RESUMO
Plants produce glutathione as a response to the intercellular redox state. Glutathione actively participates in the reactive oxygen species (ROS)-dependent signaling pathway, especially under biotic stress conditions. Most of the glutathione S-transferases (GSTs) are induced in cells during the defense response of plants not only through highly specific glutathione-binding abilities but also by participating in the signaling function. The tau class of GSTs has been reported to be induced as a response under stress conditions. Although several studies have focused on the role of the tau class of GSTs in plant-pathogen interactions, knowledge about their contribution to the response to virus inoculation is still inadequate. Therefore, in this study, the response of Atgstu19 and Atgstu24 knockout mutants to mechanical inoculation of Turnip mosaic virus (TuMV) was examined. The systemic infection of TuMV was more dynamically promoted in Atgstu19 mutants than in wild-type (Col-0) plants, suggesting the role of GSTU19 in TuMV resistance. However, Atgstu24 mutants displayed virus limitation and downregulation of the relative expression of TuMV capsid protein, accompanied rarely by TuMV particles only in vacuoles, and ultrastructural analyses of inoculated leaves revealed the lack of virus cytoplasmic inclusions. These findings indicated that Atgstu24 mutants displayed a resistance-like reaction to TuMV, suggesting that GSTU24 may suppress the plant resistance. In addition, these findings confirmed that GSTU1 and GSTU24 are induced and contribute to the susceptible reaction to TuMV in the Atgstu19-TuMV interaction. However, the upregulation of GSTU19 and GSTU13 highly correlated with virus limitation in the resistance-like reaction in the Atgstu24-TuMV interaction. Furthermore, the highly dynamic upregulation of GST and glutathione reductase (GR) activities resulted in significant induction (between 1 and 14 days post inoculation [dpi]) of the total glutathione pool (GSH + GSSG) in response to TuMV, which was accompanied by the distribution of active glutathione in plant cells. On the contrary, in Atgstu19, which is susceptible to TuMV interaction, upregulation of GST and GR activity only up to 7 dpi symptom development was reported, which resulted in the induction of the total glutathione pool between 1 and 3 dpi. These observations indicated that GSTU19 and GSTU24 are important factors in modulating the response to TuMV in Arabidopsis thaliana. Moreover, it was clear that glutathione is an important component of the regulatory network in resistance and susceptible response of A. thaliana to TuMV. These results help achieve a better understanding of the mechanisms regulating the Arabidopsis-TuMV pathosystem.
Assuntos
Arabidopsis , Potyvirus , Arabidopsis/metabolismo , Proteínas do Capsídeo/metabolismo , Dissulfeto de Glutationa/metabolismo , Glutationa Redutase/metabolismo , Doenças das Plantas/genética , Potyvirus/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Transferases/metabolismoRESUMO
Glutathione peroxidases (GPXs) are important antioxidant enzymes in animals. Plants contain GPX-like (GPXL) enzymes, which-in contrast to GPXs-contain cysteine in their active site instead of selenocysteine. Although several studies proved their importance in development and stress responses, their interaction with ethylene (ET) signalling is not known. Our aim was to investigate the involvement of AtGPXL5 in ET biosynthesis and/or signalling using Atgpxl5 mutant and AtGPXL5 cDNA-overexpressing (OX-AtGPXL5) lines. Four-day-old dark-grown Atgpxl5 seedlings had shorter hypocotyls and primary roots, while OX-AtGPXL5 seedlings exhibited a similar phenotype as wild type under normal conditions. Six-week-old OX-AtGPXL5 plants contained less H2O2 and malondialdehyde, but higher polyamine and similar ascorbate- and glutathione contents and redox potential (EGSH) than the Col-0. One-day treatment with the ET-precursor 1-aminocyclopropane-1-carboxylic acid (ACC) induced the activity of glutathione- and thioredoxin peroxidases and some other ROS-processing enzymes. In the Atgpxl5 mutants, the EGSH became more oxidised; parallelly, it produced more ethylene after the ACC treatment than other genotypes. Although the enhanced ET evolution measured in the Atgpxl5 mutant can be the result of the increased ROS level, the altered expression pattern of ET-related genes both in the Atgpxl5 and OX-AtGPXL5 plants suggests the interplay between AtGPXL5 and ethylene signalling.
