Activities Of DHAR Research Articles

Dynamic responses of germination characteristics and antioxidant systems to alfalfa (Medicago sativa) seed aging based on transcriptome

Seed aging poses a significant challenge to agronomic production and germplasm conservation. Reactive oxygen species (ROS) are highly involved in the aging process. However, dynamic response of germination characteristics and antioxidant system to seed aging are not yet very clear. This study explored the potential physiological mechanisms responsible for the reduced and rapid loss of seed vigor in alfalfa, and identified key genes regulating seed vigor. The germination percentage exhibited a decreased trend with the prolongation of aging duration. From 16 to 32 days of aging, the antioxidant enzyme activities of SOD, POD, CAT, DHAR and MDHAR declined significantly, which lead to the disruption of ROS balance and a significant increase in ROS levels, exacerbating seed aging. Based on transcriptome, 29 differentially expressed genes (DEGs) including SOD1, APX-2 and GST-7 within the ROS scavenging system showed a significantly down-regulated expression trend at aging of 16 and 24 days, indicating the abnormal function of antioxidant metabolism. Furthermore, some related genes including ATPF1B, ATPeF0C-3, NDUFS1, NDUFS3 and ND2 in the mitochondrial ETC exhibited a downturn following seed aging, which would result in the losing of seed vigor. This study has uncovered a significant array of potential target genes within the seed antioxidant system and mitochondrial ETC. These discoveries offer a wider lens for delving into the molecular regulatory mechanisms of seed aging. Further research is crucial to comprehensively elucidate the precise pathways through which these pivotal genes regulate seed vigor.

Untying arsenite tolerance mechanisms in contrasting maize genotypes attributed to NIPs-mediated controlled influx and root-to-shoot translocation, redox homeostasis and phytochelatin-mediated detoxification pathway

Contamination of ground water and soil with toxic metalloids like arsenic (As) poses a serious hazard to the global agricultural food production. One of the best ways to restrict entry of As into the food chain is selection of germplasms which accrue extremely low level of As in grains. Here, we screened diverse maize genotypes under high arsenite (100 μM AsIII) stress and identified PMI-PV-9 and PMI-PV-3 as AsIII-tolerant and -sensitive maize genotype respectively. Expression of genes associated with As uptake, vacuolar sequestration, biosynthesis of phytochelatins, root-to-shoot translocation, in vivo ROS generation, fine tuning of antioxidant defense system, DNA and membrane damage, H2O2 and superoxide anion (O2•-) levels were compared among the selected genotypes. PMI-PV-9 plants performed much better than PMI-PV-3 in terms of plant growth with no visible symptom of As toxicity. Susceptibility of PMI-PV-3 to AsIII stress may be attributed to comparatively low expression of genes involved in phytochelatins (PCs) biosynthesis. Concomitant decrease in ABCC1 expression might be another key factor for futile sequestration of AsIII into root vacuoles. Moreover, up-regulation of ZmNIP3;1 might contribute in high root-to-leaf As translocation. Substantial spike in H2O2, O2•- and MDA levels indicates that PMI-PV-3 plants have experienced more oxidative stress than PMI-PV-9 plants. Appearance of prominent deep brown and dark blue spots/stripes on leaves as revealed after DAB and NBT staining respectively suggest severe oxidative burst in PMI-PV-3 plants. Marked reduction in DHAR and MDAR activity rendered PMI-PV-3 cells to recycle ascorbate pool ineffectively, which might have exacerbated their susceptibility to AsIII stress. In a nutshell, incompetent PCs mediated detoxification system and disruption of cellular redox homeostasis owing to feeble antioxidant defence system resulting oxidative burst might be the prime reasons behind reduced performance of PMI-PV-3 plants under AsIII stress.

