Physiological and Proteomics Analyses Reveal the Mechanism of Eichhornia crassipes Tolerance to High-Concentration Cadmium Stress Compared with Pistia stratiotes

Cadmium (Cd) pollution is an environmental problem worldwide. Phytoremediation is a convenient method of removing Cd from both soil and water, but its efficiency is still low, especially in aquatic environments. Scientists have been trying to improve the ability of plants to absorb and accumulate Cd based on interactions between plants and Cd, especially the mechanism by which plants resist Cd. Eichhornia crassipes and Pistia stratiotes are aquatic plants commonly used in the phytoremediation of heavy metals. In the present study, we conducted physiological and biochemical analyses to compare the resistance of these two species to Cd stress at 100 mg/L. E. crassipes showed stronger resistance and was therefore used for subsequent comparative proteomics to explore the potential mechanism of E. crassipes tolerance to Cd stress at the protein level. The expression patterns of proteins in different functional categories revealed that the physiological activities and metabolic processes of E. crassipes were affected by exposure to Cd stress. However, when some proteins related to these processes were negatively inhibited, some analogous proteins were induced to compensate for the corresponding functions. As a result, E. crassipes could maintain more stable physiological parameters than P. stratiotes. Many stress-resistance substances and proteins, such as proline and heat shock proteins (HSPs) and post translational modifications, were found to be involved in the protection and repair of functional proteins. In addition, antioxidant enzymes played important roles in ROS detoxification. These findings will facilitate further understanding of the potential mechanism of plant response to Cd stress at the protein level.     

Temperature-independent increase in the detoxifying enzyme activity of insecticide-resistant small brown planthoppers and Drosophila

The populations of migrated small brown planthoppers (SBPHs) collected in Korea were reported to have higher insecticide resistance and percentages of viruliferous individuals than the populations of overwintering SBPHs. Therefore, the migrated SBPHs might survive after insecticide treatments and cause severe damage to rice plants. In this study, the changes in the biochemical properties of resistant SBPHs with temperature changes were investigated. The activities of detoxifying enzymes known to be involved in insecticide resistance showed no correlation with temperature, but the resistant strains had consistently higher detoxifying enzyme activities than the susceptible strains. Interestingly, the amount of reactive oxygen species (ROS) increased with temperature in all strains.Chlorantraniliprole-resistant strains of Drosophila melanogaster were examined to investigate whether the phenomena observed in the resistant SBPHs were conserved in other insects with resistance to an insecticide with different modes of action. Similar to the resistant SBPHs, the resistant Drosophila also exhibited increased amounts of ROS and detoxifying enzyme activity compared to the control Drosophila. Also in the resistant Drosophila, only the ROS showed a temperature-dependent increase.Taken together, in addition to the involvement of increased activities of detoxifying enzymes, the resistant insects also had a temperature-dependent significant increase in ROS. Thus, the development of tools to induce ROS toxicity could lead to the development of new control methods to eliminate resistant insect strains.     

Impacts of an Invasive Non-Native Annual Weed,Impatiens glandulifera,on Above- and Below-Ground Invertebrate Communities in the United Kingdom

Vegetation community composition and the above- and below-ground invertebrate communities are linked intrinsically, though few studies have assessed the impact of non-native plants on both these parts of the community together. We evaluated the differences in the above- (foliage- and ground-dwelling) and below-ground invertebrate communities in nine uninvaded plots and nine plots invaded by the annual invasive species Impatiens glandulifera, in the UK during 2007 and 2008. Over 139,000 invertebrates were identified into distinct taxa and categorised into functional feeding groups. The impact of I. glandulifera on the vegetation and invertebrate community composition was evaluated using multivariate statistics including principal response curves (PRC) and redundancy analysis (RDA). In the foliage-dwelling community, all functional feeding groups were less abundant in the invaded plots, and the species richness of Coleoptera and Heteroptera was significantly reduced. In the ground-dwelling community, herbivores, detritivores, and predators were all significantly less abundant in the invaded plots. In contrast, these functional groups in the below-ground community appeared to be largely unaffected, and even positively associated with the presence of I. glandulifera. Although the cover of I. glandulifera decreased in the invaded plots in the second year of the study, only the below-ground invertebrate community showed a significant response. These results indicate that the above- and below-ground invertebrate communities respond differently to the presence of I. glandulifera, and these community shifts can potentially lead to a habitat less biologically diverse than surrounding native communities; which could have negative impacts on higher trophic levels and ecosystem functioning.     

