RESUMO
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.
Assuntos
Quercus , Quercus/metabolismo , Antioxidantes/metabolismo , Estações do Ano , Florestas , Chuva , Folhas de Planta/metabolismo , Árvores/metabolismo , Secas , Água/metabolismoRESUMO
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.
RESUMO
Arabidopsis plants were grown from seeds at different photon flux densities (PFDs) of white light ranging from 65 to 800 µmol photons m-2 s-1. Increasing PFD brought about a marked accumulation of plastoquinone (PQ) in leaves. However, the thylakoid photoactive PQ pool, estimated to about 700 pmol mg-1 leaf dry weight, was independent of PFD; PQ accumulation in high light mostly occurred in the photochemically non-active pool (plastoglobules, chloroplast envelopes) which represented up to 75% of total PQ. The amounts of PSII reaction center (on a leaf dry weight basis) also were little affected by PFD during growth, leading to a constant PQ/PSII ratio at all PFDs. Boosting PQ biosynthesis by overexpression of a solanesyl diphosphate-synthesizing enzyme strongly enhanced the PQ levels, particularly at high PFDs. Again, this accumulation occurred exclusively in the non-photoactive PQ pool. Mutational suppression of the plastoglobular ABC1K1 kinase led to a selective reduction of the thylakoid PQ pool size to ca. 400 pmol mg-1 in a large range of PFDs, which was associated with a restriction of the photosynthetic electron flow. Our results show that photosynthetic acclimation to light intensity does not involve modulation of the thylakoid PQ pool size or the amounts of PSII reaction centers. There appears to be a fixed amount of PQ molecules for optimal interaction with PSII and efficient photosynthesis, with the extra PQ molecules being stored outside the thylakoid membranes, implying a tight regulation of PQ distribution within the chloroplasts.
Assuntos
Arabidopsis , Plastoquinona , Aclimatação , Arabidopsis/metabolismo , Transporte de Elétrons , Homeostase , Luz , Oxirredução , Fotossíntese/fisiologia , Complexo de Proteína do Fotossistema II/metabolismo , Plastoquinona/metabolismo , Tilacoides/metabolismoRESUMO
Singlet oxygen (1 O2 ) 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 1 O2 -overproducing Arabidopsis mutant (ch1) under different light regimes, we show here that the 1 O2 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 1 O2 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 1 O2 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, 1 O2 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 1 O2 , and a strong activation of the whole UPR is associated with cell death.
Assuntos
Arabidopsis/metabolismo , Retículo Endoplasmático/metabolismo , Transdução de Sinais , Oxigênio Singlete/metabolismo , Resposta a Proteínas não Dobradas , Morte Celular , Regulação da Expressão Gênica de Plantas , Luz/efeitos adversos , Estresse Fisiológico , TranscriptomaRESUMO
When exposed to unfavorable environmental conditions, plants can absorb light energy in excess of their photosynthetic capacities, with the surplus energy leading to the production of reactive oxygen species and photooxidative stress. Subsequent lipid peroxidation generates toxic reactive carbonyl species whose accumulation culminates in cell death. ß-Cyclocitral, an oxidized by-product of ß-carotene generated in the chloroplasts, mediates a protective retrograde response that lowers the levels of toxic peroxides and carbonyls, limiting damage to intracellular components. In this study, we elucidate the molecular mechanism induced by ß-cyclocitral in Arabidopsis thaliana and show that the xenobiotic detoxification response is involved in the tolerance to excess light energy. The involvement of the xenobiotic response suggests a possible origin for this pathway. Furthermore, we establish the hierarchical structure of this pathway that is mediated by the ß-cyclocitral-inducible GRAS protein SCARECROW LIKE14 (SCL14) and involves ANAC102 as a pivotal component upstream of other ANAC transcription factors and of many enzymes of the xenobiotic detoxification response. Finally, the SCL14-dependent protective mechanism is also involved in the low sensitivity of young leaf tissues to high-light stress.
Assuntos
Aldeídos/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Diterpenos/metabolismo , Estresse Fisiológico/fisiologia , Arabidopsis/efeitos dos fármacos , Proteínas de Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Inativação Metabólica , Luz , Mutação , Folhas de Planta/fisiologia , Plantas Geneticamente Modificadas , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Transativadores/genética , Transativadores/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Xenobióticos/farmacologiaRESUMO
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 ß-cyclocitral canceled the damaging effects of HNE. Conversely, in the scl14 mutant, the effects of ß-cyclocitral and HNE were additive, leading to enhanced leaf damage. These results indicate that the cellular detoxification pathway induced by the low-toxicity ß-cyclocitral targets highly toxic compounds produced during lipid peroxidation, reminiscent of a safener-type mode of action.
