. This is very distinct compared to PSI with PhQ incorporated.From the calculated and experimental spectra, we show that it is possible to distinguish between two possible orientations of PQ in the A1 protein binding site.

Chemical quenching of singlet oxygen by plastoquinols and their oxidation products in Arabidopsis.

Prenylquinols (tocochromanols and plastoquinols) serve as efficient physical and chemical quenchers of singlet oxygen (1 O2 ) formed during high light stress in higher plants. Although quenching of 1 O2 by prenylquinols has been previously studied, direct evidence for chemical quenching of 1 O2 by plastoquinols and their oxidation products is limited invivo. In the present study, the role of plastoquinol-9 (PQH2 -9) in chemical quenching of 1 O2 was studied in Arabidopsis thaliana lines overexpressing the SOLANESYL DIPHOSPHATE SYNTHASE 1 gene (SPS1oex) involved in PQH2 -9 and plastochromanol-8 biosynthesis. In this work, direct evidence for chemical quenching of 1 O2 by plastoquinols and their oxidation products is presented, which is obtained by microscopic techniques invivo. Chemical quenching of 1 O2 was associated with consumption of PQH2 -9 and formation of its various oxidized forms. Oxidation of PQH2 -9 by 1 O2 leads to plastoquinone-9 (PQ-9), which is subsequently oxidized to hydroxyplastoquinone-9 [PQ(OH)-9]. We provide here evidence that oxidation of PQ(OH)-9 by 1 O2 results in the formation of trihydroxyplastoquinone-9 [PQ(OH)3 -9]. It is concluded here that PQH2 -9 serves as an efficient 1 O2 chemical quencher in Arabidopsis, and PQ(OH)3 -9 can be considered as a natural product of 1 O2 reaction with PQ(OH)-9. The understanding of the mechanisms underlying 1 O2 chemical quenching provides information on the role of plastoquinols and their oxidation products in the response of plants to photooxidative stress.

Biosynthesis and applications of prenylquinones.

Prenylquinones are isoprenoid compounds with a characteristic quinone structure and isoprenyl tail that are ubiquitous in almost all living organisms. There are four major prenylquinone classes: ubiquinone (UQ), menaquinone (MK), plastoquinone (PQ), and rhodoquinone (RQ). The quinone structure and isoprenyl tail length differ among organisms. UQ, PQ, and RQ contain benzoquinone, while MK contains naphthoquinone. UQ, MK, and RQ are involved in oxidative phosphorylation, while PQ functions in photosynthetic electron transfer. Some organisms possess two types of prenylquinones; Escherichia coli has UQ8 and MK8, and Caenorhabditis elegans has UQ9 and RQ9. Crystal structures of most of the enzymes involved in MK synthesis have been solved. Studies on the biosynthesis and functions of quinones have advanced recently, including for phylloquinone (PhQ), which has a phytyl moiety instead of an isoprenyl tail. Herein, the synthesis and applications of prenylquinones are reviewed.

Functional Role of Fibrillin5 in Acclimation to Photooxidative Stress.

The functional role of a lipid-associated soluble protein, fibrillin5 (FBN5), was determined with the Arabidopsis thaliana homozygous fbn5-knockout mutant line (SALK_064597) that carries a T-DNA insertion within the FBN5 gene. The fbn5 mutant remained alive, displaying a slow growth and a severe dwarf phenotype. The mutant grown even under growth light conditions at 80 µmol m-2 s-1 showed a drastic decrease in electron transfer activities around PSII, with little change in electron transfer activities around PSI, a phenomenon which was exaggerated under high light stress. The accumulation of plastoquinone-9 (PQ-9) was suppressed in the mutant, and >90% of the PQ-9 pool was reduced under growth light conditions. Non-photochemical quenching (NPQ) in the mutant functioned less efficiently, resulting from little contribution by energy-dependent quenching (qE). The ultrastructure of thylakoids in the mutant revealed that their grana were unstacked and transformed into loose and disordered structures. Light-harvesting complex (LHC)-containing large photosystem complexes and photosystem core complexes in the mutant were less abundant than those in wild-type plants. These results suggest that the lack of FBN5 causes a decrease in PQ-9 and imbalance of the redox state of PQ-9, resulting in misconducting both short-term and long-term control of the input of light energy to photosynthetic reaction centers. Furthermore, in the fbn5 mutant, the expression of genes involved in jasmonic acid biosynthesis was suppressed to ≤10% of that in the wild type under both growth-light and high-light conditions, suggesting that FBN5 functions as a transmitter of 1O2 in the stroma.

