Phytochelatin Content Research Articles

Metabolic Responses to Arsenite in Rice Seedlings that Differed in Grain Arsenic Concentration

Arsenic (As) occurs naturally in the environment, and is present in all edible and nonedible plant tissues. Plants have multiple mechanisms to prevent plant injury by heavy metals such as As. These same mechanisms could be used to reduce accumulation of As in rice (Oryza sativa L.) grains. From previous study of 1765 international rice accessions, specific accessions were identified as having exceptionally high grain As concentrations (grain As accumulators) and others low grain As (grain As excluders). This study investigated As uptake, transport, and metabolism in six previously identified lines to determine which physiological responses, if any, were associated with accumulation or exclusion of As in grains. Hydroponically grown seedlings were treated with 0 (controls) or 100 μM arsenite [As(III)], and then whole seedlings were analyzed for concentrations of As plus key compounds involved in heavy metal metabolism. Both grain accumulators and grain excluders actively concentrated As within their roots, and both groups had 10‐fold higher As concentrations in roots than leaves. In response to As(III), roots of both grain excluders and grain accumulators increased in cysteine and phytochelatin (PC) production, which suggests PC sequestration of As. In contrast, only grain excluders doubled in leaf glutathione (GSH) concentration by 72 h after As(III) addition. Because PC concentrations remained constant in leaves, it appears that the additional leaf GSH in the grain excluders was not used to produce more PC but may instead be forming As‐GSH adducts, which also aid in As sequestration.

Root cell walls and phytochelatins in low-cadmium cultivar of Brassica parachinensis

‘Lvbao-701’ is a cultivar of Chinese flowering cabbage (Brassica parachinensis) that exhibits low cadmium (Cd) accumulation and high Cd tolerance. In this study, this cultivar was compared with a high-Cd accumulating cultivar, ‘Chixin-4’, to characterize the mechanisms influencing Cd accumulation in B. parachinensis. Root cell walls were isolated by dissolving the cytoplasm with an organic solvent, and root Cd and phytochelatin (PC) contents were analyzed. In addition, a PC synthase gene fragment was cloned and expressed under Cd stress conditions. The proportions of Cd bound to root cell walls were higher in the ‘Lvbao-701’ plants (68.32%–76.80%) than in the ‘Chixin-4’ plants (35.36%–54.18%) after exposure to Cd stress. The proportions of Cd bound to root cell walls measured using cell walls isolated with a non-destructive method were higher than those obtained using a conventional method that required grinding and centrifugation. Exposure to Cd stress induced the PC production and resulted in higher PC contents in the ‘Lvbao-701’ plants than in the ‘Chixin-4’ plants. Cloning and expression analysis of a B. parachinensis PC synthase cDNA fragment indicated that PC synthase gene expression was induced by Cd and occurred mainly in the roots of both ‘Lvbao-701’ and ‘Chixin-4’ plants. However, the PC synthase gene expression level was higher in the ‘Lvbao-701’ roots than in the ‘Chixin-4’ roots. Therefore, a higher abundance of Cd in the root cell walls of ‘Lvbao-701’ and up-regulated PC production in the roots are probably the main reasons why ‘Lvbao-701’ exhibits lower Cd translocation to the shoots and higher tolerance to Cd than ‘Chixin-4’.

Polyamines may influence phytochelatin synthesis during Cd stress in rice

Although the metabolism of phytochelatins and higher polyamines are linked with each other, the direct relationship between them under heavy metal stress has not yet been clarified. Two approaches were used to reveal the influence of polyamine content on cadmium stress responses, particularly with regard to phytochelatin synthesis: putrescine pre-treatment of rice plants followed by cadmium stress, and treatment with the putrescine synthesis inhibitor, 2-(difluoromethyl)ornithine combined with cadmium treatment. The results indicated that putrescine pre-treatment enhanced the adverse effect of cadmium, while the application of 2-(difluoromethyl)ornithine reduced it to a certain extent. These differences were associated with increased polyamine content, more intensive polyamine metabolism, but decreased thiol and phytochelatin contents. The gene expression level and enzyme activity of phytochelatin synthase also decreased in rice treated with putrescine prior to cadmium stress, compared to cadmium treatment alone. In contrast, the inhibition of putrescine synthesis during cadmium treatment resulted in higher gene expression level of phytochelatin synthase. The results suggest that polyamines may have a substantial influence on phytochelatin synthesis at several levels under cadmium stress in rice.

