Accumulation, subcellular localization and ecophysiological responses to copper stress in two Daucus carota L. populations

Copper accumulation, subcellular localization and ecophysiological responses to excess copper were investigated using pot culture experiments with two Daucus carota L. populations, from a copper mine and an uncontaminated field site, respectively. Significant differences of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) concentrations and antioxidant enzyme [superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX)] activities of leaves under Cu treatment were observed between the two populations. At high Cu concentrations (400 and 800 mg kg⁻¹), a significant increase in contents of MDA and H₂O₂ but a significant decrease in activities of SOD, CAT and APX were observed in uncontaminated population. Contrarily, the population from copper mine maintained a lower level of MDA and H₂O₂ but higher activities of SOD, CAT and APX. Copper accumulation in roots and shoots increased significantly with the increase of copper concentrations in soils in the two populations. No significant difference of the total Cu in roots and shoots was found between the two populations at same copper treatment. There were also no striking differences of cell wall-bound Cu and protoplasts Cu of leaves between the two populations. The difference was that Cu concentration in vacuoles of leaves was 1.5-fold higher in contaminated site (CS) population than in uncontaminated site population. Hence, more efficient vacuolar sequestration for Cu and maintaining high activities of SOD, CAT and APX in the CS population played an important role in maintaining high Cu tolerance.     

A multielement analysis of Cu induced changes in the mineral profiles of Cu sensitive and tolerant populations of Silene paradoxa L.

This work investigates the Cu induced changes in element profiles in contrasting ecotypes of Silene paradoxa L. A metallicolous copper tolerant population and a non-metallicolous sensitive population were grown in hydroponics and exposed to different CuSO₄ treatments. Shoot and root concentrations of Ca, Cu, Fe, K, Mg, Mn, Mo, Na, P, S and Zn were evaluated through ICP-OES.Results indicated that increasing the environmental Cu concentration had a population dependent effect on element profiles, shoot-to-root ratios and correlations among the elements. Generally, in the tolerant population Cu treatment induced a higher element accumulation in roots and had minimal effects on the shoot element profile, thus resulting in a progressively decreasing shoot-to-root ratio for each element. In the sensitive population element concentrations in root and shoot were much more affected and without a consistent trend. Copper treatment also affected the correlations between the elements, both in roots and shoots of the two populations, but more so in the sensitive population than in the tolerant one. Thus, Cu exposure strongly disturbed element homeostasis in the sensitive population, but barely or not in the tolerant one, probably mainly due to a higher capacity to maintain proper root functioning under Cu exposure in the latter. Differences in element profiles were also observed in the absence of toxic Cu exposure. These differences may reflect divergent population-specific adaptations to differential nutrient availability levels prevailing in the populations’ natural environments. There is no evidence of inherent side-effects of the Cu tolerance mechanism operating in the tolerant population.     

On the role of the cell wall in the phenomenon of copper tolerance in Silene paradoxa L.

The potential role of the cell wall in the phenomenon of copper tolerance in Silene paradoxa was investigated. Plants from Fenice Capanne (FC, copper tolerant population) mine waste and Colle Val D’Elsa (CVD, sensitive population) uncontaminated soil were grown in hydroponics and exposed to different CuSO₄ treatment to evaluate tolerance, accumulation and pectin concentration of the root cell wall. Physiological mechanisms relying both on avoidance and tolerance at the cellular level seemed to characterise the copper tolerance phenomenon showed by the metallicolous population of S. paradoxa. Avoidance was realised at both the symplast and the apoplast levels in the roots. Data suggested that in the tolerant population the presence of copper in the culture medium could decrease root cell wall pectin concentration and increase pectin methylation degree. Results proposed that a low cell wall ability to bind copper can probably combine to generate the tolerant/excluder phenotype of the S. paradoxa copper tolerant population.     

