Tolerance,accumulation and distribution of zinc and cadmium in hyperaccumulator Potentilla griffithii

In this study, zinc (Zn) and cadmium (Cd) tolerance, accumulation and distribution was conducted in Potentilla griffithii H., which has been identified as a new Zn hyperaccumulator found in China. Plants were grown hydroponically with different levels of Zn²⁺ (20, 40, 80 and 160 mg L^?1) and Cd²⁺ (5, 10, 20 and 40 mg L^?1) for 60 days. All plants grew healthy and attained more biomass than the control, except 40 mg L^?1 Cd treatment. Zn or Cd concentration in plants increased steadily with the increasing addition of Zn or Cd in solution. The maximum metal concentrations in roots, petioles and leaves were 14,060, 19,600 and 11,400 mg kg^?1 Zn dry weight (DW) at 160 mg L^?1 Zn treatment, and 9098, 3077 and 852 mg kg^?1 Cd DW at 40 mg L^?1 Cd treatment, respectively. These results suggest that P. griffithii has a high ability to tolerate and accumulate Cd and Zn, and it can be considered not only as Zn but also as a potential cadmium hyperaccumulator. Light microscope (LM) with histochemical method, scanning electron microscope combined with energy dispersive spectrometry (SEM-EDS) and transmission electron microscope (TEM) were used to determine the distribution of Zn and Cd in P. griffithii at tissue and cellular levels. In roots, SEM-EDS confirmed that the highest Zn concentration was found in xylem parenchyma cells and epidermal cells, while for Cd, a gradient was observed with the highest Cd concentration in rhizodermal and cortex cells, followed by central cylinder. LM results showed that Zn and Cd distributed mainly along the walls of epidermis, cortex, endodermis and some xylem parenchyma. In leaves, Zn and Cd shared the similar distribution pattern, and both were mostly accumulated in epidermis and bundle sheath. However, in leaves of 40 mg L^?1 Cd treatment, which caused the phytotoxicity, Cd was also found in the mesophyll cells. The major storage site for Zn and Cd in leaves of P. griffithii was vacuoles, to a lesser extent cell wall or cytosol. The present study demonstrates that the predominant sequestration of Zn and Cd in cell walls of roots and in vacuoles of epidermis and bundle sheath of leaves may play a major role in strong tolerance and hyperaccumulation of Zn and Cd in P. griffithii.     

Physiological mechanism of hypertolerance of cadmium in Kentucky bluegrass and tall fescue: Chemical forms and tissue distribution

Kentucky bluegrass (Poa pratensis) and tall fescue (Festuca arundinacea) are hypertolerant grasses to soil cadmium contamination. Little information is available on their tolerance mechanism. A sand culture and a hydroponic culture experiment were designed to investigate the Cd chemical form changes and its translocation in different tissues. The results showed that Kentucky bluegrass and tall fescue can tolerate 50–200 mg kg⁻¹ of soil Cd stresses and accumulate as high as 4275 and 2559 mg Cd kg⁻¹ DW, respectively, in their shoots without the loss of shoot biomass. Their Cd hypertolerance was correlated with an increase of the undissolved Cd phosphates in the leaves in both grass species, as determined by sequential solvent extraction procedures. The superior Cd tolerance of tall fescue to Kentucky bluegrass was associated with less Cd translocation into the stele of roots and less Cd transported to leaves. The pectate- and protein-integrated Cd forms may be involved in the symplastic translocation of Cd from cortex into stele, and this may lead the higher Cd concentrations in the stele of roots and then above ground leaves via long-distance transport in Kentucky bluegrass.     

The role of phytochelatins in constitutive and adaptive heavy metal tolerances in hyperaccumulator and non-hyperaccumulator metallophytes