Leaf defense system of Robinia pseudoacacia L. seedlings exposed to 3 years of elevated atmospheric CO2 and Cd-contaminated soils

Short-term exposure to elevated CO2 increases cadmium (Cd) uptake in some plant species (wheat, poplars, and willows), which triggers an increase in antioxidative system activity to deal with additional reactive oxygen species that are generated. Here, we examined leaf defenses in Robinia pseudoacacia L. seedlings exposed to elevated CO2+Cd for 3years. Three years of elevated CO2 decreased Cd uptake into leaves and the Cd content in soils and increased the pH of rhizosphere soil relative to ambient CO2. In plants exposed to Cd stress, leaf chlorophyll content was greater under elevated CO2 than under ambient CO2. Superoxide dismutase, peroxidase, and catalase activity increased, glutathione content increased, and malondialdehyde and phytochelatins contents decreased under elevated CO2+Cd relative to Cd alone. Proline, soluble sugars, flavonoids, saponins, and phenolic acids contents were greater under elevated CO2+Cd than under Cd alone, and condensed tannin content was lower. Overall, long-term elevation of CO2 enhanced the leaf defense system of R. pseudoacacia exposed to Cd by stimulating antioxidant enzyme activity, osmotic adjustment, and the production of glutathione, flavonoids and phenolic acids. Future research should focus on understanding the mechanisms involved in the decrease in Cd uptake into leaves and Cd content in soils and the increase in rhizosphere soil pH under long-term exposure to elevated CO2.

Effects of triacontanol on ascorbate-glutathione cycle in Brassica napus L. exposed to cadmium-induced oxidative stress

The ability of exogenous triacontanol (TRIA), a plant growth regulator, to reduce Cd toxicity was studied in canola (Brassica napus L.) plants. The following biological parameters were examined in canola seedlings to investigate TRIA-induced tolerance to Cd toxicity: seedling growth, chlorophyll damage and antioxidant response. In particular, TRIA application reduced Cd-induced oxidative damage, as shown by reduction of ROS content, lipoxygenase (LOX) activity and lipid peroxidation level. TRIA pretreatment increased non-enzymatic antioxidant contents (ascorbate, AsA, glutathione and GSH), phytochelatin content (PCs) and activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX), monodehydroascorbate reductase (MDHAR), dehydro ascorbate reductase (DHAR), and glutathione reductase (GR), so reducing the oxidative stress. These results clearly indicate the protective ability of TRIA to modulate the redox status through the antioxidant pathway AGC and GSH, so reducing Cd-induced oxidative stress.

Maleic acid assisted improvement of metal chelation and antioxidant metabolism confers chromium tolerance in Brassica juncea L.

Chromium (Cr) is a highly toxic environmental pollutant that negatively affects plant growth and development. Thus, remediating Cr from soil or increasing plant tolerance against Cr stress is urgent. Organic acids are recognized as agents of phytoremediation and as exogenous protectants, but using maleic acid (MA) to attain these results has not yet been studied. Therefore, our study investigated the effects of MA on Cr uptake and mitigation of Cr toxicity. We treated 8-d-old Indian mustard (Brassica juncea L.) seedlings with Cr (0.15mM and 0.3mM K2CrO4, 5 days) alone and in combination with MA (0.25mM) in a semi-hydroponic medium. Under Cr stress, plants accumulated Cr in both the roots and shoots in a dose-dependent manner, where the roots showed higher accumulation. Chromium stress reduced the growth and biomass of the Indian mustard plants by reducing water status and photosynthetic pigments, and increased oxidative damage due to generation of toxic reactive oxygen species (ROS) and methylglyoxal (MG). Chromium stress also interfered with the function of the antioxidant defense and glyoxalase systems. However, using MA in the Cr-stressed plants further increased Cr uptake in the roots, but it slightly reduced the translocation of Cr from the roots to the shoots at a lower dose of Cr and significantly at a higher dose. Moreover, MA also increased the other non-protein thiols (NPTs) containing phytochelatin (PC) content of the seedlings, which reduced Cr toxicity. Supplementing the stressed plants with MA upregulated the non-enzymatic antioxidants (ascorbate, AsA; glutathione, GSH); the activities of the enzymatic antioxidants including ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR), glutathione peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT); and the enzymes of the glyoxalase system including glyoxalase I (Gly I) and glyoxalase II (Gly II); and finally reduced oxidative damage and increased the chlorophyll content and water status as well the growth and biomass of the plants. Our findings suggested two potential uses of MA: first, enhancing phytoremediation, principally phytostabilization and second, working as an exogenous protectant to enhance Cr tolerance.

