Phytochelatin synthase of Thlaspi caerulescens enhanced tolerance and accumulation of heavy metals when expressed in yeast and tobacco

Phytochelatin synthase (PCS) is key enzyme for heavy metal detoxification and accumulation in plant. In this study, we isolated the PCS gene TcPCS1 from the hyperaccumulator Thlaspi caerulescens. Overexpression of TcPCS1 enhanced PC production in tobacco. Cd accumulation in the roots and shoots of TcPCS1 transgenic seedlings was increased compared to the wild type (WT), while Cd translocation from roots to shoots was not affected under Cd treatment. The root length of the TcPCS1 transgenic tobacco seedlings was significantly longer than that of the WT under Cd stress. These data indicate that TcPCS1 expression might increase Cd accumulation and tolerance in transgenic tobacco. In addition, the malondialdehyde content in TcPCS1 plants was below that of the wild type. However, the antioxidant enzyme activities of superoxide dismutase, peroxidase and catalase were found to be significantly higher than those of the WT when the transgenic plant was exposed to Cd stress. This suggests that the increase in PC production might enhance the Cd accumulation and thus increase the oxidative stress induced by the cadmium. The production of PCs could cause a transient decrease in the cytosolic glutathione (GSH) pool, and Cd and lower GSH concentration caused an increase in the oxidative response. We also determined TcPCS1 in Thlaspi caerulescens was regulated after exposure to various concentrations of CdCl₂ over different treatment times. Expression of TcPCS1 leading to increased Cd accumulation and enhanced metal tolerance, but the Cd contents were restrained by adding zinc in Saccharomyces cerevisiae transformants.     

Exogenous spermidine and spermine enhance cadmium tolerance of <Emphasis Type="Italic">Potamogeton malaianus</Emphasis>

In order to investigate the effects of spermidine (Spd) and spermine (Spm) on cadmium stress, the content of chlorophyll, hydrogen peroxide (H₂O₂), malondialdehyde (MDA), soluble protein and proline, the rate of O₂^·− generation, and activities of antioxidant enzymes (superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and glutathione reductase (GR)) in Potamogeton malaianus Miq. were measured. Exogenous application of Spd or Spm significantly enhanced the level of proline, retarded the loss of chlorophyll, enhanced photosynthesis, decreased the rate of O₂^·− generation and H₂O₂ content, and prevented Cd-induced lipid peroxidation. Spd and Spm also effectively maintained the balance of antioxidant enzyme activities under Cd stress; however, GR activity was found to increase only slightly in response to polyamines (PAs). The antioxidant systems, which were modified by PAs, were able to moderate the radical-scavenging system and to lessen in this way the oxidative stress. These results suggest that both Spd and Spm can enhance Cd tolerance of P. malaianus.     

Transgenic tomato plants overexpressing chloroplastic monodehydroascorbate reductase are resistant to salt- and PEG-induced osmotic stress

RNA gel hybridization showed that the expression of monodehydroascorbate reductase (MDHAR) in the wild type (WT) tomato was decreased firstly and then increased under salt- and polyethylene glycol (PEG)-induced osmotic stress, and the maximum level was observed after treatment for 12 h. WT, sense transgenic and antisense transgenic tomato plants were used to analyze the antioxidative ability to cope with osmotic stresses. After salt stress, the fresh mass (FM) and height of sense transgenic lines were greater than those of antisense lines and WT plants. Under salt and PEG treatments, sense transgenic plants showed a lower level of hydrogen peroxide (H₂O₂) and malondialdehyde (MDA), a higher net photosynthetic rate (P _N), and the maximal photochemical efficiency of PSII (F_v/F_m) compared with WT and antisense transgenic plants. Moreover, sense lines maintained higher ascorbate peroxidase (APX) activity than WT and antisense plants under salt- and PEG-induced osmotic stress. These results indicate that chloroplastic MDHAR plays an important role in alleviating photoinhibition of PSII by elevating ascorbate (AsA) level under salt- and PEG-induced osmotic stress.     