Fusarium solani Infection Depressed Photosystem Performance by Inducing Foliage Wilting in Apple Seedlings.

Fusarium fungi are soil-borne pathogens, and the pathological effects on plant photosystems remain unclear. This study aimed to deeply reveal pathological characterization in apple seedlings infected with Fusarium solani by investigating photosystems performance and interaction. Roots were immersed in conidial suspension for inoculation. Thereafter, prompt and delayed chlorophyll a fluorescence and modulated 820 nm reflection were simultaneously detected. After 30 days of infection, leaf relative water content and dry weight were remarkably decreased by 55.7 and 47.1%, suggesting that the infected seedlings were subjected to Fusarium-induced water deficit stress. PSI reaction center was more susceptible than PSII reaction center in infected seedlings due to greater decrease in the maximal photochemical efficiency of PSI than that of PSII, but PSI reaction center injury was aggravated slowly, as PSII injury could partly protect PSI by restricting electron donation. PSII donor and acceptor sides were also damaged after 20 days of infection, and the restricted electron donation induced PSII and PSI disconnection by blocking PSI re-reduction. In accordance with greater damage of PSI reaction center, PSI oxidation was also suppressed. Notably, significantly increased efficiency of electron transport from plastoquinone (PQ) to PSI acceptors (REo/ETo) after 20 days of infection suggested greater inhibition on PQ reduction than re-oxidation, and the protection for PSI acceptors might alleviate the reduction of electron transport efficiency beyond PQ upon damaged PSI reaction center. Lowered delayed fluorescence in microsecond domain verified PSII damage in infected seedlings, and elevated delayed fluorescence in sub-millisecond domain during PQ reduction process conformed to increased REo/ETo. In conclusion, F. solani infection depressed PSII and PSI performance and destroyed their coordination by inducing pathological wilting in apple seedlings. It may be a pathogenic mechanism of Fusarium to induce plant photosystems damage.

Reduction-Induced Suppression of Electron Flow (RISE) Is Relieved by Non-ATP-Consuming Electron Flow in Synechococcus elongatus PCC 7942.

Photosynthetic organisms oxidize P700 to suppress the production of reactive oxygen species (ROS) in photosystem I (PSI) in response to the lower efficiency of photosynthesis under high light and low CO2 conditions. Previously, we found a positive relationship between reduction of plastoquinone (PQ) pool and oxidation of P700, which we named reduction-induced suppression of electron flow (RISE). In the RISE model, we proposed that the highly reduced state of the PQ pool suppresses Q-cycle turnover to oxidize P700 in PSI. Here, we tested whether RISE was relieved by the oxidation of the PQ pool, but not by the dissipation of the proton gradient (ΔpH) across the thylakoid membrane. Formation of ΔpH can also suppress electron flow to P700, because acidification on the luminal side of the thylakoid membrane lowers oxidation of reduced PQ in the cytochrome b6/f complex. We drove photosynthetic electron transport using H2O2-scavenging peroxidase reactions. Peroxidase reduces H2O2 with electron donors regenerated along the photosynthetic electron transport system, thereby promoting the formation of ΔpH. Addition of H2O2 to the cyanobacterium Synechococcus elongatus PCC 7942 under low CO2 conditions induced photochemical quenching of chlorophyll fluorescence, enhanced NADPH fluorescence and reduced P700. Thus, peroxidase reactions relieved the RISE mechanism, indicating that P700 oxidation can be induced only by the reduction of PQ to suppress the production of ROS in PSI. Overall, our data suggest that RISE regulates the redox state of P700 in PSI in cooperation with ΔpH regulation.

Metabolic regulation of photosynthetic membrane structure tunes electron transfer function.