Assuntos
Peroxidação de Lipídeos , Luminescência , Aldeídos , Diterpenos , Estresse Oxidativo , Folhas de Planta , Espécies Reativas de OxigênioRESUMO
Singlet oxygen produced from triplet excited chlorophylls in photosynthesis is a signal molecule that can induce programmed cell death (PCD) through the action of the OXIDATIVE STRESS INDUCIBLE 1 (OXI1) kinase. Here, we identify two negative regulators of light-induced PCD that modulate OXI1 expression: DAD1 and DAD2, homologs of the human antiapoptotic protein DEFENDER AGAINST CELL DEATH. Overexpressing OXI1 in Arabidopsis (Arabidopsis thaliana) increased plant sensitivity to high light and induced early senescence of mature leaves. Both phenomena rely on a marked accumulation of jasmonate and salicylate. DAD1 or DAD2 overexpression decreased OXI1 expression, jasmonate levels, and sensitivity to photooxidative stress. Knock-out mutants of DAD1 or DAD2 exhibited the opposite responses. Exogenous applications of jasmonate upregulated salicylate biosynthesis genes and caused leaf damage in wild-type plants but not in the salicylate biosynthesis mutant Salicylic acid induction-deficient2, indicating that salicylate plays a crucial role in PCD downstream of jasmonate. Treating plants with salicylate upregulated the DAD genes and downregulated OXI1 We conclude that OXI1 and DAD are antagonistic regulators of cell death through modulating jasmonate and salicylate levels. High light-induced PCD thus results from a tight control of the relative activities of these regulating proteins, with DAD exerting a negative feedback control on OXI1 expression.
Assuntos
Apoptose/genética , Proteínas de Arabidopsis/genética , Arabidopsis/genética , Ciclopentanos/metabolismo , Oxilipinas/metabolismo , Fosfolipases A1/genética , Proteínas Serina-Treonina Quinases/genética , Ácido Salicílico/metabolismo , Apoptose/efeitos da radiação , Arabidopsis/citologia , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Vias Biossintéticas/efeitos dos fármacos , Vias Biossintéticas/genética , Vias Biossintéticas/efeitos da radiação , Ciclopentanos/farmacologia , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Luz , Mutação , Oxilipinas/farmacologia , Fosfolipases A1/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Folhas de Planta/citologia , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Ácido Salicílico/farmacologia , Oxigênio Singlete/metabolismoRESUMO
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 in vivo. 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 in vivo. 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.
RESUMO
The Arabidopsis vte1 mutant is devoid of tocopherol and plastochromanol (PC-8). When exposed to excess light energy, vte1 produced more singlet oxygen (1 O2 ) 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 1 O2 production and higher levels of Hydroxyplastoquinone (PQ-C), a 1 O2 -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 1 O2 scavenger and is replenished from the extrathylakoid stock.
Assuntos
Sequestradores de Radicais Livres/metabolismo , Plastoquinona/metabolismo , Oxigênio Singlete/metabolismo , Tilacoides/metabolismo , Alquil e Aril Transferases/metabolismo , Proteínas de Arabidopsis/metabolismo , Clorofila/metabolismo , Cloroplastos/metabolismo , Cromatografia Líquida de Alta Pressão , Espectroscopia de Ressonância de Spin Eletrônica , Luz , Peroxidação de Lipídeos , Estresse Oxidativo/efeitos da radiaçãoRESUMO
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.
Assuntos
Quercus/metabolismo , Mudança Climática , Desidratação , Secas , Região do Mediterrâneo , Estresse Oxidativo , Fotossíntese , Quercus/fisiologia , Fatores de TempoRESUMO
Singlet oxygen (1 O2 ) signalling in plants is essential to trigger both acclimatory mechanisms and programmed cell death under high light stress. However, because of its chemical features, 1 O2 requires mediators, and the players involved in this pathway are largely unknown. The ß-carotene oxidation product, ß-cyclocitral, is one such mediator. Produced in the chloroplast, ß-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 1 O2 signalling leading to tolerance to high light. Here, we provide evidence that MBS1 is essential for acclimation to 1 O2 and cross-talks with ß-cyclocitral to mediate transfer of the 1 O2 signal to the nucleus, leading to photoacclimation. The presented results position MBS1 downstream of ß-cyclocitral in 1 O2 signalling and suggest an additional role for MBS1 in the regulation of plant growth and development under chronic 1 O2 production.
