High Salinity Stress Research Articles

Genome-Wide Analysis of Genes Encoding Methionine-Rich Proteins in Arabidopsis and Soybean Suggesting Their Roles in the Adaptation of Plants to Abiotic Stress.

Oxidation and reduction of methionine (Met) play important roles in scavenging reactive oxygen species (ROS) and signaling in living organisms. To understand the impacts of Met oxidation and reduction in plants during stress, we surveyed the genomes of Arabidopsis and soybean (Glycine max L.) for genes encoding Met-rich proteins (MRPs). We found 121 and 213 genes encoding MRPs in Arabidopsis and soybean, respectively. Gene annotation indicated that those with known function are involved in vital cellular processes such as transcriptional control, calcium signaling, protein modification, and metal transport. Next, we analyzed the transcript levels of MRP-coding genes under normal and stress conditions. We found that 57 AtMRPs were responsive either to drought or to high salinity stress in Arabidopsis; 35 GmMRPs were responsive to drought in the leaf of late vegetative or early reproductive stages of soybean. Among the MRP genes with a known function, the majority of the abiotic stress-responsive genes are involved in transcription control and calcium signaling. Finally, Arabidopsis plant which overexpressed an MRP-coding gene, whose transcripts were downregulated by abiotic stress, was more sensitive to paraquat than the control. Taken together, our report indicates that MRPs participate in various vital processes of plants under normal and stress conditions.

Characterization of the γ-aminobutyric acid shunt pathway and oxidative damage in Arabidopsis thaliana pop 2 mutants under various abiotic stresses

In the present study, three Arabidopsis thaliana pop2 mutant lines with different T-DNA insertions in a gene coding γ-aminobutyric acid transaminase (GABA-TA) were screened for seed germination percentage, stress-induced oxidative damage, and GABA content and metabolism under various abiotic stresses including high temperature (42 °C), low temperature (4 °C), salinity (NaCl), and osmotic stress (mannitol). All mutant lines showed a decreased germination under all the stress treatments with a significant reduction in the pop2-1 and pop2-3 mutant lines. Content of GABA and MDA increased significantly in all pop2 mutants and wild type (WT) seedlings in response to all the treatments. However, content of GABA and MDA was lower in all pop2 mutants comparing to the WT under the same treatments. GABA increased already after 30 min and increased significantly after 2 h at 42 °C especially in the pop2-3 and WT seedlings. In response to the cold treatment, GABA content increased up to 4-fold compared to the control in all pop2 mutants and WT seedlings. In response to the NaCl treatment, GABA accumulated slightly in the WT and all pop2 mutants. On the contrary, GABA content increased significantly in the pop2, pop2-1, and pop2-3 mutants and WT under all mannitol treatments.

Involvement of the crustacean hyperglycemic hormone (CHH) in the physiological compensation of the freshwater crayfish Cherax quadricarinatus to low temperature and high salinity stress.

This study was aimed at determining the role of the crustacean hyperglycemic hormone (CHH) in the physiological compensation to both saline and thermal stress, in the freshwater crayfish Cherax quadricarinatus. By determining the expression of the CHH gene in the eyestalk of juvenile crayfish, we found that maximal induction of CHH was induced at high salinity (10 g/L) and low temperature (20 °C). In order to investigate the role of CHH in the physiological compensation to such stressful conditions, recombinant CHH was supplied to stressed animals. CHH-injected crayfish showed increased hemolymphatic levels of glucose, in accordance with a significant utilization of glycogen reserves from the hepatopancreas. Furthermore, CHH administration allowed stressed animals to regulate hemolymphatic sodium and potassium at more constant levels than controls. Taken together, these results suggest a relevant role of CHH in increasing the energy available intended for processes involved in the physiological compensation of C. quadricarinatus to both saline and thermal stress.

Ky-2, a Histone Deacetylase Inhibitor, Enhances High-Salinity Stress Tolerance in Arabidopsis thaliana.

