Induction Of Antioxidant Defense System Research Articles

Remediation of heavy metals by native plant species grown in Iran’s richest gold mine and study of plants’ pollution tolerance strategies

Mining is defined as an environmentally hazardous activity that releases metals and other elements to the environment. Bioremediation is a natural and sustainable technique for harnessing the potential of microorganisms and plants to remove, degrade, or stabilize pollutants from contaminated sites and enable cleanup and restoration of the environment. In the present study, following the investigation of pollution of heavy metals in soil samples collected from the Zarshuran mining area, the role of twenty native plant species in the bioaccumulation of heavy metals was evaluated. After preparation of soil and plant samples, inductively coupled plasma-mass spectroscopy (ICP-MS) was used to determine the concentrations of elements in the soil and plant samples. It was confirmed that the soil samples were highly contaminated by silver (Ag), zinc (Zn), cadmium (Cd), lead (Pb), and antimony (Sb). High amounts of Pb and Zn were accumulated in Eryngium billardieri and Astragalus rostratus. Further, these two plant species could uptake, transport, and accumulate Ag in their aerial parts and the enrichment coefficient of their shoots was more than 1. Scorzonera latifolia also had good potential to stabilise Ag, Zn, and Pb in its root. As a result, E. billardieri, A. rostratus, and S. latifolia may be suitable candidate species to clean heavy metals from soils in contaminated sites. Overall, augmentation of superoxide dismutase activity and the amounts of total phenols and flavonoids in different parts of E. billardieri and A. rostratus confirmed the induction of antioxidant defense systems in the plants (compared to the control plants) and an attempt by the plants to tolerate heavy metal pollution.

Melatonin reduces cadmium accumulation through cell wall fraction fixation capacity in cotton seedlings

Melatonin (MT) participates in plants' responses in plants’ response to cadmium (Cd) tolerance, although its work model remains elusive. Here, the function of MT in adjusting Cd accumulation in Gossypium hirsutum (CCRI12) was investigated. In this research we observed that exogenous 50 μM MT significantly reduced the Cd accumulation under 100 μМ Cd stress for 7 days of treatment in cotton seedlings. Cd markedly induced the ROS (H2O2, O2-) accumulation and elevated the expression of transporters for Cd uptake and translocation. After application of MT reduced ROS and MDA accumulation in response to high induction of antioxidant defense system; the genes encoding superoxide dismutase (GhSOD), catalase (GhCAT), and peroxidase (GhPOD), ascorbate peroxidase (GhAPX), and glutathione S-transferase (GhGST) were significantly upregulated accompanied by their level of content. Furthermore, MT down-regulated transporters for Cd uptake include Natural Resistance-Associated Macrophage Protein 1 (GhNRAMP3), Zinc/Iron-regulated transporter-like protein (GhZIP1/4/6), Heavy Metal ATPase gene (GhHMA2), and ATP-binding Cassette sub family C (GhABCC3/2). Exogenous application of MT raised the hemicelluloses, cellulose and pectin content accompanied by their expression levels, xyloglucan endotransglucosylase/hydrolase (GhXTH28/6/9/B), cellulose synthase like (GhCSLG1/2) and COBRA-like (GhCOBL4), pectin acetylesterases (GhPAE8/7), pectin methylesterases (GhPME31) which in turn increased the cell wall's binding capacity to Cd.

Insight into Recent Progress and Perspectives in Improvement of Antioxidant Machinery upon PGPR Augmentation in Plants under Drought Stress: A Review.

