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SUMMARY: With climate change, an aggravation in summer drought is expected in the Mediterranean region. To assess the impact of such a future scenario, we compared the response of Quercus pubescens, a drought‐resistant deciduous oak species, to long‐term amplified drought (AD) (partial rain exclusion in natura for 10 years) and natural drought (ND). We studied leaf physiological and physico‐chemical trait responses to ND and AD over the seasonal cycle, with a focus on chemical traits including major groups of central (photosynthetic pigments and plastoquinones) and specialized (tocochromanols, phenolic compounds, and cuticular waxes) metabolites. Seasonality was the main driver of all leaf traits, including cuticular triterpenoids, which were highly concentrated in summer, suggesting their importance to cope with drought and thermal stress periods. Under AD, trees not only reduced CO2 assimilation (−42%) in summer and leaf concentrations of some phenolic compounds and photosynthetic pigments (carotenoids from the xanthophyll cycle) but also enhanced the levels of other photosynthetic pigments (chlorophylls, lutein, and neoxanthin) and plastochromanol‐8, an antioxidant located in chloroplasts. Overall, the metabolomic adjustments across seasons and drought conditions reinforce the idea that Q. pubescens is highly resistant to drought although significant losses of antioxidant defenses and photoprotection were identified under AD. Significance Statement: Climate change will increase drought episodes, especially in the Mediterranean region. This study investigated Quercus pubescens response to long‐term amplified drought (AD) in natura, focusing on leaf physiological and physico‐chemical traits across seasons and AD. It revealed that seasonal variations influenced leaf metabolism, including leaf cuticular compounds, which are important to cope with drought and thermal stress. Also, AD led to a decrease in net photosynthesis as well as losses in antioxidant defense and photoprotection. [ABSTRACT FROM AUTHOR]

Lipid peroxidation is a primary event associated with oxidative stress in plants. This phenomenon secondarily generates bioactive and/or toxic compounds such as reactive carbonyl species (RCS), phytoprostanes, and phytofurans, as confirmed here in Arabidopsis plants exposed to photo‐oxidative stress conditions. We analyzed the effects of exogenous applications of secondary lipid oxidation products on Arabidopsis plants by luminescence techniques. Oxidative damage to attached leaves was measured by autoluminescence imaging, using a highly sensitive CCD camera, and the activity of the detoxification pathway, dependent on the transcription regulator SCARECROW‐LIKE 14 (SCL14), was monitored with a bioluminescent line expressing the firefly LUCIFERASE (LUC) gene under the control of the ALKENAL REDUCTASE (AER) gene promoter. We identified 4‐hydroxynonenal (HNE), and to a lesser extent 4‐hydroxyhexenal (HHE), as highly reactive compounds that are harmful to leaves and can trigger AER gene expression, contrary to other RCS (pentenal, hexenal) and to isoprostanoids. Although the levels of HNE and other RCS were enhanced in the SCL14‐deficient mutant (scl14), exogenously applied HNE was similarly damaging to this mutant, its wild‐type parent and a SCL14‐overexpressing transgenic line (OE:SCL14). However, strongly boosting the SCL14 detoxification pathway and AER expression by a pre‐treatment of OE:SCL14 with the signaling apocarotenoid β‐cyclocitral canceled the damaging effects of HNE. Conversely, in the scl14 mutant, the effects of β‐cyclocitral and HNE were additive, leading to enhanced leaf damage. These results indicate that the cellular detoxification pathway induced by the low‐toxicity β‐cyclocitral targets highly toxic compounds produced during lipid peroxidation, reminiscent of a safener‐type mode of action. [ABSTRACT FROM AUTHOR]

