Enhancement of cadmium effects on growth and nutrient composition of birch (Betula pendula) by buthionine sulphoximine (BSO)

Binding of Cd to non-specific metal-binding peptides (phytochelatins)in birch roots has been suggested as an explanation for toleranceto Cd toxicity in birch (Betula pendula). In the present study,the tolerance of birch roots to Cd was further investigatedby using buthionine sulphoximine (BSO) as an inhibitor of phytochelatinsynthesis. Birch seedlings, grown in nutrient solution at pH4.2, were exposed to 0 or 2 µM CdCl₂ combined with 0 or0.1 mM BSO for 6 d. Plant growth (fresh weight increase andshoot to root dry weight ratio) and the nutrient compositionin fine roots, whole roots and shoots were determined. The effectsof Cd on growth confirms the results of earlier studies on birch,suggesting a reduced shoot growth, but preserved or stimulatedroot growth. When Cd and BSO were combined, overall plant growthwas severely reduced. BSO was also shown to aggravate Cd-inducedreductions of root and shoot concentrations of K, Ca and Mgbut to impede the accumulation of Cd. The results suggest that phytochelatins participate in protectingthe root against Cd interferences with growth, possibly by restrictingCd-induced changes in the nutrient composition of the plant. Key words: Betula pendula, buthionine sulphoximine, cadmium, phytochelatins, roots, tolerance     

Regulation of phosphate influx in barley roots: Effects of phosphate deprivation and reduction of influx with provision of orthophosphate

Barley plants were grown in nutrient solutions, which were maintained at either 0 (-P) or 15 μ M orthophosphate (+P). After 11 days phosphate influx into the intact roots of the -P plants began to increase by comparison with +P plants. During this period differences became apparent between the treatments in absolute growth rates, as well as in the root:shoot ratios. Phosphate influx in the -P plants continued to increase as a function of time, to a maximum value of 2.4 μmol (g fresh wt)^-1h^-1 at 16 days after germination. This rate was 6 times higher than influx values for +P plants of the same age. During the period of enhanced uptake phosphate was strongly correlated (r²= 0.77) with root organic phosphate concentration. – The enhancement of inorganic phosphate influx into intact roots of -P plants was rapidly reduced by the provision of 15 μ M orthophosphate. Typically, within 4 h of exposure to this concentration of phosphate, influx values fell from 1.80 ± 0.20 to 0.75 ± 0.03 μmol (g fresh wt)^-1 h^-1, while inorganic phosphate concentrations of the roots increased from 0.12 to 1.15 μmol (g fresh wt)^-1 during the same period. Hill plots of the influx data obtained during this period, treating root inorganic phosphate as an inhibitor of influx, gave Hill coefficients close to 2. The rapidity of the reduction of influx associated with increased root inorganic phosphate together with the Hill plot data provide evidence for an allosteric inhibition of influx by internal inorganic phosphate.     

Effect of rhizosphere and plant-related factors on the cadmium uptake by maize (Zea mays L.)

The characteristics of 12- and 24-day-old maize plants (Zea mays L. cv MB862) related to Cd absorption were investigated with respect to the influence of Cd concentration in the plant organs, to plant age, absorption time and competition with micronutrients. Despite high Cd concentrations in the nutrient solution, hydroponically cultivated maize did not seem to be affected by Cd toxicity, except for the highest Cd level (100 μmol L⁻¹). There was on average five times more Cd in roots than in shoots and the Cd root to shoot ratio increased with increasing Cd concentration in the nutrient solution. No significant differences were observed between influx measured for 2 h in the middle of the day light period and for a full day period. Plants with different internal Cd concentrations showed similar root absorption characteristics of this metal. The root Cd influxes were three times higher in solutions with low micronutrient contents than in the solutions with higher micronutrient contents, and almost three times higher in 12-day-old roots than in 24-day-old roots. The root Cd influx was linearly related to its concentration in the solution, showing no saturable component. Our results suggest a non-specific and unregulated transport of Cd into the maize root symplast. They also indicate a regulation of the Cd translocation from root to shoot, as well as dependence of parameters of Cd root absorption on plant and rhizosphere conditions which should be taken into account for Cd uptake modelling.     