Assuntos
Arabidopsis , Arabidopsis/metabolismo , Etilenos/metabolismo , Glutationa/metabolismo , Glutationa Peroxidase/genética , Glutationa Peroxidase/metabolismo , Peróxido de Hidrogênio/metabolismo , Isoenzimas/genética , Isoenzimas/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
Glutathione transferases (GSTs) play a crucial role in detoxification processes due to the fact of their glutathione (GSH) conjugating activity, and through glutathione peroxidase or dehydroascorbate reductase (DHAR) activities, they influence the redox state of GSH and ascorbate (AsA). The plant-specific tau (GSTU) group is the largest class of Arabidopsis GSTs, and their members are involved in responses to different abiotic stresses. We investigated the effect of salt stress on two-week-old Arabidopsis thaliana wild-type (Col-0), Atgstu19 and Atgstu24 mutant plants after applying 150 mM NaCl for two days. The Atgstu19 seedlings had lower GST activity and vitality both under control conditions and after salt stress than the wild-type, but the level of total ROS was similar to the Col-0 plants. The GST activity of the knockout Atgstu24 mutant was even higher under control conditions compared to the Col-0 plants, while the ROS level and its vitality did not differ significantly from the wild-type. Analysis of the AtGSTU expression pattern revealed that the mutation in a single AtGSTU gene was accompanied by the up- and downregulation of several other AtGSTUs. Moreover, elevated AsA and GSH levels, an altered GSH redox potential and increased DHAR and glutathione reductase activities could help to compensate for the mutation of AtGSTU genes. The observed changes in the mutants suggest that the investigated isoenzymes influence the redox homeostasis under control conditions and after NaCl treatment in Arabidopsis seedlings. These data indicate for the first time the more general role of a temporary shift of redox status as part of GST mechanisms and regulation.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Glutationa Transferase/genética , Estresse Salino , Arabidopsis/fisiologia , Proteínas de Arabidopsis/metabolismo , Glutationa Transferase/metabolismo , Mutação , Oxirredução , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismoRESUMO
Salt tolerance mechanisms of halophyte Petrosimonia triandra, growing in its natural habitat in Cluj County, Romania, were investigated via biomass, growth parameters, water status, ion content, photosynthetic and antioxidative system efficiency, proline accumulation and lipid degradation. Two sampling sites with different soil electrical conductivities were selected: site 1: 3.14 dS m-1 and site 2: 4.45 dS m-1. Higher salinity proved to have a positive effect on growth. The relative water content did not decline severely, Na+ and K+ content of the roots, stem and leaves was more, and the functions of the photosynthetic apparatus and photosynthetic pigment contents were not altered. The efficiency of the antioxidative defence system was found to be assured by coordination of several reactive oxygen species scavengers. The presence of higher salinity led to accumulation of the osmolyte proline, while degradation of membrane lipids was reduced. As a whole, P. triandra evolved different adaptational strategies to counteract soil salinity, including morphological and physiological adaptations, preservation of photosynthetic activity, development of an efficient antioxidative system and accumulation of the osmotic compound, proline.
RESUMO
Although the participation of glutathione transferases (GSTs) in light-dependent pathways and the circadian changes in the whole detoxification system have been studied, there are fewer results regarding the exact daily fluctuation of GSTs. In the present study, it was demonstrated that light up-regulated, while dark period decreased the plant GST activity and the expression of the selected tau group GST genes in tomato. These findings provide additional information on our current knowledge on the circadian rhythm of GSTs in plants and could help in further defining detoxification processes.