Using the gamma-glutamylcysteine synthetase inhibitor, L-buthionine-[S,R]-sulphoximine (BSO), the role for phytochelatins (PCs) was evaluated in Cu, Cd, Zn, As, Ni, and Co tolerance in non-metallicolous and metallicolous, hypertolerant populations of Silene vulgaris (Moench) Garcke, Thlaspi caerulescens J.&C. Presl., Holcus lanatus L., and Agrostis castellana Boiss. et Reuter. Based on plant-internal PC-thiol to metal molar ratios, the metals' tendency to induce PC accumulation decreased in the order As/Cd/Cu > Zn > Ni/Co, and was consistently higher in non-metallicolous plants than in hypertolerant ones, except for the case of As. The sensitivities to Cu, Zn, Ni, and Co were consistently unaffected by BSO treatment, both in non-metallicolous and hypertolerant plants, suggesting that PC-based sequestration is not essential for constitutive tolerance or hypertolerance to these metals. Cd sensitivity was considerably increased by BSO, though exclusively in plants lacking Cd hypertolerance, suggesting that adaptive cadmium hypertolerance is not dependent on PC-mediated sequestration. BSO dramatically increased As sensitivity, both in non-adapted and As-hypertolerant plants, showing that PC-based sequestration is essential for both normal constitutive tolerance and adaptive hypertolerance to this metalloid. The primary function of PC synthase in plants and algae remains elusive.     

Lead,zinc, cadmium hyperaccumulation and growth stimulation in Arabis paniculata Franch

Metal hyperaccumulation is of great interest in recent years because of its potential application for phytoremediation of heavy metal contaminated soils. In this study, a field survey and a hydroponic experiment were conducted to study the accumulation characteristics of lead (Pb), zinc (Zn) and cadmium (Cd) in Arabis paniculata Franch., which was found in Yunnan Province, China. The field survey showed that the wild population of A. paniculata was hyper-tolerant to extremely high concentrations of Pb, Zn and Cd, and could accumulate in shoots an average level of 2300 mg kg^?1 dry weight (DW) Pb, 20,800 mg kg^?1 Zn and 434 mg kg^?1 Cd, with their translocation factors (TFs) all above one. Under the hydroponic culture, stimulatory effects of Pb, Zn and Cd on shoot dry biomass were noted from 24 to 193 μM Pb, 9 to 178 μM Cd and all Zn supply levels in nutrient solution, while the effects were not obvious in the roots. Chlorophyll concentrations in Pb, Zn and Cd treatments showed an inverted U-shaped pattern, consistent with the change of plant biomass. Pb, Zn and Cd concentrations in the shoots and roots increased sharply with increasing Pb, Zn and Cd supply levels. They reached > 1000 mg kg^?1 Pb, 10,000 mg kg^?1 Zn and 100 mg kg^?1 Cd DW in the 24 μM Pb, 1223 μM Zn and 9 μM Cd treatment, respectively, in which the plants grew healthy and did not show any symptoms of phytotoxicity. The TFs of Zn were basically higher than one and the amount of Zn taken by shoots ranged from 78.7 to 90.4% of the total Zn. However, the TFs of Pb and Cd were well below one, and 55.0–67.5% of total Pb and 57.8–83.5% of total Cd was accumulated in the shoots. These results indicate that A. paniculata has a strong ability to tolerate and hyperaccumulate Pb, Zn and Cd. Meanwhile, suitable levels of Pb, Zn and Cd could stimulate the biomass production and chlorophyll concentrations of A. paniculata. Thus, it provides a new plant material for understanding the mechanisms of stimulatory effect and co-hyperaccumulation of multiple heavy metals.     

Responses of non-protein thiols to Cd exposure in Cd hyperaccumulator Arabis paniculata Franch

We investigated the responses of phytochelatins (PCs), glutathione (GSH) and other non-protein thiols in Cd hyperaccumulator Arabis paniculata after Cd exposure. Applying γ-glutamylcysteine synthetase (γ-ECS) inhibitor, l-buthionine-sulfoximine (BSO), the roles of PCs in Cd tolerance and Cd accumulation in A. paniculata were evaluated. Plants were exposed to four Cd concentrations (0, 50, 100 and 250 μM) for different times (2w or 3w) with and without BSO. Overall, Cd exposure had little impact on plant biomass after 2w or 3w of growth except at the highest Cd level. A. paniculata tolerated ≤100 μM Cd with up to 1127 mg kg^?1 Cd in the shoots and 5624 mg kg^?1 Cd in the roots after 3w of Cd exposure. Cd exposure induced formation of PCs and three unknown thiols in the roots, but none were detected in the shoots. BSO had no significant effect on Cd sensitivity in plants though it reduced Cd accumulation in the roots. In addition, the molar ratio of PCs:Cd, which ranged from 0.7 to 1.3 after exposing to 50–100 μM Cd without BSO in the roots, was close to the value expected for PC-mediated Cd sequestration in plants. Those data indicate that GSH and PCs did not contribute to Cd tolerance in the shoots and Cd transport from the root to shoot in A. paniculata, but they may play an important role in Cd accumulation and Cd complexation in the roots of A. paniculata.     