Cadmium toxicity and translocation in rice seedlings are reduced by hydrogen peroxide pretreatment

A hydroponic experiment was carried out to study the role of hydrogen peroxide (H₂O₂) in enhancing tolerance and reducing translocation of cadmium (Cd) in rice seedlings. Plant growth (length and biomass of shoot and root) was significantly repressed by Cd exposure. However, pretreatment with 100 μM H₂O₂ for 1d mitigated Cd stress by inducing enzyme activities for antioxidation (e.g., superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPX), ascorbate peroxidase (APX)) and detoxification (e.g., glutathione S-transferase (GST)) as well as by elevating contents of reduced glutathione (GSH) and ascorbic acid (AsA). As a result, H₂O₂ and malondialdehyde (MDA) content decreased in plants and the seedling growth was less inhibited. On the other hand, H₂O₂ pretreatment decreased Cd concentration in shoots, thus lowered the ratio of Cd concentration in shoots and roots (S/R), indicating that H₂O₂ may affect Cd distribution in rice seedlings. The improved Cd tolerance is partly due to an enhanced antioxidative system that efficiently prevents the accumulation of H₂O₂ during Cd stress. Increased Cd sequestration in rice roots may contribute to the decline of Cd translocation.     

Overexpression of chloroplastic monodehydroascorbate reductase enhanced tolerance to temperature and methyl viologen‐mediated oxidative stresses

A tomato (Lycopersicon esculentum Mill.) monodehydroascorbate reductase gene (LeMDAR) was isolated. The LeMDAR–green fluorescence protein (GFP) fusion protein was targeted to chloroplast in Arabidopsis mesophyll protoplast. RNA and protein gel blot analyses confirmed that the sense‐ and antisense‐ LeMDAR were integrated into the tomato genome. The MDAR activities and the levels of reduced ascorbate (AsA) were markedly increased in sense transgenic lines and decreased in antisense transgenic lines compared with wild‐type (WT) plants. Under low and high temperature stresses, the sense transgenic plants showed lower level of hydrogen peroxide (H₂O₂), lower thiobarbituric acid reactive substance (TBARS) content, higher net photosynthetic rate (P_n), higher maximal photochemical efficiency of PSII (F_v/F_m) and fresh weight compared with WT plants. The oxidizable P700 decreased more obviously in WT and antisense plants than that in sense plants at chilling temperature under low irradiance. Furthermore, the sense transgenic plants exhibited significantly lower H₂O₂ level, higher ascorbate peroxidase (APX) activity, greater P_n and F_v/F_m under methyl viologen (MV)‐mediated oxidative stresses. These results indicated that overexpression of chloroplastic MDAR played an important role in alleviating photoinhibition of PSI and PSII and enhancing the tolerance to various abiotic stresses by elevating AsA level.     

Effect of salicylic acid potentiates cadmium-induced oxidative damage in <Emphasis Type="Italic">Oryza sativa</Emphasis> L. leaves

In the present investigation, we studied the possible potentiating effect of salicylic acid (SA) under Cd toxicity in Oryza sativa L. leaves. Cd treatments for 24 h reduced the shoot length, dry biomass and total chlorophyll content followed by high Cd accumulation in shoots. About 16 h presoaking with SA resulted in partial protection against Cd, as observed by minor changes in length, biomass and total chlorophyll. SA priming resulted in low Cd accumulation. Enhanced thiobarbituric acid reactive substances (TBARS), hydrogen peroxide (H₂O₂) and superoxide anion (O₂ ⁻) content were seen when Cd was applied alone, while under SA priming the extent of TBARS, H₂O₂ and O₂ ⁻ were significantly low, suggesting SA-regulated protection against oxidative stress. The antioxidant enzymes like Catalase (CAT), guaiacol peroxidase (GPx), glutathione reductase (GR) and superoxide dismutase (SOD) showed varied activities under Cd alone. CAT activity increased after Cd treatment, followed by a decline in GPX and GR activity. SOD also declined at the highest concentrations with an initial increase. Under SA-priming conditions, the efficiency of the antioxidant enzymes was significantly elevated. GPx and SOD activity showed significant increase in activity. The ascorbate activity increased after Cd treatment, followed by a decline in glutathione under SA-free condition. SA priming showed gradual increase in these non-enzymic antioxidants. Our results indicate that Cd-induced oxidative stress can be regulated by SA.     