Assuntos
Ritmo Circadiano/efeitos da radiação , Glutationa Transferase/metabolismo , Luz , Fotoperíodo , Proteínas de Plantas/metabolismo , Solanum lycopersicum/efeitos da radiação , Regulação Enzimológica da Expressão Gênica/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Glutationa Transferase/genética , Solanum lycopersicum/enzimologia , Solanum lycopersicum/genética , Proteínas de Plantas/genética , Fatores de TempoRESUMO
Arabidopsis thaliana contains 54 soluble glutathione transferases (GSTs, EC 2.5.1.18), which are thought to play major roles in oxidative stress responses, but little is known about the function of individual isoenzymes. The role of AtGST phi 9 (GSTF9) in the salt- and salicylic acid response was investigated using 2-week-old Atgstf9 and wild type (Wt) plants. Atgstf9 mutants accumulated more ascorbic acid (AsA) and glutathione (GSH) and had decreased glutathione peroxidase (GPOX) activity under control conditions. Treatment of 2-week-old seedlings with 10â»7 M salicylic acid (SA) for 48 h resulted in elevated H2O2level and enhanced GST activity in Atgstf9 plants, 10â»5 M SA treatment enhanced the malondialdehyde and dehydroascorbate contents compared to Wt. 50 and 150 mM NaCl increased the GST activity, AsA and GSH accumulation in Atgstf9 seedlings more pronounced than in Wt plants. We found that the Atgstf9 mutants had altered redox homeostasis under control and stress conditions, in which elevated AsA and GSH levels and modified GST and GPOX activities may play significant role. The half-cell potential values calculated from the concentration of GSH and GSSG indicate that this GST isoenzyme has an important role in the salt stress response.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Glutationa Transferase/metabolismo , Estresse Oxidativo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Ácido Ascórbico/metabolismo , Glutationa/metabolismo , Glutationa Transferase/genética , Plântula/enzimologia , Plântula/genéticaRESUMO
Glutathione transferases (GSTs) are one of the most versatile multigenic enzyme superfamilies. In our experiments, the involvement of the genotype-specific induction of GST genes and glutathione- or redox-related genes in pathways regulating salt-stress tolerance was examined in tomato cultivars (Solanum lycopersicum Moneymaker, Mobil, and Elán F1). The growth of the Mobil plants was adversely affected during salt stress (100 mM of NaCl), which might be the result of lowered glutathione and ascorbate levels, a more positive glutathione redox potential (EGSH), and reduced glutathione reductase (GR) and GST activities. In contrast, the Moneymaker and Elán F1 cultivars were able to restore their growth and exhibited higher GR and inducible GST activities, as well as elevated, non-enzymatic antioxidant levels, indicating their enhanced salt tolerance. Furthermore, the expression patterns of GR, selected GST, and transcription factor genes differed significantly among the three cultivars, highlighting the distinct regulatory mechanisms of the tomato genotypes during salt stress. The correlations between EGSH and gene expression data revealed several robust, cultivar-specific associations, underscoring the complexity of the stress response mechanism in tomatoes. Our results support the cultivar-specific roles of distinct GST genes during the salt-stress response, which, along with WRKY3, WRKY72, DREB1, and DREB2, are important players in shaping the redox status and the development of a more efficient stress tolerance in tomatoes.
RESUMO
Glutathione peroxidases (GPXs) are non-heme peroxidases catalyzing the reduction of H2O2 or organic hydroperoxides to water or corresponding alcohols using glutathione (GSH) or thioredoxin (TRX) as a reducing agent. In contrast to animal GPXs, the plant enzymes are non-seleno monomeric proteins that generally utilize TRX more effectively than GSH but can be a putative link between the two main redox systems. Because of the substantial differences compared to non-plant GPXs, use of the GPX-like (GPXL) name was suggested for Arabidopsis enzymes. GPX(L)s not only can protect cells from stress-induced oxidative damages but are crucial components of plant development and growth. Due to fine-tuning the H2O2 metabolism and redox homeostasis, they are involved in the whole life cycle even under normal growth conditions. Significantly new mechanisms were discovered related to their transcriptional, post-transcriptional and post-translational modifications by describing gene regulatory networks, interacting microRNA families, or identifying Lys decrotonylation in enzyme activation. Their involvement in epigenetic mechanisms was evidenced. Detailed genetic, evolutionary, and bio-chemical characterization, and comparison of the main functions of GPXs, demonstrated their species-specific roles. The multisided involvement of GPX(L)s in the regulation of the entire plant life ensure that their significance will be more widely recognized and applied in the future.