Phytochelatins and monothiols in salt marsh plants and their relation with metal tolerance

Phytochelatins (PCs) and monothiols and their relation with trace element concentrations were studied in three plant species from two Portuguese salt marshes. Belowground tissues showed always higher element concentrations, while enhanced values of monothiols were found in aboveground biomass. Glutathione was usually the most abundant monothiol. The concentration of total PCs was higher in leaves or stems than in roots of Halimione portulacoides and Sarcocornia perennis, while in Spartina maritima the highest concentrations were reported in large roots. PC2 was synthesized by all tissues and species and was higher in large roots of S. maritima. PC4 and PC5 were in high levels in small roots of S. maritima. PC2 was positively correlated with As, Zn and Pb. Although being the first evidence of PCs and monothiols in these species under natural conditions, our results do not point to a simple relationship with elements concentrations, suggesting a complex mechanism involved.

Arsenic speciation in the phloem exudates of rice and its role in arsenic accumulation in rice grains

Arsenic (As) speciation in the phloem sap of rice plants and its role in As accumulation in rice grains remain largely uncharacterized. In the present study, we tested As chemical species in the phloem exudates of rice treated with arsenate [As(V)], arsenite [As(III)], monomethylarsonic acid [MMA(V)], or dimethylarsinic acid [DMA(V)]. As(V) was the main species (58%) in the phloem exudates of As(V)-exposed rice, whereas As(III) predominated (69%) in As(III)-exposed rice. A large proportion of As(V) (41–45%) was observed in the phloem exudates when rice was treated with methylated As species. High concentrations of phytochelatins were detected in the phloem exudates when the As(V) treatment level was increased. The role of phloem transport was analyzed by applying a ±stem-girdling treatment to the rice plants, limiting phloem transport to the grain in rice pulsed with As(III), As(V), MMA(V), or DMA(V). The findings of the present study indicate that organic As is more mobile than inorganic As during phloem transport. Phloem transport accounted for 54% of As(III), 56% of As(V), 100% of MMA(V), and 89% of DMA(V) transport to the grain. The total As concentration and As(III) percentage in rice phloem and grain were significantly affected by increasing the phosphate concentration in the medium.

Impact of glutathione metabolism on zinc homeostasis in Saccharomyces cerevisiae.

Zinc is a crucial mineral for all organisms as it is an essential cofactor for the proper function of a plethora of proteins and depletion of zinc causes oxidative stress. Glutathione is the major redox buffering agent in the cell and therefore important for mitigation of the adverse effects of oxidative stress. In mammalian cells, zinc deficiency is accompanied by a glutathione depletion. In the yeast Saccharomyces cerevisiae, the opposite effect is observed: under low zinc conditions, an elevated glutathione concentration is found. The main regulator to overcome zinc deficiency is Zap1p. However, we show that Zap1p is not involved in this glutathione accumulation phenotype. Furthermore, we found that in glutathione-accumulating strains also the metal ion-binding phytochelatin-2, which is an oligomer of glutathione, is accumulated. This increased phytochelatin concentration correlates with a lower free zinc level in the vacuole. These results suggest that phytochelatin is important for zinc buffering in S. cerevisiae and thus explains how zinc homeostasis is connected with glutathione metabolism.

γ-aminobutyric acid (GABA) confers chromium stress tolerance in Brassica juncea L. by modulating the antioxidant defense and glyoxalase systems.

Chromium (Cr) toxicity is hazardous to the seed germination, growth, and development of plants. γ-aminobutyric acid (GABA) is a non-protein amino acid and is involved in stress tolerance in plants. To investigate the effects of GABA in alleviating Cr toxicity, we treated eight-d-old mustard (Brassica juncea L.) seedlings with Cr (0.15 and 0.3 mM K2CrO4, 5 days) alone and in combination with GABA (125 µM) in a semi-hydroponic medium. The roots and shoots of the seedlings accumulated Cr in a dose-dependent manner, which led to an increase in oxidative damage [lipid peroxidation; hydrogen peroxide (H2O2) content; superoxide (O2•-) generation; lipoxygenase (LOX) activity], methylglyoxal (MG) content, and disrupted antioxidant defense and glyoxalase systems. Chromium stress also reduced growth, leaf relative water content (RWC), and chlorophyll (chl) content but increased phytochelatin (PC) and proline (Pro) content. Furthermore, supplementing the Cr-treated seedlings with GABA reduced Cr uptake and upregulated the non-enzymatic antioxidants (ascorbate, AsA; glutathione, GSH) and the activities of the enzymatic antioxidants including ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR), glutathione peroxidase (GPX), superoxide dismutase (SOD), catalase (CAT), glyoxalase I (Gly I), and glyoxalase II (Gly II), and finally reduced oxidative damage. Adding GABA also increased leaf RWC and chl content, decreased Pro and PC content, and restored plant growth. These findings shed light on the effect of GABA in improving the physiological mechanisms of mustard seedlings in response to Cr stress.