Influx and efflux of K(+) in sunflower roots after transfer between solutions with different K(+) concentrations

It was investigated whether K(+) efflux, like K(+) influx, is affected when roots are transferred between solutions with different K(+) concentrations. Sunflower plants (Hehanthus annuus L. cv. Uniflorus) were grown on complete nutrient solutions with 0.1, 1.0, 10 or 25 mM K(+) . This produced plants with K(+) concentrations in the roots varying between 9 and 110 μmol (g fresh weight)(-1) . At the beginning of the experiments the plants were transferred to an (86) Rb-labelled experimental solution initially containing 0.1 mM K(+) . At intervals during 6.5 h samples were removed from the solution and analyzed for K(+) and radioactivity. Based on the analyses K(+) ((86) Rb) influx, K(+) net uptake and K(+) efflux could be computed. In'low K(+) 'roots, K(+) ((86) Rb) influx and K(+) net uptake agreed, suggesting a very low K(+) efflux. This was contrary to'high K(+) 'roots, where K(+) efflux was initially higher than K(+) ((86) Rb) influx. After about 4 h, K(+) efflux declined to a low value also in these roots. When 2-4-dinitrophenol was included in the experimental solution, K(+) ((86) Rb) influx was generally depressed, whereas K(+) efflux was high throughout the experiment and directly proportional to the K(+) status of the roots. Our hypothesis is that after transfer of'high K(+) 'roots to a solution with low K(+) concentration, the K(+) efflux from the vacuoles of root cells transiently increases, until a new electrochemical equilibrium is attained.     

Root morphology and cadmium uptake kinetics of the cadmium-sensitive rice mutant

To understand the physiological mechanism that confers Cd sensitivity, root morphology and Cd uptake kinetics of the Cd-sensitive mutant and wild type rice were investigated. The root length, root surface area, and root number of mutant rice decreased more significantly with increasing Cd concentration in growth media compared with the wild type rice. The uptake kinetics for ¹⁰⁹Cd²⁺ in roots of both the mutant and wild type rice were characterized by a rapid linear phase during the first 6 h and a slower linear phase during the subsequent period. Concentration-dependent Cd²⁺ influx in both species could be characterized by the Michaelis-Menten equation, with similar apparent K_m values for mutant and wild type rice (2.54 and 2.37 μM, respectively). However, the V_max for Cd²⁺ influx in mutant root cells was nearly 2-fold higher than that for wild type rice, indicating that enhanced absorption into the root is one of the mechanisms involved in Cd sensitivity in mutant rice.     

Longitudinal variation in cadmium influx in intact first order lateral roots of sunflower (Helianthus annuus. L)

Aims

Contamination of sunflower (Helianthus annuus L.) by cadmium (Cd) is a concern for food and feed safety as this species accumulates Cd to a greater extent than other crops. We examined the relationships between root architecture and Cd²⁺ uptake by roots.

Methods

We determined and mathematically modelled the longitudinal variation of Cd²⁺ influx in first order roots of sunflower grown in hydroponics by using short-term exposure to ¹⁰⁹Cd-labelled solutions (0.8 to 500 nM). Thereafter, by taking into account the longitudinal variation of the influx, we simulated the uptake of Cd²⁺ for 24 h by cohorts of roots characterised by various architectural characteristics.

Results

Cd²⁺ influx at the root tip was on average 2.9 times that of the basal region close to the taproot. The simulations indicated that the total Cd²⁺ uptake by root cohorts mainly depends on 1/ the root diameter and the number of roots, 2/ the value of the Cd²⁺ influx at the basal region 3/ the stronger influx at the root tip.

Conclusion

Considering a higher Cd²⁺ influx at the root tip may be important to understand the relationship between root architecture and Cd²⁺ uptake by the root system.     