RESUMO
Reactive oxygen species (ROS), antioxidants and their reduction-oxidation (redox) states all contribute to the redox homeostasis, but glutathione is considered to be the master regulator of it. We aimed to understand the relationship between the redox potential and the diverse glutathione transferase (GST) enzyme family by comparing the stress responses of two tomato cultivars (Solanum lycopersicum 'Moneymaker' and 'Ailsa Craig'). Four-week-old plants were treated by two concentrations of mannitol, NaCl and salicylic acid. The lower H2O2 and malondialdehyde contents indicated higher stress tolerance of 'Moneymaker'. The redox status of roots was characterized by measuring the reduced and oxidized form of ascorbate and glutathione spectrophotometrically after 24 h. The redox potential of 'Ailsa Craig' was more oxidized compared to 'Moneymaker' even under control conditions and became more positive due to treatments. High-throughput quantitative real-time PCR revealed that besides overall higher expression levels, SlGSTs were activated more efficiently in 'Moneymaker' due to stresses, resulting in generally higher GST and glutathione peroxidase activities compared to 'Ailsa Craig'. The expression level of SlGSTs correlated differently, however Pearson's correlation analysis showed usually strong positive correlation between SlGST transcription and glutathione redox potential. The possible redox regulation of SlGST expressions was discussed.
Assuntos
Peróxido de Hidrogênio , Solanum lycopersicum , Antioxidantes , Glutationa/metabolismo , Solanum lycopersicum/metabolismo , Oxirredução , Estresse Oxidativo , Ácido SalicílicoRESUMO
The detoxification of harmful metabolites can determine the effectiveness of plant stress responses. Scavenging some of these toxic stress by-products through the reduced form of glutathione is catalysed by members of the glutathione transferase (GST) enzyme superfamily. The involvement of these enzymes was studied in the model organism Brachypodium distachyon (L.)P.Beauv. Bd21 and in its derivative Bd21-3, a more drought tolerant line. Osmotic stress treatment resulted in a decrease in the water potential of both Brachypodium genotypes, the difference between the control and treated plant's ψw decreased by the last sampling day in Bd21-3, suggesting some degree of adaptation to the applied osmotic stress. Increased GST activity revealed a severe defence reaction against the harmful imbalance of the redox environment. Screening for the gene sequences led to the identification of 91 full-length or partial GST sequences. Although purple false brome has a relatively small genome, the number of identified GST genes was almost as high as the number predicted in wheat. The estimation of GST expression showed stress-induced differences: higher expression levels or the fast induction of BdGSTF8, BdGSTU35 and BdGSTU42 gene products presumably indicate a strong detoxification under osmotic stress.
Assuntos
Brachypodium , Secas , Glutationa Transferase , Pressão Osmótica , TriticumRESUMO
The tau (U) and phi (F) classes of glutathione transferase (GST) enzymes reduce the glutathione (GSH) pool using GSH as a co-substrate, thus influence numerous redox-dependent processes including hormonal and stress responses. We performed detailed analysis of the redox potential and reactive oxygen species levels in longitudinal zones of 7-day-old roots of Arabidopsis thaliana L. Col-0 wild type and Atsgtf8 and Atgstu19 insertional mutants. Using redox-sensitive cytosolic green fluorescent protein (roGFP2) the redox status of the meristematic, transition, and elongation zones was determined under control and salt stress (3-hour of 75 or 150 mM NaCl treatment) conditions. The Atgstu19 mutant had the most oxidized redox status in all root zones throughout the experiments. Using fluorescent dyes significantly higher superoxide radical (O2-) levels was detected in both Atgst mutants than in the Col-0 control. Salt treatment resulted in the highest O2- increase in the Atgstf8 root, while the amount of H2O2 elevated most in the case of Atgstu19. Moreover, vitality decreased in Atgstu19 roots more than in wild type under salt stress. Our results indicate that AtGSTF8 and especially the AtGSTU19 proteins function in the root fine-tuning the redox homeostasis both under control and salt stress conditions.
Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/fisiologia , Glutationa Transferase/fisiologia , Meristema/fisiologia , Raízes de Plantas/fisiologia , Arabidopsis/enzimologia , Arabidopsis/metabolismo , Homeostase , Peróxido de Hidrogênio/metabolismo , Meristema/metabolismo , Oxirredução , Raízes de Plantas/enzimologia , Raízes de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Estresse Salino , Superóxidos/metabolismoRESUMO
Drought resistance of bean landraces was compared in order to select genotypes with either high morphological or high biochemical-physiological plasticity. The lines in the former group exhibited fast reduction in fresh and dry mass, decreased the water potential in primary leaves after irrigation withdrawal and the biomass mobilized from the senescent primary leaves was allocated into the roots. These genotypes had high frequency of primary leaf abscission under water stress. The genotypes with plasticity at the biochemical level maintained high water potential and photochemical efficiency, i.e. effective quantum yield, high photochemical (qP) and low non-photochemical (NPQ) quenching in primary leaves under drought stress. While superoxide dismutase activity was not influenced by the drought and the genotype, catalase activity increased significantly in the primary leaves of the genotypes with efficient biochemical adaptation. Lines with high morphological plasticity exhibited higher quaiacol peroxidase activity under drought. Proline may accumulate in both cases, thus it may be a symptom of protein degradation or a successful osmotic adaptation. On the basis of contrasting responses, the genetic material cannot be screened for a large-scale breeding program by a single physiological parameter but by a set of the methods presented in this work.
Assuntos
Aclimatação , Clorofila/metabolismo , Fabaceae/metabolismo , Estresse Oxidativo , Água/metabolismo , Fluorescência , Genótipo , FenótipoRESUMO
The activity and expression of glutathione transferases (GSTs) depend on several less-known endogenous and well-described exogenous factors, such as the developmental stage, presence, and intensity of different stressors, as well as on the absence or presence and quality of light, which to date have received less attention. In this review, we focus on discussing the role of circadian rhythm, light quality, and intensity in the regulation of plant GSTs. Recent studies demonstrate that diurnal regulation can be recognized in GST activity and gene expression in several plant species. In addition, the content of one of their co-substrates, reduced glutathione (GSH), also shows diurnal changes. Darkness, low light or shade mostly reduces GST activity, while high or excess light significantly elevates both the activity and expression of GSTs and GSH levels. Besides the light-regulated induction and dark inactivation of GSTs, these enzymes can also participate in the signal transduction of visible and UV light. For example, red light may alleviate the harmful effects of pathogens and abiotic stressors by increasing GST activity and expression, as well as GSH content in leaves of different plant species. Based on this knowledge, further research on plants (crops and weeds) or organs and temporal regulation of GST activity and gene expression is necessary for understanding the complex regulation of plant GSTs under various light conditions in order to increase the yield and stress tolerance of plants in the changing environment.
RESUMO
Plants have divergent defense mechanisms against the harmful effects of heavy metals present in excess in soils and groundwaters. Poplars (Populus spp.) are widely cultivated because of their rapid growth and high biomass production, and members of the genus are increasingly used as experimental model organisms of trees and for phytoremediation purposes. Our aim was to investigate the copper and zinc stress responses of three outstanding biomass producer bred poplar lines to identify such transcripts of genes involved in the detoxification mechanisms, which can play an important role in the protection against heavy metals. Poplar cuttings were grown hydroponically and subjected to short-term (one week) mild and sublethal copper and zinc stresses. We evaluated the effects of the applied heavy metals and the responses of plants by detecting the changes of multiple physiological and biochemical parameters. The most severe cellular oxidative damage was caused by 30µM copper treatment, while zinc was less harmful. Analysis of stress-related transcripts revealed genotype-specific differences that are likely related to alterations in heavy metal tolerance. P. deltoides clones B-229 and PE 19/66 clones were clearly more effective at inducing the expression of various genes implicated in the detoxification process, such as the glutathione transferases, metallothioneins, ABC transporters, (namely PtGSTU51, PxMT1, PdABCC2,3), while the P. canadensis line M-1 accumulated more metal, resulting in greater cellular oxidative damage. Our results show that all three poplar clones are efficient in stress acclimatization, but with different molecular bases.