Bioaccumulation and detoxification mechanisms for lead uptake identified in Rhus chinensis Mill. seedlings

A greenhouse experiment was conducted to assay the bioaccumulation and tolerance characteristics of Rhus chinensis Mill. to lead (Pb). The effects of exposing R. chinensis Mill seedlings to increasing Pb concentrations (0, 250, 500, 100mgkg-1) in the soil were assessed by measuring Pb accumulation, subcellular distribution, ultrastructure, photosynthetic characteristics, antioxidative enzyme activity, malondialdehyde content, and phytochelatin content. The majority of Pb taken up by R. chinensis Mill was associated with the cell wall fraction in the roots, where the absorption of Ca increased to maintain cell wall stability, and Pb deposits were found in the intercellular space or in the cell wall structures. In leaves, Pb was primarily stored in the cell wall, while it was compartmentalized into the vacuolar structures in the stem. Pb concentrations adversely affected the morphology of Rhus chinensis Mill cellular substructures. Furthermore, increased Peroxidase (POD) and catalase (CAT) activity was observed in plants grown in Pb-amended soil, and this may have led to reduced ROS to maintain the function of the membrane. Changes in phytochelatin levels (PCs) that were observed in Pb treated plants suggest that PCs formed complexes with Pb in the cytoplasm to reduce Pb2+ toxicity in the metabolically active cellular compartment. This mechanism may allow for the plant to accumulate higher concentrations of toxic Pb and survive for a longer period of time. Our study provides a better understanding of how Rhus chinensis Mill detoxifies Pb.

Phytochelatin synthesis in response to Hg uptake in aquatic plants near a chlor-alkali factory

The effects of mercury (Hg) released from a chlor-alkali factory in aquatic plants along the Ebro River basin (NE Spain) were analysed considering the phytochelatins (PCn) and their isoforms content in these plants. These compounds were analyzed using HPLC with amperometric detection, and the macrophytes species Ceratophyllum demersum and Myriopyllum spicatum were collected in two sampling campaigns, autumn and spring, respectively. To correlate the PCn content in macrophytes with the Hg contamination, analysis of total Hg (THg) content in plants and suspended particulate matter, as well as the dissolved-bioavailable fraction of Hg in water measured by the diffusive gradient in thin film (DGT) technique were done. The results confirm the presence of PC2-Ala in extracts of C. demersum and PC2-desGly in M. spicatum, and the concentration of these thiol compounds depends clearly on the distance between the hot spot and the downstream sites: the higher the levels are, the closer the hot spot is. Since most of the Hg is hypothesized to be associated with SPM and transported downstream, our results of the DGT suggest that trace amounts of Hg in water can be released as free metal ions yielding a certain accumulation in plants (reaching the ppb level) that are enough for activation of induction of PCs. A few PCs species have been determined, at different seasons, indicating that they can be used as good indicators of the presence of bioavailable Hg in aquatic media throughout the year.

Phytochelatin 2 accumulates in roots of the seagrass Enhalus acoroides collected from sediment highly contaminated with lead.

Phytochelatins (PCs), the heavy metal-binding peptides of plants, play a main function in heavy metal detoxification. In this study, Enhalus acoroides samples collected at six distinct seagrass beds from the coast of Khanh Hoa province, Viet Nam, were evaluated for their PCs. The contents of different PCs in each organ including leaf, rhizome, and root were determined by using HPLC analysis. Significant differences of PC2 contents among specific organs and their relation were tested by ANOVA, Tukey test, and Pearson's correlation. The results showed that higher PC2, appearance of PC3 and a strong correlation between PC2 and Pb concentration were found in the root organ collected from a Pb contaminated area. We conclude that high Pb in the sediment induce high PC2 and PC3 production in the root. This first report on in situ detection of PCs of seagrass encourages future investigation on the ability to use seagrass for phytoremediation and as a bioindicator of heavy metals based on PC contents.