Root morphology, root-hair development and rhizosheath formation on perennial grass seedlings is influenced by soil acidity

Cadmium (Cd) exerts a detrimental effect on the metabolism of plants, whereas selenium (Se) may protect them against various stressors through its antioxidative activity. In this in vitro study we investigated the impact of Se (2 µM Na₂SeO₄) on the growth, nutrient (P, S, K, Ca, Mg, B, Mn, Fe and Zn) concentrations and cell integrity of rape (Brassica napus oleifera) and two wheat (Triticum aestivum) genotypes subjected to Cd stress (600 µM CdCl₂). Rape accumulated both Cd and Se more than did wheat. In all plants, Cd markedly reduced the biomass, enhanced lipid peroxidation and diminished plasmalemma fluidity. A drop in the K uptake and the reduced plasmalemma permeability diminished the K efflux from the leaf cells. In contrast, Cd elevated S concomitantly with Zn, indicating an activity of detoxifying SH groups and SOD isoenzymes. When added alone, Se promoted the growth of all plants, it enhanced the accumulation of S, but the impact on other nutrients remained minor. In Cd-stressed plants, Se tended to counterbalance the Cd-induced changes in nutrients, it also reduced the lipid peroxidation and exerted positive effects on the cell membrane stability. The Cd stress and the protective role of Se were most evident in rape. The Finnish wheat genotype was less tolerant to Cd than the Polish one.     

Lipid peroxidation and antioxidative response in Arabidopsis thaliana exposed to cadmium and copper

Formation of lipid hydroperoxides, malondialdehyde (MDA) and hydroxyalkenals (HAEs), membrane damages and antioxidative response of plants expressed as changes in glutathione S-transferase activity (GST) and anthocyanin accumulation were studied in Arabidopsis thaliana (L.) Heynh cv. Columbia plants treated for 7 days with various concentrations: 5, 25, 50, 100 μM Cd and Cu. Increased lipid hydroperoxide content was metal concentration-dependent. The level of MDA + HAE was elevated in Cd- and Cu- treated plants, but it was metal concentration-dependent under Cu stress. Electrolyte leakage measurements showed a larger membrane damage under Cu- than Cd-treatment. In Cu-stressed plants, GST activity was always enhanced in comparison with control, while in plants exposed to Cd it dropped slightly at lower metal concentrations; but at 100 μM Cd it was even higher than in plants treated with the same Cu concentration. Anthocyanin accumulation was considerably higher under Cu than Cd stress. Both lipid peroxidation and antioxidative response was stronger in Cu- than Cd-treated Arabidopsis thaliana plants. Various mechanisms of defense against the lipid peroxidation products, depending on the metal type, are discussed.     

Paraquat-induced oxidative stress response during amphibian early embryonic development

In addition to the endogenous production of reactive oxygen species (ROS) as a result of normal development, amphibian external development often forces embryos to deal with oxidative stress-producing agents present in the environment. Embryos should therefore develop protective systems to reduce ROS toxicity and achieve successful development. The present work was aimed to characterize the effects produced by the widespread-used ROS-generator pesticide Paraquat during early embryonic development in the toad Chaunus arenarum, as well as to get insights into the defense response elicited by amphibian embryos. The approach consisted in generating a sharp and brief oxidative stress condition early during embryonic development to stimulate the cellular mechanisms involved in ROS-antioxidant response. Results revealed that Paraquat-treatment reduced the ability of embryos to develop normally, leading to arrests of development and severe malformations such as tail abnormalities, abdominal edema, reduced head development and curved dorsal structures. Although Paraquat effects were morphologically evident from gastrula stage on, alterations such as chromatin condensation were observed even at blastula stage by histological examinations. Regarding detoxifying enzymes, a significant induction of Mn-superoxide dismutase activity was detected at stages beyond gastrula in embryos surviving Paraquat treatment, suggesting a major role of this enzyme in the antioxidant response during early embryonic development.     

Overexpression of the glutamine synthetase gene modulates oxidative stress response in rice after exposure to cadmium stress

Key message

Overexpression of OsGS gene modulates oxidative stress response in rice after exposure to cadmium stress. Our results describe the features of transformants with enhanced tolerance to Cd and abiotic stresses.