Summary: Singlet oxygen (1O2) is a by‐product of photosynthesis that triggers a signalling pathway leading to stress acclimation or to cell death. By analyzing gene expressions in a 1O2‐overproducing Arabidopsis mutant (ch1) under different light regimes, we show here that the 1O2 signalling pathway involves the endoplasmic reticulum (ER)‐mediated unfolded protein response (UPR). ch1 plants in low light exhibited a moderate activation of UPR genes, in particular bZIP60, and low concentrations of the UPR‐inducer tunicamycin enhanced tolerance to photooxidative stress, together suggesting a role for UPR in plant acclimation to low 1O2 levels. Exposure of ch1 to high light stress ultimately leading to cell death resulted in a marked upregulation of the two UPR branches (bZIP60/IRE1 and bZIP28/bZIP17). Accordingly, mutational suppression of bZIP60 and bZIP28 increased plant phototolerance, and a strong UPR activation by high tunicamycin concentrations promoted high light‐induced cell death. Conversely, light acclimation of ch1 to 1O2 stress put a limitation in the high light‐induced expression of UPR genes, except for the gene encoding the BIP3 chaperone, which was selectively upregulated. BIP3 deletion enhanced Arabidopsis photosensitivity while plants treated with a chemical chaperone exhibited enhanced phototolerance. In conclusion, 1O2 induces the ER‐mediated UPR response that fulfils a dual role in high light stress: a moderate UPR, with selective induction of BIP3, is part of the acclimatory response to 1O2, and a strong activation of the whole UPR is associated with cell death. Significance Statement: Singlet oxygen, produced from photosystem II under conditions of excess light energy, triggers the endoplasmic reticulum‐mediated unfolded protein response (UPR), with different UPR levels inducing different physiological responses to light stress. Moderate activation of UPR participates in the acclimation to singlet oxygen, while strong activation leads to cell death. [ABSTRACT FROM AUTHOR]

Summary: Stressful environmental conditions lead to the production of reactive oxygen species in the chloroplasts, due to limited photosynthesis and enhanced excitation pressure on the photosystems. Among these reactive species, singlet oxygen (1O2), which is generated at the level of the PSII reaction center, is very reactive, readily oxidizing macromolecules in its immediate surroundings, and it has been identified as the principal cause of photooxidative damage in plant leaves. The two β‐carotene molecules present in the PSII reaction center are prime targets of 1O2 oxidation, leading to the formation of various oxidized derivatives. Plants have evolved sensing mechanisms for those PSII‐generated metabolites, which regulate gene expression, putting in place defense mechanisms and alleviating the effects of PSII‐damaging conditions. A new picture is thus emerging which places PSII as a sensor and transducer in plant stress resilience through its capacity to generate signaling metabolites under excess light energy. This review summarizes new advances in the characterization of the apocarotenoids involved in the PSII‐mediated stress response and of the pathways elicited by these molecules, among which is the xenobiotic detoxification. [ABSTRACT FROM AUTHOR]

Abstract: The capacity of a Quercus pubescens forest to resist recurrent drought was assessed on an in situ experimental platform through the measurement of a large set of traits (ecophysiological and metabolic) studied under natural drought (ND) and amplified drought (AD) induced by partial rain exclusion. This study was performed during the third and fourth years of AD, which correspond to conditions of moderate AD in 2014 and harsher AD in 2015, respectively. Although water potential (Ψ) and net photosynthesis (Pn) were noticeably reduced under AD in 2015 compared to ND, trees showed similar growth and no oxidative stress. The absence of oxidative damage could be due to a strong accumulation of α‐tocopherol, suggesting that this compound is a major component of the Q. pubescens antioxidant system. Other antioxidants were rather stable under AD in 2014, but slight changes started to be observed in 2015 (carotenoids and isoprene) due to harsher conditions. Our results indicate that Q. pubescens could be able to cope with AD, for at least 4 years, likely due to its antioxidant system. However, growth decrease was observed during the fifth year (2016) of AD, suggesting that this resistance could be threatened over longer periods of recurrent drought. [ABSTRACT FROM AUTHOR]

Abstract: The Arabidopsis vte1 mutant is devoid of tocopherol and plastochromanol (PC‐8). When exposed to excess light energy, vte1 produced more singlet oxygen (1O2) and suffered from extensive oxidative damage compared with the wild type. Here, we show that overexpressing the solanesyl diphosphate synthase 1 (SPS1) gene in vte1 induced a marked accumulation of total plastoquinone (PQ‐9) and rendered the vte1 SPS1oex plants tolerant to photooxidative stress, indicating that PQ‐9 can replace tocopherol and PC‐8 in photoprotection. High total PQ‐9 levels were associated with a noticeable decrease in 1O2 production and higher levels of Hydroxyplastoquinone (PQ‐C), a 1O2‐specific PQ‐9 oxidation product. The extra PQ‐9 molecules in the vte1 SPS1oex plants were stored in the plastoglobules and the chloroplast envelopes, rather than in the thylakoid membranes, whereas PQ‐C was found almost exclusively in the thylakoid membranes. Upon exposure of wild‐type plants to high light, the thylakoid PQ‐9 pool decreased, whereas the extrathylakoid pool remained unchanged. In vte1 and vte1 SPS1oex plants, the PQ‐9 losses in high light were strongly amplified, affecting also the extrathylakoid pool, and PQ‐C was found in high amounts in the thylakoids. We conclude that the thylakoid PQ‐9 pool acts as a 1O2 scavenger and is replenished from the extrathylakoid stock. [ABSTRACT FROM AUTHOR]