Phosphorus mediation of cadmium stress in two mangrove seedlings Avicennia marina and Kandelia obovata differing in cadmium accumulation

Mangrove ecosystems are vulnerable to environmental threats. In order to elucidate the effect of phosphorus (P) on cadmium (Cd) tolerance and physiological responses in mangroves under Cd stress, a mangrove specie with salt exclusion Kandelia obovata and a specie with salt secretion Avicennia marina were compared in a hydroponic experiment. The results showed that most Cd was accumulated in mangrove roots and that P addition induced Cd immobilisation in them. Cd stress significantly increased malonaldehyde content, whereas P significantly decreased malonaldehyde in mangroves. Phosphorus positively regulated the photosynthetic pigment, proline content and synthesis of non-protein thiols, glutathione and phytochelatins in the leaves under Cd stress conditions. The results suggest different adaptive strategies adopted by two mangroves in a complex environment and A. marina showed a stronger Cd tolerance than K. obovata. The study provides a theoretical basis for P mediated detoxification of Cd in mangrove plants.

Arsenic-induced stress activates sulfur metabolism in different organs of garlic (Allium sativum L.) plants accompanied by a general decline of the NADPH-generating systems in roots

Arsenic (As) contamination is a major environmental problem which affects most living organisms from plants to animals. This metalloid poses a health risk for humans through its accumulation in crops and water. Using garlic (Allium sativum L.) plants as model crop exposed to 200μM arsenate, a comparative study among their main organs (roots and shoots) was made. The analysis of arsenic, glutathione (GSH), phytochelatins (PCs) and lipid peroxidation contents with the activities of antioxidant enzymes (catalase, superoxide dismutase, ascorbate-glutathione cycle), and the main components of the NADPH-generating system, including glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH), NADP-malic enzyme (NADP-ME) and NADP-isocitrate dehydrogenase (NADP-ICDH) was carried out. Data showed a correlation among arsenic accumulation in the different organs, PCs content and the antioxidative response, with a general decline of the NADPH-generating systems in roots. Overall, our results demonstrate that there are clear connections between arsenic uptake, increase of their As-chelating capacity in roots and a decline of antioxidative enzyme activities (catalase and the ascorbate peroxidase) whose alteration provoked As-induced oxidative stress. Thus, the data suggest that roots act as barrier of arsenic mediated by a prominent sulfur metabolism which is characterized by the biosynthesis of high amount of PCs.

Phytochelatins and Phenol Content in Barley Seedlings Exposed to Al Toxicity and Primed with Ascorbic Acid and Salicylic Acid

Aluminium (Al) is an essential element to constrain crop production in acid soil all over the world. Root growth inhibition is the primary visible symptom of Al toxicity in plants. It causes oxidative damage in plant cell as a result of excess production of harmful oxygen radicals. Cell provokes antioxidant system against oxidative damage to protect the cell. In this experiment, the performance of non-enzymatic antioxidant like phytochelatin (PC) and phenol against Al toxicity in two barley varieties (RD2052 and RD2552) were evaluated. PC content increased in both shoot and root, whereas phenolic content decreased with increased Al concentration, but priming with two plant growth regulators like ascorbic acid (AA) and salicylic acid (SA) successfully alleviated Al stress and noticed less increased PC and less decreased phenolic content compared with unprimed barley varieties (RD2052 and RD2552). All data were prepared by one-way analysis of variance (ANOVA).

Phytochelatin synthesis in Dunaliella salina induced by arsenite and arsenate under various phosphate regimes

This study investigated the dynamic variations in thiol compounds, including cysteine (Cys), glutathione (GSH), and phytochelatins (PCs), in Dunaliella salina samples exposed to arsenite [As(III)] and arsenate [As(V)] under various phosphate (PO43−) regimes. Our results showed that GSH was the major non-protein sulfhydryl compound in D. salina cells. As(III) and As(V) induced PC syntheses in D. salina. PC2, PC3, and PC4 were all found in algal cells; the PC concentrations decreased gradually while exposed to As for 3 d. The synthesis of PC2–3 was significantly affected by As(III) and As(V) concentrations in the cultures. More PCs were detected in the As(V)-treated algal cells compared with the As(III) treatment. PC levels increased with As(III)/As(V) amount in the medium, but remained stable after 112μgL−1 As(V) exposure. In contrast, significant (p<0.001) positive correlations were observed between PC synthesis and intracellular As(III) content or As accumulation in As(III)-treated algal cells during the 72-h exposure. PO43− had a significant influence on the PC synthesis in algal cells, irrespective of the As-treated species. Reductions in As uptake and subsequent PC synthesis by D. salina were observed as the PO43− concentration in the growth medium increased. L-Buthionine sulfoximine (BSO) differentially influenced PC synthesis in As-treated D. salina under different extracellular PO43− regimes. Overall, our data demonstrated that the production of GSH and PCs was affected by PO43− and that these thiols played an important role in As detoxification by D. salina.

