Enhanced root-to-shoot translocation of cadmium in the hyperaccumulating ecotype of Sedum alfredii

Sedum alfredii (Crasulaceae) is the only known Cd-hyperaccumulating species that are not in the Brassica family; the mechanism of Cd hyperaccumulation in this plant is, however, little understood. Here, a combination of radioactive techniques, metabolic inhibitors, and fluorescence imaging was used to contrast Cd uptake and translocation between a hyperaccumulating ecotype (HE) and a non-hyperaccumulating ecotype (NHE) of S. alfredii. The K(m) of (109)Cd influx into roots was similar in both ecotypes, while the V(max) was 2-fold higher in the HE. Significant inhibition of Cd uptake by low temperature or metabolic inhibitors was observed in the HE, whereas the effect was less pronounced in the NHE. (109)Cd influx into roots was also significantly decreased by high Ca in both ecotypes. The rate of root-to-shoot translocation of (109)Cd in the HE was >10 times higher when compared with the NHE, and shoots of the HE accumulated dramatically higher (109)Cd concentrations those of the NHE. The addition of the metabolic inhibitor carbonyl cyanide m-chlorophenylhydrazone (CCCP) resulted in a significant reduction in Cd contents in the shoots of the HE, and in the roots of the NHE. Cd was distributed preferentially to the root cylinder of the HE but not the NHE, and there was a 3-5 times higher Cd concentration in xylem sap of the HE in contrast to the NHE. These results illustrate that a greatly enhanced rate of root-to-shoot translocation, possibly as a result of enhanced xylem loading, rather than differences in the rate of root uptake, was the pivotal process expressed in the Cd hyperaccumulator HE S. alfredii.     

Root Morphology and Zn2+ Uptake Kinetics of the Zn Hyperaccumulator of Sedum alfredii Hance

Root morphology and Zn2 uptake kinetics of the hyperaccumulating ecotype (HE) and nonhyperaccumulating ecotype (NHE) of Sedum alfredii Hance were investigated using hydroponic methods and the radiotracer flux technique. The results indicate that root length, root surface area, and root volume of NHE decreased significantly with increasing Zn2 concentration in growth media, whereas the root growth of HE was not adversely affected, and was even promoted, by 500 μmol/L Zn2 . The concentrations of Zn2 in both ecotypes of S. alfredii were positively correlated with root length, root surface area and root volumes, but no such correlation was found for root diameter. The uptake kinetics for 65Zn2 in roots of both ecotypes of S. alfredii were characterized by a rapid linear phase during the first 6 h and a slower linear phase during the subsequent period of investigation. The concentration-dependent uptake kinetics of the two ecotypes of S. alfredii could be characterized by the Michaelis-Menten equation, with the Vmax for 65Zn2 influx being threefold greater in HE compared with NHE, indicating that enhanced absorption into the root was one of the mechanisms involved in Zn hyperaccumulation. A significantly larger Vmax value suggested that there was a higher density of Zn transporters per unit membrane area in HE roots.     

Root adaptations to cadmium-induced oxidative stress contribute to Cd tolerance in the hyperaccumulator Sedum alfredii

Short-term responses of Sedum alfredii roots to Cd exposure was compared in Cd hyperaccumulator (HE) and nonhyperaccumulating ecotype (NHE). Cadmium exposure significantly inhibited root elongation and induced loss of plasma membrane integrity and lipid peroxidation of roots tips in the NHE, whereas these effects were much less pronounced in the HE plants. A strong accumulation of reactive oxygen species with increasing Cd concentration was noted in the NHE root tips, but not in HE. After Cd exposure, a dose-dependent decrease in oxidized glutathione and marked increase in reduced glutathione and non-protein thiols were observed in root tips of HE, but were not seen in the NHE plants. These results suggest that the HE tolerates high Cd in the environment through the differential adaptations against Cd-induced oxidative stress.     