Summary: Prenylquinols (tocochromanols and plastoquinols) serve as efficient physical and chemical quenchers of singlet oxygen (1O2) formed during high light stress in higher plants. Although quenching of 1O2 by prenylquinols has been previously studied, direct evidence for chemical quenching of 1O2 by plastoquinols and their oxidation products is limited in vivo. In the present study, the role of plastoquinol‐9 (PQH2‐9) in chemical quenching of 1O2 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 1O2 by plastoquinols and their oxidation products is presented, which is obtained by microscopic techniques in vivo. Chemical quenching of 1O2 was associated with consumption of PQH2‐9 and formation of its various oxidized forms. Oxidation of PQH2‐9 by 1O2 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 1O2 results in the formation of trihydroxyplastoquinone‐9 [PQ(OH)3‐9]. It is concluded here that PQH2‐9 serves as an efficient 1O2 chemical quencher in Arabidopsis, and PQ(OH)3‐9 can be considered as a natural product of 1O2 reaction with PQ(OH)‐9. The understanding of the mechanisms underlying 1O2 chemical quenching provides information on the role of plastoquinols and their oxidation products in the response of plants to photooxidative stress. [ABSTRACT FROM AUTHOR]

Singlet oxygen (1O2) signalling in plants is essential to trigger both acclimatory mechanisms and programmed cell death under high light stress. However, because of its chemical features, 1O2 requires mediators, and the players involved in this pathway are largely unknown. The β-carotene oxidation product, β-cyclocitral, is one such mediator. Produced in the chloroplast, β-cyclocitral induces changes in nuclear gene expression leading to photoacclimation. Recently, the METHYLENE BLUE SENSITIVITY protein MBS has been identified as a key player in 1O2 signalling leading to tolerance to high light. Here, we provide evidence that MBS1 is essential for acclimation to 1O2 and cross-talks with β-cyclocitral to mediate transfer of the 1O2 signal to the nucleus, leading to photoacclimation. The presented results position MBS1 downstream of β-cyclocitral in 1O2 signalling and suggest an additional role for MBS1 in the regulation of plant growth and development under chronic 1O2 production. [ABSTRACT FROM AUTHOR]

Carotenoids are known to play important roles in plants as antioxidants, accessory light-harvesting pigments, and attractants for pollinators and seed dispersers. A new function for carotenoids has recently emerged, which relates to the response of plants to environmental stresses. Reactive oxygen species, especially singlet oxygen, produced in the chloroplasts under stress conditions, can oxidize carotenoids leading to a variety of oxidized products, including aldehydes, ketones, endoperoxides and lactones. Some of those carotenoid derivatives, such as volatile β-cyclocitral, derived from the oxidation of β-carotene, are reactive electrophile species that are bioactive and can induce changes in gene expression leading to acclimation to stress conditions. This review summarizes the current knowledge on the non-enzymatic oxidation of carotenoids, the bioactivity of the resulting cleavage compounds and their functions as stress signals in plants. [ABSTRACT FROM AUTHOR]

Lipocalins are a group of multifunctional proteins, recognized as carriers of small lipophilic molecules, which have been characterized in bacteria and animals. Two true lipocalins have been recently identified in plants, the temperature-induced lipocalin ( TIL) and the chloroplastic lipocalin ( CHL), the expression of which is induced by various abiotic stresses. Each lipocalin appeared to be specialized in the responses to specific stress conditions in Arabidopsis thaliana, with AtTIL and AtCHL playing a protective role against heat and high light, respectively. The double mutant AtCHL KO × AtTIL KO deficient in both lipocalins was more sensitive to temperature, drought and light stresses than the single mutants, exhibiting intense lipid peroxidation. AtCHL deficiency dramatically enhanced the photosensitivity of mutants ( vte1, npq1) affected in lipid protection mechanisms (tocopherols, zeaxanthin), confirming the role of lipocalins in the prevention of lipid peroxidation. Seeds of the AtCHL KO × AtTIL KO double mutant were very sensitive to natural and artificial ageing, and again this phenomenon was associated with the oxidation of polyunsaturated lipids. The presented results show that the Arabidopsis lipocalins AtTIL and AtCHL have overlapping functions in lipid protection which are essential for stress resistance and survival. [ABSTRACT FROM AUTHOR]