The photosynthetic chloroplast thylakoid membrane of higher plants is a complex three-dimensional structure that is morphologically dynamic on a timescale of just a few minutes. The membrane dynamics are driven by the phosphorylation of light-harvesting complex II (LHCII) by the STN7 kinase, which controls the size of the stacked grana region relative to the unstacked stromal lamellae region. Here, I hypothesise that the functional significance of these membrane dynamics is in controlling the partition of electrons between photosynthetic linear and cyclic electron transfer (LET and CET), which determines the ratio of NADPH/ATP produced. The STN7 kinase responds to the metabolic state of the chloroplast by sensing the stromal redox state. A high NADPH/ATP ratio leads to reduction of thioredoxin f (TRXf), which reduces a CxxxC motif in the stromal domain of STN7 leading to its inactivation, whereas a low NADPH/ATP ratio leads to oxidation of TRXf and STN7 activation. Phosphorylation of LHCII leads to smaller grana, which favour LET by speeding up diffusion of electron carriers plastoquinone (PQ) and plastocyanin (PC) between the domains. In contrast, dephosphorylation of LHCII leads to larger grana that slow the diffusion of PQ and PC, leaving the PQ pool in the stroma more oxidised, thus enhancing the efficiency of CET. The feedback regulation of electron transfer by the downstream metabolism is crucial to plant fitness, since perturbations in the NADPH/ATP ratio can rapidly lead to the inhibition of photosynthesis and photo-oxidative stress.

Maizew3disrupts homogentisate solanesyl transferase(ZmHst) and reveals a plastoquinone-9 independent path for phytoene desaturation and tocopherol accumulation in kernels.

Maize white seedling 3 (w3) has been used to study carotenoid deficiency for almost 100years, although the molecular basis of the mutation has remained unknown. Here we show that the w3 phenotype is caused by disruption of the maize gene for homogentisate solanesyl transferase (HST), which catalyzes the first and committed step in plastoquinone-9 (PQ-9) biosynthesis in the plastid. The resulting PQ-9 deficiency prohibits photosynthetic electron transfer and eliminates PQ-9 as an oxidant in the enzymatic desaturation of phytoene during carotenoid synthesis. As a result, light-grown w3 seedlings are albino, deficient in colored carotenoids and accumulate high levels of phytoene. However, despite the absence of PQ-9 for phytoene desaturation, dark-grown w3 seedlings can produce abscisic acid (ABA) and homozygous w3 kernels accumulate sufficient carotenoids to generate ABA needed for seed maturation. The presence of ABA and low levels of carotenoids in w3 nulls indicates that phytoene desaturase is able to use an alternate oxidant cofactor, albeit less efficiently than PQ-9. The observation that tocopherols and tocotrienols are modestly affected in w3 embryos and unaffected in w3 endosperm indicates that, unlike leaves, grain tissues deficient in PQ-9 are not subject to severe photo-oxidative stress. In addition to identifying the molecular basis for the maize w3 mutant, we: (1) show that low levels of phytoene desaturation can occur in w3 seedlings in the absence of PQ-9; and (2) demonstrate that PQ-9 and carotenoids are not required for vitamin E accumulation.

Surprise: The classic white seedling 3 mutant in maize lacks plastoquinone-9 but can still make carotenoids.

Surprise: The classic white seedling 3 mutant in maize lacks plastoquinone-9 but can still make carotenoids.

High anthocyanin accumulation in poinsettia leaves is accompanied by thylakoid membrane unstacking, acting as a photoprotective mechanism, to prevent ROS formation

In poinsettia (Euphorbia pulcherrima Willd. ex Klotzsch.), green leaves turn reddish and then red, due to vacuolar anthocyanin accumulation. Reddish leaves accumulate anthocyanins mainly in the adaxial (upper) epidermis, and less in the mesophyll cells, while red, in both adaxial and abaxial (lower) epidermides, and the adjacent mesophyll cells. In green leaves, the photoprotective mechanism of non-photochemical quenching (NPQ) is sufficient under low light (LL), but not under high light (HL), which results in a more reduced redox state of the plastoquinone (PQ) pool compared to reddish. In red leaves, higher anthocyanin accumulation is accompanied by unstacking of thylakoid membranes that results in loss of photosystem II (PSII) complexes and undetectable hydrogen peroxide (H2O2). Superoxide dismutase (SOD) activity is enhanced in the reddish compared to green leaves, while it decreases significantly in the red ones. Anthocyanin accumulation was significantly correlated to the redox state of the PQ pool and the higher accumulation in reddish compared to green leaves was responsible for the diminished H2O2 production, since ascorbate peroxidase (APX) activity remained unchanged in all leaves. We suggest that H2O2 production regulated by the redox state of the PQ pool induces anthocyanin biosynthesis that is accompanied by thylakoid membrane unstacking and loss of PSII complexes, serving as a photoprotective mechanism to HL, preventing the formation of reactive oxygen species (ROS).

Nanobrass CuZn Nanoparticles as Foliar Spray Nonphytotoxic Fungicides.