Abstract

Glutamine synthetase (GS) (EC 6.3.1.2) is an enzyme that plays an essential role in the metabolism of nitrogen by catalyzing the condensation of glutamate and ammonia to form glutamine. Exposure of plants to cadmium (Cd) has been reported to decrease GS activity in maize, pea, bean, and rice. To better understand the function of the GS gene under Cd stress in rice, we constructed a recombinant pART vector carrying the GS gene under the control of the CaMV 35S promoter and OCS terminator and transformed using Agrobacterium tumefaciens. We then investigated GS overexpressing rice lines at the physiological and molecular levels under Cd toxicity and abiotic stress conditions. We observed a decrease in GS enzyme activity and mRNA expression among transgenic and wild-type plants subjected to Cd stress. The decrease, however, was significantly lower in the wild type than in the transgenic plants. This was further validated by the high GS mRNA expression and enzyme activity in most of the transgenic lines. Moreover, after 10 days of exposure to Cd stress, increase in the glutamine reductase activity and low or no malondialdehyde contents were observed. These results showed that overexpression of the GS gene in rice modulated the expression of enzymes responsible for membrane peroxidation that may result in plant death.     

Cadmium stress-induced oxidative stress and role of nitric oxide in rice (Oryza sativa L.)

Cadmium (Cd) is a potential environmental phytotoxicant. The generation of reactive oxygen species (ROS) due to Cd stress is responsible for the induction of oxidative stress in plants. On the other hand, SNP, a NO donor is known to have effect on Cd-induced oxidative stress in plants. We evaluated the effect of NO on the regulation of Cd stress in the rice (Oryza sativa L.) variety MSE-9. Cd treatment was given in the form of 50, 100 and 200 ??M, whereas for interaction study, 100 ??M of Cd and 100 ??M of SNP were used. The result showed that Cd-induced oxidative stress in MSE-9 by generating ROS. However, when SNP was given with Cd stress, it was seen that SNP treatment regulated the stress metabolism in rice seedlings under Cd toxicity by generating NO. It can be said that the SNP in combination with Cd treatment might possess the way to protect rice seedlings under Cd stress.     

Shoot endophytic plant growth-promoting bacteria reduce cadmium toxicity and enhance switchgrass (<Emphasis Type="Italic">Panicum virgatum</Emphasis> L.) biomass

The aims of the study were to increase the biomass and to alleviate the deleterious effects of cadmium (Cd) in the switchgrass cultivars (Panicum virgatum L.) Alamo and Cave-in-Rock (CIR) under cadmium (Cd) stress using Cd-tolerant shoot endophytic plant growth-promoting bacteria (PGPB). Four shoot endophytic bacterial strains, viz. Bc09, So23, E02, and Oj24, were isolated from the above-ground parts of plants grown in a Cd-polluted soil and were successfully identified by 16S rRNA gene sequencing as Pseudomonas grimontii, Pantoea vagans, Pseudomonas veronii, and Pseudomonas fluorescens, respectively. These four strains were adapted to high CdCl₂ concentrations as they had higher Cd uptake capacities. In addition, they possessed a huge amount of growth regulatory activities e.g., indole acetic acid production, 1-aminocyclopropane-1-carboxylic acid deaminase (ACCD) activity, and phosphate solubilization. Growth particularly the height and biomass of both cultivars increased significantly in response to PGPB inoculation in the 20 µM CdCl₂ stress. The shoot biomass of the PGPB-inoculated Alamo was higher than the CIR under Cd stress. Interestingly, the level of Cd inside PGPB-inoculated plant tissues and the translocation factors were lower compared with the noninoculated Cd control plants. CIR plants exhibited higher Cd content than Alamo plants. Through confocal microscopy, green fluorescence was observed in roots and leaf tissues 2 days after the inoculation of green fluorescent protein (GFP)-labeled bacteria in Alamo, which confirmed the successful colonization of bacteria inside the plant tissues. These shoot endophytic PGPB and switchgrass interactions are useful for the sustainable biomass production of bioenergy crop in a Cd-contaminated environment.     