Adaptation to environmental stress requires genome-wide changes in gene expression. Histone modifications are involved in gene regulation, but the role of histone modifications under environmental stress is not well understood. To reveal the relationship between histone modification and environmental stress, we assessed the effects of inhibitors of histone modification enzymes during salinity stress. Treatment with Ky-2, a histone deacetylase inhibitor, enhanced high-salinity stress tolerance in Arabidopsis. We confirmed that Ky-2 possessed inhibition activity towards histone deacetylases by immunoblot analysis. To investigate how Ky-2 improved salt stress tolerance, we performed transcriptome and metabolome analysis. These data showed that the expression of salt-responsive genes and salt stress-related metabolites were increased by Ky-2 treatment under salinity stress. A mutant deficient in AtSOS1(Arabidopis thaliana SALT OVERLY SENSITIVE 1), which encodes an Na(+)/H(+)antiporter and was among the up-regulated genes, lost the salinity stress tolerance conferred by Ky-2. We confirmed that acetylation of histone H4 at AtSOS1 was increased by Ky-2 treatment. Moreover, Ky-2 treatment decreased the intracellular Na(+)accumulation under salinity stress, suggesting that enhancement of SOS1-dependent Na(+)efflux contributes to increased high-salinity stress tolerance caused by Ky-2 treatment.

Increase in wheat production through management of abiotic stresses : A review

About 9% of area on earth is under crops out of which 91% is under various stresses. On an average, about 50% yield losses are due to abiotic stresses mostly due to high temperature (20%), low temperature (7%), salinity (10%), drought (9%) and other abiotic stresses (4%). As there is no scope for increasing area under agriculture, the increased productivity from these stressed land is a must to meet the ever increasing demand. Further, the severity of abiotic stresses is likely to increase due to changing climate leading to adverse effect on crops. Therefore, abiotic stresses like drought, salinity, sodicity, acidity, water logging, heat, nutrient toxicities/ deficiencies etc need to be effectively addressed through adoption of management practices like tillage and planting options, residue management, sowing time, stress tolerant cultivars, irrigation scheduling and integrated nutrient management to conserve natural resources, mitigating their adverse effect and sustainable wheat production.

Sucrose metabolism gene families and their biological functions.

Sucrose, as the main product of photosynthesis, plays crucial roles in plant development. Although studies on general metabolism pathway were well documented, less information is available on the genome-wide identification of these genes, their expansion and evolutionary history as well as their biological functions. We focused on four sucrose metabolism related gene families including sucrose synthase, sucrose phosphate synthase, sucrose phosphate phosphatase and UDP-glucose pyrophosphorylase. These gene families exhibited different expansion and evolutionary history as their host genomes experienced differentiated rates of the whole genome duplication, tandem and segmental duplication, or mobile element mediated gene gain and loss. They were evolutionarily conserved under purifying selection among species and expression divergence played important roles for gene survival after expansion. However, we have detected recent positive selection during intra-species divergence. Overexpression of 15 sorghum genes in Arabidopsis revealed their roles in biomass accumulation, flowering time control, seed germination and response to high salinity and sugar stresses. Our studies uncovered the molecular mechanisms of gene expansion and evolution and also provided new insight into the role of positive selection in intra-species divergence. Overexpression data revealed novel biological functions of these genes in flowering time control and seed germination under normal and stress conditions.

Effect of pre-chilling and environmental factors on breaking seed dormancy and germination of three foxtail species

The effect of wet and dry pre-chilling duration, pH, osmotic stress, salt stress and planting depth on seed germination and seedling emergence of three foxtail species (Setaria glauca, S. verticillata and S. viridis) was investigated in a series of laboratory and greenhouse experiments. Both wet and dry pre-chilling for 45 days promoted seed germination of S. glauca compared with the control. Pre-chilling was not significantly effective in seed dormancy breaking of S. viridis and S. verticillata. The maximum germination of foxtails (S. verticillata, S. viridis) was obtained when seeds were treated with pH 7 buffer solution. Increasing of osmotic and salt stress decreased seed germination of foxtails. Setaria verticillata seed germination was more tolerant than those of S. glauca and S. viridis to high water stress condition. Setaria glauca and S. verticillata seed germination were more tolerant to high salinity stress than S. viridis. Seedling emergence decreased with increasing the burial depth and no germination observed at 8 cm soil depth.