Agriculture has a lot of responsibility as the rise in the world’s population demands more food requirements. However, more than one type of biotic and abiotic stress continually impacts agricultural productivity. Drought stress is a major abiotic stress that significantly affects agricultural productivity every year as the plants undergo several morphological, biochemical, and physiological modifications, such as repressed root and shoot growth, reduced photosynthesis and transpiration rate, excessive production of reactive oxygen species (ROS), osmotic adjustments, and modified leaf senescence regulating and stress signaling pathways. Such modifications may permanently damage the plants; therefore, mitigation strategies must be developed. The use of drought resistant crop cultivars is more expensive and labor-intensive with few advantages. However, exploiting plant growth promoting rhizobacteria (PGPR) is a proven alternative with numerous direct and indirect advantages. The PGPR confers induced systemic tolerance (IST) mechanisms in plants in response to drought stress via multiple mechanisms, including the alteration of root architecture, maintenance of high relative water content, improvement of photosynthesis rate, production of phytohormones, exopolysaccharides, ACC deaminase, carotenoids and volatiles, induction of antioxidant defense system, and alteration in stress-responsive gene expression. The commercial application of PGPR as bioinoculants or biostimulants will remain contingent on more robust strain selection and performance under unfavorable environmental conditions. This review highlights the possible mechanisms of PGPR by activating the plant adaptive defense systems for enhancing drought tolerance and improving overall growth and yield.

Effect of heat stress on oxidative damage and antioxidant defense system in white clover (Trifolium repens L.)

This study leads to advances in the field of heat tolerance among different plant species. We concluded that a coordinated, increased antioxidant defense system enabled white clover to reduce heat-induced oxidative damage. The rise in global ambient temperature has a wide range of effects on plant growth, and, therefore, on the activation of various molecular defenses before the appearance of heat damage. Elevated temperatures result in accelerated generation of reactive oxygen species (ROS), causing an imbalance between ROS production and the ability of scavenging systems to detoxify and remove the reactive intermediates. The aim of this study was to determine the role of antioxidant defense systems in the alleviation of heat stress (HS) consequences in white clover (Trifolium repens L.), which is cultivated worldwide. We evaluated how temperature and time parameters contribute to the thermotolerance of white clover at different growth stages. We revealed HS protection in white clover from 37 to 40°C, with 40°C providing the greatest protection of 3-day-old seedlings and 28-day-old adult plants. Heat-provoked oxidative stress in white clover was confirmed by substantial changes in electrolyte leakage, malondialdehyde (MDA), and chlorophyll content, as well as superoxide anion (O2·-) and hydrogen peroxide (H2O2) production. Furthermore, superoxide dismutase (SOD) and ascorbate peroxidase (APX) as well as a high level of GSH non-enzymatic antioxidant were the most responsive, and were associated with acquired thermotolerance through the regulation of ROS generation.We demonstrated, by studying protoplast transient gene expression, direct genetic evidence of endogenous antioxidant-related genes that confer HS tolerance in white clover. Our present study clearly establishes that oxidative stress ensues from HS, which triggers the induction of antioxidant defense systems for ROS scavenging in white clover.

Physiological response of diverse halophytes to high salinity through ionic accumulation and ROS scavenging

Salt stress induced modulations in different ionic ratios and ROS system were studied in ten halophytic species, namely Atriplex lentiformis, Tamarix aphylla, Sporobolus marginatus, Suaeda nudiflora, Urochondra setulosa, Arundo donax, Aeluropus lagopoides, Heliotropium ramossimum, Atriplex nummularia, Leptachloa fusca at salinity level of ECe ∼ 30 dSm−1 (≈300 mM NaCl) to explore their possible role in salt tolerance ability of these halophytes. These halophytes were categorized for their salt tolerance levels based on the ratios of Na+/K+, Na+/Ca2+, Na+/Cl− and Na + Cl/K + Ca. Variable responses were observed among all halophytes where Atriplex lentiformis had lowest leaf Na+/K+ (0.44) which is one of the best indicator of salt tolerance, Heliotropium ramossimum had lowest Na+/Ca2+ and Na+/Cl− (0.97 and 0.18), whereas Sporobolus marginatus had lowest Na + Cl/K + Ca (0.79). Specific enzymes activities of ascorbate peroxidase (APX), superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX) were also assessed to get better comprehension of the ROS scavenging system under salinity in these halophytes. Urochondra setulosa showed highest APX and SOD activity followed by Atriplex lentiformis. Most efficient enzyme in degrading hydrogen peroxide i.e. CAT showed highest activity in Suaeda nudiflora followed by Atriplex nummularia and Urochondra setulosa, whereas Atriplex nummularia and Atriplex lentiformis showed higher POX activity. Significant variability in H2O2 and MDA content was also observed. These results possibly suggest higher inbuilt genetic potential of these halophytes to combat high salinity induced oxidative stress via higher antioxidant activities. Novelty statement: Halophytic plant adopt different strategies to cope up with the toxic ions and our studies show that the induction of antioxidant defense system to scavenge ROS, alongwith structural modifications in terms of lipid peroxidation and compartmentalization of toxic ions are the main strategies for tighter control of ion fluxes in the studied halophytes.