Assuntos
Metais Pesados/toxicidade , Populus/genética , Populus/fisiologia , Estresse Fisiológico/efeitos dos fármacos , Análise de Variância , Biodegradação Ambiental/efeitos dos fármacos , Células Clonais , Cobre/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas , Inativação Metabólica/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos dos fármacos , Malondialdeído/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/metabolismo , Populus/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Espécies Reativas de Nitrogênio/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Estresse Fisiológico/genética , Água/metabolismo , Zinco/metabolismoRESUMO
The plant glutathione peroxidase (GPX) family consists of multiple isoenzymes with distinct subcellular locations which exhibit different tissue-specific expression patterns and environmental stress responses. Contrary to most of their counterparts in animal cells, plant GPXs contain cysteine instead of selenocysteine in their active site and while some of them have both glutathione peroxidase and thioredoxin peroxidase functions, the thioredoxin regenerating system is much more efficient in vitro than the glutathione system. At present, the function of these enzymes in plants is not completely understood. The occurrence of thiol-dependent activities of plant GPX isoenzymes suggests that - besides detoxification of H2O2 and organic hydroperoxides - they may be involved in regulation of the cellular redox homeostasis by maintaining the thiol/disulfide or NADPH/NADP(+) balance. GPXs may represent a link existing between the glutathione- and the thioredoxin-based system. The various thiol buffers, including Trx, can affect a number of redox reactions in the cells most probably via modulation of thiol status. It is still required to identify the in vivo reductant for particular GPX isoenzymes and partners that GPXs interact with specifically. Recent evidence suggests that plant GPXs does not only protect cells from stress induced oxidative damage but they can be implicated in plant growth and development. Following a more general introduction, this study summarizes present knowledge on plant GPXs, highlighting the results on gene expression analysis, regulation and signaling of Arabidopsis thaliana GPXs and also suggests some perspectives for future research.
Assuntos
Antioxidantes/metabolismo , Glutationa Peroxidase/metabolismo , Desenvolvimento Vegetal , Plantas/enzimologia , Estresse Fisiológico , Modelos BiológicosRESUMO
The role of salicylic acid (SA) in the control of abscisic acid (ABA) biosynthesis is controversial although both plant growth regulators may accumulate in tissues under abiotic and biotic stress conditions. Hardening of tomato plants to salinity stress with 10(-4)M SA ("high SA") resulted in an up-regulation of ABA biosynthesis genes, zeaxanthin epoxidase (SlZEP1), 9-cis-epoxycarotenoid dioxygenase (SlNCED1) and aldehyde oxidases (SlAO1 and SlAO2) in the roots and led to ABA accumulation both in root and leaf tissues. In plants pre-treated with lower concentration of SA (10(-7)M, "low SA"), the up-regulation of SlNCED1 in the roots promoted ABA accumulation in the root tissues but the hormone concentration remained at control level in the leaves. Salt stress induced by 100mM NaCl reduced the transcript abundance of ABA biosynthetic genes and inhibited SlAO activity in plants hardened with "high SA", but the tissues maintained root ABA level over the untreated control. The combined effect of "high SA" and ABA under salt stress led to partially recovered photosynthetic activity, reduced ethylene production in root apices, and restored root growth, which is one of the main features of salt tolerance. Unlike "high SA", hardening with "low SA" had no influence on ethylene production, and led to reduced elongation of roots in plants exposed to 100mM NaCl. The up-regulation of carotenoid cleavage dioxygenases SlCCD1A and SlCCD1B by SA, which produce apocarotenoids, may open new pathways in SA sensing and signalling processes.
Assuntos
Ácido Abscísico/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Proteínas de Plantas/genética , Ácido Salicílico/farmacologia , Cloreto de Sódio/farmacologia , Solanum lycopersicum/efeitos dos fármacos , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Proteínas de Plantas/metabolismo , Tolerância ao Sal , Estresse Fisiológico/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacosRESUMO
Salicylic acid (SA) applied exogenously is a potential priming agent during abiotic stress. In our experiments, the priming effect of SA was tested by exposing Arabidopsis thaliana (L.) Heynh. plants to 2-week-long 10-9-10-5 M SA pretreatments in a hydroponic medium, followed by 1 week of 100mM NaCl stress. The levels of reactive oxygen species and H2O2, changes in antioxidant enzyme activity and the expression of selected glutathione transferase (GST) genes were investigated. Although 10-9-10-7 M SA pretreatment insufficiently induced defence mechanisms during the subsequent salt stress, 2-week pretreatments with 10-6 and 10-5 M SA alleviated the salinity-induced H2O2 and malondialdehyde accumulation, and increased superoxide dismutase, guaiacol peroxidase, GST and glutathione peroxidase (GPOX) activity. Our results indicate that long-term 10-6 and 10-5 M SA treatment mitigated the salt stress injury in this model plant. Enhanced expression of AtGSTU19 and AtGSTU24 may be responsible for the induced GST and GPOX activity, which may play an important role in acclimation. Modified GST expression suggested altered signalling in SA-hardened plants during salt stress. The hydroponic system applied in our experiments proved to be a useful tool for studying the effects of sequential treatments in A. thaliana.