Expression and distribution of neuropeptides in the nervous system of the crab Carcinus maenas and their roles in environmental stress.

Environmental fluctuations, such as salinity, impose serious challenges to marine animal survival. Neuropeptides, signaling molecules involved in the regulation process, and the dynamic changes of their full complement in the stress response have yet to be investigated. Here, a MALDI-MS-based stable isotope labeling quantitation strategy was used to investigate the relationship between neuropeptide expression and adaptability of Carcinus maenas to various salinity levels, including high (60 parts per thousand [p.p.t.]) and low (0 p.p.t.) salinity, in both the crustacean pericardial organ (PO) and brain. Moreover, a high salinity stress time course study was conducted. MS imaging (MSI) of neuropeptide localization in C. maenas PO was also performed. As a result of salinity stress, multiple neuropeptide families exhibited changes in their relative abundances, including RFamides (e.g. APQGNFLRFamide), RYamides (e.g. SSFRVGGSRYamide), B-type allatostatins (AST-B; e.g. VPNDWAHFRGSWamide), and orcokinins (e.g. NFDEIDRSSFGFV). The MSI data revealed distribution differences in several neuropeptides (e.g. SGFYANRYamide) between color morphs, but salinity stress appeared to not have a major effect on the localization of the neuropeptides.

Overexpression of OsLEA4 enhances drought, high salt and heavy metal stress tolerance in transgenic rice (Oryza sativa L.)

Late embryogenesis abundant (LEA) proteins are involved in tolerance to drought, cold and high salinity in many different organisms. In this study, real-time PCR was used to analyze the expression pattern of the OsLEA4 gene in rice, which showed that OsLEA4 was expressed in different organ tissues during different development stages of rice. The expression levels of OsLEA4 in the leaves during the tillering stage and leaves and panicles during the heading stage, the filling stage and the full ripe stage were dramatically increased. Moreover, based on seed germination, growth status and physiological indices, the overexpression of OsLEA4 in transgenic rice plants conferred increased resistance to drought, salt and heavy metal stresses compared with the wild type (WT) plants. After drought, high salinity and heavy metal stresses, the increase in the malondialdehyde content in transgenic plants was less than that in WT, and the increase in the free proline and soluble sugar contents in transgenic plants were greater than those of the WT plants. These results indicated that the OsLEA4 protein contributes to drought, salt and heavy metal tolerance in rice plants. Thus, OsLEA4 is a potential candidate gene for plant genetic modification.

Isolation and expression analysis of 18 CsbZIP genes implicated in abiotic stress responses in the tea plant (Camellia sinensis)

Basic leucine zipper (bZIP) transcription factors (TFs) play essential roles in regulating stress processes in plants. Despite the economic importance of this woody crop, there is little information about bZIP TFs in tea plants. In this study, 18 bZIP genes were isolated from the tea plant (Camellia sinensis) and named sequentially from CsbZIP1 to CsbZIP18. According to the phylogenetic classification as in Arabidopsis, the CsbZIP genes spanned ten subgroups (Group A, B, C, D, E, F, H, I, S and K) of bZIP TFs. When analyzed for organ specific expression, all CsbZIP genes were found to be ubiquitously expressed in roots, stems, leaves and flowers. Expression analysis of CsbZIP genes in response to four abiotic stresses showed that in leaves, 9, 9, 15 and 11 CsbZIPs have 2-fold greater variation in transcript abundance under cold, exogenous ABA, high salinity and dehydration conditions, respectively. In roots, 5, 12, 14 and 11 CsbZIPs were differentially expressed under conditions of cold, exogenous ABA, high salinity and dehydration stresses. Moreover, CsbZIP genes in Groups F, H, S and K exhibited several folds up-and/or down-regulation against the above four stresses. Notably, CsbZIP18 of group K showed significant up-regulation in response to these same stresses, suggesting a vital functional role in stress response. Together, these findings increase our knowledge of bZIP TFs in the tea plant and suggest the significance of CsbZIP genes in plant abiotic responses.