Effects of zinc deficiency and zinc supplementation on homocysteine levels and related enzyme expression in rats

Methionine synthase (MS) and betaine-homocysteine methyltransferase (BHMT) are both zinc (Zn)-dependent methyltransferases and involved in the methylation of homocysteine. The objective of this study was to investigate the effects of dietary Zn supply on homocysteine levels and expression of the two enzymes in growing rats. Male weanling Sprague-Dawley rats were assigned randomly to four dietary groups (n = 8/group) for 3 weeks: Zn deficient (ZD; <1 mg Zn/kg); Zn control (ZC; 30 mg Zn/kg); Zn supplemented (ZS; 300 mg Zn/kg); pair fed (PF; 30 mg Zn/kg) to the ZD group. Serum and femur Zn concentrations were 83% and 58% lower in ZD, and 49% and 62% higher in ZS compared to ZC (P < 0.001), respectively. The ZD rats had lower feed intake (37%), body weight gains (45%), liver (43%) and kidney (31%) weights than those of ZC (P < 0.001), but these parameters in ZD were not significantly different from the PF controls. Serum homocysteine concentrations were 65% higher in ZD compared to PF (P < 0.05), and there was no significant difference in serum folate levels between ZD and PF groups. The mRNA expression of liver and kidney MS was 57% and 38% lower in ZD than PF (P < 0.001), respectively. Hepatic and renal BHMT mRNA levels were not altered in ZD compared to controls. The aforementioned measurements were not significantly different between ZS and ZC groups, except Zn levels. These results demonstrated that homocysteine homeostasis appeared to be disturbed by Zn deficiency but not Zn supplementation, and elevated serum homocysteine might be due to reduced expression of MS during Zn deficiency.     

Accumulation of copper in the kidney of pigs fed high dietary zinc is due to metallothionein expression with minor effects on genes involved in copper metabolism

A study was conducted to determine the effect of high dietary zinc (Zn) oxide on trace element accumulation in various organs with special emphasis on the kidney. A total of 40 weaned piglets were allocated into two groups with 16 and 24 piglets each receiving a diet containing normal (NZn; 100 mg Zn/kg) or high (HZn; 2,100 mg Zn/kg) Zn concentration, respectively. After two weeks, eight piglets from each treatment were killed and organ samples were taken. Eight piglets from the remaining 16 pigs fed HZn diets were changed to NZn diets (CZn). All remaining piglets were killed after another two weeks for organ sampling. Trace element concentration was determined in the jejunum, liver, kidney, pancreas, bone (metacarpal IV), spleen, lung, thymus, tonsils and lymph nodes of jejunum, ileum and colon. Kidney mRNA expression of Zn transporter ZnT1 and ZIP4, genes involved in Cu metabolism (Ctr1, Atox1, SOD1, ATP7A, CCS, CP) and divalent metal ion transport (DMT1) and binding (MT-1a, MT-2b, MT-3) were determined. The Zn concentration in jejunum, liver, pancreas tissue and metacarpal IV was higher (P < 0.05) in HZn group compared with NZn and CZn groups. Trace element concentration in organs of CZn pigs was similar to those fed NZn diets. Zn concentration in muscle, lung and lymphatic organs as thymus, tonsils, spleen and lymph nodes of jejunum, ileum and colon did not differ between the groups. Zn and Cu were positively correlated (R = 0.67; P < 0.05) in the kidney. No significant differences for Cu chaperones, Cu transporters and Cu-dependent factors were determined despite decreased expression of Atox1 after two weeks and increased Ctr1 expression over time in the HZn group. Expression of MT-1a, MT-2b and MT-3 were significantly higher in HZn fed pigs with most pronounced effects for MT-1a > MT-2b > MT-3. Gene expression of MTs in pigs fed CZn diets did not differ from pigs fed NZn diets. The data suggest that high dietary Zn feeding in pigs leads to Cu co-accumulation in the kidney of pigs with minor effect on genes relevant for Cu metabolism. In addition, the organ Zn and Cu accumulation is reversible after two weeks of withdrawal of high dietary Zn.     