Cd accumulation in roots and shoots of durum wheat: the roles of transpiration rate and apoplastic bypass

Cadmium is readily taken up from soils by plants, depending on soil chemistry, and variably among species and cultivars; altered transpiration and xylem transport and/or translocation in the phloem could cause this variation in Cd accumulation, some degree of which is heritable. Using Triticum turgidum var. durum cvs Kyle and Arcola (high and low grain Cd accumulating, respectively), the objectives of this study were to determine if low-concentration Cd exposure alters transpiration, to relate transpiration to accumulation of Cd in roots and shoots at several life stages, and to evaluate the role of apoplastic bypass in the accumulation of Cd in shoots. The low abundance isotope (106)Cd was used to probe Cd translocation in plants which had been exposed to elemental Cd or were Cd-na?ve; apoplastic bypass was monitored using the fluorescent dye PTS (8-hydroxy-1,3,6-pyrenetrisulphonate). Differential accumulation of Cd by 'Kyle' and 'Arcola' could be partially attributed to the effect of Cd on transpiration, as exposure to low concentrations of Cd increased mass flow and concomitant Cd accumulation in 'Kyle'. Distinct from this, exposure of 'Arcola' to low concentrations of Cd reduced translocation of Cd from roots to shoots relative to root accumulation of Cd. It is possible, but not tested here, that sequestration mechanisms (such as phytochelatin production, as demonstrated by others) are the genetically controlled difference between these two cultivars that results in differential Cd accumulation. These results also suggest that apoplastic bypass was not a major pathway of Cd transport from the root to the shoot in these plants, and that most of the shoot Cd resulted from uptake into the stele of the root via the symplastic pathway.     

Identification of high levels of phytochelatins, glutathione and cadmium in the phloem sap of Brassica napus. A role for thiol-peptides in the long-distance transport of cadmium and the effect of cadmium on iron translocation

Phytochelatins (PCs) are glutathione-derived peptides that function in heavy metal detoxification in plants and certain fungi. Recent research in Arabidopsis has shown that PCs undergo long-distance transport between roots and shoots. However, it remains unknown which tissues or vascular systems, xylem or phloem, mediate PC translocation and whether PC transport contributes to physiologically relevant long-distance transport of cadmium (Cd) between shoots and roots. To address these questions, xylem and phloem sap were obtained from Brassica napus to quantitatively analyze which thiol species are present in response to Cd exposure. High levels of PCs were identified in the phloem sap within 24 h of Cd exposure using combined mass spectrometry and fluorescence HPLC analyses. Unexpectedly, the concentration of Cd was more than four-fold higher in phloem sap compared to xylem sap. Cadmium exposure dramatically decreased iron levels in xylem and phloem sap whereas other essential heavy metals such as zinc and manganese remained unchanged. Data suggest that Cd inhibits vascular loading of iron but not nicotianamine. The high ratios [PCs]/[Cd] and [glutathione]/[Cd] in the phloem sap suggest that PCs and glutathione (GSH) can function as long-distance carriers of Cd. In contrast, only traces of PCs were detected in xylem sap. Our results suggest that, in addition to directional xylem Cd transport, the phloem is a major vascular system for long-distance source to sink transport of Cd as PC–Cd and glutathione–Cd complexes.     

Root Morphology and Zn^2＋ Uptake Kinetics of the Zn Hyperaccumulator of Sedum alfredii Hance

Root morphology and Zn^2＋ uptake kinetics of the hyperaccumulating ecotype （HE） and nonhyperaccumulating ecotype （NHE） of Sedum alfredii Hance were investigated using hydroponic methods and the radiotracer flux technique. The results indicate that root length, root surface area, and root volume of NHE decreased significantly with increasing Zn^2＋ concentration in growth media, whereas the root growth of HE was not adversely affected, and was even promoted, by 500μmol/L Zn^2＋. The concentrations of Zn^2＋ in both ecotypes of S. alfredii were positively correlated with root length, root surface area and root volumes, but no such correlation was found for root diameter. The uptake kinetics for ^65Zn^2＋ in roots of both ecotypes of S. alfredii were characterized by a rapid linear phase during the first 6 h and a slower linear phase during the subsequent period of investigation. The concentration-dependent uptake kinetics of the two ecotypes of S. alfredii could be characterized by the Michaelis-Menten equation, with the Vmax for ^65Zn^2＋ influx being threefold greater in HE compared with NHE, indicating that enhanced absorption into the root was one of the mechanisms involved in Zn hyperaccumulation. A significantly larger Vmax value suggested that there was a higher density of Zn transporters per unit membrane area in HE roots.     