重金属镉对水稻根毛细胞钾离子吸收过程的影响

重金属镉进入植物体后会引起一系列的毒害反应,然而迄今为止有关镉毒害的机制并不明了。本研究综合使用了膜电势测定、非损伤微测和膜片钳等电生理学实验技术,检测了镉对水稻根毛细胞钾离子吸收过程的影响。研究结果显示,外源施用50μmol·L-1 CdCl2能明显造成水稻根毛细胞膜的去极化,抑制根毛细胞质膜内向K＋通道活性,同时诱导外向K＋通道开放,导致根系内K＋外渗,进而降低了水稻根部K＋含量。通过上述影响,镉扰乱了水稻根系对钾离子的吸收过程,造成钾元素缺失,成为植物体镉毒害作用的机理之一。     

Potassium and phosphate uptake of cucumber plants at different ammonia supply

Summary Experiments on cucumber plants grown in nutrient solution were conducted in order to study long and short time effects of ammonia on growth, nutrient element uptake and respiration of roots.Shoot yield and potassium concentration in tissue of plants treated 18 days with varied ammonia concentration were decreased. However, it was not assumed that K deficiency caused the yield reduction. The ammonia effect on K content was more pronounced in roots than in shoots.The decreased K concentration of plant tissue was linked to a diminished ability of plant roots to absorb potassium. The maximum rate of potassium uptake was lowered by ammonia during both, long- and short-time treatment. The results indicated that the NH₃ influence on potassium uptake was due to effects on metabolism and permeability of roots because changes of K uptake rate occurred immediately after starting the NH₃ treatment. Furthermore, it is shown that ammonia inhibited respiration of roots.During the short-time treatment net potassium efflux of roots was observed at higher NH₃ concentrations. The extent of K efflux depended on K concentration of both, root tissue and nutrient solution.Pretreating the plants for 12 hours with ammonia also resulted a decline in K uptake rate. However, plant roots subjected to ammonia concentrations up to 0.09 mM completely recovered during 24 hours after removing the NH₃ treatment whereas at higher NH₃ concentrations only a partial recovery occurred.Furthermore, it was shown that ammonia also influenced P uptake by plant roots.     

Cadmium uptake kinetics and plants factors of shoot Cd concentration

Background and aims

Accumulation of Cd in the shoots of plants grown on Cd contaminated soils shows considerable variation. A previous preliminary experiment established that one major reason for this variation was the rate of Cd influx into the roots (mol Cd cm^?2 root s^?1). However, this experiment did not distinguish between solubilization of soil Cd on the one hand and difference in Cd uptake kinetics on the other. The main objectives of the present study were thus to characterize Cd uptake kinetics of plants continuously exposed to Cd concentrations similar to those encountered in soils. Furthermore we determined the factors responsible for differences in shoot Cd concentration such as net Cd influx, root area-shoot dry weight ratio, shoot growth rate and proportion of Cd translocated to the shoot.

Materials and methods

Maize, sunflower, flax and spinach were grown in nutrient solution with five constant Cd concentrations varying from 0 to 1.0 μmol?L^?1. Root and shoot parameters as well as Cd uptake were determined at two harvest dates and from these data Cd net influx and shoot growth rates were calculated.

Results and conclusions

Cadmium uptake kinetics, i.e. the net Cd influx vs. Cd solution concentration followed a straight line. Its slope is the root absorbing power, α, $ \left( {\alpha ={{{\mathrm{Cd}\;\mathrm{net}\;\mathrm{influx}}} \left/ {{\mathrm{Cd}\;\mathrm{solution}\;\mathrm{concentration}}} \right.}} \right) $ . The α values of spinach and flax were about double that of maize and sunflower (5?×?10^?6?cm?s^?1 vs. 2.5?×?10^?6?cm?s^?1). Spinach and flax had a 3–5 times higher shoot Cd concentration than maize and sunflower. The difference in shoot Cd concentration was partly due to the higher Cd influx but also to a higher translocation of Cd from root to shoot and also to a slower shoot growth rate.     