Cadmium tolerance in Thlaspi caerulescens: I. Growth parameters,metal accumulation and phytochelatin synthesis in response to cadmium

A population of the metallophyte, Thlaspi caerulescens, originating from a Cd–Pb–Zn old mining and smelter site at Plombières (Belgium) was studied. T. caerulescens was cultivated hydroponically to investigate Cd uptake and tolerance. Cd was added to Hoagland’s medium at concentration range from 5 to 500 μM. The plants could tolerate 500 μM Cd in the solution showing only minor visible symptoms of toxicity but with a 32% decrease in fresh weight. After 14 days at 500 μM, Cd content in roots and shoots was 707 and 602 mg kg⁻¹ of dry weight (d.w.), respectively. Application of Cd to hydroponically cultivated T. caerulescens induced the accumulation of PCs in plant roots and shoots. Buthionine sulfoximine (BSO) application almost completely reduced (by 98–100%) the accumulation of PCs without simultaneous increase in plants sensitivity to Cd. These results suggest a minor if any role of PCs in tolerance to Cd of the studied population of T. caerulescens in hydroponics. On the other hand, no PC accumulation was detected either in T. caerulescens plants growing in their natural environment at Plombierès or in plants growing in their native soil in a greenhouse. These results suggest that naturally selected tolerance in T. caerulescens population from Plombières is not associated with enhanced PCs synthesis.     

Accumulation of zinc and cadmium and localization of zinc in Picris divaricata Vant.

Picris divaricata Vant., a plant species native to subtropical China, was recently identified as the first Cd/Zn hyperaccumulator from Asteraceae. P. divaricata was grown from wild collected seed for 4 months in a series of pH adjusted test soils with added Zn levels 0–7000 mg kg⁻¹ and Cd levels 0–150 mg kg⁻¹. Plants did not hyperaccumulate Zn (threshold >3000 μg g⁻¹) and weakly hyperaccumulated Cd with little or no dose–response.P. divaricata has multicellular simple trichomes concentrated on the leaf margins and midrib. X-ray analysis showed that Zn was concentrated in larger trichomes and epidermal cells adjacent to the trichome but virtually absent in other leaf tissues. Within the trichomes, Zn was localized in ovate spots around the tips of individual cells. These tips and other locations in the trichome cell contained black electron dense material when examined with transmission electron microscopy, some of which was identified as SiO₂. Silicon and Mn were concentrated in the same areas as Zn. Si has been previously associated with alleviating Zn, Mn and Cd toxicity. Our results support this observation and further investigation is warranted.Calcium and P were concentrated in the distal tips of trichomes, similar to patterns previously observed for calcicole plants grown in elevated Ca soils. Overall, nonsecretory trichomes from many plant families may have a common origin as tissues adapted to handle a variety of environmental metals.     

Evaluation of zinc tolerance and accumulation potential of the coastal shrub Limoniastrum monopetalum (L.) Boiss.

The coastal shrub Limoniastrum monopetalum is capable of growth in soil containing extremely high concentrations of heavy metals. A greenhouse experiment was conducted in order to investigate the effects of a range of Zn concentrations (0–130 mmol l⁻¹) on growth and photosynthetic performance, by measuring relative growth rate, total leaf area, plant height, gas exchange, chlorophyll fluorescence parameters and photosynthetic pigment concentrations. We also determined the total zinc, nitrogen, phosphorus, sulphur, calcium, magnesium, sodium, potassium, iron and copper concentrations in the plant tissues. The study species demonstrated hypertolerance to Zn stress, since survival was recorded with leaf concentrations of up to 1700 mg Zn kg⁻¹ dry mass when treated with 130 mmol Zn l⁻¹. L. monopetalum exhibited little overall effects on photosynthetic function at Zn levels of up to 90 mmol l⁻¹. At greater external Zn concentration, plant growth was negatively affected, due in all probability to the recorded decline in net photosynthetic rate, which may be linked to the adverse effect of the metal on photosynthetic electron transport. Growth parameters were virtually unaffected by leaf tissue concentrations as high as 1400 mg Zn kg⁻¹ dry mass thus indicating that this species could play an important role in the phytoremediation of Zn-polluted areas.     