Zinc tolerance and accumulation in metallicolous and nonmetallicolous populations of Arabidopsis halleri (Brassicaceae)

Zinc tolerance was investigated in five populations of Arabidopsis halleri (syn.: Cardaminopsis halleri ) raised from seeds collected from contaminated and uncontaminated sites. Tolerance was measured by determining the concentration which inhibited root growth (EC₁₀₀). A. halleri populations from contaminated and uncontaminated sites were found to be Zn-tolerant compared with the Zn-nontolerant species Arabidopsis thaliana and A. lyrata subsp. petraea . At very high Zn concentrations, populations of A. halleri from uncontaminated sites were slightly less Zn-tolerant than those from contaminated sites. These observations support the hypothesis that in A. halleri , Zn tolerance is largely a constitutive property. One population from an uncontaminated site and one population from a contaminated site were studied for Zn uptake. Zinc content was measured in shoots and roots using a colorimetric test under laboratory conditions. The results showed that whatever their origin, individuals from both populations are Zn accumulators compared with the nonaccumulator species A. thaliana . Moreover, the population from the uncontaminated area accumulated Zn in its shoots and roots more quickly than the population from the contaminated site. These results suggest that, in A. halleri , Zn accumulation to very high concentration is a constitutive property.     

Differential invertase activity and root growth between Cu-tolerant and non-tolerant populations in Kummerowia stipulacea under Cu stress and nutrient deficiency

There has been no study on key enzymes in sucrose cleavage in metallophyte plants so far, which may be crucial for the plants’ root growth and heavy-metal tolerance maintenance. Here, we tested the hypothesis that the roots of copper tolerant plants should manifest a higher activity of acid invertases that are rate-limiting in sucrose catabolism than non-tolerant plants both for supporting growth and for their maintaining tolerance under Cu stress. Two populations of Kummerowia stipulacea, one from an ancient waste heap at a Cu mine, and the other from a non-contaminated site, were used in the experiments. The plants were grown in 1/2-fold (control) or 1/20-fold (nutrient deficiency) Hoagland’ solution, with (Cu stress) or without (control) 10 μmol/L Cu²⁺. Plants from the mine proved to be of Cu tolerance. Cu exposure had a stronger inhibition on root growth and thus resulting in a lower root/shoot ratio in the plants of non-mine population compared to the mine population. Cu exposure showed a stronger inhibition of acid invertase activity of Cu non-tolerant plants than Cu-tolerant plants, while neutral/alkaline invertase was insensitive to Cu. A positive correlation between the activity of acid invertases and the root growth and root/shoot ratio was observed. The results indicated an important role of acid invertases in governing root growth and root/shoot biomass allocation in the plants of mine population. The results also suggested that the higher activities in acid invertases of mine population plants might at least partly associate with the plants’ Cu tolerance, and their higher activities in acid invertases in turn played an role in maintenance of the Cu tolerance by supplying carbon and energy for tolerance mechanisms. In addition, the results showed evidence that neutral/alkaline invertase might play a role in compensating for the depression in sucrose catabolism due to Cu-induced inhibition in acid invertases.     

Tolerance and bioaccumulation of copper in Phragmites australis and Zea mays

The effects of copper on the growth, tolerance indices, mineral composition (N, P, K, Fe, Zn and Mn) and metal uptake of reed (Phragmites australis [Cav. Trin. ex Steudel]) and maize (Zea mays L.) were investigated in hydroponic experiments at copper concentrations ranging from 0.5 to 157 M Cu. A reduction in root length was shown to be a good indicator of copper toxicity, concentrations of 15.7 and 78.7 M Cu inhibiting root growth in maize and reed, respectively. The reed was significantly more tolerant of copper than maize and at 7.85 M Cu (external concentration), reed can be described as a Cu tolerant plant, and maize as a Cu non-tolerant species. As a result of Cu toxicity, the concentrations of macronutrients N, P and K decreased in both shoot and root of maize, while the concentrations were hardly affected in reed tissues. Fe concentration increased in shoots and roots of maize and in roots of reed with increasing Cu treatments, leading to highly significant (p<0.01) linear relationships between tissue Fe and Cu concentrations. The bioconcentration factor (BCF) of Cu was higher in roots than in shoots of both plant species, ranging from 612 to 1592 in reed for the Cu treatments tested. In the roots of maize, BCF of Cu increased from 349 to 1931 when increasing Cu in nutrient solution from 7.85 M to 78.5 M. Therefore, reed could be useful in wastewater treatments for the removal of Cu. However, the use of reed in phytoextraction of Cu from contaminated soils is limited by the low accumulation rate in shoots and although reed can be more efficient than maize for Cu phytoextraction, harvesting the full biomass, including roots, may be required.     