Uptake and long-distance transport of phosphate,potassium and chloride in relation to internal ion concentrations in barley: evidence of non-allosteric regulation

The extent to which uptake and transport of either phosphate, potassium or chloride are controlled by the concentration of these ions within the root, perhaps through an allosteric mechanism, was investigated with young barley plants in nutrient solution culture. Plants were grown with their roots divided between two containers, such that a single seminal root was continuously supplied with all the required nutrient ions, while the remaining four or five seminal roots were either supplied with the same solution (controls) or, temporarily, a solution lacking a particular nutrient ion (nutrient-deficient treatment). Compared with controls, there was a marked stimulation of uptake and transport of labelled ions by the single root following 24 h or more of nutrient dificiency to the remainder of the root system. This stimulation, which comprised an increased transport to the shoot and, for all ions except Cl^-, increased transport to the remainder of the root system, took place without appreciable change in the concentration of particular ions within the single root. However, nutrient deficiency quickly caused a lower concentration of ions in the shoot and the remaining roots. The results are discussed in relation to various mechanisms, proposed in the literature, by which the coordination of ion uptake and transport may be maintained within the plant. We suggest that under our conditions any putative allosteric control of uptake and transport by root cortical cells was masked by an alternative mechanism, in which ion influx appears to be regulated by ion efflux to the xylem, perhaps controlled by the concentration of particular ions recycled in the phloem to the root from the shoot.     

Effects of aluminium on growth and kinetics of K+(86Rb) uptake in two cultivars of wheat (Triticum aestivum) with different sensitivity to aluminium

Two cultivars of wheat (Triticum aestivum L. cvs Kadett and WW 20299) were grown for 9 days with 20% relative increase in nutrient supply per day at pH 4.1. Aluminium at 50 μ M retarded the growth of roots more than that of shoots in both cultivars, thus decreasing the root/shoot ratio. The inhibition was largest in WW 20299. With long term Al treatment (9 days), K_m for K⁺(⁸⁶Rb) influx increased five times in both cultivars and V_max decreased in WW 20299. Efflux of K⁺(⁸⁶Rb) was little affected. When the roots were treated with aluminium for two days, only relative growth rate of roots was retarded, while growth of shoots was unaffected and influx of K⁺(⁸⁶Rb) adjusted to the actual K⁺ demand of the plants. It is concluded that the effects of aluminium on K⁺ uptake in these wheat cultivars are not primary factors contributing to aluminium sensitivity. However, in soil with Al the demand for a comparatively high concentration of K⁺ to maintain an adequate K⁺ uptake rate, in combination with a slow growth rate of the roots, may secondarily lead to K⁺ deficiency in the plants.     

Effect of potassium status on K+ (Rb)+ uptake and transport in sunflower roots

Young sunflower plants ( Helianthus annuus L. cv. Halcón), grown in nutrient solution at two K⁺ levels (0.25 and 2.5 m M ) were used to study the effect of K⁺ content in the root on uptake and transport of K⁺ to the exuding stream of decapitated plants. Roots of plants grown in low K⁺ gave higher exudation flux, higher K⁺ concentration in exudate and higher K⁺ flux than high K⁺ roots. After 6 h of uptake the K⁺ flux in low K⁺ roots was about three times that in high K⁺ roots. When the roots were kept in a nutrient solution in which Rb⁺ replaced K⁺, low K⁺ roots exuded much more Rb⁺ than K⁺ after the first 2 h, whereas high K⁺ roots exuded about similar amounts of K⁺ and Rb⁺. In intact plants grown at three different K⁺ levels (0.1, 1.0 and 10.0 m M ), there was an inverse relationship between the K⁺ level in the nutrient solution and the Rb⁺ accumulated in the roots or transported to the shoot. The results suggest that the transport of ions from xylem parenchyma to stele apoplast may be controlled by ions coming down from the shoot in sieve tubes.     

Hyperaccumulation of cadmium by roots, bulbs and shoots of garlic (Allium sativum L.)

The effects of cadmium chloride concentration on root, bulb and shoot growth of garlic (Allium sativum L.), and the uptake and accumulation of Cd2+ by garlic roots, bulbs and shoots were investigated. The range of cadmium chloride (CdCl2 x 2.5H2O) concentrations was 10(-6) - 10(-2) M. Cadmium stimulated root length at lower concentrations (10(-6) - 10(-5) M) significantly (P < 0.005) during the entire treatment period. The seedlings exposed to 10(-3) - 10(-2) M Cd exhibited substantial growth reduction (P < 0.005), but did not develop chlorosis. Garlic has considerable ability to remove Cd from solutions and accumulate it. The Cd content in roots of garlic increased with increasing solution concentration of Cd2+. The roots in plants exposed to 10(-2) M Cd accumulated a large amount of Cd. approximately 1,826 times the control. The Cd contents in roots of plants treated with 10(-3), 10(-4), 10(-5) and 10(-6) M Cd were approximately 114, 59, 24 and 4 times the control, respectively. However, the plants transported only a small amount of Cd to their bulbs and shoots and concentrations in these tissues were low.     