Phytotoxicity of green synthesized silver nanoparticles on Camelina sativa L.

Due to the increased production and release of silver nanoparticles (AgNPs) in the environment, the concerns about the possibility of toxicity and oxidative damage to plant ecosystems should be considered. In the present study, the effects of different concentrations of AgNPs (0, 0.5, 1, 2, 3 and 4g/L) synthesized using the extract of camelina (Camelina sativa) leaves on the growth and the biochemical traits of camelina seedlings were investigated. The results showed that AgNPs significantly increased Ag accumulation in the roots and shoots which decreased the growth and photosynthetic pigments of camelina seedlings. The highest decrease in the height and total dry weight was observed by 53.1 and 61.8% under 4g/L AgNPs, respectively over control plants. AgNPs application over 2g/L enhanced the accumulation of proline, malondialdehyde, hydrogen peroxide and methylglyoxal, and up-regulated the activity of antioxidant enzymes (superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) and glyoxalase (glyoxalase I and II) system which indicates oxidative stress induction in camelina seedlings. Moreover, AgNPs reduced ASA and GSH contents and increased DHA and GSSG contents, hence disrupting the redox balance. These results showed that AgNPs at 4g/L had the most toxic effects on the camelina growth. Therefore, increasing oxidative stress markers and the activity of antioxidant enzymes and enzymes involved in glyoxalase system indicated the oxidative stress induced by AgNPs treatments over 2g/L as well as the induction of antioxidant defense systems to combat AgNPs-induced oxidative stress.

Water deficit-induced oxidative stress and differential response in antioxidant enzymes of tolerant and susceptible tea cultivars under field condition

Tea (Camellia sinensis (L) O. Kuntze) sustainable production is threatened by frequent drought. Tolerance to drought is a complex phenotypic response, which is controlled by the interactive properties of the plant’s genetic makeup, environment and its growing stage. The present investigation aimed to identify the differential responses of antioxidative enzymes, non-enzymatic antioxidants and status of osmolytes in tea plants under water deficit condition with respect to gradually depleting soil moisture content (SMC) and leaf relative water content (RWC). We examined the consequences of water deficit on the physiological characteristics and biochemical compounds of C. sinensis leaves. Our results indicated that the tolerant genotypes maintained favorable water relations, relatively higher photosynthetic pigments and reduced membrane injury by lowering hydrogen peroxide (H2O2) accumulation compared to susceptible cultivars. The relatively lower efficiency of superoxide dismutase (SOD), ascorbate peroxidase (APX) and peroxidase (POX) in susceptible cultivars under subsequent water deficit stress make them more prone to magnified oxidative damage than the tolerant cultivars. Furthermore, a substantial enhancement in the accumulation of osmolytes (proline and glycine betaine) has a great contribution in tolerance during drought. These results provide the insight into the basic mechanisms necessary for induction of antioxidant defense system and accumulation of osmolytes under water deficit conditions.

Exogenous melatonin alleviates cadmium uptake and toxicity in apple rootstocks.