Differences of Cu uptake and acid phosphatase activities of two Elsholtzia haichowensis Sun populations

Two Elsholtzia haichowensis Sun populations, one from Cu mine sites (Tonglushan) and the other from uncontaminated sites (Hong'an) in Hubei Province, China, were investigated in hydroponic experiments for the differences of Cu accumulation and ecophysiological responses to Cu under Cu treatment. The results showed that a significant increase in malondialdehyde (MDA) concentration in roots was observed in Hong'an population, whereas no significant change was observed in Tonglushan population. The root pressure exudates (ml) decreased significantly in Hong'an population, but increased significantly in Tonglushan population at high Cu concentration (≥ 20 μ mol/L) treatment. Cu concentration in roots of Hong'an population was significantly higher than that in Tonglushan population. For example, Cu concentration in Hong'an population was 3 times and 4 times higher than that in Tonglushan population after 20 μ mol/L Cu treatment for one day and three days, respectively. Cu concentration in Hong'an population was 20 times and 5 times higher than that in Tonglushan population after 80 μ mol/L Cu treatment for one day and three days, respectively. The intracellular and secreted acid phosphatase (APase) activities in roots were significantly higher in Tonglushan population than in Hong'an one. The intracellular APase activities in Tonglushan population were 3 times higher than those in Hong'an population after 80 μ mol/L Cu treatment for three days and five days, respectively. The root-secreted APase activities in Tonglushan population were 1.6 times and 1.8 times higher than those in Hong'an population after 80 μ mol/L Cu treatment for three days and five days, respectively. In conclusion, excessive Cu caused serious oxidative damage in roots of Hong'an population, but not in Tonglushan population. The high intracellular and root-excreted APase activities in Tonglushan population might play an important role in reducing Cu accumulation in roots and maintaining normal phosphorus physiological metabolism in root cells under Cu stress.     

Leaf endophyte Neotyphodium coenophialum modifies mineral uptake in tall fescue

Neotyphodium coenophialum (Morgan-Jones and Gams) Glenn, Bacon and Hanlin, a fungal endophyte found primarily in shoots of tall fescue (Festuca arundinacea Shreb.), can modify rhizosphere activity in response to phosphorus (P) deficiency. In a controlled environment experiment, two cloned tall fescue genotypes (DN2 and DN4) free (E-) and infected (E+) with their naturally occurring endophyte strains were grown in nutrient solutions at low P (3.1 ppm) or high P (31 ppm) concentrations for 21 d. Endophyte infection increased root dry matter (DM) of DN4 by 21% but did not affect root DM of DN2. Under P deficiency, shoot and total DM were not affected by endophyte but relative growth rate was greater in E+ than E- plants. In high P nutrient solution, E+ plants produced 13% less (DN2) or 29% more (DN4) shoot DM than E- plants. Endophyte affected mineral concentrations in roots more than in shoots. Regardless of P concentration in nutrient solution, E+ DN2 accumulated more P, Ca, Zn and Cu but less K in roots than E- plants. When grown in high P nutrient solution, concentrations of Fe and B in roots of E+ DN2 plants were reduced compared with those of E- plants. Concentrations of P, Ca and Cu in roots of DN4 were less, but K was greater in E+ than E- plants. In shoots, E+ DN2 had greater concentrations of Fe and Cu than E- DN2, regardless of P concentration in nutrient solution. Genotype DN4 responded to endophyte infection by reducing B concentration in shoots. Nutrient uptake rates were affected by endophyte infection in plants grown in low P nutrient solution. A greater uptake rate of most nutrients and their transport to shoots was observed in DN2, but responses of DN4 were not consistent. Results suggest that endophyte may elicit different modes of tall fescue adaptation to P deficiency. This revised version was published online in June 2006 with corrections to the Cover Date.     

Selective adaptation to copper of populations ofAgrostis tenuis andFestuca rubra (Poaceae)

Populations ofAgrostis Tenuis andFestuca rubra, tolerant and non-tolerant towards Zn and Pb, were studied in regard to their adaptability by natural selection, towards a third metal (Cu) and the time required for the appearance of such tolerance. It was found that Zn and Pb tolerant and non-tolerant populations are likely to select the character of Cu-tolerance within the course of only one generation. More specifically, Zn and Pb tolerant populations show a better adaptability compared with non-tolerant ones. The amount of tolerance to copper obtained by artificial selection is comparable with that of populations growing naturally on copper mines.     