C4 plants display higher cyclic electron transport activity than C3 plants. This activity is suggested to be important for the production of ATPs required for C4 metabolism., To understand the process by which photosystem I ( PSI) cyclic electron transport was promoted during C4 evolution, we conducted comparative analyses of the functionality of PSI cyclic electron transport among members of the genus Flaveria, which contains several C3, C3- C4 intermediate, C4-like and C4 species., The abundance of NDH- H, a subunit of NADH dehydrogenase-like complex, increased markedly in bundle sheath cells with the activity of the C4 cycle. By contrast, PROTON GRADIENT REGULATION5 ( PGR5) and PGR5- LIKE1 increased in both mesophyll and bundle sheath cells in C4-like Flaveria palmeri and C4 species. Grana stacks were drastically reduced in bundle sheath chloroplasts of C4-like F. palmeri and C4 species; these species showed a marked increase in PSI cyclic electron transport activity., These results suggest that both the expression of proteins involved in PSI cyclic electron transport and changes in thylakoid structure contribute to the high activity of cyclic electron flow in NADP-malic enzyme-type C4 photosynthesis. We propose that these changes were important for the establishment of C4 photosynthesis from C3- C4 intermediate photosynthesis in Flaveria. [ABSTRACT FROM AUTHOR]

Summary Plants, like almost all living organisms, spontaneously emit photons of visible light. We used a highly sensitive, low-noise cooled charge coupled device camera to image spontaneous photon emission (autoluminescence) of plants. Oxidative stress and wounding induced a long-lasting enhancement of plant autoluminescence, the origin of which is investigated here. This long-lived phenomenon can be distinguished from the short-lived chlorophyll luminescence resulting from charge recombinations within the photosystems by pre-adapting the plant to darkness for about 2 h. Lipids in solvent were found to emit a persistent luminescence after oxidation in vitro, which exhibited the same time and temperature dependence as plant autoluminescence. Other biological molecules, such as DNA or proteins, either did not produce measurable light upon oxidation or they did produce a chemiluminescence that decayed rapidly, which excludes their significant contribution to the in vivo light emission signal. Selective manipulation of the lipid oxidation levels in Arabidopsis mutants affected in lipid hydroperoxide metabolism revealed a causal link between leaf autoluminescence and lipid oxidation. Addition of chlorophyll to oxidized lipids enhanced light emission. Both oxidized lipids and plants predominantly emit light at wavelengths higher than 600 nm; the emission spectrum of plant autoluminescence was shifted towards even higher wavelengths, a phenomenon ascribable to chlorophyll molecules acting as luminescence enhancers in vivo. Taken together, the presented results show that spontaneous photon emission imaged in plants mainly emanates from oxidized lipids. Imaging of this signal thus provides a simple and sensitive non-invasive method to selectively visualize and map patterns of lipid oxidation in plants. [ABSTRACT FROM AUTHOR]

Methionine oxidation to methionine sulfoxide (MetSO) is reversed by two types of methionine sulfoxide reductases (MSRs), A and B, specific to MetSO S- and R-diastereomers, respectively. Two MSRB isoforms, MSRB1 and MSRB2, are present in chloroplasts of Arabidopsis thaliana. To assess their physiological role, we characterized Arabidopsis mutants knockout for the expression of MSRB1, MSRB2 or both genes. Measurements of MSR activity in leaf extracts revealed that the two plastidial MSRB enzymes account for the major part of leaf peptide MSR capacity. Under standard conditions of light and temperature, plants lacking one or both plastidial MSRBs do not exhibit any phenotype, regarding growth and development. In contrast, we observed that the concomitant absence of both proteins results in a reduced growth for plants cultivated under high light or low temperature. In contrast, double mutant lines restored for MSRB2 expression display no phenotype. Under environmental constraints, the MetSO level in leaf proteins is higher in plants lacking both plastidial MSRBs than in Wt plants. The absence of plastidial MSRBs is associated with an increased chlorophyll a/b ratio, a reduced content of Lhca1 and Lhcb1 proteins and an impaired photosynthetic performance. Finally, we show that MSRBs are able to use as substrates, oxidized cpSRP43 and cpSRP54, the two main components involved in the targeting of Lhc proteins to the thylakoids. We propose that plastidial MSRBs fulfil an essential function in maintaining vegetative growth of plants during environmental constraints, through a role in the preservation of photosynthetic antennae. [ABSTRACT FROM AUTHOR]