Phytomelatonin maintained chromium toxicity induced oxidative burst in Brassica juncea L. through improving antioxidant system and gene expression

Chromium (Cr) contamination in soils reduces crop yields and poses a remarkable risk to human and plant system. The main objective of this study was to observe the protective mechanisms of exogenously applied melatonin (Mel- 0.05, 0.1, and 0.15 μM) in seedlings of Brassica juncea L. under Cr (0.2 mM) stress. This was accomplished by analysing the plant's morpho-physiological, biochemical, nuclear, membrane, and cellular characteristics, as well as electrolyte leakage. Superoxide, malondialdehyde, and hydrogen peroxide increased with Cr toxicity. Cr also increased electrolyte leakage. Seedlings under Cr stress had 86.4% more superoxide anion and 27.4% more hydrogen peroxide. Electrolyte leakage increased 35.7% owing to Cr toxicity. B. juncea L. cells with high radical levels had membrane and nuclear damage and decreased viability. Besides this, the activities of the antioxidative enzymes, as POD, APOX, SOD, GST, DHAR, GPOX and GR also elevated in the samples subjected to Cr toxicity. Conversely, the activity of catalase was downregulated due to Cr toxicity. In contrast, Mel reduced oxidative damage and conserved membrane integrity in B. juncea seedlings under Cr stress by suppressing ROS generation. Moreover, the activity of antioxidative enzymes that scavenge reactive oxygen species was substantially upregulated by the exogenous application of Mel. The highest concentration of Mel (Mel c- 0.15 μM) applied showed maximum ameliorative effect on the toxicity caused by Cr. It causes alleviation in the activity of SOD, CAT, POD, GPOX, APOX, DHAR, GST and GR by 51.32%, 114%, 26.44%, 48.91%, 87.51%, 149%, 42.30% and 40.24% respectively. Histochemical investigations showed that Mel increased cell survival and reduced ROS-induced membrane and nuclear damage. The findings showed that Mel treatment upregulated several genes, promoting plant development. Its supplementation decreased RBOH1 gene expression in seedling sunder stress. The results supported the hypothesis that Mel concentrations reduce Cr-induced oxidative burst in B. juncea.

Modification in the ascorbate-glutathione cycle leads to a better acclimation to high light in the rose Bengal resistant mutant of Nannochloropsis oceanica

Understanding how to adapt outdoor cultures of Nannochloropsis oceanica to high light (HL) is vital for boosting productivity. The N. oceanica RB2 mutant, obtained via ethyl methanesulfonate mutagenesis, was chosen for its tolerance to Rose Bengal (RB), a singlet oxygen (1O2) generator. Compared to the wild type (WT), the RB2 mutant showed higher resilience to excess light conditions. Analyzing the ascorbate-glutathione cycle (AGC), involving ascorbate peroxidases (APX, EC 1.11.1.11), dehydroascorbate reductase (DHAR, EC 1.8.5.1), and glutathione reductase (GR, EC 1.8.1.7), in the RB2 mutant under HL stress provided valuable insights. At 250 μmol photon m−2 s−1 (HL), the WT strain displayed superoxide anion radicals (O2▪-) and hydrogen peroxide (H2O2) accumulation, increased lipid peroxidation, and cell death compared to normal light (NL) conditions (50 μmol photon m−2 s−1). The RB2 mutant didn't accumulate O2▪- and H2O2 after HL exposure, and exhibited increased APX, DHAR, and GR activities and transcript levels compared to WT and remained consistent after HL treatment. Although the RB2 mutant had a smaller ascorbate (AsA) pool than the WT, its ability to regenerate dehydroascorbate (DHA) increased post HL exposure, indicated by a higher AsA/DHA ratio. Additionally, under HL conditions, the RB2 mutant displayed an improved glutathione (GSH) regeneration rate (GSH/GSSG ratio) without changing the GSH pool size. Remarkably, H2O2 or menadione (a O2▪- donor) treatment induced cell death in the WT strain but not in the RB2 mutant. These findings emphasize the essential role of AGC in the RB2 mutant of Nannochloropsis in handling photo-oxidative stress.

Nickel toxicity mitigation by supplementation of acetylcholine in wheat: Growth, photosynthetic and antioxidant activities