Diversity and heavy metal tolerance of endophytic fungi from six dominant plant species in a Pb–Zn mine wasteland in China

The diversity and metal tolerance of endophytic fungi from six dominant plant species in a Pb–Zn mine wasteland in Yunnan, China were investigated. Four hundred and ninety-five endophytic fungi were isolated from 690 tissue segments. The endophytic fungal colonization extent and isolation extent ranged from 59 % to 75 %, and 0.42–0.93, respectively, and a positive correlation was detected between them. Stems harboured more endophytic fungi than leaves in each plant species, and the average colonization extent of stems was 82 %, being significantly higher than that of leaves (47 %) (P ≤ 0.001, chi-square test). The fungi were identified to 20 taxa in which Phoma, Alternaria and Peyronellaea were the dominant genera and the relative frequencies of them were 39.6 %, 19.0 % and 20.4 %, respectively. Metal tolerance test showed that 3.6 mM Pb²⁺ or 11.5 mM Zn²⁺ exhibited the greatest toxicity to some isolates and they did not grow on the metal-amended media. In contrast, some isolates were growth stimulated in the presence of tested metals. The isolates of Phoma were more sensitive to Zn²⁺ than the isolates of Alternaria and Peyronellaea. However, the sensitivity of isolates to Pb²⁺ was not significantly different among Phoma, Alternaria, Peyronellaea and other taxa (P > 0.05, chi-square test). Our results suggested that fungal endophyte colonization in Pb–Zn polluted plants is moderately abundant and some isolates have a marked adaptation to Pb²⁺ and Zn²⁺ metals, which has a potential application in phytoremediation in this area.     

Earthworms as bioindicator of metals (Zn,Fe, Mn,Cu, Pb and Cd) in soils: Is metal bioaccumulation affected by their ecological category?

The importance of earthworms in metal pollution monitoring is widely recognized in terrestrial ecosystems. Metal bioaccumulation by soil-dwelling earthworms can be used as an ecological indicator of metal availability in soils. In this study, we quantify the level of DTPA extractable metals in casts and tissues of earthworms (endogeic: Metaphire posthuma (Vaillant) and anecic: Lampito mauritii Kinberg) and ingesting soils, collected form cultivated land, urban garden and sewage soils. Soil and worm casts collected from sewage and cultivated land showed the greater metal concentrations. The concentration of Zn, Fe, Pb and Mn in earthworm casts was in the order: sewage soil > cultivated land > urban garden, while for Cu and Cd the order was cultivated land > sewage soil > urban garden. Data suggested that the level of DTPA extractable metals was higher than that of surrounding soils. We got close relationships between metal concentration in worm tissues and surrounding soils: Zn (r² = 0.94 and 0.89, P < 0.01 for both), Fe (r² = 0.95 and 0.97, P < 0.01 for both), Cu (r² = 0.93 and 0.96, P < 0.01), Pb (0.63, P < 0.01 and 0.57, P > 0.05), and Cd (r² = 0.15, P > 0.01 and 0.75, P < 0.01), respectively, for M. posthuma and L. mauritii. The study clearly indicates that earthworms have efficient potentials for bioaccumulation of metals in their tissues which can be used as an ecological indicator of soil contaminations. A species-specific metal accumulation pattern was observed in studied earthworms. Comparatively, endogeic showed the higher metal contents in their tissues than anecic (t-test: P < 0.05); collected form different habitats studied. Data suggested that species-specific feeding behaviour, earthworm niche structure, ecological category of inhabiting earthworm and even horizontal distribution of contaminants in soil layers are some major determinant for metal accumulation patterns in soil dwelling earthworms. The difference in burrowing patterns can influence the patterns of metal bioaccumulations between endogeic and anecic, although other factors are also contributory. Further more detailed study is still required to elaborate the proposed hypothesis.     

Characterization and efficacy of Muscodor cinnamomi in promoting plant growth and controlling Rhizoctonia root rot in tomatoes

Muscodor cinnamomi was selected and investigated for its in vitro ability to produce indole-3-acetic acid (IAA) to solubilize different toxic metal (Ca, Co, Cd, Cu, Pb, Zn)-containing insoluble minerals and tolerance to metals, herbicides and an insecticide. The results indicated that this fungus is able to produce IAA (45.36 ± 2.40 μg ml⁻¹) in liquid media. This phytohormone stimulated coleoptile elongation, and increased seed germination and root elongation of tested plants. The metal tolerance and solubilizing ability depended on the type of insoluble minerals. M. cinnamomi showed the highest growth tolerance on Ca-containing media at 150 mM, followed by Zn-containing media at 100 mM and Cd-containing media at 10 mM. This fungus tolerated the three herbicides (2,4-d-dimethylammonium, glyphosate and paraquat dichloride) and an insecticide (methomyl) at the recommended dosages for field application. Moreover, M. cinnamomi completely controlled Rhizoctonia solani AG-2 root rot in tomato plants, and increased root length, shoot dry weight and root dry weight. This is the first report of in vitro IAA production, solubilization of insoluble metal minerals, and tolerance to herbicides, an insecticide and metals as well as the plant growth promoting ability of M. cinnamomi.     