Lipocalins are small ligand-binding proteins with a simple tertiary structure that gives them the ability to bind small, generally hydrophobic, molecules. Recent studies have shown that animal lipocalins play important roles in the regulation of developmental processes and are involved in tolerance to oxidative stress. Plants also possess various types of lipocalins, and bioinformatics analyses have predicted that some lipocalin members may be present in the chloroplast. Here we report the functional characterization of the Arabidopsis thaliana chloroplastic lipocalin AtCHL. Cellular fractionation showed that AtCHL is a thylakoid lumenal protein. Drought, high light, paraquat and abscisic acid treatments induce AtCHL transcript and protein accumulation. Under normal growth conditions, knockout (KO) and over-expressing (OEX) lines do not differ from wild-type plants in terms of phenotype and photosynthetic performance. However, KO plants, which do not accumulate AtCHL, show more damage upon photo-oxidative stress induced by drought, high light or paraquat. In contrast, a high level of AtCHL allows OEX plants to cope better with these stress conditions. When exposed to excess light, KO plants display a rapid accumulation of hydroxy fatty acids relative to the wild-type, whereas the lipid peroxidation level remains very low in OEX plants. The increased lipid peroxidation in KO plants is mediated by singlet oxygen and is not correlated with photo-inhibition of the photosystems. This work provides evidence suggesting that AtCHL is involved in the protection of thylakoidal membrane lipids against reactive oxygen species, especially singlet oxygen, produced in excess light. [ABSTRACT FROM AUTHOR]

Non-ribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs) are necessary for the production of a variety of secondary metabolites, such as siderophores involved in iron acquisition. In response to iron limitation, the cyanobacterium Anabaena sp. strain PCC 7120 synthesizes several siderophores. The chromosome of this organism contains a large gene cluster of 76 kb with 24 open-reading frames from all2658 to all2635, including those that encode seven NRPSs and two PKSs. The function of this gene cluster was unknown, and one possibility could be the synthesis of siderophores. These genes were indeed activated under conditions of iron limitation. One mutant, MΔ41–49, bearing a large deletion of 43.4 kb in this gene cluster, synthesized considerably less siderophores and contained less iron as compared with the wild type. Its growth rate was similar to the wild type in the presence of iron, but was reduced when iron became limiting. Two other mutants, MΔ44–45 and MΔ47–49, lacking either all2644 and all2645, or all2647, all2648 and all2649 respectively, produced more siderophores than MΔ41–49, but less than the wild type. These genes were also activated under oxidative stress conditions to which MΔ41–49 was highly sensitive, consistent with the importance of iron in oxidative stress response. We propose that this gene cluster is involved in the synthesis of siderophores in Anabaena sp. PCC 7120 and plays an important role in defence against oxidative stress. [ABSTRACT FROM AUTHOR]

Arabidopsis ( Arabidopsis thaliana) plants were grown in a hydroponic culture system for 7 to 14 d in the absence or presence of 75 µM Cd or 75 µM Cu. The Cu treatment resulted in visual leaf symptoms, together with anthocyanin accumulation and loss of turgor. Pronounced lipid peroxidation, which was detected by autoluminescence imaging and malondialdehyde titration, was observed in Cu-treated leaves. The Cd treatment also resulted in loss of leaf pigments but lipid peroxidation and oxidative stress were less pronounced than in the leaves exposed to Cu. Analysis of low-molecular-weight chloroplast and cytosolic antioxidants (ascorbate, glutathione, tocopherols, carotenoids) and antioxidant enzymes (thiol-based reductases and peroxidases) revealed relatively few responses to metal exposure. However, there was a marked increase in vitamin E ( α-tocopherol) in response to Cd and Cu treatments. Ascorbate increased significantly in Cu-exposed leaves. Other antioxidants either remained stable or decreased in response to metal stress. Transcripts encoding enzymes of the vitamin E biosynthetic pathway were increased in response to metal exposure. In particular, VTE2 mRNA was enhanced in Cu- and Cd-treated plants, while VTE5 and hydroxylpyruvate dioxygenase (HPPD) mRNAs were only up-regulated in Cd-treated plants. Consistent increases in HPPD transcripts and protein were observed. The vitamin E-deficient ( vte1) mutant exhibited an enhanced sensitivity towards both metals relative to the wild-type (WT) control. Unlike the vte1 mutants, which showed enhanced lipid peroxidation and oxidative stress in the presence of Cu or Cd, the ascorbate-deficient ( vtc2) mutant showed WT responses to metal exposure. Taken together, these results demonstrate that vitamin E plays a crucial role in the tolerance of Arabidopsis to oxidative stress induced by heavy metals such as Cu and Cd. [ABSTRACT FROM AUTHOR]