Influence of exogenous treatment of acetylcholine (ACh; 5 and 10 µM) was studied on the growth, photosynthesis and nitrate reductase activity in wheat under nickel (500 µM Ni) stress. Plant height, fresh and dry weight of shoot showed a decline of 36%, 23% and 40% respectively while as increased by 35%, 44% and 47% due to 10 µM ACh over control. Treatment of ACh, at both concentrations, alleviated the decline with obvious effects at higher (10 µM) concentrations. Treatment of ACh resulted in increased content of total chlorophylls, carotenoids and photosynthesis, intercellular carbon dioxide, transpiration and PSII activity, which were however significantly reduced by Ni stress. Exogenous application of ACh alleviated the Ni stress mediated decline in the activity of nitrate reductase. Wheat plants treated with Ni showed increased production of hydrogen peroxide resulting in increased lipid peroxidation and electrolyte leakage, however treatment of ACh reduced these parameters significantly reflecting in alleviation of oxidative damage. Activity of antioxidant enzymes assayed (SOD, CAT, APX, DHAR and GR) and the content of ascorbic acid and reduced glutathione increased significantly in ACh treated plants under normal and Ni stress. Activities of SOD, CAT, APX, DHAR and GR was increased by 162%, 108%, 70%, 107% and 87% respectively in Ni + 10 µM ACh treated plants. In addition, the content of phenols and flavonoids increased due to application of ACh. Content of nitrogen, potassium and calcium increased due to ACh and also alleviated the decline induced due to Ni stress while as the content of Ni was reduced in plants treated with ACh.

Pseudomonas stutzeri improves the tolerance of Lemna minor to Cu(OH)2 nanopesticide by regulating the uptake of copper, antioxidant defense mechanisms, and the expression of metacaspase-1, chlorophyllase, and stress-responsive genes

This study investigated the effect of Pseudomonas stutzeri inoculation on Lemna minor treated with Cu(OH)2 nanopesticide (NP). The results showed that P. stutzeri inoculation increased the relative growth rate (RGR) in NP-treated plants. Although chlorophyll and carotenoid contents decreased significantly in NP-treated plants, P. stutzeri inoculation led to an increase in chlorophyll and carotenoid contents in NP-treated plants. Copper (Cu) content increased with increasing NP concentration, but it decreased significantly in the presence of P. stutzeri. NP treatment caused increased H2O2 and TBARS levels, as well as proline levels. However, P. stutzeri inoculation led to decreased H2O2 and TBARS levels and increased SOD, POX, GST, GR, GPX, and DHAR activities. The expression of genes encoding SOD, GST, metacaspase-1, and chlorophyllase was upregulated by NP treatment alone. Additionally, when plants were inoculated with P. stutzeri, the expression of these genes was further enhanced. In conclusion, P. stutzeri inoculation had a positive effect on the growth and antioxidant system of L. minor treated with NP as it enhanced RGR, increased chlorophyll and carotenoid contents, and decreased Cu content and oxidative stress. These findings suggested that P. stutzeri has the potential to promote aquatic plant growth and counteract the negative impacts of NP on these plants.

Inhibition of wound healing in fresh-cut potatoes by ascorbic acid is associated with control of the levels of reactive oxygen species and the AsA-GSH cycle

This study aimed to investigate the regulatory mechanisms of ascorbic acid (AsA) treatment on reactive oxygen species (ROS) and the ascorbate-glutathione (AsA-GSH) cycle during wound healing of fresh-cut potatoes (FCPes). AsA treatment at two concentrations (5 and 10 g L−1) was used to measure changes in H2O2 and O2.− contents, antioxidant enzymes of SOD, POD, and CAT activities, StPOD-related genes expression levels, and the AsA-GSH cycle metabolism activity. The results showed that AsA treatment could reduce the contents of H2O2 and O2.−, downregulate the expressions of suberization-associated StPOD genes, decrease the activities of SOD, POD, MDHAR, and DHAR, induce the activities of CAT, APX, GR, and GPX, as well as maintain high levels of AsA and GSH and AsA/(AsA + DHA) and GSH/(GSH + GSSG) ratios. In summary, exogenous AsA treatment delayed the wound-healing process and maintained the quality of FCPes by reducing ROS production, inducing antioxidant enzyme activities, and enhancing the AsA-GSH cycle metabolism activity, thus improving the overall antioxidant capacity and balancing the generation of ROS and the antioxidant defense system.

Melatonin enhanced low-temperature combined with low-light tolerance of pepper (Capsicum annuum L.) seedlings by regulating root growth, antioxidant defense system, and osmotic adjustment.