Cellular sequestration of cadmium in the hyperaccumulator plant species Sedum alfredii

Spatial imaging of cadmium (Cd) in the hyperaccumulator Sedum alfredii was investigated in vivo by laser ablation inductively coupled plasma mass spectrometry and x-ray microfluorescence imaging. Preferential Cd accumulation in the pith and cortex was observed in stems of the Cd hyperaccumulating ecotype (HE), whereas Cd was restricted to the vascular bundles in its contrasting nonhyperaccumulating ecotype. Cd concentrations of up to 15,000 μg g(-1) were measured in the pith cells, which was many fold higher than the concentrations in the stem epidermis and vascular bundles in the HE plants. In the leaves of the HE, Cd was mainly localized to the mesophyll and vascular cells rather than the epidermis. The distribution pattern of Cd in both stems and leaves of the HE was very similar to calcium but not zinc, irrespective of Cd exposure levels. Extended x-ray absorption fine structure spectroscopy analysis showed that Cd in the stems and leaves of the HE was mainly associated with oxygen ligands, and a larger proportion (about 70% in leaves and 47% in stems) of Cd was bound with malic acid, which was the major organic acid in the shoots of the plants. These results indicate that a majority of Cd in HE accumulates in the parenchyma cells, especially in stems, and is likely associated with calcium pathways and bound with organic acid (malate), which is indicative of a critical role of vacuolar sequestration of Cd in the HE S. alfredii.     

Zinc compartmentation in root, transport into xylem, and absorption into leaf cells in the hyperaccumulating species of Sedum alfredii Hance

Sedum alfredii Hance can accumulate Zn in shoots over 2%. Leaf and stem Zn concentrations of the hyperaccumulating ecotype (HE) were 24- and 28-fold higher, respectively, than those of the nonhyperaccumulating ecotype (NHE), whereas 1.4-fold more Zn was accumulated in the roots of the NHE. Approximately 2.7-fold more Zn was stored in the root vacuoles of the NHE, and thus became unavailable for loading into the xylem and subsequent translocation to shoot. Long-term efflux of absorbed ⁶⁵Zn indicated that ⁶⁵Zn activity was 6.8-fold higher in shoots but 3.7-fold lower in roots of the HE. At lower Zn levels (10 and 100 μM), there were no significant differences in ⁶⁵Zn uptake by leaf sections and intact leaf protoplasts between the two ecotypes except that 1.5-fold more ⁶⁵Zn was accumulated in leaf sections of the HE than in those of the NHE after exposure to 100 μM for 48 h. At 1,000 μM Zn, however, approximately 2.1-fold more Zn was taken up by the HE leaf sections and 1.5-fold more ⁶⁵Zn taken up by the HE protoplasts as compared to the NHE at exposure times >16 h and >10 min, respectively. Treatments with carbonyl cyanide m-chlorophenylhydrazone (CCCP) or ruptured protoplasts strongly inhibited ⁶⁵Zn uptake into leaf protoplasts for both ecotypes. Citric acid and Val concentrations in leaves and stems significantly increased for the HE, but decreased or had minimal changes for the NHE in response to raised Zn levels. These results indicate that altered Zn transport across tonoplast in the root and stimulated Zn uptake in the leaf cells are the major mechanisms involved in the strong Zn hyperaccumulation observed in S. alfredii H.     