Cloning and characterization of the pepper CaPAO gene for defense responses to salt-induced leaf senescence.

BackgroundPheophorbide a oxygenase (PAO) is an important enzyme in the chlorophyll catabolism pathway and is involved in leaf senescence. It opens the porphyrin macrocycle of pheophorbide a and finally forms the primary fluorescent chlorophyll catabolite. Previous studies have demonstrated the function of PAO during cell death. However, the characterizaton of PAO during leaf senescence induced by environmental factors is not well understood.MethodsHomology-based cloning and RACE techniques were used to obtain the full-length cDNA of the CaPAO gene. CaPAO expression was determined by quantitative real-time PCR. Function of CaPAO gene were studied using virus-induced gene silencing and transgenic techniques with tobacco plants (Nicotiana tabacum).ResultsA novel PAO gene CaPAO was isolated from pepper (Capsicum annuum L.). The full-length CaPAO cDNA is comprised of 1838 bp, containing an open reading frame of 1614 bp, and encodes a 537 amino acid protein. This deduced protein belongs to the Rieske-type iron-sulfur superfamily, containing a conserved Rieske cluster. CaPAO expression, as determined by quantitative real-time PCR, was higher in leaves than roots, stems and flowers. It was upregulated by abscisic acid, methyl jasmonate and salicylic acid. Moreover, CaPAO was significantly induced by high salinity and osmotic stress treatments and also was regulated by Phytophthora capsici. The virus-induced gene silencing technique was used to silence the CaPAO gene in pepper plants. After 3 days of high salt treatment, the chlorophyll breakdown of CaPAO-silenced pepper plants was retarded. RD29A promoter-inducible expression vector was constructed and transferred into tobacco plant. After 7 days of salt treatment, the leaves of transgenic plants were severely turned into yellow, the lower leaves showed necrotic symptom and chlorophyll content was significantly lower than that in the control plants.ConclusionsThe expression of CaPAO gene was induced in natural senescence and various stresses. The CaPAO gene may be related to defense responses to various stresses and play an important role in salt-induced leaf senescence.

Hormetic effect of 60Co gamma radiation on tolerance to salinity and temperature stress in Haliotis discus discus

Low levels of radiation increase productivity and environmental resistance in a variety of organisms by hormesis. Here, we investigated the effects of different dose levels of 60Co gamma radiation (GR) on tolerance to high temperature (HT) and low salinity (LS) stress and immunological responses in the commercially consumed abalone, Haliotis discus discus. Although H. discus discus exhibited a significantly lower survival rate (SR) at a high level of GR (25Gy) than the control (0Gy), an increased SR and higher cardiovascular hemocyte counts were exhibited at 20Gy. Furthermore, enhanced tolerance to HT (30°C) and LS (25psu) stress, and increased lysozyme activity, were exhibited by the GR-treated H. discus discus groups. Although similar respiratory burst activity after 24h of stress exposure was exhibited by all of the groups, significantly enhanced superoxide dismutase activity was exhibited in the GR-treated abalone groups under LS stress. Our results indicate that GR-treated H. discus discus exhibit hormetic responses to 60CO gamma radiation against environmental stressors because of increased innate immune responses. Statement of relevanceIt is considered that our study will provide beneficial information about potential use of gamma-radiation techniques for the environmentally-strong abalone production in aquaculture industry.

Cotton mitogen-activated protein kinase4 (GhMPK4) confers the transgenic Arabidopsis hypersensitivity to salt and osmotic stresses