Melatonin (MT) is an important biologically active hormone that plays a vital role in plant growth and development. In particular, it has been investigated for its roles in abiotic stress management. In this study, pepper seedlings were subjected to low-temperature combined with low-light stress (LL) (15/5°C, 100 μmol m-2s-1) prior to a foliar spray of 200mM MT for 168h to investigate the protective role of MT in pepper seedlings. Our results demonstrated that LL stress negatively affected root growth, and accelerated the accumulation of reactive oxygen species (ROS), including H2O2 and , changed the osmolytes contents, and antioxidative system. However, these were reversed by exogenous MT application. MT effectively promoted the root growth as indicated by significant increase in root length, surface area, root volume, tips, forks, and crossings. In addition, MT reduced the burst of ROS and MDA contents induced by LL, enhanced the activities of SOD, CAT, POD, APX, DHAR, and MDHAR resulted by upregulated expressions of CaSOD, CaPOD, CaCAT, CaAPX, CaDHAR, and CaMDHAR, and elevated the contents of AsA and GSH, declined DHA and GSSH contents, which prevented membrane lipid peroxidation and protected plants from oxidative damages under LL stress. Furthermore, seedlings treated with MT released high contents of soluble sugar and soluble protein in leave, which might enhance LL tolerance by maintaining substance biosynthesis and maintaining cellular homeostasis resulted by high levels of osmolytes and carbohydrate in the cytosol. Our current findings confirmed the mitigating potential of MT application for LL stress by promoting pepper root growth, improving antioxidative defense system, ascorbate-glutathione cycle, and osmotic adjustment.

The Involvement of Antioxidant Enzyme System, Nitrogen Metabolism and Osmoregulatory Substances in Alleviating Salt Stress in Inbred Maize Lines and Hormone Regulation Mechanisms.

Salt stress inhibited the growth of maize. B46 and NC236 were chosen as materials and NaCl concentrations (0, 55, 110, 165, and 220 mmol L−1) were set. We found the activities of SOD, POD, CAT, APX, GR, MDHAR, and DHAR decreased under NaCl stress. Compared with NC236, the contents of AsA and GSH, AsA/DHA and GSH/GSSG of B46 decreased. The content of O2−, H2O2, MDA, and EL of B46 increased. The contents of NO3− and NO2− decreased, while the content of NH4+ increased under high NaCl concentration. The activities of NR and NiR decreased, while the activities of GS and GOGAT increased first and then decreased. For B46 and NC236, the maximum of NADH-GDH and NAD-GDH appeared at 165 and 110 mmol L−1 NaCl concentration, respectively. Compared with B46, and the GOT and GPT activities of NC236 increased first and then decreased. With the increase of NaCl concentration, the contents of proline, soluble protein, and soluble sugar were increased. The Na+ content of B46 and NC236 increased, and the K+ content and K+/Na+ decreased. Compared with NC236, B46 had higher IAA content in leaf, higher Z + ZR content in leaf and root, and lower ABA content in leaf and root.

Towards Better Grafting: SCoT and CDDP Analyses for Prediction of the Tomato Rootstocks Performance under Drought Stress

This study aims to predict the behavior of different tomato rootstocks under drought stress conditions. SCoT and CDDP analyses were employed to characterize the genetic relatedness among a commercial drought-sensitive tomato hybrid (cv. Bark) and four wild tomato accessions (LA2711, LA1995, LA3845, and LA4285) known for their tolerance to adverse conditions. The Bark plants were grafted onto the aforementioned wild accessions and self-grafted as control, and then the behavior of all graft unions was followed under normal and drought stress conditions. Our results showed a general genotype-dependent better growth and yield of heterografts than autografts under all growth conditions. Furthermore, clustering analysis based on growth, yield quantity and quality traits, and the leaf content of minerals, ABA, GA3, and proline, in addition to the activity of APX, POD, and DHAR reflected the same grouping pattern of the studied rootstocks exhibited by SCoT and CDDP. The identical grouping pattern supports the utilization of SCoT and CDDP as a robust screening tool helpful to predict the physiological and agronomical behavior of grafting on different tomato rootstocks. Furthermore, grafting could be a cost-efficient alternative method to improve drought tolerance in sensitive tomato genotypes.

Defensive capabilities of contrasting sorghum genotypes against Atherigona soccata (Rondani) infestation.