Plasma levels of trace elements and exercise induced stress hormones in well-trained athletes

This study analyzed the variation and relationship of several trace elements, metabolic substrates and stress hormones activated by exercise during incremental exercise. Seventeen well-trained endurance athletes performed a cycle ergometer test: after a warm-up of 10 min at 2.0 W kg⁻¹, the workload was increased by 0.5 W kg⁻¹ every 10 min until exhaustion. Prior diet, activity patterns, and levels of exercise training were controlled, and tests timed to minimize variations due to the circadian rhythm. Oxygen uptake, blood lactate concentration, plasma ions (Zn, Se, Mn and Co), serum glucose, non-esterified fatty acids (NEFAs) and several hormones were measured at rest, at the end of each stage and 3, 5 and 7 min post-exercise. Urine specific gravity was measured before and after the test, and participants drank water ad libitum.Significant differences were found in plasma Zn and Se levels as a function of exercise intensity. Zn was significantly correlated with epinephrine, norepinephrine and cortisol (r = 0.884, P < 0.01; r = 0.871, P < 0.01; and r = 0.808, P = 0.05); and Se showed significant positive correlations whit epinephrine and cortisol (r = 0.743, P < 0.05; and r = 0.776, P < 0.05). Neither Zn nor Se levels were associated with insulin or glucagon, and neither Mn nor Co levels were associated with any of the hormones or substrate metabolites studied. Further, while Zn levels were found to be associated only with lactate, plasma Se was significantly correlated with lactate and glucose (respectively for Zn: r = 0.891, P < 0.01; and for Se: r = 0.743, P < 0.05; r = 0.831, P < 0.05).In conclusion, our data suggest that there is a positive correlation between the increases in plasma Zn or Se and stress hormones variations induced by exercise along different submaximal intensities in well-hydrated well-trained endurance athletes.     

Plasma trace elements levels are not altered by submaximal exercise intensities in well-trained endurance euhydrated athletes

The purpose of this study was to assess the effect of relative exercise intensity on various plasma trace elements in euhydrated endurance athletes.Twenty-seven well-trained endurance athletes performed a cycloergometer test: after a warm-up of 10 min at 2.0 W kg⁻¹, workload increased by 0.5 W kg⁻¹ every 10 min until exhaustion. Oxygen uptake, blood lactate concentration ([La⁻]_b), and plasma ions (Zn, Se, Mn and Co) were measured at rest, at the end of each stage, and 3, 5 and 7 min post-exercise. Urine specific gravity (U_SG) was measured before and after the test, and subjects drank water ad libitum. Fat oxidation (FAT_OXR), carbohydrate oxidation (CHO_OXR), energy expenditure from fat (EE_FAT), from carbohydrates (EE_CHO) and total EE (EE_T) were estimated using stoichiometric equations. A repeated measure (ANOVA) was used to compare plasma ion levels at each exercise intensity level. The significance level was set at P < 0.05.No significant differences were found in U_SG between, before, and after the test (1.014 ± 0.004 vs. 1.014 ± 0.004 g cm⁻³) or in any plasma ion level as a function of intensity. There were weak significant correlations of Zn (r = 0.332, P < 0.001) and Se (r = 0.242, P < 0.01) with [La⁻]_b, but no relationships were established between [La⁻]_b, VO₂, FAT_OXR, CHO_OXR, EE_FAT, EE_CHO, or EE_T and plasma ion levels.Acute exercise at different submaximal intensities in euhydrated well-trained endurance athletes does not provoke a change in plasma trace element levels, suggesting that plasma volume plays an important role in the homeostasis of these elements during exercise.     

Chloride salinity reduces cadmium accumulation by the Mediterranean halophyte species Atriplex halimus L.