The early light-induced proteins (ELIPs) belong to the multigenic family of pigment-binding light-harvesting complexes. ELIPs accumulate transiently and are believed to play a protective role in plants exposed to high levels of light. Constitutive expression of the ELIP2 gene in Arabidopsis resulted in a marked reduction of the pigment content of the chloroplasts, both in mature leaves and during greening of etiolated seedlings. The chlorophyll loss was associated with a decrease in the number of photosystems in the thylakoid membranes, but the photosystems present were fully assembled and functional. A detailed analysis of the chlorophyll-synthesizing pathway indicated that ELIP2 accumulation downregulated the level and activity of two important regulatory steps: 5-aminolevulinate synthesis and Mg-protoporphyrin IX (Mg-Proto IX) chelatase activity. The contents of glutamyl tRNA reductase and Mg chelatase subunits CHLH and CHLI were lowered in response to ELIP2 accumulation. In contrast, ferrochelatase activity was not affected and the inhibition of Heme synthesis was null or very moderate. As a result of reduced metabolic flow from 5-aminolevulinic acid, the steady state levels of various chlorophyll precursors (from protoporphyrin IX to protochlorophyllide) were strongly reduced in the ELIP2 overexpressors. Taken together, our results indicate that the physiological function of ELIPs could be related to the regulation of chlorophyll concentration in thylakoids. This seems to occur through an inhibition of the entire chlorophyll biosynthesis pathway from the initial precursor of tetrapyrroles, 5-aminolevulinic acid. We suggest that ELIPs work as chlorophyll sensors that modulate chlorophyll synthesis to prevent accumulation of free chlorophyll, and hence prevent photooxidative stress. [ABSTRACT FROM AUTHOR]

The 2-cysteine peroxiredoxins (2-Cys-Prxs) are antioxidants that reduce peroxides through a thiol-based mechanism. During catalysis, these ubiquitous enzymes are occasionally inactivated by the substrate-dependent oxidation of the catalytic cysteine to the sulfinic acid (-SO2H) form, and are reactivated by reduction by sulfiredoxin (Srx), an enzyme recently identified in yeast and in mammal cells. In plants, 2-Cys-Prxs constitute the most abundant Prxs and are located in chloroplasts. Here we have characterized the unique Srx gene in Arabidopsis thaliana ( AtSrx) from a functional point of view, and analyzed the phenotype of two AtSrx knockout ( AtSrx–) mutant lines. AtSrx is a chloroplastic enzyme displaying sulfinic acid reductase activity, as shown by the ability of the recombinant AtSrx to reduce the overoxidized 2-Cys-Prx form in vitro, and by the accumulation of the overoxidized Prx in mutant lines lacking Srx in vivo. Furthermore, AtSrx mutants exhibit an increased tolerance to photooxidative stress generated by high light combined with low temperature. These data establish that, as in yeast and in mammals, plant 2-Cys-Prxs are subject to substrate-mediated inactivation reversed by Srx, and suggest that the 2-Cys-Prx redox status and sulfiredoxin are parts of a signaling mechanism participating in plant responses to oxidative stress. [ABSTRACT FROM AUTHOR]

Abstract: The products of the isiAB operon are a chlorophyll antenna protein (IsiA) and flavodoxin (IsiB), which accumulate in cyanobacteria grown under iron starvation conditions. Here we show that strong light triggers de-repression of isiAB transcription and leads to IsiA and flavodoxin accumulation under iron replete conditions. Genetic deletion of isiAB resulted in a photosensitive phenotype, with accumulation of reactive oxygen species and cell bleaching in high light, while the flavodoxin-deficient isiB null mutant expressing isiA was phototolerant. We conclude that IsiA protects cyanobacteria from photooxidative stress. IsiA is the first example of a chlorophyll antenna protein outside the extended LHC family that is induced transiently by high light and that fulfills a photoprotective role. [Copyright &y& Elsevier]

A psaFJ-null mutant of Synechocystis sp. strain PCC 6803 was characterised. As opposed to similar mutants in chloroplasts of green algae, electron transfer from plastocyanin to photosystem 1 was not affected. Instead, a restraint in full chain photosynthetic electron transfer was correlated to malfunction of photosystem 1 at its stromal side. Our hypothesis is that absence of PsaF causes oxidative stress, which triggers the induction of the ‘iron stress inducible’ operon isiAB. Products are the IsiA chlorophyll-binding protein (CP43′) and the isiB gene product flavodoxin. Supporting evidence was obtained by similar isiAB induction in wild type cells artificially exposed to oxidative stress. [Copyright &y& Elsevier]