Plants are equipped with a wide range of defensive mechanisms such as morphophysiological, biochemical, molecular, and hormonal signaling for protecting against insect-pest infestation. The infestation of a devastating pest shoot fly [Atherigona soccata (Rodani)] at seedling stage causes huge loss of sorghum crop productivity. In morphophysiological screening ICSV700, ICSV705, and IS18551 have been categorized as resistant, PSC-4 moderately resistant, SL-44 and SWARNA as susceptible. The present study focused on the role of defensive gene expression and its products viz: superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), glutathione reductase (GR), polyphenol oxidase (PPO), phenyl alanine ammonia lyase (PAL), responsive enzymes, and metabolites restoring redox status in sorghum plants against shoot fly infestation. In both leaf and stem tissue of sorghum genotypes,shoot fly infestation induced SOD, APX, DHAR, GR, PAL, and PPO activities while CAT activity was significantly declined at 15 and 21days after emergence (DAE). IS18551 with resistant behavior showed upregulation of SOD, GR, APX, and DHAR along with accumulation of ascorbate, glutathione enhancing redox status of the plant during shoot fly infestation at later stage of infestation. While SWARNA with susceptible response exhibited enhanced activity of phenylpropanoid pathway enzymes PAL and PPO which in turn increased the levels of secondary metabolites like o-dihydroxyphenol and other phenols deterring the insect to attack the plant. The qRT-PCR data predicted that stress-responsive genes were initially unregulated in SWARNA; however, at 21 DAE, multifold higher expression of SOD, CAT, APX, and PPO (24.8-, 37.2-, 21.7-, and 17.9-fold respectively) in 1S18551 indicates the resistance behavior of this genotype against insect infestation owing to sustainable development capability.

Kinetin mitigates Cd-induced damagesto growth, photosynthesis and PS II photochemistry of Trigonella seedlings by up-regulating ascorbate-glutathione cycle.

Cytokinins (CKs) plays a key role in plant adaptation over a range of different stress conditions. Here, we analyze the effects of a cytokinin (i.e., kinetin, KN) on the growth, photosynthesis (rate of O2 evolution), PS II photochemistry and AsA–GSH cycle in Trigonella seedlings grown under cadmium (Cd) stress. Trigonella seeds were sown in soil amended with 0, 3 and 9 mg Cd kg-1 soil, and after 15 days resultant seedlings were sprayed with three doses of KN, i.e.,10 μM (low, KNL), 50 μM (medium, KNM) and 100 μM (high, KNH); subsequent experiments were performed after 15 days of KN application, i.e., 30 days after sowing. Cadmium toxicity induced oxidative damage as shown by decreased seedling growth and photosynthetic pigment production (Chl a, Chl b and Car), rates of O2-evolution, and photochemistry of PS II of Trigonella seedlings, all accompanied by an increase in H2O2 accumulation. Supplementation with doses of KN at KNL and KNM significantly improved the growth and photosynthetic activity by reducing H2O2 accumulation through the up-regulation AsA–GSH cycle. Notably, KNL and KNM doses stimulated the rate of enzyme activities of APX, GR and DHAR, involved in the AsA–GSH cycle thereby efficiently regulates the level of AsA and GSH in Trigonella grown under Cd stress. The study concludes that KN can mitigate the damaging effects of Cd stress on plant growth by maintaining the redox status (>ratios: AsA/DHA and GSH/GSSG) of cells through the regulation of AsA-GSH cycle at 10 and 50 μM KN under Cd stress conditions. At 100 μM KN, the down-regulation of AsA-GSH cycle did not support the growth and PS II activity of the test seedlings.

Melatonin alleviates imidacloprid phytotoxicity to cucumber (Cucumis sativus L.) through modulating redox homeostasis in plants and promoting its metabolism by enhancing glutathione dependent detoxification

Melatonin (Mel), a powerful antioxidant that has the ability to regulate physiological and biochemical processes in plants under abiotic stresses. However, its roles in pesticide detoxification is poorly understood. Herein, selecting leaf spraying insecticide imidacloprid (IMD) as the model, we demonstrated the detoxification mechanism underlying root pretreatment of Mel on IMD in cucumber. IMD treatment affected the primary light conversion efficiency of photosystem II (Fv/Fm), reduced the quantum yield, and increased hydrogen peroxide and superoxide anions contents as well as the levels of membrane lipid peroxidation, indicating that excessive IMD treatment induces oxidative stress. Nonetheless, by increasing the appropriate levels of exogenous Mel, the photosynthesis of cucumber under IMD treatment reached the control levels, effectively removing reactive oxygen species. Furthermore, the content and ratio of ascorbate (AsA) and glutathione (GSH) were decreased under IMD treatment; Mel treatment enhanced the AsA/DHA and GSH/GSSG ratios, as well as the activities of MDHAR, DHAR and GR, suggesting that Mel could alleviate oxidative stress of cucumber treated with IMD by regulating the ascorbic acid-glutathione cycle. Importantly, IMD degradation rate and glutathione S-transferase (GST) activity increased after Mel treatment. The levels of transcripts encoding antioxidant enzymes GPX and GST (GST1,2 and 3) were also increased, indicating that Mel accelerated IMD degradation. These results suggest that Mel plays an important role in the detoxification of IMD by promoting GST activity and transcription and the AsA-GSH cycle, thus providing an approach for plants to reduce IMD residue through the plant's own detoxification mechanism.