Overexpression of a peroxiredoxin gene from <Emphasis Type="Italic">Tamarix hispida</Emphasis>, <Emphasis Type="Italic">ThPrx1</Emphasis>, confers tolerance to oxidative stress in yeast and Arabidopsis

Peroxiredoxins (Prxs) are ubiquitous thiol-specific antioxidant enzymes that are critically involved in cell defense and protect cells from oxidative damage. In this study, a putative Type II Prx (ThPrx1) was identified and characterized from Tamarix hispida. The expression of ThPrx1 is highly induced in response to hydrogen peroxide (H₂O₂) and methyl viologen (MV) stresses. When expressed ectopically, ThPrx1 showed enhanced tolerance against oxidative stress in yeast and Arabidopsis. In addition, transgenic Arabidopsis plants overexpressing ThPrx1 displayed improved seedling survival rates and increased root growth and fresh weight gain under H₂O₂ and MV treatments. Moreover, transgenic Arabidopsis plants showed decreased accumulation of H₂O₂, superoxide (O₂^??) and malondialdehyde (MDA), increased superoxide dismutase (SOD) activity compared to wild-type (WT) plants under oxidative stress. Moreover, transgenic plants maintained higher photosynthesis efficiency and lower electrolyte leakage rates than that of WT plants under stress conditions. These results clearly indicated that ThPrx1 plays an important role in cellular redox homeostasis under stress conditions, leading to the maintenance of membrane integrity and increased tolerance to oxidative stress.     

Salicylic acid increases tolerance to oxidative stress induced by hydrogen peroxide accumulation in leaves of cadmium-exposed flax (Linum usitatissimum L.)

The aim of this study is to investigate the impacts of exogenous salicylic acid (SA) pretreatments on hydrogen peroxide (H₂O₂) accumulation, protein oxidation, and H₂O₂-scavenging enzymes in leaves of Cd-treated flax seedlings. Cd-enhanced H₂O₂ levels were related to increased activities of guaiacol peroxidase (POX, EC 1.11.1.7) and ascorbate peroxidase (APX, EC 1.11.1.11), and were independent of changes in catalase (CAT, EC 1.11.1.6) and superoxide dismutase (SOD, EC 1.15.1.1) activities. In control flax seedlings, exogenous SA pretreatments inhibited the activity of CAT, resulted in an enhanced production of H₂O₂ suggesting that SA requires H₂O₂ to initiate an oxidative stress. However, although leaves of Cd-free flax seedlings pretreated with SA accumulated in vivo H₂O₂ by 1.2-fold compared with leaves of Cd-only exposed ones; the damage to growth and proteins after the exposure to Cd was significantly less, indicating that SA can regulate the Cd-induced oxidative stress. Moreover, the Cd-treated seedlings primed with SA exhibited a higher level of total antioxidant capacities and increased activities of H₂O₂-detoxifying enzymes.     

The Responses of Pro‐ and Antioxidative Systems to Cold‐hardening and Pathogenesis Differ in Triticale (x Triticosecale Wittm.) Seedlings Susceptible or Resistant to Pink Snow Mould (Microdochium nivale Fr., Samuels & Hallett)

Two winter triticale (x Triticosecale Wittm.) cultivars, Magnat (susceptible to pink snow mould) and Hewo (relatively resistant), were used in a model system to test the effect of prehardening and different cold‐hardening regimes on pro‐ and antioxidative activity in seedling leaves. The concentration of hydrogen peroxide and the activity of total superoxide dismutase, catalase, peroxidase and ascorbic peroxidase were analysed spectrophotometrically. As there has been no previous analysis of the pro/antioxidative reaction of cereals to Microdochium nivale infection has been undertaken to‐date, this is the first in the series describing our results. We confirmed that both exposure to abiotic stress of low temperature and subsequent low light intensity, as well as biotic stress of M. nivale infection, change the pro‐ and antioxidative activity in model plants. Genotypes differed substantially in their hydrogen peroxide content: susceptible cv. Magnat generally showed higher levels during all the experiments. This result can lead to the conclusion that cv. Magnat is also more susceptible to low temperature and low light intensity than cv. Hewo. Simultaneous measurements of antioxidative activity indicated that the increased activity of catalases and peroxidases and the consequent lower H₂O₂ level are correlated with a higher resistance to low temperature, low light intensity and pink snow mould in triticale seedlings. The higher H₂O₂ level observed in the susceptible line is likely to be derived from the imbalance of reactive oxygen species production and consumption in this genotype under stress conditions.     