Cotton (Gossypium hirsutum) as one of the most important economic crop in the world is often suffering from biotic and abiotic stresses during its growth seasons. Mitogen-activated protein kinase (MAPK) cascades participate in signal transduction of extracellular stimuli and regulate multiple biotic and abiotic stress responses in plants. In this study, a MAPK gene (GhMPK4) belonging to the group B of MAPK family was identified in cotton. Quantitative RT-PCR analysis showed that expression of GhMPK4 was induced by high salinity and osmotic stresses. Overexpression of GhMPK4 in Arabidopsis significantly enhanced the transgenic plants’ sensitivity to salt and osmotic stresses and exogenous abscisic acid (ABA). Under NaCl, mannitol and ABA treatments, the rates of seed germination and cotyledon expansion/greening of the GhMPK4 overexpression transgenic lines were remarkably declined compared with those of wild type, and roots of the GhMPK4 overexpression transgenic seedlings were shorter than those of wild type. Chlorophyll and proline contents in leaves of the GhMPK4 transgenic lines were obviously lower than those in wild type. Growth status of the GhMPK4 transgenic plants was worse than that of wild type when being subjected to drought and salinity stresses. Furthermore, the expression levels of the stress-related genes were altered in the transgenic plants under NaCl, mannitol and ABA treatments, compared with those in wild type. Taken the data together, it is suggested that GhMPK4 as a negative regulator may participate in response to salt and osmotic stresses and ABA signaling through affecting the expression of the stress-related genes in plants.

Responses of Germlings to Acute Environmental Stress

The responses of Sargassum thunbergii germlings to high temperature, low salinity, desiccation, combined thermal and osmotic stress (35 ºC combined with 12 psu), anthracene, and eutrophication were examined. Probit regression analysis results showed that the median lethal time (LT50) values of high temperature decreased with the increase in temperature. The 24 h median lethal temperature was 36.9 ºC. For salinity treatment, the LT50 value of fresh water was 47.6 h. Survival rates of germlings were over 60% when germlings were exposed to salinities ranging from 27 psu to 7 psu at a time interval of 108 h post-treatment. The LT50 values of desiccation and combined thermal and osmotic stress (35 ºC combined with 12 psu) were 7.0 h and 9.8 h, respectively. Anovas showed that germlings were inhibited by high concentrations of anthracene (5 mg L-1 and 10 mg L-1) with low survival rates of below 50% and low relative growth rates of below 1% after 25 days of culture; however, low concentrations (0.01-1 mg L-1) had no significant effects. In addition, neither severe eutrophication nor disproportionality of N/P showed any significant effect on the survival and growth of germlings. Of the environmental stresses tested, possible occurrence of high temperature of 40 ºC and combined thermal and osmotic stress directly impacted the survival of germlings, suggesting that the deterioration of S. thunbergii bed may be related more to increasing extreme climatic events.

Isolation and characterisation of cDNA encoding a wheat heavy metal‐associated isoprenylated protein involved in stress responses

In cells, metallochaperones are important proteins that safely transport metal ions. Heavy metal-associated isoprenylated plant proteins (HIPPs) are metallochaperones that contain a metal binding domain and a CaaX isoprenylation motif at the carboxy-terminal end. To investigate the roles of wheat heavy metal-associated isoprenylated plant protein (TaHIPP) genes in plant development and in stress responses, we isolated cDNA encoding the wheat TaHIPP1 gene, which contains a heavy metal-associated domain, nuclear localisation signals and an isoprenylation motif (CaaX motif). Quantitative real-time PCR analysis indicated that the TaHIPP1 gene was differentially expressed under biotic and abiotic stresses. Specifically, TaHIPP1 expression was up-regulated by ABA exposure or wounding. Additionally, TaHIPP1 over-expression in yeast (Schizosaccharomyces pombe) significantly increased the cell growth rate under Cu(2+) and high salinity stresses. The nuclear localisation of the protein was confirmed with confocal laser scanning microscopy of epidermal onion cells after particle bombardment with chimeric TaHIPP1-GFP constructs. In addition, TaHIPP1 was shown to enhance the susceptibility of wheat to Pst as determined by virus-induced gene silencing. These data indicate that TaHIPP1 is an important component in defence signalling pathways and may play a crucial role in the defence response of wheat to biotic and certain abiotic stresses.

Nicotiana sylvestris calcineurin B-like protein NsylCBL10 enhances salt tolerance in transgenic Arabidopsis.