Mitochondrial bioenergetics and enzymatic antioxidant defense differ in Paraná pine cell lines with contrasting embryogenic potential

Araucaria angustifolia is classified as a critically endangered species by the International Union for Conservation of Nature. This threat is worsened by the inefficiency of methods for ex-situ conservation and propagation. In conifers, somatic embryogenesis (SE) associated with cryopreservation is an efficient method to achieve germplasm conservation and mass clonal propagation. However, the efficiency of SE is highly dependent on genotype responsivity to the artificial stimulus used in vitro during cell line proliferation and later during somatic embryo development. In this study, we evaluated the activity of antioxidant enzymes and characterized mitochondrial functions during the proliferation of embryogenic cells of A. angustifolia responsive (SE1) and non-responsive (SE6) to the development of somatic embryos. The activities of the antioxidant enzymes GR (EC 1.6.4.2), MDHAR (EC 1.6.5.4), and POX (EC 1.11.1.7) were increased in SE1 culture, while in SE6 culture, only the activity of DHAR (EC 1.8.5.1) was significantly higher. Additionally, SE6 culture presented a higher number of mitochondria, which agreed with the increased rate of oxygen consumption compared to responsive SE1 culture; however, the mitochondrial volume was lower. Although the ATP levels did not differ, the NAD(P)H levels were higher in SE1 cells. NDs, AOX, and UCP were less active in responsive SE1 than in non-responsive cells. Our results show significant differences between SE1 and SE6 embryogenic cells regarding mitochondrial functions and antioxidant enzyme activities, which may be intrinsic to the in vitro proliferation phase of both cell lines, possessing a crucial role for the induction of in vitro maturation process.

Three metabolic pathways are responsible for the accumulation and maintenance of high AsA content in kiwifruit (Actinidia eriantha)

BackgroundActinidia eriantha is a precious material to study the metabolism and regulation of ascorbic acid (AsA) because of its high AsA content. Although the pathway of AsA biosynthesis in kiwifruit has been identified, the mechanism of AsA metabolism and regulation is still unclear. The purpose of this experiment is to reveal the AsA metabolic characteristics of A. eriantha ‘Ganmi 6’ from the molecular level, and lay a theoretical foundation for the research on the genetic improvement of kiwifruit quality.ResultsWe found that AsA reached the accumulation peak at S7 (110 DAF) during the process of fruit growth and development. The activity of GalDH, GalLDH, MDHAR and DHAR in fruit was similar to AsA accumulation trend, and both of them were significantly positively correlated with AsA content. It was speculated that GalDH and GalLDH were key enzymes in AsA biosynthesis, while MDHAR and DHAR were key enzymes in AsA regeneration cycle, which together regulated AsA accumulation in fruit. Also, we identified 98,656 unigenes with an average length of 932 bp from the transcriptome libraries using RNA-seq technology after data assembly. There were 50,184 (50.87%) unigenes annotations in four databases. Two thousand nine hundred forty-nine unigenes were enriched into the biosynthesis pathway of secondary metabolites, among which 133 unigenes involved in the AsA and aldehyde metabolism pathways, and 23 candidate genes related to AsA biosynthesis, cycling and degradation were screened out.ConclusionsConsidering gene expression levels and changes of physiological traits and related enzyme activity, we concluded that the accumulation of AsA depends mainly on the L-galactose pathway, and the D-galacturonic acid pathway and AsA recycling pathway as the secondary pathways, which co-maintain the high AsA content in fruit of A. eriantha.