Invasive herb <Emphasis Type="Italic">Impatiens glandulifera</Emphasis> has minimal impact on multiple components of temperate forest ecosystem function

Forests understories in Europe are known to generally resist invasion, though some alien plants do invade woodland communities. Here we focused on the impact of the widespread invasive annual Impatiens glandulifera, common along watercourses, but recently spreading in forests up to timberline. We investigated its impact on plant–soil feedback and ecosystem functioning. We recorded >40 variables focusing on: soil characteristics, including micro- and macro-nutrients; characteristics of litter layer and enzyme activity in litter; and richness and species composition of the forest understory. Three treatments were followed for 3 years: plots invaded by I. glandulifera; adjacent invader removal plots within the invaded area; and spatially separated uninvaded plots outside the invaded area. The effect of year-to-year variation was generally greater than that of the treatments, especially in soil and litter characteristics. Copper and boron were higher in invaded than invader removal and uninvaded plots, though in quantities that are unlikely to harm other plants. We found no effect of I. glandulifera on litter characteristics or enzyme activity. Despite almost 80% cover of I. glandulifera, we did not detect any difference in species richness and total vegetation cover between invaded and uninvaded plots. The floristic composition differed among the uninvaded, invader removal and invaded plots across 3 years. Our results indicate that the effect of I. glandulifera on the forest community studied was minor, and largely resulted from its increased shading to other plant species. In conclusion, we show how misleading the evaluation of impacts can be if based on a single season.     

Mutation of mpk6 enhances cadmium tolerance in Arabidopsis plants by alleviating oxidative stress

Background and aims

Cadmium (Cd) could activate activity of mitogen-activated protein kinase MPK6 in plants. In this study, we investigated the role of MPK6 in mediating Cd toxicity in plants.

Methods

The wild type Arabidopsis plants (WT) and the mpk6-2 mutants were subjected either 0 (Control) or 10 μM Cd treatment. Kinase activity of MPK6, nitric oxide (NO) level, Cd concentration, and oxidative stress were measured.

Results

In WT plants, Cd exposure rapidly stimulated kinase activity of MPK6. However, upon Cd exposure, mpk6-2 showed better growth than the WT. Although Cd-induced production of NO in roots was greater in WT than in mpk6-2, there was no difference in Cd concentration between the two plants. Nevertheless, the Cd-induced hydroperoxide burst, lipid peroxidation and loss of membrane integrity, were all more severe in the WT than in mpk6-2. Foliar applications of antioxidant ascorbic acid, vigorously improved the growth of both the WT and mpk6-2 under Cd exposure. Thereby the growth difference between these two plants was minimized.

Conclusions

Mutation of mpk6 enhances Cd tolerance in plants by alleviating oxidative stress, but did not affect cadmium accumulation in plants.     

Antioxidant defense system in leaves of Indian mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis>) and rape (<Emphasis Type="Italic">Brassica napus</Emphasis>) under cadmium stress

Plant species capable of hyper-accumulating heavy metals are of considerable interest for phytoremediation, and differ in their ability to accumulate metals from environment. Using two brassica species (Brassica juncea and Brassica napus), nutrient solution experiments were conducted to study variation in tolerance to cadmium (Cd) toxicity based on (1) lipid peroxidation and (2) changes in antioxidative defense system in leaves of both plants (i.e., superoxide dismutase (SOD EC 1.15.1.1), catalase (CAT EC 1.11.1.6), ascorbate peroxidase (APX EC 1.11.1.11), guaiacol peroxidase (GPX EC 1.11.1.7), glutathione reductase (GR EC 1.6.4.2), levels of phytochelatins (PCs), non-protein thiols (NP-SH), and glutathione. Plants were grown in nutrient solution under controlled environmental conditions, and subjected to increasing concentrations of Cd (0, 10, 25 and 50 μM) for 15 days. Results showed marked differences between both species. Brassica napus under Cd stress exhibited increased level of lipid peroxidation, as was evidenced by the increased malondialdehyde (MDA) content in leaves. However, in Brassica juncea treated plants, MDA content remained unchanged. In Brassica napus, with the exception of GPX, activity levels of some antioxidant enzymes involved in detoxification of reactive oxygen species (ROS), including SOD, CAT, GR, and APX, decreased drastically at high Cd concentrations. By contrast, in leaves of Brassica juncea treated plants, there was either only slight or no change in the activities of the antioxidative enzymes. Analysis of the profile of anionic isoenzymes of GPX revealed qualitative changes occurring during Cd exposure for both species. Moreover, levels of NP-SH and PCs, monitored as metal detoxifying responses, were much increased in leaves of Brassica juncea by increasing Cd supply, but did not change in Brassica napus. These results indicate that Brassica juncea plants possess the greater potential for Cd accumulation and tolerance than Brassica napus.     