Nicotiana sylvestris calcineurin B-like protein NsylCBL10 improves tolerance to high-salt stress through better maintenance of Na (+) balance. The calcineurin B-like (CBL) proteins represent a unique group of plant calcium sensors and play an important role in regulating the response of a plant cell to the stress. Although many studies have been made in Arabidopsis (Arabidopsis thaliana), rice (Oryza sativa) and poplar (Populus trichocarpa), the characterization and elucidation of the functions of CBLs in tobacco have not yet been reported. In this study, NsylCBL10, a CBL gene showing higher similarities to other CBL10 genes, was cloned from Nicotiana sylvestris. NsylCBL10 is expressed in most of the tobacco tissues, and the protein targets to the plasma membrane specifically. Over-expression of NsylCBL10 enhanced the salt tolerance of Arabidopsis wild type plants greatly, and rescued the high-salt-sensitive phenotype of Arabidopsis cbl10 mutant. The analysis of ion content indicated that over-expressing NsylCBL10 in plants is able to maintain a lower Na(+)/K(+) ratio in roots and higher Na(+)/K(+) ratio in shoots, compared with cbl10 mutant. The results suggest that NsylCBL10 might play an important role in response to high salinity stress in N. sylvestris, by keeping a better ionic homeostasis to reduce the damage of toxic ion to the plant cell.

Overexpression of a cotton (Gossypium hirsutum) WRKY gene, GhWRKY34, in Arabidopsis enhances salt-tolerance of the transgenic plants

Soil salinity is one of the most serious threats in world agriculture, and often influences cotton growth and development, resulting in a significant loss in cotton crop yield. WRKY transcription factors are involved in plant response to high salinity stress, but little is known about the role of WRKY transcription factors in cotton so far. In this study, a member (GhWRKY34) of cotton WRKY family was functionally characterized. This protein containing a WRKY domain and a zinc-finger motif belongs to group III of cotton WRKY family. Subcellular localization assay indicated that GhWRKY34 is localized to the cell nucleus. Overexpression of GhWRKY34 in Arabidopsis enhanced the transgenic plant tolerance to salt stress. Several parameters (such as seed germination, green cotyledons, root length and chlorophyll content) in the GhWRKY34 transgenic lines were significantly higher than those in wild type under NaCl treatment. On the contrary, the GhWRKY34 transgenic plants exhibited a substantially lower ratio of Na+/K+ in leaves and roots dealing with salt stress, compared with wild type. Growth status of the GhWRKY34 transgenic plants was much better than that of wild type under salt stress. Expressions of the stress-related genes were remarkably up-regulated in the transgenic plants under salt stress, compared with those in wild type. Based on the data presented in this study, we hypothesize that GhWRKY34 as a positive transcription regulator may function in plant response to high salinity stress through maintaining the Na+/K+ homeostasis as well as activating the salt stress-related genes in cells.

Genome-wide identification, classification and expression analysis of the PHD-finger protein family in Populus trichocarpa

The plant homeobox domain (PHD) proteins are widespread in eukaryotes, and play important roles in regulating chromatin and transcription. Comprehensive analyses of PHD-finger proteins have been performed in animals, but few plant PHD-finger proteins involved in growth and development have been characterized functionally. In this study, we conducted a genome-wide survey of PHD-finger proteins in Populus trichocarpa by describing the phylogenetic relationship, gene structure, and chromosomal location and microarray analyses of each predicted PHD-finger family member. We identified 73 PHD-finger genes (PtPHD1-73) and classified them into eleven subfamilies (A–K) by phylogenetic analysis. Seventy-two of the 73 genes were unevenly distributed on all 19 chromosomes, with seven segmental duplication events. Analysis of the Ka (non-synonymous substitution rate)/Ks (synonymous substitution rate) ratios suggested that the duplicated genes of the PHD-finger family mainly underwent purifying selection with restrictive functional divergence after the duplication events. Expression profiles analysis indicated that 67 PHD-finger genes were differentially expressed in various tissues. Quantitative real-time RT-PCR (qRT-PCR) analyses of nine selected PtPHD genes under high salinity, drought and cold stresses were also performed to explore their stress-related expression patterns. The results of this study provide a thorough overview of poplar PHD-finger proteins and will be valuable for further functional research of poplar PHD-finger genes to unravel their biological roles.