Responses of two kidney bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis>) cultivars to the combined stress of sulfur deficiency and cadmium toxicity

Plants suffer from combined stress of sulfur deficiency and cadmium toxicity in some agricultural lands. However, little is known about the reaction in plants, such as responses in antioxidant enzymes and non-protein thiol compounds, to such combined stress. Therefore, in this study, four treatments, S-sufficiency (T_S?Cd), S-deficiency (T_?S?Cd), Cd stress (T_S+Cd) and combined stress of S-deficiency and Cd stress (T_?S+Cd), were set up to investigate (1) the effects of sulfur deficiency or sulfur sufficiency on Cd toxicity to kidney bean cultivar seedlings and the related mechanisms, and (2) the responses of two kidney bean cultivars to combined stress of S-deficiency and Cd-tolerance. The results showed significant increases in hydrogen peroxide (H₂O₂) and malondialdehyde contents and significant increases in antioxidant enzyme (superoxide dismutase, catalase, peroxidase, and glutathione S-transferase) activities and non-protein thiol compounds (non-protein thiols, reduced glutathione, phytochelatins) synthesis in the plants in T_S+Cd and T_?S+Cd. On the tissue level, higher proportion of Cd was found to be immobilized/deposited in roots, while on the sub-cell level, higher proportion of Cd was located in cell walls and vacuole fractions with lower in cell organelles. Taken together, the results indicated that Cd detoxification was achieved by the two kidney bean cultivars through antioxidant enzyme activation, non-protein thiol compound synthesis and sub-cellular compartmentalization. In addition, the results indicated that sufficient S supply helped to relieve Cd toxicity, which is of special significance for remediation or utilization of Cd-contaminated soils as S is a plant essential nutrient.     

28-Homobrassinolide potential for oxidative interface in <Emphasis Type="Italic">Brassica juncea</Emphasis> under temperature stress

Exploration of scavenging potential of 28-homobrassinolide (28-homoBL) in mitigating the oxidative stress caused by free radicals (·O₂ ^?, H₂O₂, ·NO, OH^?) produced due to temperature stress (4, 44 °C) in Brassica juncea L. was made in the present research. Brassica juncea var. RLC-1 seeds were given pre-sowing soaking of different concentrations of 10^?9 M 28-homoBL for 8 h. Seeds were sown in bedded petri plates lined with 10 No. What’s man filter paper under controlled laboratory conditions. Temperature of 4 and 44 °C, taken as low- and high-temperature stress, suppressed membrane stability and overall growth of the seedlings, while cell death was triggered. Accumulation of malondialdehyde (MDA), hydrogen peroxide (H₂O₂), and nitric oxide (NO) was boosted which resulted in enhanced oxidative stress on the 10th day after sowing. Activity level of antioxidant enzymes viz. superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPOX), and ascorbate peroxidase (APOX) was enhanced which was ensued for up-regulation of total antioxidant potential in 10-day-old plants exposed to negative effect of temperature stress. Priming treatment of 28-homoBL at seed level helped in maintaining the growth of seedlings to higher level as compared to only stressed as well as from control double distilled water-raised seedlings. 10^?9 M 28-HBL found to be the best in enhancing the enzymatic activities of SOD, CAT, GPOX, and APOX and thus maintained antioxidant potential at higher level which accounted for alleviating oxidative stress caused due to extreme temperature stress. Dead cell formation reduced significantly in 28-homoBL-treated plants, membrane stability was upturned, while production of MDA, H₂O_2, and NO was under control. These results suggested and try to establish 28-homoBL as effective stress protector for B. juncea particularly from the oxidative damage induced by extreme temperatures.     

Arabidopsis thaliana Metallothionein, AtMT2a, Mediates ROS Balance during Oxidative Stress

Cold stress has been shown to induce the production of reactive oxygen species (ROS), which can elicit a potentially damaging oxidative burden on cellular metabolism. Here, the expression of a metallothionein gene (AtMT2a) was upregulated under low temperature and hydrogen peroxide (H₂O₂) stresses. The Arabidopsis T-DNA insertion mutant, mt2a, exhibited more sensitivity to cold stress compared to WT plants during the seed germination, and H₂O₂ levels in mt2a mutant were higher than that in WT plants during the cold stress. Synthetic GFP fused to AtMT2a was observed to be localized in cytosol. These results indicated that AtMT2a functions in tolerance against cold stress by mediating the ROS balance in the cytosol. Interestingly, mRNA level of AtMT2a was increased in seedlings of Arabidopsis cat2 mutant after cold treatment compared to WT seedlings, and overexpression of AtMT2a in cat2 could improve CAT activity under chilling stress. Moreover, the enzymatic activity of CAT in mt2a was higher than that in WT plants after cold treatment, suggesting that AtMT2a and CAT might complement each other in antioxidative process potentially in Arabidopsis. Taken together, our results provided a novel insight into the relationship between MTs and antioxidative enzymes in the ROS-scavenging system in plants.     

