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.     

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.     

Emergence and Seedling Growth of a Cultivated and a Wild Strain of Safflower under Various Amounts of Simulated Rainfall

Two separate experiments were conducted to investigate the effects of different planting depths and irrigation regimes on the emergence and seedling heights of a cultivated (Carthamus tinctorius L. cv. Arak 2811) and a wild strain (C. oxyacantha Bieb.) of safflower. In the first experiment, three planting depths (0.5, 2.0 and 3.5 cm) and four water levels (6,9,12, and 15 mm) were used. Water was applied only once at the start of the experiment. For Arak 2811, the highest emergence percentage and seedling height were obtained when seeds were planted 0.5 cm deep in the soil and irrigated with the equivalent of 12 mm of rainfall. In the second experiment, five water levels (3, 6, 9, 12 and 15 mm) were applied at 3-day intervals for a period of 25 days and the planting depth was 0.5 cm. For Arak 2811, the seedlings emerged within 4 to 7 days after the first watering while for the wild strain, the emergence started on the 4th day and continued up to the 17th day. The emergence percentage of the wild safflower was almost half of that of the cultivated strain. However, the emerged wild seedlings were much more resistant to low available water than the seedlings of the cultivated variety of safflower.     

Antioxidant defense mechanisms in response to cadmium treatments in two safflower cultivars

By using two safflower (Carthamus tinctorius L.) cultivars, Arak2811 and Goldasht, the experiments were conducted in order to study (i) the genotypic variation in cadmium (Cd) tolerance, (ii) Cd concentrations in plants, and (iii) changes in the antioxidant defense systems in leaves, including antioxidant enzymes and nonenzymatic antioxidants. Plants were grown under controlled environmental conditions and subjected to Cd treatments (0, 25, 50, 75, and 100 μM Cd) for different time periods. Cd concentrations and cultivar-dependent response to Cd were assessed. Of the two cultivars, Goldasht showed a greater sensitivity to Cd toxicity as judged from the severity of Cd toxicity symptoms on leaves, much stronger enhancement in the MDA level, and decreases in dry matter production. Increasing Cd supply markedly reduced the shoot and root dry weights in both cultivars, but at the higher Cd concentrations and longer exposure durations, this decrease was more marked in cv. Goldasht. Plants accumulated substantial amount of Cd, especially in the roots, the highest being in the roots of cv. Arak2811 at 100 μM Cd after 4 days. Cd-induced oxidative stress as was indicated by the increase in lipid peroxidation with the increase in metal concentration and exposure duration. Under different Cd stress levels, activities of antioxidant enzymes differed in the two cultivars. The results indicated that Cd tolerance of cv. Arak2811 was related to the retention of Cd in the roots and avoiding the toxic effect by activation of the antioxidant system.     

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.     

Response of miscanthus to toxic cadmium applications during the period of maximum growth

Plants of miscanthus were grown in a Cd-free solution up to 1 month before heading and then were exposed to 0, 0.75, 1.5, 2.25 and 3 mg l⁻¹ cadmium for 36 days. All cadmium levels were toxic to miscanthus. Growth response was not dose-dependent and two toxicity thresholds were identified: one between 0 and 0.75 mg l⁻¹ Cd, the other between 2.25 and 3 mg l⁻¹ Cd. The former caused a biomass decrease by about 50%, whereas the latter completely inhibited growth and disrupted the mechanisms that restricted Cd translocation to the shoot. Growth of the aerial part was affected by cadmium more than that of the hypogeal one. Cadmium did not change the N concentration of different plant parts, but markedly reduced the N uptake of the plant, the N net uptake rate (NUR) and the N net translocation rate (NTR) from the rhizome to the aerial part. These two indexes equalled zero when plants ceased to grow. Otherwise, the Cd-NUR increased with Cd supply and the Cd-NTR from rhizome to aerial part showed the highest increment when plants did not grow at all. This suggests different uptake pathways for the two elements, active for nitrogen and passive for cadmium. The Cd concentration and the Cd content markedly increased with all Cd levels, following the order roots ≫ rhizome > culms > leaves. The Cd concentration and the Cd content of aerial organs increased with Cd supply, but increments were highest between 2.25 and 3 mg l⁻¹ Cd. The highest Cd concentrations were recorded in plants grown with 3 mg l⁻¹ Cd and were 41 and 122 mg kg⁻¹, respectively, for the aerial and the hypogeal plant parts. The hypogeal plant part retained most of the cadmium taken up from solution, accounting for approximately 87% of total plant cadmium with the three lower Cd levels, and for 73% with the highest one. The maximum Cd content of the entire plant was achieved with the two higher Cd levels and was approximately 4.7 mg, while the Cd content of the aerial part was highest with 3 mg l⁻¹ Cd (1.2 mg Cd per plant) and that of the hypogeal one with 2.25 mg l⁻¹ Cd (4 mg Cd per plant). The highest aerial content achieved in this experiment was 10-fold that obtained in a previous research when small-sized plants were exposed to the same Cd level.     