Nutritional Status as the Key Modulator of Antioxidant Responses Induced by High Environmental Ammonia and Salinity Stress in European Sea Bass (Dicentrarchus labrax).

Salinity fluctuation is one of the main factors affecting the overall fitness of marine fish. In addition, water borne ammonia may occur simultaneously with salinity stress. Additionally, under such stressful circumstances, fish may encounter food deprivation. The physiological and ion-osmo regulatory adaptive capacities to cope with all these stressors alone or in combination are extensively addressed in fish. To date, studies revealing the modulation of antioxidant potential as compensatory response to multiple stressors are rather lacking. Therefore, the present work evaluated the individual and combined effects of salinity challenge, ammonia toxicity and nutritional status on oxidative stress and antioxidant status in a marine teleost, European sea bass (Dicentrarchus labrax). Fish were acclimated to normal seawater (32 ppt), to brackish water (20 ppt and 10 ppt) and to hypo-saline water (2.5 ppt). Following acclimation to different salinities for two weeks, fish were exposed to high environmental ammonia (HEA, 20 mg/L representing 50% of 96h LC50 value for ammonia) for 12 h, 48 h, 84 h and 180 h, and were either fed (2% body weight) or fasted (unfed for 7 days prior to HEA exposure). Results show that in response to decreasing salinities, oxidative stress indices such as xanthine oxidase activity, levels of hydrogen peroxide (H2O2) and lipid peroxidation (malondialdehyde, MDA) increased in the hepatic tissue of fasted fish but remained unaffected in fed fish. HEA exposure at normal salinity (32 ppt) and at reduced salinities (20 ppt and 10 ppt) increased ammonia accumulation significantly (84 h–180 h) in both feeding regimes which was associated with an increment of H2O2 and MDA contents. Unlike in fasted fish, H2O2 and MDA levels in fed fish were restored to control levels (84 h–180 h); with a concomitant increase in superoxide dismutase (SOD), catalase (CAT), components of the glutathione redox cycle (reduced glutathione, glutathione peroxidase and glutathione reductase), ascorbate peroxidase (APX) activity and reduced ascorbate (ASC) content. On the contrary, fasted fish could not activate many of these protective systems and rely mainly on CAT and ASC dependent pathways as antioxidative sentinels. The present findings exemplify that in fed fish single factors and a combination of HEA exposure and reduced seawater salinities (upto 10 ppt) were insufficient to cause oxidative damage due to the highly competent antioxidant system compared to fasted fish. However, the impact of HEA exposure at a hypo-saline environment (2.5 ppt) also defied antioxidant defence system in fed fish, suggesting this combined factor is beyond the tolerance range for both feeding groups. Overall, our results indicate that the oxidative stress mediated by the experimental conditions were exacerbated during starvation, and also suggest that feed deprivation particularly at reduced seawater salinities can instigate fish more susceptible to ammonia toxicity.

Modulation of superoxide dismutase (SOD) isozymes by organ development and high long-term salinity in the halophyte Cakile maritima.

Superoxide dismutase (SOD) activity catalyzes the disproportionation of superoxide radicals into hydrogen peroxide and oxygen. This enzyme is considered to be a first line of defense for controlling the production of reactive oxygen species (ROS). In this study, the number and type of SOD isozymes were identified in the principal organs (roots, stems, leaves, flowers, and seeds) of Cakile maritima. We also analyzed the way in which the activity of these SOD isozymes is modulated during development and under high long-term salinity (400 mM NaCl) stress conditions. The data indicate that this plant contains a total of ten SOD isozymes: two Mn-SODs, one Fe-SOD, and seven CuZn-SODs, with the Fe-SOD being the most prominent isozyme in the different organs analyzed. Moreover, the modulation of SOD isozymes, particularly CuZn-SODs, was only detected during development and under severe salinity stress conditions. These data suggest that, in C. maritima, the occurrence of these CuZn-SODs in roots and leaves plays an adaptive role since this CuZn-SOD isozyme might replace the diminished Fe-SOD activity under salinity stress to overcome this adverse environmental condition.