Genotype-Dependent Effect of Exogenous Nitric Oxide on Cd-induced Changes in Antioxidative Metabolism, Ultrastructure, and Photosynthetic Performance in Barley Seedlings (Hordeum vulgare)

A greenhouse hydroponic experiment was performed using Cd-sensitive (cv. Dong 17) and Cd-tolerant (Weisuobuzhi) barley seedlings to evaluate how different genotypes responded to cadmium (Cd) toxicity in the presence of sodium nitroprusside (SNP), a nitric oxide (NO) donor. Results showed that 5 μM Cd increased the accumulation of O₂^•−, H₂O₂, and malondialdehyde (MDA) but reduced plant height, chlorophyll content, net photosynthetic rate (P _n), and biomass, with a much more severe response in the Cd-sensitive genotype. Antioxidant enzyme activities increased significantly under Cd stress in the roots of the tolerant genotype, whereas in leaves of the sensitive genotype, superoxide dismutase (SOD) and ascorbate peroxide (APX), especially cytosol ascorbate peroxidase (cAPX), decreased after 5–15 days Cd exposure. Moreover, Cd induces NO synthesis by stimulating nitrate reductase and nitric oxide synthetase-like enzymes in roots/leaves. A Cd-induced NO transient increase in roots of the Cd-tolerant genotype might partly contribute to its Cd tolerance. Exogenous NO dramatically alleviated Cd toxicity, markedly diminished Cd-induced reactive oxygen species (ROS) and MDA accumulation, ameliorated Cd-induced damage to leaf/root ultrastructure, and increased chlorophyll content and P _n. External NO counteracted the pattern of alterations in certain antioxidant enzymes induced by Cd; for example, it significantly elevated the depressed SOD, APX, and catalase (CAT) activities in the Cd-sensitive genotype after 10- and 15-day treatments. Furthermore, NO significantly increased stromal APX and Mn-SOD activities in both genotypes and upregulated Cd-induced decrease in cAPX activity and gene expression of root/leaf cAPX and leaf CAT1 in the Cd-sensitive genotype. These data suggest that under Cd stress, NO, as a potent antioxidant, protects barley seedlings against oxidative damage by directly and indirectly scavenging ROS and helps to maintain stability and integrity of the subcellular structure.     

24-Epibrassinolide alleviated zinc-induced oxidative stress in radish (Raphanus sativus L.) seedlings by enhancing antioxidative system

This article encompasses the results on the effects of 24-epibrassinolide (EBR) on the changes in reactive oxygen species (ROS) and activities of antioxidative enzymes in radish (Raphanus sativus L.) seedlings subjected to zinc (Zn) stress. Zn toxicity resulted in significant enhancement in the level of membrane lipid peroxidation, protein oxidation, contents of hydrogen peroxide (H₂O₂) and hydroxyl radical (^·OH), the production rate of superoxide radicals (O ₂ ^·? ) and the activities of lipoxygenase and NADPH oxidase in radish seedlings indicating the induction of oxidative stress. However, Zn-mediated enhancement in indices of oxidative stress was considerably decreased by EBR treatment. EBR application enhanced the activities of catalase, superoxide dismutase, guaiacol peroxidase, glutathione peroxidase, and peroxidase in radish seedlings under Zn stress. EBR treatment reduced the activity of ascorbic acid oxidase in Zn stressed seedlings. Further, EBR application also enhanced the free proline and phenol levels under Zn stress. From the results obtained in this study, it can be inferred that EBR application alleviated oxidative damage caused by over production of ROS through the up regulation of antioxidative capacity in Zn stressed radish seedlings.     

Suppression of tomato <Emphasis Type="Italic">SlGGP</Emphasis> aggravates methyl viologen-mediated oxidative stress

Ascorbate (AsA) is an important antioxidant that can scavenge reactive oxygen species to protect plant cells against oxidative stress. Guanosine 5'-diphosphate (GDP)-L-galactose phosphorylase (GGP) is a key enzyme in the AsA biosynthetic pathway. To investigate the functions of GGP in AsA synthesis and oxidative stress tolerance in tomato, antisense lines with a reduced expression of SlGGP were obtained. Photobleaching after treatment of leaf disks with methyl viologen was more severe in transgenic lines compared to wild type (WT) plants. Moreover, compared with the WT plants, the transgenic plants showed a higher content of hydrogen peroxide, superoxide anion, malondialdehyde, as well as ion leakage, but a lower content of AsA and chlorophylls, ascorbate peroxidase activity, net photosynthetic rate, and maximal photochemical efficiency of photosystem II. Results of real-time quantitative polymerase chain reaction show that suppression of the SlGGP gene in the transgenic plants reduced their oxidative stress tolerance.     