铜处理对菠菜幼苗矿质营养吸收和细胞超微结构的影响

土壤重金属积累严重影响植物生长和生态系统平衡,探寻植物对重金属的耐性机理尤为重要.菠菜可能具有一定的耐铜性,但Cu对其矿质元素吸收、细胞超微结构等方面的耐性机理尚不明确.本研究以菠菜幼苗为研究对象,通过盆栽试验,探究不同浓度铜处理对菠菜幼苗生长、矿质元素吸收、叶片细胞超微结构等指标的影响.结果表明: 100 mg·L^-1 CuSO₄处理浓度时,菠菜幼苗根Cu²⁺积累量小于地上部,其根系生长量增加,地上部生长量稍有下降,继续增加铜处理浓度,植物体各器官生长参数均呈下降趋势.低浓度铜处理时(<400 mg·L^-1 CuSO₄),菠菜幼苗叶N、K、Ca、Mg、Fe含量增加,P含量减少;根N、P、K含量减少,Ca、Mg、Fe含量增加;叶片细胞内各细胞器清晰可见,基粒片层排列仍较为整齐,叶绿体内外膜完整.高浓度铜处理时(>600 mg·L^-1 CuSO₄),菠菜幼苗叶N含量增加,P、K、Ca、Mg、Fe含量减少;根N、P、K、Ca、Mg、Fe含量均减少;叶片细胞内叶绿体变圆,叶绿体膜变薄,基质、基粒片层变少,层堆积高度下降,细胞核解体,液泡、细胞壁中有黑色小点分布,可能是大量Cu²⁺聚集导致细胞内膨压增大所致.低浓度铜处理并未对菠菜幼苗的生长生理特性产生明显的负面影响,而高浓度铜处理并未终止菠菜幼苗的生长.说明菠菜幼苗具有一定的耐铜性.     

Role of arbuscular mycorrhizal symbiosis in root mineral uptake under CaCO3 stress

This study investigated the effects of increasing CaCO(3) concentrations (0, 5, 10, 20?mM) on arbuscular mycorrhizal (AM) symbiosis establishment as well as on chicory root growth and mineral nutrient uptake in a monoxenic system. Although CaCO(3) treatments significantly decreased root growth and altered the symbiosis-related development steps of the AM fungus Rhizophagus irregularis (germination, germination hypha elongation, root colonization rate, extraradical hyphal development, sporulation), the fungus was able to completely fulfill its life cycle. Even when root growth decreased more drastically in mycorrhizal roots than in non-mycorrhizal ones in the presence of high CaCO(3) levels, the AM symbiosis was found to be beneficial for root mineral uptake. Significant increases in P, N, Fe, Zn and Cu concentrations were recorded in the mycorrhizal roots. Whereas acid and alkaline phosphatase enzymatic activities remained constant in mycorrhizal roots, they were affected in non-mycorrhizal roots grown in the presence of CaCO(3) when compared with the control.     

Effect of zinc fertilization on the mineral nutrition of rices differing in tolerance to zinc deficiency

Summary The effects of four Zn levels on the electrochemical and chemical properties of the soil solution, and on the growth and mineral nutrition of two rice varieties (IR26 and IR34) differing in tolerance to Zn deficiency were studied in the greenhouse using Zn-deficient soils from two locations. A similar experiment was conducted in culture solution to check how Zn addition affects translocation of other nutrients.In both soil and culture solution, plant Zn concentrations alone was not enough to account for varietal tolerance to Zn deficiency. Comparison of nutrient to Zn and shoot to root ratios of nutrients was more useful in determining the possible mechanism of varietal tolerance. IR 34 appeared to tolerate the disorder due to its lower Zn requirement, more efficient Zn translocation and ability to maintain lower Fe/Zn, Cu/Zn, Mg/Zn and P/Zn ratios in the shoot than the more susceptible variety, IR26. This was shown to be due to decreased translocation of Fe, Mg and P to shoots and decreased absorption of Cu by the root in IR34 in culture solution studies. Adding Zn further reduces translocation or absorption of these nutrients and depending on the nutrient supply of the soil, could cause deficiencies or mineral imbalances, especially of Fe, Cu, and P.These observed varietal differences regarding Zn requirement and the interaction of Zn with absorption and translocation of plant nutrients necessitates revision of recommendations for Zn fertilization. There is an inevitable need for Zn application in severely Zn-deficient soils regardless of rice variety. But on marginally Zn-deficient soils especially those low in Fe, Cu, or P, Zn fertilization is not advisable when resistant rice varieties are used.     