To examine the potential roles of melatonin in Cd uptake, accumulation and detoxification in Malus plants, we exposed two different apple rootstocks varying greatly in Cd uptake and accumulation to either 0 or 30 μM Cd together with 0 or 100 μM melatonin. Cd stress stimulated endogenous melatonin production to a greater extent in the Cd-tolerant M. baccata than in the Cd-susceptible M. micromalus ‘qingzhoulinqin’. Melatonin application attenuated Cd-induced reductions in growth, photosynthesis, and enzyme activity, as well as ROS and MDA accumulation. Melatonin treatment more effectively restored photosynthesis, photosynthetic pigments, and biomass in Cd-challenged M. micromalus ‘qingzhoulinqin’ than in Cd-stressed M. baccata. Exogenous melatonin lowered root Cd2+ uptake, reduced leaf Cd accumulation, decreased Cd translocation factors (Tfs), and increased root, stem, and leaf melatonin contents in both Cd-exposed rootstocks. Melatonin application increased both antioxidant concentrations and enzyme activities to scavenge Cd-induced ROS. Exogenous melatonin treatment altered the mRNA levels of several genes regulating Cd uptake, transport, and detoxification including HA7, NRAMP1, NRAMP3, HMA4, PCR2, NAS1, MT2, ABCC1, and MHX. Taken together, these results suggest that exogenous melatonin reduced aerial parts Cd accumulation and mitigated Cd toxicity in Malus plants, probably due to the melatonin-mediated Cd allocation in tissues, and induction of antioxidant defense system and transcriptionally regulated key genes involved in detoxification.

Exogenous methyl salicylate alleviates senescent spotting by enhancing the activity of antioxidative ascorbate-glutathione cycle in harvested ‘Sucrier’ bananas

Senescent peel spotting, a physiological disorder, develops during the latter stage of ripening which coincides with the best eating quality of ‘Sucrier’ banana fruit. It is a major constraint for banana growers and traders. The aims of this study were to evaluate the regulatory roles of methyl salicylate (MeSA) on the senescent spotting and the activity of antioxidative ascorbate glutathione (ASA-GSH) cycle in ‘Sucrier’ banana during storage. ‘Sucrier’ bananas (ripening stage 3–4) were immersed in 0 (control) and 2 mM MeSA for 30 min, then air dried and stored at 25 ± 1 °C for 6 d. After treatment, peel spotting, reactive oxygen species (ROS) production (hydrogen peroxide and hydroxyl radical contents), oxidative membrane damage (malondialdehyde and protein carbonyl contents and electrolyte leakage), enzymatic and non-enzymatic components of ASA-GSH cycle were determined. It was shown that the symptoms of peel spotting in the control group was observed on day 2 and the severity increased continuously throughout storage. The occurrence of peel spotting coincided with the marked increase in the ROS production and oxidative membrane damage. The activities of ASA-GSH cycle including ascorbate peroxidase, dehydroascorbate reductase, monodehydroascorbate reductase and glutathione reductase activities as well as ascorbate (ASA) and reduced glutathione (GSH) contents declined with an increase in the severity of peel spotting. However, MeSA treatment caused an overall increase in the activities of enzymatic and non-enzymatic antioxidants as well as ASA/dehydroascorbate and GSH/oxidized glutathione ratios for up to 5–6 d of storage. The increased activity of ASA-GSH cycle was also associated with the decreases in ROS levels, oxidative membrane damage and senescent spotting development, indicating that MeSA treatment could reduce senescent spotting of ‘Sucrier’ bananas during storage by enhancing the activity of ASA-GSH cycle leading to the induction of antioxidant defense system to overcome ROS production, oxidative damage and fruit senescence.

Upregulation of Znf179 acetylation by SAHA protects cells against oxidative stress

The accumulation of reactive oxygen species (ROS) commonly occurs during normal aging and during some acute/chronic progressive disorders. In order to avoid oxidative damage, scavenging of these radicals is important. Previously, we identified zinc finger protein 179 (Znf179) as a neuroprotector that increases antioxidant enzymes against superoxide radicals. However, the molecular mechanisms involved in the activation and regulation of Znf179 remain unresolved. Here, by performing sequence alignment, bioinformatics analysis, immunoprecipitation using two specific acetyl-lysine antibodies, and treatment with the histone deacetylase (HDAC) inhibitor SAHA, we determined the lysine-specific acetylation of Znf179. Furthermore, we investigated Znf179 interaction with HDACs and revealed that peroxide insult induced a dissociation of Znf179-HDAC1/HDAC6, causing an increase in Znf179 acetylation. Importantly, HDAC inhibition by SAHA further prompted Znf179 hyperacetylation, which promoted Znf179 to form a transcriptional complex with Sp1 and increased antioxidant gene expression against oxidative attack. In summary, the results obtained in this study showed that Znf179 was regulated by HDACs and that Znf179 acetylation was a critical mechanism in the induction of antioxidant defense systems. Additionally, HDAC inhibitors may have therapeutic potential for induction of Znf179 acetylation, strengthening the Znf179 protective functions against neurodegenerative processes.