Summary Biochemical and genetic studies have established that the light-harvesting chlorophyll proteins (LHCPs) of the photosystems use the cpSRP (chloroplast signal recognition particle) pathway for their targeting to thylakoids. Previous analyses of single cpSRP mutants, chaos and ffc, deficient in cpSRP43 and cpSRP54, respectively, have revealed that half of the LHCPs are still integrated into the thylakoid membranes. Surprisingly, the effects of both mutations are additive in the double mutant ffc/chaos described here. This mutant has pale yellow leaves at all stages of growth and drastically reduced levels of all the LHCPs except Lhcb 4. Although the chloroplasts have a normal shape, the thylakoid structure is affected by the mutation, probably as a consequence of reduction of all the LHCPs. ELIPs (early light-inducible proteins), nuclear-encoded proteins related to the LHCP family and inducible by light stress, were also drastically reduced in the double mutant. However, proteins targeted by other chloroplastic targeting pathways (ΔpH, Sec and spontaneous pathways) accumulated to similar levels in the wild-type and the double mutant. Therefore, the near total loss of LHCPs and ELIPs in the double mutant suggests that cpSRP is the predominant, if not exclusive, targeting pathway for these proteins. Phenotypic analysis of the double mutant, compared to the single mutants, suggests that the cpSRP subunits cpSRP43 and cpSRP54 contribute to antenna targeting in an independent but additive way. [ABSTRACT FROM AUTHOR]

We investigated the role of non-photochemical energy quenching (NPQ) in cold acclimation in potato. We first analyzed the expression of the PsbS gene, which encodes a PSII subunit involved in NPQ, during chilling treatment in two potato species, a cold-tolerant Solanum sogarandinum and a cold-sensitive Solanum tuberosum (cv. Cisa). In in vitro plantlets, a transient transcript accumulation was observed after 1 h in the light at room temperature in both species, and this light-induced PsbS transcript accumulation was strongly amplified at 4°C. Nuclear run-off transcription experiments indicated that this increase likely originates from a higher transcriptional activity of PsbS gene. In phytotron-grown plants, chilling treatment was shown also to result in a substantial increase in PsbS mRNA level. However, no change in protein abundance was noticed in either Solanum species. PSII photochemistry and photosynthetic electron transport were severely decreased in S. tuberosum plants at low temperature, while both activities were only slightly affected in S. sogarandinum. NPQ was substantially reduced in both species during chilling stress. These results indicate that neither PsbS nor NPQ are involved in acclimation of S. sogarandinum to low temperature. In contrast, the level of two other plastid proteins, one related to thioredoxins, CDSP32, and the other homologous to plastid lipid-associated proteins, CDSP34 (for chloroplastic drought-induced stress proteins of 32 and 34 kDa, respectively), was higher at low temperature in the cold-tolerant species. This result is discussed in relation to the potential roles of CDSPs in the protection of photosynthetic structures. [ABSTRACT FROM AUTHOR]

The salt-regulated accumulation of flavodoxin encoded by the isiB gene and its possible function were investigated in the cyanobacterium Synechocystis sp. strain PCC 6803. In Northern blot experiments, a slight increase of the isiB-specific mRNA was observed in salt-shocked and salt-acclimated cells. High levels of flavodoxin protein were detected in cells acclimated to 342 mM NaCl. In order to analyze the function of flavodoxin in cyanobacterial salt acclimation, an insertion null mutant of isiB was constructed. It was possible to adapt this mutant to raised salt concentrations and, as expected, to low iron contents. Salt-acclimated cells of wild type (WT) Synechocystis display increased activity of photosystem I (PSI), primarily used for increased cyclic electron transport capacity (Jeanjean et al. 1993, Plant Cell Physiol 34: 1073–1079). In salt-acclimated cells of the flavodoxin null mutant, the level of cyclic electron flow was lower than in wild type cells. It was concluded that flavodoxin plays a role as an alternative electron carrier, used for cyclic electron flow in salt-treated Synechocystis cells. [ABSTRACT FROM AUTHOR]