Influence of aluminium on phosphorus and calcium localization in roots of beech (Fagus sylvatica)

Beech plants ( Fagus sylvatica L. provenance Maramures) were grown in nutrient solution at low pH (4.2) and exposed to different concentrations of AlCl₃. Uptake and leakage of Ca²⁺(⁴⁵Ca²⁺) and H²PO₄-(³²P) were studied. A high external aluminium concentration (1.0m M ) reduced the uptake and export to the shoot of both calcium and phosphate, while 0.1 m M Al increased the phosphorus level in the roots. To determine the impact of aluminium on the localization of calcium and phosphate, leakage of the elements from both intact plants and plants frozen prior to the leakage experiment was studied. The leakage of Ca²⁺ from intact plants was not affected by prior exposure to 0.1 m M Al. Freezing of the beech plants before the leakage experiment increased leakage of calcium slightly more from roots of control plants than for roots exposed to 0.1 m M Al, indicating that even low concentrations of alminium may impede the influx of calcium across the plasma membrane in the roots. The patterns of Ca²⁺ leakage from roots previously exposed to 1.0 m M Al indicated that very little Ca²⁺ was located extracellularly. The extracellular fraction of phosphate increased with increasing Al concentration in the nutrient solution. Low Al concentration (0.1 m M ) only reduced the intracellular phosphate concentration to a minor extent, while 1.0 m M Al profoundly decreased it. It is concluded that 0.1 m M AlCl₃ has a limited effect upon the localization of Ca²⁺ and phosphate in the roots. At higher levels of Al, 0.1–1.0 m M , there is a more dramatic change in nutrient localization in the free space and uptake over the plasma membrane.     

Inward-Rectifying K+ Channels in Root Hairs of Wheat (A Mechanism for Aluminum-Sensitive Low-Affinity K+ Uptake and Membrane Potential Control)

K+ is the most abundant cation in cells of higher plants, and it plays vital roles in plant growth and development. Extensive studies on the kinetics of K+ uptake in roots have shown that K+ uptake is mediated by at least two transport mechanisms, one with a high and one with a low affinity for K+. However, the precise molecular mechanisms of K+ uptake from soils into root epidermal cells remain unknown. In the present study we have pursued the biophysical identification and characterization of mechanisms of K+ uptake into single root hairs of wheat (Triticum aestivum L.), since root hairs constitute an important site of nutrient uptake from the soil. These patch-clamp studies showed activation of a large inward current carried by K+ ions into root hairs at membrane potentials more negative than -75 mV. This K+ influx current was mediated by hyperpolarization-activated K+-selective ion channels, with a selectivity sequence for monovalent cations of K+ > Rb+ [almost equal to] NH4+ >> Na+ [almost equal to] Li+ > Cs+. Kinetic analysis of K+ channel currents yielded an apparent K+ equilibrium dissociation constant (Km) of [almost equal to]8.8 mM, which closely correlates to the major component of low-affinity K+ uptake. These channels did not inactivate during prolonged stimulation and would thus enable long-term K+ uptake driven by the plasma membrane proton-extruding pump. Aluminum, which is known to inhibit cation uptake at the root epidermis, blocked these inward-rectifying K+ channels with half-maximal current inhibition at [almost equal to]8 [mu]M free Al3+. Aluminum block of K+ channels at these Al3+ concentrations correlates closely to Al3+ phytotoxicity. It is concluded that inward-rectifying K+ channels in root hairs can function as both a physiologically important mechanism for low-affinity K+ uptake and as regulators of membrane potential. The identification of this mechanism is a major step toward a detailed molecular characterization of the multiple components involved in K+ uptake, transport, and membrane potential control in root epidermal cells.     