RESUMO
Auxin autotrophic and heterotrophic tobacco callus lines were grown on MS medium with or without 100 mmol/L NaCl and growth and some of the stress-related activities, such as GPX, SOD, CAT, GST, GSH-PX, as well as the concentration of ethylene and H2O2, were measured and compared with each other. The auxin autotrophic calli grew slower, however, on the NaCl-containing medium the growth rate was higher than that of the heterotrophic cultures after two weeks of culturing. The stress-related ethylene production was lower in the autotrophic cultures and, contrary to the heterotrophic tissues, its level did not change significantly upon NaCl treatment. The guaiacol peroxidase (GPX) activities were higher in the autotrophic tissues in all cell fractions regardless of the presence of NaCl. Treated with NaCl, the GPX activities elevated in the soluble and covalently-bound fractions in the heterotrophic calli, but were not further increased in the autotrophic line. SOD and CAT activities were higher in the heterotrophic tissues, and were increased further by 100 mmol/L NaCl treatment. The GST and GSH-PX activities were higher in the autotrophic line, which might explain their enhanced stress tolerance. In the autotrophic tissues, the elevated antioxidant activities led to reduced levels of H2O2 and malondialdehyde; under mild NaCl stress, these levels decreased further. The lower growth rate and the effective protection against NaCl stress-induced oxidative damage of the autotrophic line can be explained by the cell wall-bound peroxidase and GSH-PX activities in the auxin autotrophic tissues. Their maintained growth rate indicates that the autotropic cultures were more resistant to exogenous H2O2.
Assuntos
Glutationa Peroxidase/metabolismo , Ácidos Indolacéticos/fisiologia , Nicotiana/fisiologia , Estresse Oxidativo/fisiologia , Reguladores de Crescimento de Plantas/fisiologia , Glutationa Transferase/metabolismo , Peróxido de Hidrogênio/metabolismo , Malondialdeído/metabolismo , Sódio/metabolismo , Cloreto de Sódio/farmacologia , Nicotiana/efeitos dos fármacos , Nicotiana/enzimologiaRESUMO
A family tree of the multifunctional proteins, glutathione transferases (GSTs, EC 2.5.1.18) was created in Solanum lycopersicum based on homology to known Arabidopsis GSTs. The involvement of selected SlGSTs was studied in salt stress response of tomato primed with salicylic acid (SA) or in un-primed plants by real-time qPCR. Selected tau GSTs (SlGSTU23, SlGSTU26) were up-regulated in the leaves, while GSTs from lambda, theta, dehydroascorbate reductase and zeta classes (SlGSTL3, SlGSTT2, SlDHAR5, SlGSTZ2) in the root tissues under salt stress. Priming with SA exhibited a concentration dependency; SA mitigated the salt stress injury and caused characteristic changes in the expression pattern of SlGSTs only at 10(-4) M concentration. SlGSTF4 displayed a significant up-regulation in the leaves, while the abundance of SlGSTL3, SlGSTT2 and SlGSTZ2 transcripts were enhanced in the roots of plants primed with high SA concentration. Unexpectedly, under high salinity the SlDHAR2 expression decreased in primed roots as compared to the salt-stressed plants, however, the up-regulation of SlDHAR5 isoenzyme contributed to the maintenance of DHAR activity in roots primed with high SA. The members of lambda, theta and zeta class GSTs have a specific role in salt stress acclimation of tomato, while SlGSTU26 and SlGSTF4, the enzymes with high glutathione conjugating activity, characterize a successful priming in both roots and leaves. In contrast to low concentration, high SA concentration induced those GSTs in primed roots, which were up-regulated under salt stress. Our data indicate that induction of GSTs provide a flexible tool in maintaining redox homeostasis during unfavourable conditions.