不同生态型东南景天对土壤中Cd的生长反应及吸收积累的差异性

 采用盆栽方法研究了两种生态型东南景天(Sedum alfredii)对土壤中不同含量Cd(即对照, 12.5, 25, 50, 100, 200, 300, 400 mg&;#8226;kg^－1)的生 长反应、吸收和积累Cd的差异性。结果表明,土壤添加重金属Cd后,矿山生态型东南景天生长正常,地上部和根系Cd含量随着土壤中Cd含量的 增加而增加,在400 mg&;#8226;kg^－1 Cd处理下含量分别高达2 900和500 mg&;#8226;kg^－1,其地上部显著大于根部;然而,土壤添加Cd后,非矿山生态型东 南景天的生长受到抑制,地上部和根部的生物量显著降低。当土壤Cd含量为50～100 mg&;#8226;kg^－1 时,非矿山生态型东南景天的地上部和根系Cd含 量随着土壤中Cd含量的增加而增加,而且根系Cd含量则大于地上部。当土壤Cd≤50 mg&;#8226;kg^－1时,矿山生态型东南景天根系Cd含量比非矿山生态 型高 ,但当土壤Cd≥100 mg&;#8226;kg^－1,两者之间无显著差异;然而,但在同一Cd处理水平下,矿山生态型东南景天地上部Cd含量总是高于非矿山 生态型。这些结果表明,矿山生态型东南景天有很强的忍耐和吸收土壤Cd的能力,再次证明其为一种Cd超积累植物。     

Xylem loading process is a critical factor in determining Cd accumulation in the shoots of Solanum melongena and Solanum torvum

Cadmium (Cd) concentration in eggplant (Solanum melongena) fruits can be drastically reduced by grafting them with Solanum torvum rootstock. We thus examined the characteristics of Cd absorption in roots and Cd translocation from roots to shoots between S. melongena and S. torvum over 7 days using a hydroponic culture. Although there is no significant difference in Cd concentration in the roots of S. melongena and S. torvum, Cd concentration in the shoots and xylem sap was higher in S. melongena than in S. torvum. By evaluating symplastic Cd absorption in roots, using enriched isotopes ¹¹³Cd and ¹¹⁴Cd, and measuring the kinetics in xylem loading, we characterized Cd absorption and translocation for S. torvum (low Cd translocation) and S. melongena (high Cd translocation). A concentration-dependent study in roots indicated that K_m values were almost the same for species, but the V_max value was 1.5-fold higher in S. melongena than in S. torvum. A concentration-dependent study in xylem loading indicated that V_max was almost the same, but K_m values were approximately 7-fold higher in S. torvum compared to S. melongena. These results, together, suggest that the affinity for Cd in the xylem loading process is a critical factor for determining the different Cd concentrations in the shoots between both plants under low Cd concentration conditions. In addition, a metabolic inhibitor, carbonyl cyanide-m-chloro-phenyl-hydrazone (CCCP) inhibited Cd absorption and translocation from roots to shoots in both plants. This suggests that Cd absorption in roots and Cd translocation from roots to shoots via the xylem loading process, under low Cd concentration conditions, are partly mediated by an active energy-dependent process in both plants.     

Mechanisms of Cadmium Mobility and Accumulation in Indian Mustard

Indian mustard (Brassica juncea L.), a high biomass crop plant, accumulated substantial amounts of cadmium, with bioaccumulation coefficients (concentration of Cd in dry plant tissue/concentration in solution) of up to 1100 in shoots and 6700 in roots at nonphytotoxic concentrations of Cd (0.1 [mu]g/mL) in solution. This was associated with a rapid accumulation of phytochelatins in the root, where the majority of the Cd was coordinated with sulfur ligands, probably as a Cd-S4 complex, as demonstrated by x-ray absorption spectroscopy. In contrast, Cd moving in the xylem sap was coordinated predominantly with oxygen or nitrogen ligands. Cd concentrations in the xylem sap and the rate of Cd accumulation in the leaves displayed similar saturation kinetics, suggesting that the process of Cd transport from solution through the root and into the xylem is mediated by a saturable transport system(s). However, Cd translocation to the shoot appeared to be driven by transpiration, since ABA dramatically reduced Cd accumulation in leaves. Within leaves, Cd was preferentially accumulated in trichomes on the leaf surface, and this may be a possible detoxification mechanism.     