Involvement of salicylic acid in cold priming-induced freezing tolerance in wheat plants

Freezing is one of the most damaging abiotic stress factors. Cold priming has been shown to enhance freezing tolerance in wheat. However, the underlying mechanisms are unclear. Salicylic acid (SA) is an important signal molecule involved in plant responses to abiotic and biotic stresses. This study aims to investigate the role of SA in cold priming-induced freezing tolerance in wheat plants. The results showed that the expression of gene encoding phenylalanine ammonia-lyase (PAL, a key enzyme involved in SA biosynthesis) and the level of endogenous SA (both free and conjugated) were significantly up-regulated following cold priming treatment. The role of SA in cold priming-induced freezing tolerance was further explored using the donor and inhibitor of endogenous SA. l-α-aminooxy-β-phenylpropionic acid (AOPP, a specific inhibitor of PAL) pretreatment reversed the increment of SA level induced by cold priming treatment, attenuated the positive effects of cold priming on antioxidant capacity (as indicated by the contents of MDA, ASA and GSH, and the activities of APX, GR, DHAR and MDHAR) and cold-responsive genes expression (such as AOX1a, IRI2 and HSP70) under the later freezing stress as well as the freezing tolerance (as indicated by the photochemical activity, electrolyte leakage and biomass). The application of exogenous SA could obviously counteract the negative effects of AOPP. Findings of the present study suggested that the roles of SA in cold priming-induced freezing tolerance in wheat might involve regulation of antioxidant capacity and cold-responsive gene expression.

Effects of salt stress on antioxidant and ascorbate glutathione cycle enzyme activities in Pokkali rice varieties – Vytilla 1-9

The enzymatic and non-enzymatic antioxidant levels in the released salt tolerant Pokkali, (vytilla, VTL 1-9) varieties were studied under different NaCl concentrations (0-150 mM NaCl). The specific activity of superoxide dismutase (SOD), catalase (CAT) and ascorbate-glutathione cycle enzymes and non-enzymatic antioxidants like superoxide (O2-), hydrogen peroxide (H2O2), malondialdehyde (MDA), glutathione (GSH) and ascorbic acid (AsA) was determined in plants exposed to salt stress. IR-28 was used as positive control and the VTL varieties were used as negative control. The H2O2 and superoxide (O2-) contents were higher in IR-28 at all the applied concentrations of NaCl. The VTL varieties without salt treatment did not evoke any response substantiating the role of salt priming in antioxidant signalling. The MDA contents were higher in the positive and negative control. MDA content was reduced in the NaCl treated VTL varieties. In the positive and negative control varieties, the quantity of ascorbate and glutathione contents were lesser and upregulated in salt treated VTL varieties. Highest H2O2 content was observed in 150 mM NaCl treatment. The H2O2 contents decreased with the increase in all concentrations of NaCl and lowest H2O2 contents was observed in VTL-1 and highest in VTL-2 and VTL-8 treated with 150 mM NaCl. Superoxide contents varied in all the nine varieties depending on the salt concentration. The SOD levels in all the varieties showed a positive correlation with the superoxide and H2O2 content. Lesser quantities of SOD, CAT and the ascorbate - glutathione cycle enzymes were expressed in the positive and negative control. The increased NaCl concentration (25-150 mM) upregulated antioxidant and ascorbate-glutathione cycle enzymes in the VTL varieties. The APX activity was lower in the control and salt treated plants. The GR activity increased linearly in all the varieties with respect to salt concentrations. The MDHAR and DHAR activities showed marginally linear increase, with all concentrations of NaCl. The APX activity was similar or lower to MDHAR activity while DHAR activity was similar to MDHAR activity. The results of the present study reveals the higher levels of enzymatic and non-enzymatic antioxidants under salt stress reflect the salt tolerance potential of pokkali varieties mediated by the up regulation of ROS scavenging enzymes.

Effect of antimony in soils of an Sb mine on the photosynthetic pigments and antioxidant system of Dittrichia viscosa leaves.