Inorganic nanoparticles (NPs) have been proposed as alternative fertilizers to suppress plant disease and increase crop yield. However, phytotoxicity of NPs remains a key factor for their massive employment in agricultural applications. In order to investigate new effective, nonphytotoxic, and inexpensive fungicides, in the present study CuZn bimetallic nanoparticles (BNPs) have been synthesized as antifungals, while assessment of photosystem II (PSII) efficiency by chlorophyll fluorescence imaging analysis is utilized as an effective and noninvasive phytotoxicity evaluation method. Thus, biocompatible coated, nonoxide contaminated CuZn BNPs of 20 nm crystallite size and 250 nm hydrodynamic diameter have been prepared by a microwave-assisted synthesis. BNPs' antifungal activity against Saccharomyces cerevisiae was found to be enhanced compared to monometallic Cu NPs. Reactive oxygen species (ROS) formation and photosystem II (PSII) functionality at low light (LL) and high light (HL) intensity were determined on tomato plants sprayed with 15 and 30 mg L-1 of BNPs for the evaluation of their phytotoxicity. Tomato leaves sprayed with 15 mg L-1 of BNPs displayed no significant difference in PSII functionality at LL, while exposure to 30 mg L-1 of BNPs for up to 90 min resulted in a reduced plastoquinone (PQ) pool that gave rise to H2O2 accumulation, initiating signaling networks and regulating acclimation responses. After 3 h of exposure to 30 mg L-1 of BNPs, PSII functionality at LL was similar to control, indicating nonphytotoxic effects. Meanwhile, exposure of tomato leaves either enhanced (15 mg L-1) or did not have any significant effect (30 mg L-1) on PSII functionality at HL, attributed to the absence of semiconducting oxide phases and photochemical toxicity-reducing modifications. The use of chlorophyll fluorescence imaging analysis is recommended as a tool to monitor NPs behavior on plants.

Photosystem I with benzoquinone analogues incorporated into the A1 binding site.

Time-resolved FTIR difference spectroscopy has been used to study photosystem I (PSI) particles with three different benzoquinones [plastoquinone-9 (PQ), 2,6-dimethyl-1,4-benzoquinone (DMBQ), 2,3,5,6-tetrachloro-1,4-benzoquinone (Cl4BQ)] incorporated into the A1 binding site. If PSI samples are cooled in the dark to 77K, the incorporated benzoquinones are shown to be functional, allowing the production of time-resolved (P700+A1--P700A1) FTIR difference spectra. If samples are subjected to repetitive flash illumination at room temperature prior to cooling, however, the time-resolved FTIR difference spectra at 77K display contributions typical of the P700 triplet state (3P700), indicating a loss of functionality of the incorporated benzoquinones, that occurs because of double protonation of the incorporated benzoquinones. The benzoquinone protonation mechanism likely involves nearby water molecules but does not involve the terminal iron-sulfur clusters FA and FB. These results and conclusions resolve discrepancies between results from previous low-temperature FTIR and EPR studies on similar PSI samples with PQ incorporated.

Zinc isotope fractionation by Emiliania huxleyi cultured across a range of free zinc ion concentrations

AbstractZinc (Zn) isotope fractionation by the coccolithophore Emiliania huxleyi (a Tasman Sea isolate) was investigated by culturing it across a range of free Zn2+ ion concentrations (1.2 pmol L−1 to 3.2 nmol L−1), which span the natural range observed in the global oceans. Across the range of free Zn2+ ion concentrations tested, the amount of bio‐available Zn did not have any appreciable influence on the specific growth rate or cell morphology of E. huxleyi. However, a suite of secondary physiological indicators reflected changes in Zn bioavailability. An increase in the photosynthetic efficiency of photosystem II (Fv/Fm) was observed with increasing free Zn2+ ion concentration. The time constants for electron transport from QA to QB and from QB to the plastoquinone (PQ) pool decreased as the free Zn2+ ion concentration increased, reflecting enhanced photosynthetic electron transport. The effect of Zn bioavailability on photosynthetic physiology was speculated to be due to increased activity of carbonic anhydrase with increasing Zn bioavailability. Zinc uptake by E. huxleyi resulted in Zn isotope fractionation and the Δ66Zncells‐media ranged between −0.6‰ and −0.2‰. The major finding from this work is that the lighter Zn isotope is taken up preferentially by this coccolithophore, making the dissolved Zn composition of the surrounding seawater isotopically heavier.