A slow water stress over several days was imposed on tobacco plants (Nicotiana tabacum L. var. Xanthi) by withholding water from the soil. Photosynthesis was measured in leaves from those water-stressed plants by the photoacoustic method. Slow drought induced marked changes in the photoacoustic signals, which were largely similar to those observed previously in leaves subjected to rapid desiccation in air (over 3-4 h), reflecting two simultaneous changes: 1) Modification of the heat and oxygen diffusion characteristics of the leaves due to changes in their anatomical structure [shown by the change in the slope of the plot of the oxygen (AOX) to photothermal signal (APT) ratio vs the square root of the modulation frequency]; 2) Inhibition of gross photosynthesis measured by the extrapolation of the AOX/APT ratio to zero frequency. However, in contrast to rapid water stress in detached leaves, where it was shown that mainly the oxidizing side of photosystem II (PS II) was damaged, we found a slower and more complex phenomenology having largely biphasic kinetics. During the first 6 days, there was a strong reduction in the photochemical energy storage, but the inhibition of oxygen evolution was relatively mild. The Emerson enhancement in state 1 dropped considerably, indicating lowering of the apparent absorption cross-section of PS II. Fluorescence measurements suggest that PS II reaction center itself maybe the primary site of the damage. PS I activity, judged by cytochrome f photooxidation, remained largely intact. The subsequent days were associated with a further spectacular decrease in the oxygen evolution quantum yield with both photosystems damaged. The photochemical energy storage continued to decrease further. The Emerson enhancement ratio of the remaining activities in both state 1 and 2 showed a marked increase, indicating the reestablishment of a strong imbalance in the distribution of excitation energy within the photochemical apparatus in favor of PS II. All the photoacoustic changes observed in response to drought were completely reversible within 2-3 days upon rewatering of the soil. [ABSTRACT FROM AUTHOR]

Tadmor is a Syrian barley landrace that has adapted to semi-arid environments. Its leaves are pale green because of a 30% decrease in the chlorophyll and the carotenoid content of the chloroplasts (leading to a 7·5% decrease in light absorption) compared with barley genotypes that are not adapted to harsh Mediterranean climatic conditions (e.g. Plaisant). This difference in pigment content was attenuated during growth of the plants in strong light, but was strongly amplified when strong light was combined with a high growth temperature. The low pigment content of Tadmor leaves was not associated with significant changes in the pigment distribution between the photosystems or between the reaction centres of the photosystems and their associated chlorophyll antennae. No significant difference in the photosynthetic activity (O2 production per unit absorbed light) was observed between Tadmor and Plaisant. The conversion of violaxanthin to zeaxanthin in strong light and its reversal in darkness were much faster and operated at a higher capacity in Tadmor leaves compared with Plaisant leaves, resulting in an increased photostability of photosystem II in the former leaves. The accelerated xanthophylls interconversion in the Syrian landrace was associated with, and possibly related to, an increased fluidity of the thylakoid membranes. The lipid peroxide level was lower in Tadmor compared with Plaisant. In contrast, no difference was found in the non-photochemical quenching of chlorophyll fluorescence between the two barley genotypes. The data indicate that the pale green Syrian landrace is equipped to survive excessive irradiance through a passive reduction of the light absorptance of its leaves, which mitigates the heating effects of strong light, and through the active protection of its photochemical apparatus by a rapid xanthophyll cycling. [ABSTRACT FROM AUTHOR]

Ionising γ radiation produces reactive oxygen species by water radiolysis, providing an interesting model approach for studying oxidative stress in plants. Three‐week old plants of Arabidopsis thaliana were exposed to a low dose rate (25 mGy h−1) of γ radiation for up to 21 days. This treatment had no effect on plant growth and morphology, but it induced chronic oxidation of lipids which was associated with an accumulation of reactive carbonyl species (RCS). However, contrary to lipid peroxidation, lipid RCS accumulation was transient only, being maximal after 1 day of irradiation and decreasing back to the initial level during the subsequent days of continuous irradiation. This indicates the induction of a carbonyl‐metabolising process during chronic ionising radiation. Accordingly, the γ‐radiation treatment induced the expression of xenobiotic detoxification‐related genes (AER, SDR1, SDR3, ALDH4, and ANAC102). The transcriptomic response of some of those genes (AER, SDR1, and ANAC102) was deregulated in the tga256 mutant affected in three TGAII transcription factors, leading to enhanced and/or prolonged accumulation of RCS and to a marked inhibition of plant growth during irradiation compared to the wild type. These results show that Arabidopsis is able to acclimate to chronic oxidative stress and that this phenomenon requires activation of a carbonyl detoxification mechanism controlled by TGAII. This acclimation did not occur when plants were exposed to an acute γ radiation stress (100 Gy) which led to persistent accumulation of RCS and marked inhibition of plant growth. This study shows the role of secondary products of lipid peroxidation in the detrimental effects of reactive oxygen species. [ABSTRACT FROM AUTHOR]