Apigenin pretreatment enhances growth and salinity tolerance of rice seedlings

Soil salinity is a limiting factor in rice production. Since flavonoids present in most plant tissues play multiple roles in plant-environment interactions, in this study, we focused on the contribution of flavone aglycone (Apigenin) to the adaptation of salinity-sensitive rice cultivar ‘Koshihikari,’ to salinity stress, for the first time. Rice seeds were soaked in Apigenin solution (10 ppm) for 24 h, then air-dried and grown hydroponically under 50 mM NaCl for 14 days. Apigenin pretreatment improved the growth of rice seedlings by enhancing shoot elongation and dry mass accumulation under both unstressed and NaCl-stress conditions, compared with that in the non-pretreated seedlings. Apigenin pretreatment significantly reduced Na+ accumulation in the salinity-stressed seedlings, and helped to maintain a lower Na+/K+ ratio in all plant organs, compared with that in the non-pretreated seedlings, possibly by regulating the expression of some important Na+ transporter-encoding genes (OsHKT2;1, OsCNGC1, OsSOS1). Higher levels of lipid peroxidation and hydrogen peroxide (H2O2) concentrations were observed in the shoots of the salinity-stressed seedlings; however, lower levels of lipid peroxidation and H2O2 concentration were detected in the Apigenin-treated seedlings. Apigenin pretreatment was associated with the induction of the rice antioxidant defense system represented by the induced activities of the antioxidant enzymes Catalase (CAT) and Ascorbate peroxidase (APX) in the roots, as well as by increased accumulation of the non-enzymatic antioxidants carotenoids and flavonoids in the shoots, relative to that in the untreated seedlings, under salinity stress conditions. Together, these results suggest that Apigenin pretreatment can alleviate the damaging effects of salinity on rice seedlings, presumably by regulating selective ion uptake by the roots and translocation to the shoots, thereby maintaining higher K+/Na+ ratios critical for normal plant growth under salinity stress, and by triggering the induction of the antioxidant defense system.

Synergistic effects of plant defense elicitors and Trichoderma harzianum on enhanced induction of antioxidant defense system in tomato against Fusarium wilt disease

Plant defense against their pathogens can be induced by a complex network of different inducers. The present study investigates the synergistic effect of Trichoderma harzianum, exogenous salicylic acid (SA) and methyl jasmonate (MeJA) over the response and regulation of the antioxidant defense mechanisms and lipid peroxidation in tomato plants against Fusarium wilt disease. In the present work, tomato plants were infected by Fusarium oxysporum f. sp. lycopersici 3 days after inoculated with T. harzianum and/or sprayed daily for 3 days with chemical inducers (SA and MeJA). Plants were analysed at 0, 24, 48, 72 and 96 h after inoculation with Fusarium oxysporum f. sp. lycopersici. Infection of tomato plants by pathogen led to strong reduction in the dry weight of roots and shoots with the enhanced concentration of H2O2 and varying degree of lipid peroxidation. Concurrently, exogenous SA, when applied with pathogen greatly enhanced H2O2 content as well as activities of antioxidant enzymes except catalase (CAT) and ascorbate peroxidase (APx). The pathogen challenged plants pretreated with T. harzianum and MeJA together exhibited less lipid peroxidation and as well as the elevated level of ascorbic acid and enhanced activities of antioxidant enzymes. All applied treatments protected tomato seedlings against Fusarium wilt disease but the percentage of protection was found higher in plants pretreated with the combination of T. harzianum and chemical inducers.