Effects of exogenous nitric oxide on photosynthesis,antioxidative ability,and mineral element contents of perennial ryegrass under copper stress

A hydroponics experiment was conducted to test the effects of sodium nitroprusside (SNP, a donor of NO) supplied with different concentrations on copper (Cu) toxicity in ryegrass seedlings (Lolium perenne L.). Excess Cu (200 µM) reduced chlorophyll content, resulting a decrease in photosynthesis. Cu stress induced the production of hydrogen peroxide (H₂O₂) and superoxide anion (O₂^{? ?}), leading to malondialdehyde (MDA) accumulation. Furthermore, activities of antioxidant enzymes in Cu-treated seedlings such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were decreased. In addition, Cu stress inhibited the uptake of K, Mg, Fe, and Zn and increased Ca content in roots. Moreover, in leaves of Cu-stressed seedlings, K, Fe, and Zn contents were decreased and the contents of Ca and Mg were not affected significantly. In Cu-treated seedlings, Cu concentration in roots was higher than in leaves. Addition of 50, 100, 200 µM SNP in Cu-mediated solutions increased chlorophyll content and photosynthesis, improved antioxidant enzyme activities, reduced Cu-induced oxidative damages, kept intracellular ion equilibrium under Cu stress, increased Cu concentration in roots and inhibited Cu accumulation in leaves. In particular, addition of 100 µM SNP had the best effect on promoting growth of ryegrass seedlings under Cu stress. However, the application of 400 µM SNP had no obvious alleviating effect on Cu toxicity in ryegrass seedlings.     

Cadmium-induced changes in antioxidant enzyme activities in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L. cv. Dongjin)

We studied how the relationship between cadmium (Cd) toxicity and oxidative stress influenced the growth, photosynthetic efficiency, lipid peroxidation, and activity of ntioxidative enzymes in the roots and leaves of rice(Oryza sativa L Dongjin). Plants were exposed to Cd for 21 d. Both seedling growth and photosynthetic efficiency decreased gradually with increasing cadmium concentrations. Lipid peroxidation increased slowly in both roots and leaves, causing oxidative stress. However, each tissue type responded differently to Cd concentrations with regard to the induction/ inhibition of antioxidative enzymes. The activity of Superoxide dismutase (SOD) increased in both roots and leaves. Ascorbate peroxidase (APX) activity increased in leaves treated with up to 0.25 μM Cd, then decreased gradually at higher concentrations. In contrast, APX activity in roots increased and remained constant between 0.25 and 25 μM Cd. Enhanced peroxidase (POD) activity was recorded for treatments with up to 25/M Cd, gradually decreasing at higher concentrations in the leaves but remaining unchanged in the roots. Catalase (CAT) activity increased in the roots, but decreased in the leaves, whereas the activity of glutathione reductase (GR) was enhanced in both roots and leaves, where it remained elevated at higher Cd concentrations. These results suggest that rice seedlings tend to cope with free radicals generated by Cd through coordinated, enhanced activities of the antioxidative enzymes involved in detoxification.     

Effect of Cd on growth, photosynthetic gas exchange, and chlorophyll fluorescence of wild and Cd-sensitive mutant rice

Growth, photosynthetic gas exchange, and chlorophyll fluorescence characteristics were investigated in wild type (WT) and Cd-sensitive mutant rice (Oryza sativa L.) plants using 50 μM Cd treatment for 12 d followed by a 3-d recovery. Under Cd stress, net dry mass and pigment contents were significantly lower in the mutant plants than in the WT. The mutant had lower net photosynthetic rate (P _N), transpiration rate (E), and stomatal conductance (g _s) than WT rice, however, it had higher intercellular CO₂ concentration (C _i), indicating that non-stomatal factors accounted for the inhibition of P _N. Maximal photochemical efficiency of photosystem 2 (F_v/F_m), effective quantum yield of PS2 (Φ_PS2), and photochemical quenching (q_P) decreased much in the mutant under Cd stress. Cd content in roots and leaves of the mutant was significantly higher than those in the WT. Hence Cd toxicity was associated with the marked increases in Cd contents of plant tissue. After the recovery for 3 d, the WT rice had higher capacity to recover from Cd injury than the mutant.