Quantitative Relationship between Cadmium Uptake and the Kinetics of Phytochelatin Induction by Cadmium in a Marine Diatom.

Heavy metals activate the synthesis of phytochelatins (PCs), while the induced PCs might affect metal uptake via chelating intracellular free metals. However, the relationship of PCs to metal uptake is poorly understood. In this study, we examined the kinetics of cadmium (Cd) accumulation and the synthesis of PCs in a marine diatom, Thalassiosira weissflogii, under different irradiance levels. Irradiance alone could not change the concentrations of PCs in the Cd-free treatments, while higher irradiance accelerated the induction of intracellular PCs at the same [Cd2+] level. PC-SH (2 × PC2 + 3 × PC3 + 4 × PC4) was bound with Cd at a stoichiometric ratio of 2 to 49 in our short-term uptake experiments, indicating that PC induction is sufficient to serve as the first line of defense against Cd stress. A positive linear correlation between the induction rate of PCs and the Cd uptake rate was observed, while the ratio of the PC content to intracellular Cd varied greatly when the irradiance was increased several fold. Because metal uptake has been successfully used in predicting acute metal toxicity, our findings are helpful for understanding the role of PCs in metal detoxification and developing PCs as biomarkers for metal sensitivity.

The arbuscular mycorrhizal fungus Rhizophagus irregularis differentially regulates the copper response of two maize cultivars differing in copper tolerance

Arbuscular mycorrhiza can increase plant tolerance to heavy metals. The effects of arbuscular mycorrhiza on plant metal tolerance vary depending on the fungal and plant species involved. Here, we report the effect of the arbuscular mycorrhizal fungus Rhizophagus irregularis on the physiological and biochemical responses to Cu of two maize genotypes differing in Cu tolerance, the Cu-sensitive cv. Orense and the Cu-tolerant cv. Oropesa. Development of the symbiosis confers an increased Cu tolerance to cv. Orense. Root and shoot Cu concentrations were lower in mycorrhizal than in non-mycorrhizal plants of both cultivars. Shoot lipid peroxidation increased with soil Cu content only in non-mycorrhizal plants of the Cu-sensitive cultivar. Root lipid peroxidation increased with soil Cu content, except in mycorrhizal plants grown at 250mg Cu kg−1soil. In shoots of mycorrhizal plants of both cultivars, superoxide dismutase, ascorbate peroxidase, catalase and glutathione reductase activities were not affected by soil Cu content. In Cu-supplemented soils, total phytochelatin content increased in shoots of mycorrhizal cv. Orense but decreased in cv. Oropesa. Overall, these data suggest that the increased Cu tolerance of mycorrhizal plants of cv. Orense could be due to an increased induction of shoot phytochelatin biosynthesis by the symbiosis in this cultivar.

Changes in subcellular distribution and antioxidant compounds involved in Pb accumulation and detoxification in Neyraudia reynaudiana

The effects of increasing concentrations of lead (Pb) on Pb accumulation, subcellular distribution, ultrastructure, photosynthetic characteristics, antioxidative enzyme activity, malondialdehyde content, and phytochelatin contents were investigated in Neyraudia reynaudiana seedlings after a 21-day exposure. A Pb analysis at the subcellular level showed that the majority of Pb in the roots was associated with the cell wall fraction, followed by the soluble fraction. In contrast, the majority of the Pb in the leaves was located in the soluble fraction based on transmission electron microscopy and energy dispersive X-ray analyses. Furthermore, high Pb concentrations adversely affected N. reynaudiana cellular structure. The changes in enzyme activity suggested that the antioxidant system plays an important role in eliminating or alleviating Pb toxicity, both in the roots and leaves of N. reynaudiana. Additionally, the phytochelatin contents in the roots and leaves differed significantly between Pb-spiked treatments and control plants. Our results provide strong evidence that cell walls restrict Pb uptake into the protoplasm and establish an important protective barrier. Subsequent vacuolar compartmentalization in leaves could isolate Pb from other substances in the cell and minimize Pb toxicity in other organelles over time. These results also demonstrated that the levels of antioxidant enzymes and phytochelatin in leaves and roots are correlated with Pb toxicity. These detoxification mechanisms promote Pb tolerance in N. reynaudiana.