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.     

Silicon mitigates cadmium inhibitory effects in young maize plants

The influence of silicon on the growth of maize plants cultivated in hydroponics in the presence of cadmium (5 μM) was investigated. Four different treatments were used: Control (C), Cadmium (Cd), Silicon (Si) and Cadmium plus Silicon (Cd + Si). The Si concentration was 35 mM. Thirteen-day-old plants were harvested. Growth parameters (length of primary seminal root, leaf area of first and second fully developed leaves, fresh and dry weight of below- and above-ground parts of the plants), and Cd concentration and total amount of Cd in the below- and above-ground parts were determined. In roots, the development of the endodermal barrier was observed by fluorescent staining with Fluorol yellow 088.Inhibitory effects of Cd on plant growth were observed. Silicon treatment in the absence of Cd had positive effects on most of observed growth parameters compared with the control. Moreover, Si in the Cd + Si treatment improved all growth parameters compared with the cadmium treatment. Silicon increased the cell-wall extensibility both in Si and Cd + Si treatments when compared with the control. Alleviation of the Cd-inhibitory effect on maize plants by Si was not due to exclusion of Cd from the plant; in contrast, Cd concentration in below- and above-ground plant parts and the total amount of Cd per plant were significantly higher in the Cd + Si plants than in the Cd treatment. The increased Cd content in Cd + Si plants was correlated with the development of the endodermis; during the second stage of endodermal development, suberin lamellae were formed at a greater distance from the root apex in the Cd + Si than in the Cd treatment. Silicon itself did not influence the development of suberin lamellae in the maize roots compared with the control.     

Role of iron plaque in Cd uptake by and translocation within rice (Oryza sativa L.) seedlings grown in solution culture

A hydroponics culture experiment was conducted to investigate the effect of iron plaque on Cd uptake by and translocation within rice seedlings grown under controlled growth chamber conditions. Rice seedlings were pre-cultivated for 43 days and then transferred to nutrient solution containing six levels of Fe (0, 10, 30, 50, 80 and 100 mg L⁻¹) for 6 days to induce different amounts of iron plaque on the root surfaces. Seedlings were then exposed to solution containing three levels of Cd (0, 0.1 and 1.0 mg L⁻¹) for 4 days. In order to differentiate the uptake capability of Cd by roots with or without iron plaque, root tips (white root part without iron plaque) and middle root parts (with iron plaque) of pre-cultivated seedlings treated with 0, 30 and 50 mg L⁻¹ Fe were exposed to ¹⁰⁹Cd for 24 h. Reddish iron plaque gradually became visible on the surface of rice roots but the visual symptoms of the iron plaque on the roots differed among treatments. In general, the reddish color of the iron plaque became darker with increasing Fe supply, and the iron plaque was more homogeneously distributed all along the roots. The Fe concentrations increased significantly with increasing Fe supply regardless of Cd additions. The Cd concentrations in dithionite–citrate–bicarbonate (DCB)-extracts and in shoots and roots were significantly affected by Cd and Fe supply in the nutrient solution. The Cd concentrations increased significantly with increasing Cd supply in the solution and were undetectable when no Cd was added. The Cd concentrations in DCB-extracts with Fe supplied tended to be higher than that at Fe₀ at Cd_0.1, and at Cd_1.0, DCB-Cd with Fe supplied was significantly lower. Cd concentrations in roots and shoots decreased with increasing Fe supply at both Cd additions. The proportion of Cd in DCB-extracts was significantly lower than in roots or shoots. Compared to the control seedlings without Fe supply, the radioactivity of ¹⁰⁹Cd in shoots of seedlings treated with Fe decreased when root tips were exposed to ¹⁰⁹Cd and did not change significantly when middle parts of roots were exposed. Our results suggest that root tissue rather than iron plaque on the root surface is a barrier to Cd uptake and translocation within rice plants, and the uptake and translocation of Cd appear to be related to Fe nutritional levels in the plants.     

Long-term effects of exogenous silicon on cadmium translocation and toxicity in rice (Oryza sativa L.)