Effects of Low Root Temperature on Ion Uptake and Ion Translocation in Wheat

Roots of wheat seedlings (Triticum aestivum L. cv. Weibulls Starke) were cooled (+1°C) for 24 h while the shoots were kept at 25°C. The treatment induced an increased water deficit in the leaves. Fresh weight, dry weight, and the uptake and distribution of potassium and calcium were measured before and after cooling. Growth, measured both as fresh weight and dry weight increase, was reduced during the cold treatment. Afterwards (at 20°C), growth recovered to nearly pre-stress rates. Analysis of the potassium fluxes in and out of the roots by ⁸⁶Rb techniques showed that influx, and to a lesser extent efflux, were inhibited at low temperature. The result was a net potassium uptake rate of one-third that of unstressed plants. After the cooling period the potassium influx increased to the rate of control plants. The potassium efflux increased to one and one-half times the rate of unstressed wheat so that net uptake was negative. The increase in potassium efflux was explained by a higher permeability of the root cell membranes after cooling. The net uptake of calcium was reduced to one-third by root cooling. Contrary to potassium uptake, calcium uptake increased under post-stress conditions, partly due to a low efflux rate. During root cooling there was a redistribution of dry matter from the leaves down towards the lower part of the shoot. Afterwards the original distribution of dry matter was reestablished. The net flow of potassium and calcium followed a similar pattern as dry matter, suggesting a growth-regulated flow.     

Alkali grass resists salt stress through high [K+] and an endodermis barrier to Na+

In order to understand the salt-tolerance mechanism of alkali grass (Puccinellia tenuiflora) compared with wheat (Triticum aestivum L.), [K(+)] and [Na(+)] in roots and shoots in response to salt treatments were examined with ion element analysis and X-ray microanalysis. Both the rapid K(+) and Na(+) influx in response to different NaCl and KCl treatments, and the accumulation of K(+) and Na(+) as the plants acclimated to long-term stress were studied in culture- solution experiments. A higher K(+) uptake under normal and saline conditions was evident in alkali grass compared with that in wheat, and electrophysiological analyses indicated that the different uptake probably resulted from the higher K(+)/Na(+) selectivity of the plasma membrane. When external [K(+)] was high, K(+) uptake and transport from roots to shoots were inhibited by exogenous Cs(+), while TEA (tetraethylammonium) only inhibited K(+) transport from the root to the shoot. K(+) uptake was not influenced by Cs(+) when plants were K(+) starved. It was shown by X-ray microanalysis that high [K(+)] and low [Na(+)] existed in the endodermal cells of alkali grass roots, suggesting this to be the tissue where Cs(+) inhibition occurs. These results suggest that the K(+)/Na(+) selectivity of potassium channels and the existence of an apoplastic barrier, the Casparian bands of the endodermis, lead to the lateral gradient of K(+) and Na(+) across root tissue, resulting not only in high levels of [K(+)] in the shoot but also a large [Na(+)] gradient between the root and the shoot.     

Effect of cadmium on iron uptake in cucumber roots: A Mössbauer-spectroscopic study

The uptake and accumulation of iron in cucumber roots exposed to cadmium were investigated with Fe sufficient and deficient cucumber plants using Mössbauer spectroscopy, Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and ferric chelate reductase activity measurements. Both Fe sufficient and Fe deficient plants were applied. In the case of Fe sufficient cucumber roots grown in nutrient solution with 10 μM Cd no changes were found in the occurrence of Fe species (mostly hydrous ferric oxides and ferric-carboxylate complexes) compared to the control where no Cd was added. In the Fe deficient roots pretreated with 0, 0.1, 1, 10 and 100 μM Cd for 3 h then supplied also with 0.5 mM ⁵⁷Fe-citrate for 30 min, Fe^II was identified in a hexaaqua complex form. The relative amount of Fe^II was decreasing simultaneously with increasing Cd concentration, while the relative occurrence of Fe^III species and total Fe concentration were increasing. The results support the inhibitory effect of Cd on Fe-chelate reduction. Although the reductase activity at 10 and 100 μM Cd treatment was lower than in the iron sufficient control plants, Fe^II could be identified by Mössbauer spectroscopy whereas in the Fe sufficient control, this form was below detection limit. These data demonstrate that the influx and the reoxidation of Fe^II was decreased by Cd, consequently, they refer to the competition of Cd²⁺ and Fe²⁺ during the membrane transport and the inhibition of the reoxidation process.