Comparison of cadmium and copper effect on phenolic metabolism, mineral nutrients and stress-related parameters in Matricaria chamomilla plants

Cadmium and copper uptake and its consequence for activity of selected enzymes of phenolic metabolism, phenolic acids accumulation, quantity of mineral nutrients and stress-related parameters in Matricaria chamomilla plants exposed to 60 μM and 120 μM for 7 days has been studied. Cu content in the above-ground biomass was ca. 10-fold lower compared to Cd and amount of Cd in the methanol-soluble fraction was lower than in the water-soluble fraction. “Intra-root” Cd represented 68% and 63% of total Cd content at 60 μM and 120 μM, but no difference was observed in Cu-exposed roots. Cu excess had more pronounced effect on shikimate dehydrogenase, cinnamyl alcohol dehydrogenase, polyphenol oxidase and ascorbate peroxidase activity mainly in the roots. Among eight detected benzoic acid derivatives and four cinnamic acid derivatives, the latter were preferentially accumulated in response to Cd excess. Content of salicylic acid increased in all variants. Amount of superoxide was elevated in both the rosettes (preferentially by Cu) and roots (preferentially by Cd). Accumulation of Ca and Mg was not affected by excess of metals, while potassium decreased in both the rosettes and roots (Cu caused stronger depletion). Amount of Fe increased in the roots in response to both metals (more expressively in Cu-treated ones). Present study using other metabolic parameters (and supplementing our previous studies) has confirmed higher Cu toxicity for chamomile plants, to support its strong pro-oxidant properties. These observations as complex metabolic responses are discussed.     

The effects of toxic metals,content of nutrients and inoculation with mycorrhizal fungi on the level of phenolics in roots and growth of Scots pine seedlings

The effects of two substrates and several strains of mycorrhizal fungi on the content of soluble phenolics in roots and growth of Scots pine seedlings was investigated. The first substrate was fertile and contaminated with copper, zinc and lead, whereas the second one displayed nutrient deficiency. The dry weights of needles, trunks, roots and the total biomass were higher in groups of seedlings inoculated with mycorrhizal fungi on fertile and polluted substrate. Inoculation of pine seedlings on this substrate resulted in a decrease in concentration of phenolics in roots and except for seedlings inoculated with Laccaria laccata negatively influenced the above-ground part: root ratio.     

Heavy metal impact on bacterial biomass based on DNA analyses and uptake by wild plants in the abandoned copper mine soils

The metals contamination in surface soils and their accumulation in wild plants from the abandoned Burra and Kapunda copper mines located in South Australia were assessed, and the predominant bacterial diversity in the contaminated surface soils from these two abandoned copper mine sites were evaluated through polymerase chain reaction–denaturing gradient gel electrophoresis (PCR-DGGE) analysis. The results showed the average concentration of Cu in soils was 3821.59 mg/kg while wild plants accumulated up to 173.44 mg/kg. The concentration of Cu in shoots of spear grass (Stipa uitida) and berry saltbush (Afriplex semibaccata) was higher than that of roots. The concentration of total and extractable As, Cd, Cu and Pb in soils slightly correlated with of these elements in the corresponding wild plants. The toxicity of Cu in heavily contaminated soils impacted on the quantities of specific microbial populations and no significant change in the microbial diversity of highly contaminated soils.     

Copper phytotoxicity affects root elongation and iron nutrition in durum wheat (Triticum turgidum durum L.)

This work investigated how copper (Cu) phytotoxicity affected iron (Fe) nutrition and root elongation in hydroponically grown durum wheat (Triticum turgidum durum L., cv Acalou) in order to establish the critical level of Cu concentration in roots above which significant Cu phytotoxicity occurs. This was assessed at two levels of Fe supply (2 and 100 μM). Severe symptoms of Cu phytotoxicity were observed at Cu²⁺ concentration above 1 μM, i.e. interveinal chlorosis symptoms and global root growth alteration. Total root Cu concentration of about 100, 150 and 250–300 mg kg^?1 corresponded to 10%, 25% and 50% reduction in root elongation, respectively. Copper and Fe concentrations as well as amounts of Cu and Fe accumulated in shoots varied inversely which suggested an antagonism between Cu and Fe leading to Fe deficiency. In addition, the root-induced release of complexing compounds increased significantly with increasing Cu concentration in nutrient solution and was positively correlated with Cu uptake without significant difference between the two Fe treatments (high and low Fe supply). This work suggests that total root Cu concentration might be a simple, sensitive indicator of Cu rhizotoxicity. It also indicated that Cu phytotoxicity which may have resulted in Fe deficiency and significant increase in root-induced release of complexing compounds (presumably phytosiderophores) was independent of the level of Fe supply provided that the threshold values of phytotoxicity were based on the free Cu-ion concentration.     