Assuntos
Aclimatação , Aldeídos/farmacologia , Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Diterpenos/farmacologia , Oxigênio Singlete/farmacologia , Aclimatação/efeitos da radiação , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/efeitos da radiação , Proteínas de Arabidopsis/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Proteínas de Fluorescência Verde/metabolismo , Luz , Mutação/genética , Fenótipo , Fotossíntese/efeitos da radiação , Folhas de Planta/fisiologia , Folhas de Planta/efeitos da radiação , Estresse Fisiológico/efeitos da radiaçãoRESUMO
Singlet oxygen (¹O2) is a reactive oxygen species that can function as a stress signal in plant leaves leading to programmed cell death. In microalgae, ¹O2-induced transcriptomic changes result in acclimation to ¹O2. Here, using a chlorophyll b-less Arabidopsis thaliana mutant (chlorina1 [ch1]), we show that this phenomenon can also occur in vascular plants. The ch1 mutant is highly photosensitive due to a selective increase in the release of ¹O2 by photosystem II. Under photooxidative stress conditions, the gene expression profile of ch1 mutant leaves very much resembled the gene responses to ¹O2 reported in the Arabidopsis mutant flu. Preexposure of ch1 plants to moderately elevated light intensities eliminated photooxidative damage without suppressing ¹O2 formation, indicating acclimation to ¹O2. Substantial differences in gene expression were observed between acclimation and high-light stress: A number of transcription factors were selectively induced by acclimation, and contrasting effects were observed for the jasmonate pathway. Jasmonate biosynthesis was strongly induced in ch1 mutant plants under high-light stress and was noticeably repressed under acclimation conditions, suggesting the involvement of this hormone in ¹O2-induced cell death. This was confirmed by the decreased tolerance to photooxidative damage of jasmonate-treated ch1 plants and by the increased tolerance of the jasmonate-deficient mutant delayed-dehiscence2.
Assuntos
Aclimatação/efeitos da radiação , Arabidopsis/genética , Luz , Mutação , Oxigenases/genética , Oxigênio Singlete/metabolismo , Aclimatação/genética , Acetatos/farmacologia , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Vias Biossintéticas/efeitos dos fármacos , Vias Biossintéticas/genética , Vias Biossintéticas/efeitos da radiação , Clorofila/metabolismo , Ciclopentanos/metabolismo , Ciclopentanos/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Peroxidação de Lipídeos/efeitos da radiação , Análise de Sequência com Séries de Oligonucleotídeos , Oxirredução/efeitos da radiação , Oxigenases/metabolismo , Oxilipinas/metabolismo , Oxilipinas/farmacologia , Complexo de Proteína do Fotossistema II/genética , Complexo de Proteína do Fotossistema II/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Folhas de Planta/genética , Folhas de Planta/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transcriptoma/efeitos dos fármacos , Transcriptoma/genética , Transcriptoma/efeitos da radiaçãoRESUMO
The fungal elicitor cryptogein triggers a light-dependent hypersensitive response in tobacco (Nicotiana tabacum). To assess the effect of light on this nonhost resistance in more detail, we studied various aspects of the response under dark and light conditions using the tobacco-cryptogein experimental system. Here, we show that light drastically alters the plant's transcriptional response to cryptogein, notably by dampening the induction of genes involved in multiple processes, such as ethylene biosynthesis, secondary metabolism, and glutathione turnover. Furthermore, chlorophyll fluorescence measurements demonstrated that quantum yield and functioning of the light-harvesting antennae decreased simultaneously, indicating that photoinhibition underlies the observed decreased photosynthesis and that photooxidative damage might be involved in the establishment of the altered response. Analysis of the isomer distribution of hydroxy fatty acids illustrated that, in the light, lipid peroxidation was predominantly due to the production of singlet oxygen. Differences in (reduced) glutathione concentrations and the rapid development of symptoms in the light when cryptogein was coinfiltrated with glutathione biosynthesis inhibitors suggest that glutathione might become a limiting factor during the cryptogein-induced hypersensitive response in the dark and that this response might be modified by an increased antioxidant availability in the light.
Assuntos
Proteínas Fúngicas/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Nicotiana/efeitos dos fármacos , Vias Biossintéticas , Resistência à Doença , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Glutationa Transferase/metabolismo , Glutationa Transferase/fisiologia , Glicosiltransferases/metabolismo , Glicosiltransferases/fisiologia , Oxilipinas/metabolismo , Doenças das Plantas/genética , Nicotiana/microbiologia , Nicotiana/efeitos da radiaçãoRESUMO
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.
Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/fisiologia , Lipocalinas/fisiologia , Estresse Fisiológico , Arabidopsis/genética , Arabidopsis/efeitos da radiação , Proteínas de Arabidopsis/genética , Secas , Temperatura Alta , Luz , Peroxidação de Lipídeos , Lipocalinas/genética , Estresse Oxidativo , Sementes/genética , Sementes/fisiologia , Sementes/efeitos da radiaçãoRESUMO
Lipid droplets are ubiquitous cellular structures in eukaryotes and are required for lipid metabolism. Little is currently known about plant lipid droplets other than oil bodies. Here, we define dual roles for chloroplast lipid droplets (plastoglobules) in energy and prenylquinone metabolism. The prenylquinones--plastoquinone, plastochromanol-8, phylloquinone (vitamin K(1)), and tocopherol (vitamin E)--are partly stored in plastoglobules. This work shows that NAD(P)H dehydrogenase C1 (NDC1) (At5g08740), a type II NAD(P)H quinone oxidoreductase, associates with plastoglobules. NDC1 reduces a plastoquinone analog in vitro and affects the overall redox state of the total plastoquinone pool in vivo by reducing the plastoquinone reservoir of plastoglobules. Finally, NDC1 is required for normal plastochromanol-8 accumulation and is essential for vitamin K(1) production.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Cloroplastos/enzimologia , Cromanos/metabolismo , Lipídeos/química , NADH NADPH Oxirredutases/metabolismo , Plastoquinona/metabolismo , Quinona Redutases/metabolismo , Quinonas/metabolismo , Tocoferóis/metabolismo , Vitamina E/análogos & derivados , Vitamina K 1/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Cloroplastos/genética , Regulação da Expressão Gênica de Plantas , Medições Luminescentes , Mutação/genética , NADH NADPH Oxirredutases/genética , Fotossíntese , Transporte Proteico , Vitamina E/metabolismoRESUMO
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.
Assuntos
Metabolismo dos Lipídeos/fisiologia , Peróxidos Lipídicos/análise , Medições Luminescentes/métodos , Arabidopsis/genética , Arabidopsis/metabolismo , Clorofila/metabolismo , Técnicas In Vitro , Medições Luminescentes/instrumentação , Mutação , Oxirredução , Estresse Oxidativo , FótonsRESUMO
Plants, like most living organisms, spontaneously emit photons of visible light. This ultraweak endogenous chemiluminescence is linked to the oxidative metabolism, with lipid peroxidation constituting a major source of photons in plants. We imaged this signal using a very sensitive cooled CCD camera and analysed its spectral characteristics using bandpass interference filters. In vitro oxidation of lipids induced luminescence throughout the visible spectrum (450−850 nm). However, luminescence in the red spectral domain (>640 nm) occurred first, then declined in parallel with the appearance of the emission in the blue-green (<600 nm). This temporal separation suggests that the chemical species emitting in the blue-green are secondary products, possibly deriving from the red light-emitting species. This conversion did not seem to occur in planta because spontaneous chemiluminescence from plant tissues (leaves, roots) occurred only in the red/far-red light domain (>640 nm), peaking at 700−750 nm. The spectrum of plant chemiluminescence was independent of chlorophyll. The in vivo signal was modulated by cellular detoxification mechanisms and by changes in the concentration of singlet oxygen in the tissues, although the singlet oxygen luminescence bands did not appear as major bands in the spectra. Our results indicate that the intensity of endogenous chemiluminescence from plant tissues is determined by the balance between the formation of luminescent species through secondary reactions involving lipid peroxide-derived intermediates, including singlet oxygen, and their elimination by metabolizing processes. The kinetic aspects of plant chemiluminescence must be taken into account when using the signal as an oxidative stress marker.
RESUMO
Because they are highly unsaturated, plant lipids are sensitive to oxidation and constitute a primary target of reactive oxygen species. Therefore, quantification of lipid peroxidation provides a pertinent approach to evaluating oxidative stress in plants. Here, we describe a simple method to measure upstream products of the peroxidation of the major polyunsaturated fatty acids in plants, namely, linolenic acid (C18:3) and linoleic acid (C18:2). The method uses conventional HPLC with UV detection to measure hydroxy C18:3 and C18:2 after reduction of their respective hydroperoxides. The described experimental approach requires low amounts of plant material (a few hundred milligrams), monitors oxidation of both membrane and free fatty acids, and can discriminate between enzymatic and non-enzymatic lipid peroxidation.
Assuntos
Ácidos Graxos Insaturados , Ácidos Graxos , Cromatografia Líquida de Alta Pressão , Peroxidação de Lipídeos , Oxirredução , Espécies Reativas de OxigênioRESUMO
Guanosine pentaphosphate and tetraphosphate (together referred to as ppGpp) are hyperphosphorylated nucleotides found in bacteria and the chloroplasts of plants and algae. In plants and algae artificial ppGpp accumulation can inhibit chloroplast gene expression, and influence photosynthesis, nutrient remobilization, growth, and immunity. However, it is so far unknown whether ppGpp is required for abiotic stress acclimation in plants. Here, we demonstrate that ppGpp biosynthesis is necessary for acclimation to nitrogen starvation in Arabidopsis. We show that ppGpp is required for remodeling the photosynthetic electron transport chain to downregulate photosynthetic activity and for protection against oxidative stress. Furthermore, we demonstrate that ppGpp is required for coupling chloroplastic and nuclear gene expression during nitrogen starvation. Altogether, our work indicates that ppGpp is a pivotal regulator of chloroplast activity for stress acclimation in plants.