The role of acetone in the [omim][BF4]-mediated adverse effects on tissues of mussels, human lymphocytes and the fruit fly Drosophila melanogaster

The present study investigated [omim][BF4]-mediated adverse effects on biological models widely used in toxicological studies. Specifically, mussels of the genus Mytilus, human lymphocytes and fruit flies of the species Drosophila melanogaster, were exposed to [omim][BF4] at concentrations ranging from micro- to milligrams per liter, with or without the presence of acetone as a carrier solvent and thereafter [omim][BF4]-mediated adverse effects were analyzed appropriately (stress indices, such as lipid peroxidation byproducts, acetylcholinesterase/AChE activity and micronucleus/MN formation frequency, in mussel gills, Cytokinesis Block Micronucleus/CBMN assay and SMART test in human lymphocytes and fruit flies respectively). LC–MS–TOF analysis was also performed for elucidating [omim][BF4] mode of action in the presence of the carrier solvent. The results showed the toxic potential of [omim][BF4], as well as acetone’s ability to attenuate [omim][BF4]-mediated toxicity in almost all cases, probably due to the significant effect of acetone on the hydrophilic-lipophilic character and the viscosity of [omim][BF4], as well as its interaction and permeability on the cell membranes. The slight involvement of acetone in the attenuation of [omim][BF4]-mediated genotoxic effects on D. melanogaster could be due to species feeding experimental conditions, thus favoring the induction of antioxidant defense system against the [omim][BF4]-mediated effects in all cases.

Early and efficient induction of antioxidant defense system in Mytilus galloprovincialis embryos exposed to metals and heat stress

The present study aims to elucidate the stress response of early life stages of Mytilus galloprovincialis to the combine effects of selected metals and elevated temperature.For this purpose, we investigated the response of a large panel of oxidative stress markers such as catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST) activities and lipid peroxidation (thiobarbituric acid reactive substrates (TBARS) concentration) and metallothionein accumulation (MT) as well as selected gene transcription level and metal accumulation in mussels larvae exposed to a sub-lethal concentration of Cu (9.54µg/L), Ag (2.55µg/L) and mixture of the two metals (Cu (6.67µg/L)+Ag (1.47µg/L)) along with a temperature gradient (18, 20 and 22°C) for 48h.Cu and Ag applied as single or mixture were differentially accumulated in mussel larvae according to the exposure temperature. Sod, cat, gst and mt-10 gene transcription levels showed an important increase in larvae exposed to Cu, Ag or to the mix compared to the control condition at 18°C. The same pattern but with higher induction levels was recorded in larvae co-exposed to metals at 20°C. At 22°C, a significant decrease in mRNA abundance of cat, gst and sod and a significant up-regulation of mts targets (mt10 and mt20) were observed. Resultssuggest that co-exposure to metals and moderate elevated temperature (20 and 22°C) significantly increased the antioxidant enzyme activities of catalase (CAT), and glutathione-S-transferase (GST) and caused an increase of metal and metallothionein concentrations. In contrast, no significant change in lipid peroxidation products measured as TBARS content was observed indicating a protective response of anti-oxidative system.This study provides first evidences of the early and efficient protective response of antioxidant defense mechanisms in mussel's early life stages facing in multi stressors situations.

Abscisic Acid-Induced H2O2 Accumulation Enhances Antioxidant Capacity in Pumpkin-Grafted Cucumber Leaves under Ca(NO3)2 Stress.

With the aim to clarifying the role of the ABA/H2O2 signaling cascade in the regulating the antioxidant capacity of grafted cucumber plants in response to Ca(NO3)2 stress, we investigated the relationship between ABA-mediated H2O2 production and the activities of antioxidant enzymes in the leaves of pumpkin-grafted cucumber seedlings. The results showed that both ABA and H2O2 were detected in pumpkin-grafted cucumber seedlings in response to Ca(NO3)2 treatment within 0.5 h in the leaves and peaked at 3 and 6 h after Ca(NO3)2 treatment, respectively, compared to the levels under control conditions. The activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), and peroxidase (POD) in pumpkin-grafted cucumber leaves gradually increased over time and peaked at 12 h of Ca(NO3)2 stress. Furthermore, in the leaves of pumpkin-grafted cucumber seedlings, the H2O2 generation, the antioxidant enzyme activities and the expression of SOD, POD and cAPX were strongly blocked by an inhibitor of ABA under Ca(NO3)2 stress, but this effect was eliminated by the addition of exogenous ABA. Moreover, the activities and gene expressions of these antioxidant enzymes in pumpkin-grafted leaves were almost inhibited under Ca(NO3)2 stress by pretreatment with ROS scavengers. These results suggest that the pumpkin grafting-induced ABA accumulation mediated H2O2 generation, resulting in the induction of antioxidant defense systems in leaves exposed to Ca(NO3)2 stress in the ABA/H2O2 signaling pathway.