Transpiration inhibition by stored xylem sap from well-watered maize plants

There is increasing evidence that a chemical signal exists in xylem sap of plants subjected to water deficits which influences physiological responses in plant shoots. An important method of studying this signal is the transpiration response of excised leaves exposed to xylem sap collected from plants. However, Munns et al [Plant, Cell & Environment 16, 867–877] cautioned that transpiration inhibition is observed when xylem sap collected from wheat and barley is stored before determining physiological activity. The objective of the study reported here was to determine if transpiration inhibition develops in maize sap collected from well-watered plants when the sap is stored under various conditions. It was found that storage of maize sap collected from well-watered plants for only 1 d at -20°C resulted in the development of substantial transpiration inhibition in bioassay leaves. Storage of sap at 4°C resulted in the development of the effect after 2 weeks, while storage at ?86°C showed only small transpiration inhibition after 3 weeks. The major source of the transpiration inhibition was the development of a substance in the stored sap that resulted in physical blockage of the transpiration stream in bioassay leaves. However, a small signal component may also have developed in the stored sap. Because of the possibility of ionic activity under freezing conditions at ?20°C, calcium was studied for its potential involvement in the transpiration inhibition. However, the calcium concentrations found to inhibit transpiration were nearly an order of magnitude larger than the calcium concentrations observed in xylem sap.     

Rice is more efficient in arsenite uptake and translocation than wheat and barley

Rice is efficient at arsenic (As) accumulation, thus posing a potential health risk to humans and animals. Arsenic bioavailability in submerged paddy soil is enhanced due to mobilisation of arsenite, but rice may also have an inherently greater ability to take up and translocate arsenite than other cereal crops. To test this hypothesis, rice, wheat and barley were exposed to 5 µM arsenate or arsenite for 24 h. Arsenic uptake and distribution, and As speciation in the xylem sap and nutrient solution were determined. Regardless of the As form supplied to plants, rice accumulated more As in the shoots than wheat or barley. Arsenite uptake by rice was double of that by wheat or barley, whereas arsenate uptake was similar between rice and wheat and approximately a third smaller in barley. The efficiency of As translocation from roots to shoots was greater when plants were supplied with arsenite than with arsenate, and in both treatments rice showed the highest translocation efficiency. Arsenite was the main species of As (86–97%) in the xylem sap from arsenite-treated plants of all three species. In the arsenate-treated plants, 84%, 45% and 63% of As in the xylem sap of rice, wheat and barley, respectively, was arsenite. Arsenite efflux to the external medium was also observed in all three plant species exposed to arsenate. The results show that rice is more efficient than wheat or barley in arsenite uptake and translocation, probably through the highly efficient pathway for silicon.     

Genotypic variation in safflower (Carthamus spp.) cadmium accumulation and tolerance affected by temperature and cadmium levels

Soil pollution is a world-wide problem, with heavy metals being a major part of the concern. To investigate the effect of temperature on cadmium (Cd) uptake and translocation, as well as Cd tolerance in wild and cultivated species of safflower, a hydroponic experiment was conducted under controlled conditions. The responses of four wild genotypes (Isfahan, Arak, Azari, and Shiraz) and four cultivated genotypes (AC-Sterling, 2811, Saffire, and C111) of safflower to nine levels of CdCl₂ (0, 0.5, 1, 5, 10, 20, 50, 100, and 500 μM) in solution were examined under two temperatures (18 and 23 °C). Cadmium sensitivity was determined using the Weibull model on the total dry weight of the plants. Cadmium uptake and translocation were analyzed on 1 μM Cd treated plants. Results revealed that safflower genotypes differed in terms of uptake, translocation, and tolerance to Cd, with AC-Sterling and Arak indicating the most and the least tolerance to Cd, respectively. Relative Cd uptake and Cd concentration in roots and shoots increased with an increase in temperature in all genotypes, with the exception of AC-Sterling. Net accumulation of Cd via root increased with an increase in temperature for the wild Azari and the cultivated 2811, Saffire, and C111, though it decreased for the rest of genotypes. Cadmium translocation to shoots significantly increased with increased temperature in all genotypes. Cadmium translocation from roots to shoots in cultivated genotypes was significantly greater than in wild genotypes. Root Cd concentration in wild genotypes was significantly greater than in cultivated genotypes. It seems that wild and cultivated species of safflower differ in their response to Cd. Furthermore, temperature may affect the plant's tolerance to Cd, probably through accompanying changes in Cd uptake and translocation from root to shoot.     