Most nutrient solution studies on the interactions between silicon (Si) and cadmium (Cd) are short term. Here we reported a long-term experiment in which rice (Oryza sativa L.) was cultured for 105 days and harvested at four different growth stages to measure biomass accumulation and Cd uptake and distribution in shoots and roots. Exogenous Si increased shoot biomass by 61–238% and root biomass by 48–173% when the culture solution was free of Cd. When 2 μmol L^?1 Cd was added, Si supply increased shoot and root biomass by 125–171% and by 100–106% compared to the zero-Si treatment. Increasing the Cd concentration to 4 μmol L^?1 decreased the beneficial effects of Si on root and shoot biomass. Silicon supply decreased shoot Cd concentrations by 30–50% and Cd distribution ratio in shoot by 25.3–46%, compared to the treatment without Si supply. Additionally, lower Si supply or more serious Cd stress would lead to roots with bigger biomass and higher Si concentration. Energy-dispersive X-ray microanalysis showed that both Si and Cd accumulated synchronously in the border and middle of phytoliths of the shoots. We conclude that Si enhances plant growth and decreases Cd accumulation in shoots and thereby helps to lower the potential risks of food contamination.     

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.     

Influence of water temperature and waterborne cadmium toxicity on growth performance and metallothionein–cadmium distribution in different organs of Nile tilapia,Oreochromis niloticus (L.)

Cadmium (Cd) is believed to be one of the most abundant and ubiquitously distributed toxins in the aquatic system. This metal is released to the aquatic environment from both anthropogenic sources, such as industrial, agricultural and urban effluents as well as natural sources, such as rocks and soils. Otherwise, the temperature increase of water bodies, which has been observed due to global climatic changes, has been shown to increase Cd toxicity for several aquatic animal species including fish. In the present study, Nile tilapia, Oreochromis niloticus (L.), (26.0±0.38 g) were reared at 20, 24, 28, or 32 °C and exposed to 0.0 or 0.5 mg Cd/L for 8 weeks to investigate effects of water temperature, Cd toxicity and their interaction on fish performance as well as metallothionein (MT) and Cd distribution in different fish organs. It was found that fish reared in Cd-free group at 28 °C showed the optimum growth and feed intake, while Cd-exposed fish showed low growth and feed intake irrespective to water temperature. A synergetic relationship between water temperature and Cd toxicity was observed where Cd toxicity increased as water temperature increased and the worse growth was obtained in Cd-exposed fish reared at 32 °C. Additionally, the highest Cd residues in different fish organs were detected in Cd-exposed fish reared at 32 °C. Similarly, MT concentrations in different fish organs increased as water temperature increased especially in Cd-exposed fish groups. A high positive correlation between MT and Cd concentrations in fish organs was detected. The distribution of MT and Cd levels was in the order of liver>kidney>gills>muscles. The present study revealed that the optimum water temperature suitable for Nile tilapia growth is 28 °C. Additionally, Cd exposure had a deteriorate effect on the growth and health of Nile tilapia. This hazardous effect increased as water temperature increased. Further, liver and kidney were the prime sites of Cd accumulation, while Cd load in the muscles was the lowest as compared to the other investigated organs.     

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.     

Cadmium distribution and its effects on molybdate-containing hydroxylases in Phragmites australis

The influence of cadmium (Cd) on physiological and biochemical parameters was studied to elucidate the mechanism of Cd resistance in Phragmites australis. Cadmium concentrations in roots, stems and leaves increased with exogenous Cd concentration, but Cd content in roots was much higher than in shoots. X-ray microanalysis was used to reveal compartments in which Cd accumulated in root cortex. Cadmium concentrations followed a gradient with the sequence: intercellular space > cell wall > vacuole > cytoplasm, indicating that most Cd was immobilized in the apoplast or sequestered into the vacuolar lumen. Sequential extraction of various Cd chelates revealed that more than half of extractable Cd was bound to proteins, whereas 26% was bound to organic acids. Cd-binding protein fractions were found in the roots after gel filtration chromatography, among which a polypeptide with an apparent molecular mass of 14 kDa bound Cd most avidly. One newly synthesized polypeptide of low molecular mass (1 kDa) appeared under Cd pollution, whereas a prominent fraction of 72 kDa disappeared. Four aldehyde oxidase (AO) isoenzyme activities increased significantly in roots under Cd pollution. Cd stress also enhanced xanthine dehydrogenase (XDH) activities in roots. Two AO polypeptides of different molecular sizes were detected in the roots by Western blot assay. The abundance of the 160 kDa subunit correlated with Cd stress, but the amount of the 90 kDa polypeptide did not change under Cd treatment. Enhanced abscisic acid (ABA) contents were observed in roots of P. australis exposed to Cd. The involvement of Cd distribution in plant tissues and subcellular compartments and of AO and XDH enzymatic activities in the acclimation mechanism of P. australis to Cd pollution is discussed herein.     