Soil salinity usually increases bioavailability of Cd on heavy metal polluted soils but its impact on Cd absorption and accumulation by plants remains largely unknown. Plants from the halophyte species Atriplex halimus were therefore exposed for 12 and 14 days to nutrient solution containing 50 μM CdCl₂ in the presence of NaCl, KCl or NaNO₃ 50 mM. Most Cd present in solution remained as Cd–EDTA and salinity had no impact on Cd speciation. Chloride salinity (NaCl and KCl) reduced Cd accumulation in shoots and roots while NaNO₃ increased Cd accumulation in leaves. More than 30% of accumulated Cd was found at the leaf surface and accumulated in trichomes but all tested salts decreased the proportion of excreted Cd. Cadmium induced a decrease in the leaf water content. External NaCl and KCl mitigated the deleterious impact of Cd by inducing osmotic adjustment while NaNO₃ and synthesis of protecting compounds such as soluble sugars and glycinebetaine. Free polyamines (putrescine, spermidine and spermine) increased in response to Cd, Cd + NaCl and Cd + KCl while only putrescine increased in response to Cd + NaNO₃. Proline exhibited maximal concentration in the leaves of Cd + NaCl and Cd + KCl-treated plants and was correlated with osmotic adjustment. Our results suggest that chloride salinity improved the resistance of A. halimus to Cd toxicity both by decreasing the absorption of heavy metal and by improving tissular tolerance through an increase in the synthesis of osmoprotective compounds.     

Effect of NaCl on growth and Cd accumulation of halophyte Spartina alterniflora under CdCl2 stress

A study quantifying the effect of NaCl on growth and Cd accumulation of Spartina alterniflora subjected to Cd stress was conducted. Seedlings were cultivated in the presence of 1 or 3 mM Cd alone, or combined with NaCl (50 or 100 mM). The results showed that NaCl magnified the phytotoxicity of moderate Cd stress (1 mM Cd) on plants due to reduced levels of plant biomass, plant height, and chlorophyll a + b, while no synergistic effects were recorded under severe Cd stress (3 mM Cd). Proline and Ca^2 + accumulated along with additional NaCl under moderate Cd stress, instead of reduced or unchanged levels under severe Cd stress owing to different adoption strategies caused by NaCl under different Cd stresses. NaCl reduced the oxidative stress in Cd-treated plants through increasing levels of antioxidative enzymes (catalase (CAT) and peroxidase (POD)) under moderate Cd stress. With NaCl addition, Cd^2 + contents in S. alterniflora increased and reduced under moderate and severe Cd stress, respectively. However, total Cd^2 + amounts increased with increasing NaCl concentration due to biological dilution. NaCl improved the increase of Cd^2 + translocation factor (TF) under moderate Cd stress, indicating that NaCl might improve Cd^2 + uptake and translocation from roots to shoots, and enhance the phytoextraction of S. alterniflora on Cd; while phytostabilization of Cd under severe Cd stress may be possible due to the reduced TF. Thus, NaCl alleviated phytotoxicity caused by Cd stress through improved management of osmotic solutes and oxidative status, and affected Cd accumulations in S. alterniflora differently under moderate and severe Cd stresses.     

Subcellular distribution and chemical forms of cadmium in Bechmeria nivea (L.) Gaud.

Bechmeria nivea (L.) Gaud. (Ramie) is a promising species for Cd phytoextraction with large biomass and fast growth rate. Nevertheless, little information is available on its tolerance mechanisms towards Cd. Determination of Cd distribution and chemical speciation in ramie is essential for understanding the mechanisms involved in Cd accumulation, transportation and detoxification. In the present study, ramie plants were grown in hydroponics with increasing Cd concentrations (0, 1, 3, 7 mg l^?1). The subcellular distribution and chemical forms of Cd in different tissues were determined after 20 days exposure to this metal. To assess the effect of Cd uptake on plant performance, nitrate reductase activity in leaves and root activity were analyzed during the entire experimental period. Increased Cd level in the medium caused a proportional increase in Cd uptake, and the highest Cd concentration occurred in roots, followed by stems and leaves. Subcellular fractionation of Cd-containing tissues indicated that about 48.2–61.9% of the element was localized in cell walls and 30.2–38.1% in soluble fraction, and the lowest in cellular organelles. Cd taken up by ramie rapidly equilibrated among different chemical forms. Results showed that the greatest amount of Cd was found in the extraction of 1 M NaCl and 2% HAC, and the least in residues in all test tissues. In roots, the subdominant amount of Cd was extracted by d-H₂O and 80% ethanol, followed by 0.6 M HCl. While in stems and leaves, the amount of 0.6 M HCl-extractable Cd was comparable with that extracted by 80% ethanol or d-H₂O. 1 mg l^?1 Cd stimulated nitrate reductase activity in leaves and root activity, while a concentration-dependent inhibitory effect was observed with increasing Cd concentration, particularly at 7 mg l^?1 Cd. It could be suggested that the protective mechanisms evolved by ramie play an important role in Cd detoxification at relatively low Cd concentrations (below 3 mg l^?1 Cd) but become restricted to maintain internal homeostasis with higher Cd stress.     