Do increases in xylem sap pH and/or ABA concentration mediate stomatal closure following nitrate deprivation?

Stomatal conductance (g(s)) of pepper (Capsicum annuum L.) plants decreased during the second photoperiod (day 2) after withholding nitrate (N). Stomatal closure of N-deprived plants was not associated with a decreased shoot water potential (Psi(shoot)); conversely Psi(shoot) was lower in N-supplied plants. N deprivation transiently (days 2 and 3) alkalized (0.2-0.3 pH units) xylem sap exuded from de-topped root systems under root pressure, and xylem sap expressed from excised shoots by pressurization. The ABA concentration of expressed sap increased 3-4-fold when measured on days 2 and 4. On day 2, leaves detached from N-deprived and N-supplied plants showed decreased transpiration rates when fed an alkaline (pH 7) artificial xylem (AX) solution, independent of the ABA concentration (10-100 nM) supplied. Thus changes in xylem sap composition following N deprivation can potentially close stomata. However, the lower transpiration rate of detached N-deprived leaves relative to N-supplied leaves shows that factors residing within N-deprived leaves also mediate stomatal closure, and that these factors assume greater importance as the duration of N deprivation increases.     

Phosphorus transport to the xylem and its regulation by water flow

Summary The effects of water flow on phosphorus uptake by roots and on its subsequent translocation to shoots were separated by giving short-term pulses of ³²P-labelled nutrient to intact tomato plants. At the end of a 5 min pulse, all the ³²P taken up by the plants was confined to the roots. Only about half of this ³²P was later translocated to shoots; there was very little translocation after 4 hours.Experiments after long-term labelling showed that only a small part of the total P in the root is readily translocated to shoots. This P appears to be in part of the symplast and contributes about 75% of the P transported to the xylem sap. The rest is presumably derived by leakage from vacuoles.A slow rate of water flow reduced both uptake into the symplast and the translocation to the shoots of P which had already been absorbed by the roots. This was conclusively demonstrated by giving a ³²P pulse before reducing the rate of water flow; ³²P not translocated to shoots was partly retained by the roots and partly lost to the external solution. Water flow also accelerates transport to the xylem of previously-absorbed P in excised roots.It is concluded that the major effect of water flow on phosphorus transport to shoots occurs after phosphorus uptake by the roots, probably during radial transport to the xylem.     

Export of Abscisic Acid, 1-Aminocyclopropane-1-Carboxylic Acid,Phosphate, and Nitrate from Roots to Shoots of Flooded Tomato Plants (Accounting for Effects of Xylem Sap Flow Rate on Concentration and Delivery)

We determined whether root stress alters the output of physiologically active messages passing from roots to shoots in the transpiration stream. Concentrations were not good measures of output. This was because changes in volume flow of xylem sap caused either by sampling procedures or by effects of root stress on rates of whole-plant transpiration modified concentrations simply by dilution. Thus, delivery rate (concentration x sap flow rate) was preferred to concentration as a measure of solute output from roots. To demonstrate these points, 1-aminocyclopropane-1-carboxylic acid (ACC), abscisic acid, phosphate, nitrate, and pH were measured in xylem sap of flooded and well-drained tomato (Lycopersicon esculentum Mill., cv Ailsa Craig) plants expressed at various rates from pressurized detopped roots. Concentrations decreased as sap flow rates were increased. However, dilution of solutes was often less than proportional to flow, especially in flooded plants. Thus, sap flowing through detopped roots at whole-plant transpiration rates was used to estimate solute delivery rates in intact plants. On this basis, delivery of ACC from roots to shoots was 3.1-fold greater in plants flooded for 24 h than in well-drained plants, and delivery of phosphate was 2.3-fold greater. Delivery rates of abscisic acid and nitrate in flooded plants were only 11 and 7%, respectively, of those in well-drained plants.