Thiol metabolism play significant role during cadmium detoxification by Ceratophyllum demersum L.

In the present study, the level of thiols and activity of related enzymes were investigated in coontail (Ceratophyllum demersum L.) plants to analyze their role in combating the stress caused upon exposure to cadmium (Cd; 0–10 μM) for a duration up to 7 d. Plants showed the maximum accumulation of 1293 μg Cd g^?1 dw after 7 d at 10 μM. Significant increases in the level of total non-protein thiols (NP-SH) including phytochelatins (PCs) as well as upstream metabolites of the PC biosynthetic pathway, cysteine and glutathione (GSH) were observed. In addition, significant increases in the activities of cysteine synthase (CS), glutathione-S-transferase (GST), glutathione reductase (GR), as well as in vitro activation of phytochelatin synthase (PCS), were noticed in response to Cd. In conclusion, under Cd stress, plants adapted to a new metabolic equilibrium of thiols through coordinated synthesis and consumption to combat Cd toxicity and to accumulate it.     

镉暴露对黑斑蛙精巢ROS的诱导及其蛋白质氧化损伤作用机理

重金属镉对精巢发育、呼吸及神经系统信号转导等途径均有不良影响,被认为是造成两栖动物种群数量急剧下降的重要原因之一。然而,有关镉对精巢损伤的分子机理还不清楚。通过对镉暴露后的黑斑蛙精巢活性氧自由基(ROS)、蛋白质羰基(PCO)以及DNA蛋白质交联(DPC)等指标的系统分析,探讨了镉对精巢毒害的分子作用机理。随镉浓度的增加,黑斑蛙精巢细胞线粒体ROS随镉暴露浓度的增加而升高,0.5、1.0 mg/L镉染毒组与对照组比较有显著性差异(P<0.05);精巢组织PCO和DPC也随镉暴露浓度的增加而逐渐上升,且均呈明显的浓度-效应关系。结果表明:镉诱导机体产生ROS,进而导致蛋白质氧化损伤以及DNA损伤,说明精巢组织ROS的产生是镉致雄性生殖毒效应机制的重要因素之一。     

Drought stress induces oxidative stress and the antioxidant defense system in ascorbate-deficient vtc1 mutants of Arabidopsis thaliana

Drought stress has a negative impact on plant cells and results in the generation of reactive oxygen species (ROS). To increase our understanding of the effects of drought stress on antioxidant processes, we investigated the response of the ascorbate-deficient Arabidopsis thaliana vtc1 mutant to drought stress. After drought stress, vtc1 mutants exhibited increases in several oxidative parameters, including H₂O₂ content and the production of thiobarbituric acid reactive substances. Decreases in chlorophyll content and chlorophyll fluorescence parameters were also observed. The vtc1 mutants had higher total glutathione than did wild-type (WT) plants after 48 h of drought stress. A reduced ratio of glutathione/total glutathione and an increased ratio of dehydroascorbate/total ascorbate were observed in the vtc1 mutants compared with the WT plants. In addition, the activities of enzymes that are responsible for ROS scavenging, including superoxide dismutase, catalase, and ascorbate peroxidase, were decreased in the vtc1 mutants compared with the WT plants. Similar reductions in activity in the vtc1 mutant were observed for the enzymes that are responsible for the regeneration of ascorbate and glutathione, including monodehydroascorbate reductase, dehydroascorbate reductase, and glutathione reductase. These results suggest that low intrinsic ascorbate and impaired ascorbate–glutathione cycling in the vtc1 mutant induced a decrease in the reduced form of ascorbate, which enhanced sensitivity to drought stress.     

Cadmium activates Arabidopsis MPK3 and MPK6 via accumulation of reactive oxygen species

Cadmium (Cd) is a non-essential toxic heavy metal that influences normal growth and development of plants. However, the molecular mechanisms by which plants recognize and respond to Cd remain poorly understood. We show that, in Arabidopsis, Cd activates the mitogen-activated protein kinases, MPK3 and MPK6, in a dose-dependent manner. Following treatment with Cd, these two MAPKs exhibited much higher activity in the roots than in the leaves, and pre-treatment with the reactive oxygen species (ROS) scavenger, glutathione, effectively inhibited their activation. These results suggest that the Cd sensing signaling pathway uses a build-up of ROS to trigger activation of Arabidopsis MPK3 and MPK6.