Biochemical and physiological characteristics of tropical mung bean (Vigna radiata L.) cultivars against chronic ozone stress: an insight to cultivar-specific response.

Surface-level ozone (O3) has been regarded as one of the most significant phytotoxic pollutants worldwide. Investigations addressing adverse impacts of elevated O3 on mung bean (Vigna radiata L.), an important leguminous crop of the Indian subcontinent, are still limited. The present study analyzed the differences on the foliar injury, reactive oxygen species (ROS) generation, antioxidative defense system, physiology, and foliar protein profile of two tropical mung bean cultivars (HUM-2 and HUM-6) exposed to elevated O3 under near-natural conditions. Both cultivars were negatively affected by the pollutant, but the response was cultivar-specific. Results revealed that elevated O3 induced higher levels of ROS (O2 (·-) and H2O2) and lipid peroxidation leading to greater foliar injury in HUM-2 compared to HUM-6. Photosynthetic pigments, photosynthetic rate, stomatal conductance, and photochemical efficiency reduced under elevated O3 exposure and the extent of reduction was higher in HUM-2. Principal component analysis revealed that photosynthetic performance and quantum yield were drastically affected in HUM-2 as compared to HUM-6. Activities of antioxidative enzymes were also stimulated, suggesting generation of oxidative stress under elevated O3. HUM-6 showed higher induction of antioxidative enzymes than HUM-2. One-dimensional gel electrophoresis analysis showed drastic reductions in the abundantly present ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) large and small subunits and the decrease was higher in HUM-2. Altogether, results suggested that higher accumulation of ROS and limited induction of antioxidant defense system led to more leaf injury and impairment of photosynthesis in HUM-2 than HUM-6 depicting its higher sensitivity towards elevated O3.

Arsenate induced differential response in rice genotypes

To study the differential response in two rice genotypes (PB1 and IR-64), hydroponically grown 14 days old plants were exposed to 50, 150 and 300µM As(V) for 24 and 96h. Accumulation of As was not significantly higher in PB1 variety except at higher concentration (300μM) and duration (96h), but up regulation of gene transcripts were higher as compared to IR-64. Inhibition in seed germination, root–shoot length, chlorophyll and protein content was observed in both varieties with increasing concentration and exposure time. PB1 variety was found more capable to detoxify As(V) through induction of antioxidant defense system and other stress related parameters (cysteine, proline content). SDS-PAGE and semi quantitative RT-PCR analysis showed significant changes in protein profile and gene expression analysis. The results suggests that various studied parameters and transcripts accumulation showed a combinatorial type of tolerance mechanism in PB1 variety to provide better protection against As(V) stress.

Long-term dietary exposure to low concentration of dichloroacetic acid promoted longevity and attenuated cellular and functional declines in aged Drosophila melanogaster

Dichloroacetic acid (DCA), a water disinfection by-product, has attained emphasis due to its prospect for clinical use against different diseases including cancer along with negative impact on organisms. However, these reports are based on the toxicological as well clinical data using comparatively higher concentrations of DCA without much of environmental relevance. Here, we evaluate cellular as well as organismal effects of DCA at environmentally and mild clinically relevant concentrations (0.02-20.0 μg/ml) using an established model organism, Drosophila melanogaster. Flies were fed on food mixed with test concentrations of DCA for 12-48 h to examine the induction of reactive oxygen species (ROS) generation, oxidative stress (OS), heat shock genes (hsps) and cell death along with organismal responses. We also examined locomotor performance, ROS generation, glutathione (GSH) depletion, expression of GSH-synthesizing genes (gclc and gclm), and hsps at different days (0, 10, 20, 30, 40, 50) of the age in flies after prolonged DCA exposure. We observed mild OS and induction of antioxidant defense system in 20.0 μg/ml DCA-exposed organism after 24 h. After prolonged exposure to DCA, exposed organism exhibited improved survival, elevated expression of hsp27, gclc, and gclm concomitant with lower ROS generation and GSH depletion and improved locomotor performance. Conversely, hsp27 knockdown flies exhibited reversal of the above end points. The study provides evidence for the attenuation of cellular and functional decline in aged Drosophila after prolonged DCA exposure and the effect of hsp27 modulation which further incites studies towards the therapeutic application of DCA.