Antimony is a toxic element whose concentration in soil and water has been rising due to anthropogenic activities. This study focuses on its accumulation in leaves of Dittrichia viscosa growing in soils of an abandoned Sb mine, and the effect on oxidant/antioxidant systems and photosynthetic efficiency. The results showed leaves to have a high Sb accumulation capacity. The amount of total chlorophyll decreased depending on Sb concentration and of carotenoids increased slightly, with a consequent increase in carotenoid/chlorophyll ratio. Photosynthetic efficiency was unaffected. The amount of O 2 .- rose, although there was no increase in cell membrane damage, with lipid peroxidation levels being similar to normal. This response may be due to considerable increases that were observed in total phenolics, PPO activity, and enzymatic antioxidant system. SOD, POX, and DHAR activities increased in response to increased Sb amounts in leaves. The ascorbate/glutathione cycle was also affected, with strong increases observed in all of its components, and consequent increases in total contents of the ascorbate and glutathione pools. However, the ratio between reduced and oxidized forms declined, reflecting an imbalance between the two, especially that between GSH and GSSG. Efficient detoxification of Sb may take place either through increases in phenolics, carotenoids, and components of the glutathione-ascorbate cycle or through the enzymatic antioxidant system. Since Dittrichia viscosa accumulates large amounts of Sb without suffering oxidative damage, it could be used for phytoremediation.

Effects of Antimony on Reactive Oxygen and Nitrogen Species (ROS and RNS) and Antioxidant Mechanisms in Tomato Plants.

This research studies the effects that Sb toxicity (0.0, 0.5, and 1.0 mM) has on the growth, reactive oxygen and nitrogen species, and antioxidant systems in tomato plants. Sb is accumulated preferentially in the roots, with little capacity for its translocation to the leaves where the concentration is much lower. The growth of the seedlings is reduced, with alteration in the content in other nutrients. There is a decrease in the content of Fe, Mg, and Mn, while Cu and Zn increase. The contents in chlorophyll a and b decrease, as does the photosynthetic efficiency. On the contrary the carotenoids increase, indicating a possible action as antioxidants and protectors against Sb. The phenolic compounds do not change, and seem not to be involved in the defense response of the tomato against the stress by Sb. The water content of the leaves decreases while that of proline increases in response to the Sb toxicity. Fluorescence microscopy images and spectrofluorometric detection showed increases in the production of O2.–, H2O2, NO, and ONOO–, but not of nitrosothiols. The Sb toxicity induces changes in the SOD, POX, APX, and GR antioxidant activities, which show a clear activation in the roots. In leaves, only the SOD and APX increase. The DHAR activity is inhibited in roots but undergoes no changes in the leaves, as is also the case for the POX and GR activities. Ascorbate increases while GSH decreases in the roots. The total AsA + DHA content increases in the roots, but the total GSH + GSSG content decreases, while neither is altered in the leaves. Under Sb toxicity increases the expression of the SOD, APX, and GR genes, while the expression of GST decreases dramatically in roots but increases in leaves. In addition, an alteration is observed in the pattern of the growth of the cells in the elongation zone, with smaller and disorganized cells. All these effects appear to be related to the ability of the Sb to form complexes with thiol groups, including GSH, altering both redox homeostasis and the levels of auxin in the roots and the quiescent center.

Transcriptomic analysis reveals the changes of energy production and AsA-GSH cycle in oat embryos during seed ageing

Deterioration during seed storage generally causes seed vigour declining. However, the mechanism of deterioration occurred still not clear. Seeds and embryos of oat (Avena sativa L.) were selected to analyze the relation of physiological and metabolic reactions with DEGs by using RNA-seq. Oat seed vigour declined during seeds aged 0 day (CK), 16 days (CD16) and 32 days (CD32). The changes of MDA and H2O2 contents, antioxidant enzymes activities of APX, DHAR, MDHAR and GR related with AsA-GSH cycle in embryos illustrated that seed vigour declined to the minimum at CD32. Transcriptomic analysis showed a total of 11335 and 8274 DEGs were identified at CD16 and CD32 compared with CK respectively, of which 4070 were overlapped. When seed vigour declined to the moderate level (CD16), the accumulation of H2O2 caused by the inhibition of complex I in ETC could be alleviated with AsA-GSH cycle. RNA-seq and qRT-PCR results both showed alternative oxidase in alternate respiratory pathway was upregulated which would maintain seed respiration. However, as seed vigour was at the lowest level (CD32), blocked ETC caused by down-regulation of complex III, including Ubiquinol-cytochrome C reductase complex 14kD subunit and Ubiquinol-cytochrome C reductase, UQCRX/QCR9 like, were more seriously and H2O2 scavenging was limited by the inactive AsA-GSH cycle. It could be suggested that the function of AsA-GSH would play a key role for regulating the physiological responses of ETC in embryos during seed ageing. These results would provide an insight into embryo for the transcriptomic information during oat seed ageing.