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.     

Combined expression of the Arabidopsis metallothionein MT2b and the heavy metal transporting ATPase HMA4 enhances cadmium tolerance and the root to shoot translocation of cadmium and zinc in tobacco

We expressed the AtMt2b and AtHMA4 genes under the 35S cauliflower mosaic virus promoter simultaneously in Nicotiana tabacum (SR1), using leaf disc transformation. A single AtMT2b tobacco T2 line was used for re-transformation with AtHMA4 to obtain the double transformant. Cadmium (Cd) and zinc (Zn) tolerance, uptake and translocation were measured in the double transformant, and compared to untransformed (‘wild type’) tobacco and single gene transformants. The double transformant exhibited enhanced Cd-tolerance, enhanced Cd and Zn root to shoot transport, but unaltered Zn tolerance and Cd and Zn uptake, compared with wild type.The single transformant lines did not show significant phenotypes. Our results suggest that the phenotypes of the double transformant are due to synergistic interaction between the transgenes. Except for Cd tolerance, the phenotypes were moderate for Cd and Zn root to shoot transport, which may be due to use of the 35S promotor, resulting in incorrect tissue-specificity.     

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.     

Comparisons of cadmium tolerance and accumulation at seedling stage in wheat varieties grown in different decades in China

Cadmium (Cd) tolerance and accumulation in wheat varieties were investigated at seedling stage under a controlled environmental condition. The 46 leading wheat varieties cultivated from the 1950s to 2000s in China were treated at the three-leaf stage with a 50 μM CdCl₂ solution for 24 days. Growth and photosynthesis parameters were measured and the Cd-tolerance index (ratio of a given parameter under Cd treatment to that of the control) was determined. Cd accumulation in shoots and roots and Cd translocation were also determined. It was found that Cd tolerance and accumulation of these wheat varieties varied over the different decades. Principal components analysis (PCA) showed that wheat varieties in 1950s and 1980s were tolerant while varieties from the last decade were sensitive to Cd stress. Wheat varieties in 1960s and 1970s were particularly tolerant to Cd stress for the parameters of shoot height, secondary root numbers, net photosynthesis and transpiration rate while the varieties in the 1990s were sensitive to Cd stress for shoot dry weight and root dry weight. Comparing each decade to the average Cd translocation ratio from the roots to the shoots for the whole period, the varieties from the 1950s and 1960s had a higher translocation ratio, while varieties in the 1970s were below that average. Varieties from the 1980s to 2000s showed an average translocation ratio. Using cluster analysis (CA), Shannongfu 63, Yangmai 1 and Yangmai 158 were the most Cd-tolerant varieties in which Cd translocation ratio were low, and Yumai 18 and Huaimai 20 were the most Cd-sensitive varieties in which Cd translocation ratio were high. The results indicating that wheat varieties from different decades were different in Cd tolerance and accumulation, and could be useful for breeding wheat for Cd stress tolerance.     

Arsenate uptake and translocation in seedlings of two genotypes of rice is affected by external phosphate concentrations

Two genotypes of rice (Oryza sativa L.), 94D-54 and 94D-64 were used to investigate the formation of iron plaque controlled by different phosphorus (P) concentrations and the effect of iron plaque on arsenate uptake in a hydroponic experiment. External P concentrations from 10 to 50 μM caused a marked decrease in dithionite-citrate-bicarbonate (DCB)–Fe concentrations for both genotypes, but further increases from 50 to 300 μM only resulted in small decrease. Arsenic (As) concentrations in DCB-extracts were determined by the amounts of iron plaque and the adsorption capacity of As by iron plaque, and both controlled by external P concentrations. At 10 μM external P, genotype 94D-54 had higher Fe, As and P concentrations in DCB-extracts than genotype 94D-64, but the difference disappeared with increasing P concentrations. Increasing P concentrations decreased the percentages of As distributed in iron plaque from around 70 to 10%, and increased the percentages of As in roots and shoots gradually from around 20 to 60% for toots and from 5 to nearly 35% for shoots, respectively. Moreover, P concentration increased the molar ratio of shoot-to-root As, from 0.05 to nearly 0.2, indicating P concentration may promote As translocation from roots to shoots.