Morphological, physiological and oxidative stress markers during acclimatization and field transfer of micropropagated Tuberaria major plants

Tuberaria major (Willk.) P. Silva and Rozeira is a critically-endangered rock rose species endemic to Portugal. Because the species needs to be preserved, this study evaluated the morphological and physiological traits of micropropagated T. major plants during acclimatization and field transfer. There were no significant differences between wild and micropropagated plants in the field, although the latter underwent significant changes during acclimatization. Leaf pubescence and leaf mass per area increased during acclimatization whereas the chlorophyll content and chlorophyll/carotenoid ratio declined to eventually match those of wild plants. Stomatal conductance (gs) and transpiration rates (E) also declined substantially during acclimatization, thus preventing uncontrolled wilting. Photosynthetic rate (PN) was initially negative but increased during the later stages of acclimatization. Maximum quantum yield of PSII (Fv/Fm) remained constant at 0.78–0.85, showing that the plants were healthy and unstressed. PSII quantum efficiency (ϕPSII) was initially low but increased during acclimatization along with photosynthetic performance as the energy partitioning in PSII was adjusted. This was balanced by the decline in non-regulated energy dissipation (ϕNO) from an initially high value. Electrolyte leakage and malondialdehyde content remained constant at similar levels in both groups of plants, but H2O2 levels were higher in the field, perhaps indicating the early induction of antioxidant defense systems. The present study shows that T. major has enough phenotypic plasticity to adapt to changing environments and that the procedure described herein can be used for the restoration and preservation of this species.

Reactive oxygen species, ascorbate–glutathione pool, and enzymes of their metabolism in drought-sensitive and tolerant indica rice (Oryza sativa L.) seedlings subjected to progressing levels of water deficit

Water deficit for rice is a worldwide concern, and to produce drought-tolerant varieties, it is essential to elucidate molecular mechanisms associated with water deficit tolerance. In the present study, we investigated the differential responses of nonenzymatic antioxidants ascorbate (AsA), glutathione (GSH), and their redox pool as well as activity levels of enzymes of ascorbate-glutathione cycle in seedlings of drought-sensitive rice (Oryza sativa L.) cv. Malviya-36 and drought-tolerant cv. Brown Gora subjected to water deficit treatment of -1.0 and -2.1MPa for 24-72h using PEG-6000 in sand cultures. Water deficit caused increased production of reactive oxygen species such as O2[Symbol: see text](-), H2O2, and HO[Symbol: see text] in the tissues, and the level of production was higher in the sensitive than the tolerant cultivar. Water deficit caused reduction in AsA and GSH and decline in their redox ratios (AsA/DHA and GSH/GSSG) with lesser decline in tolerant than the sensitive seedlings. With progressive level of water deficit, the activities of monodehydroascorbate reductase, dehydroascorbate reductase, ascorbate peroxidase (APX), and glutathione transferase increased in the seedlings of both rice cultivars, but the increased activity levels were higher in the seedlings of drought-tolerant cv. Brown Gora compared to the sensitive cv. Malviya-36. Greater accumulation of proline was observed in stressed seedlings of tolerant than the sensitive cultivar. In-gel activity staining of APX revealed varying numbers of their isoforms and their differential expression in sensitive and tolerant seedlings under water deficit. Results suggest that an enhanced oxidative stress tolerance by a well-coordinated cellular redox state of ascorbate and glutathione in reduced forms and induction of antioxidant defense system by elevated activity levels of enzymes of ascorbate-glutathione cycle is associated with water deficit tolerance in rice.