Relative bioavailabilities of organic zinc sources with different chelation strengths for broilers fed diets with low or high phytate content

A six-day experiment was conducted to estimate the relative bioavailability values (RBV) of zinc (Zn) in three organic sources (oZn) with different chelation strengths compared to inorganic ZnSO₄ (iZn) for broilers fed a low or high phytate diet. A total of 1080, one-d-old male broiler chicks were randomly assigned to one of 18 dietary treatments (six replicates cages of ten chicks per cage) in a completely randomized design involving a 2 × 2 × 4 factorial arrangement with two levels of added phytate (0 or 10 g phytate as sodium phytate/kg), two levels of added Zn (30 or 60 mg/kg) and four Zn sources (iZn and three oZn sources) plus one low and one high phytate control treatments without Zn addition. The three oZn sources consisted of (1) Zn amino acid with weak chelation strength (ZnAA-L, formation quotient Q_f = 6.6, containing 119 g Zn/kg), (2) Zn proteinate with moderate chelation strength (ZnPRO-M, Q_f = 30.7, containing 133 g Zn/kg) or (3) Zn proteinate with strong chelation strength (ZnPRO-H, Q_f = 944.0, containing 186 g Zn/kg). Chicks were harvested at 6 days of age and pancreas metallothionein (MT) mRNA expression was used to estimate Zn RBV. Pancreas MT mRNA expression increased (P<0.01) as dietary Zn level increased. Chicks fed high phytate diets had lower (P<0.05) MT mRNA expression than chicks fed low phytate diets. Based on multiple linear regression slope ratios with ZnSO₄ set at 1.00, the RBV of ZnAA-L, ZnPRO-M and ZnPRO-H were 1.01, 1.28 and 0.70, respectively, for low phytate diets, and 1.05, 1.39 and 0.92, respectively, for high phytate diets. The slope for the oZn source with moderate chelation strength differed (P<0.05) from iZn and the other two oZn sources. The RBV of ZnAA-L, ZnPRO-M and ZnPRO-H under the high phytate diet increased by 0.04, 0.11 and 0.22, respectively, compared to those under the low phytate diet. Results indicate that the oZn sources with moderate or strong chelation strength offer partial or complete resistance to interference from high dietary phytate during digestion; and the oZn with moderate chelation strength had a greater RBV with both low and high phytate diets than iZn or oZn sources with weak or strong chelation strength.     

Response of native grasses and Cicer arietinum to soil polluted with mining wastes: Implications for the management of land adjacent to mine sites

Mine tailings are an environmental problem in Southern Spain because wind and water erosion of bare surfaces results in the dispersal of toxic metals over nearby urban or agricultural areas. Revegetation with tolerant native species may reduce this risk. We grew two grasses, Lygeum spartum and Piptatherum miliaceum, and the crop species Cicer arietinum (chickpea) under controlled conditions in pots containing a mine tailings mixed into non-polluted soil to give treatments of 0%, 25%, 50%, 75% and 100% mine tailings. We tested a neutral (pH 7.4) mine tailings which contained high concentrations of Cd, Cu, Pb and Zn. Water-extractable metal concentrations increased in proportion to the amount of tailings added. The biomass of the two grasses decreased in proportion to the rate of neutral mine-tailing addition, while the biomass of C. arietinum only decreased in relation to the control treatment. Neutron radiography revealed that root development of C. arietinum was perturbed in soil amended with the neutral tailings compared to those of the control treatment, despite a lack of toxicity symptoms in the shoots. In all treatments and for all metals, the plants accumulated higher concentrations in the roots than in shoots. The highest concentrations occurred in the roots of P. miliaceum (2500 mg kg^?1 Pb, 146 mg kg^?1 Cd, 185 mg kg^?1 Cu, 2700 mg kg^?1 Zn). C. arietinum seeds had normal concentrations of Zn (70–90 mg kg^?1) and Cu (6–9 mg kg^?1). However, the Cd concentration in this species was ～1 mg kg^?1 in the seeds and 14.5 mg kg^?1 in shoots. Consumption of these plant species by cattle and wild fauna may present a risk of toxic metals entering the food chain.