Mycorrhizal infection of an Agrostis capillaris population on a copper contaminated soil

To investigate the possible natural development of heavy metal tolerance in VA-mycorrhizal fungi, plants of Agrostis capillaris from an uncontaminated, a copper-contaminated and a zinc/cadmium-contaminated area were examined for VA-mycorrhizal infection. During a period of 5 years (1987 to 1991) the plants of the copper-tolerant population were hardly infected, whereas the population on the uncontaminated soil showed a mean infection of nearly 60% and the zinc/cadmium-tolerant population of 40%. A detailed analysis of the surroundings of the copper-enriched site revealed the presence of VA-mycorrhizal fungi and a negative correlation between the infection rate of A. capillaris and the copper content of the soil. In contrast to the copper-contaminated soil, the abundant presence of VA-mycorrhizal fungi in the area contaminated by zinc and cadmium indicates that these fungi have evolved a zinc and cadmium tolerance and that they may play a role in the zinc and cadmium tolerance of A. capillaris.     

Effects of copper on growth, radial oxygen loss and root permeability of seedlings of the mangroves Bruguiera gymnorrhiza and Rhizophora stylosa

Purpose

Mangrove wetlands have experienced significant contaminant input such as copper (Cu), aggravated by rapid urban development. This study aimed to investigate the possible function of root permeability in metal detoxification.

Methods

Pot trials were conducted to evaluate the responses of root permeability in relation to metal (Cu) exposure in seedlings of two mangroves: Bruguiera gymnorrhiza and Rhizophora stylosa.

Results

Copper inhibited plant growth and root permeability of the two species significantly (due to decreases in root porosity, thickening of exodermis and increases in lignification), leading to a significant reduction in radial oxygen loss (ROL). A negative correlation between soil Cu and ROL from root tip was also observed. The observed metal uptake by excised roots further indicated that increased lignification would directly prevent excessive Cu from further entering into the roots.

Conclusions

In summary, the two mangroves reacted to Cu by producing an impermeable barrier in roots. Such an inducible barrier to ROL is likely to be an adaptive strategy against Cu toxicity. This study reveals new evidence of a structural adaptive strategy for metal tolerance by mangrove plants.     

Copper Uptake and Its Effect on Metal Distribution in Root Growth Zones of Commelina communis Revealed by SRXRF

To explore the copper uptake mechanisms by the Cu-tolerant plant Commelina communis, the contents of Cu and other metals (including Fe, Zn, and Mn) in roots were detected using atomic absorption spectrometer under transporter inhibitors, partial element deficiency, or Cu excess treatments, while distribution characters of Cu and other metals in root growth zones were investigated by synchrotron radiation X-ray fluorescence spectroscopy (SRXRF). Cu uptake was inhibited by the uncoupler DNP and P-type ATPase inhibitor Na₃VO₄, not by the Ca²⁺ ion channel inhibitor LaCl₃, suggesting that Cu could probably be assimilated actively by root and be related with P-type ATPase, but not through Ca²⁺ ion channel. Fe or Zn deficiency could enhance Cu uptake, while 100 μM Cu inhibited Fe, Zn, and Mn accumulation in roots significantly. Metal distribution under 100 μM Cu treatment was investigated by SRXRF. High level of Cu was found in the root meristem, and higher Cu concentrations were observed in the vascular cylinder than those in the endodermis, further demonstrating the initiative Cu transport in the root of C. communis. Under excess Cu stress, most Fe was located in the epidermis, and Fe concentrations in the endodermis were higher than those in the vascular cylinder, suggesting Cu and Fe competition not only in the epidermal cells but also for the intercellular and intracellular transport in roots. Zn was present in the meristem and the vascular cylinder similar to Cu. Cu and Zn showed a similar pattern. Mn behaves as Zn does, but not like Fe.