Molybdenum (Mo) increases endogenous phenolics,proline and photosynthetic pigments and the phytoremediation potential of the industrially important plant Ricinus communis L. for removal of cadmium from contaminated soil

Cadmium (Cd) in agricultural soil negatively affects crops yield and compromises food safety. Remediation of polluted soil is necessary for the re-establishment of sustainable agriculture and to prevent hazards to human health and environmental pollution. Phytoremediation is a promising technology for decontamination of polluted soil. The present study investigated the effect of molybdenum (Mo) (0.5, 1.0 and 2.0 ppm) on endogenous production of total phenolics and free proline, plant biomass and photosynthetic pigments in Ricinus communis plants grown in Cd (25, 50 and 100 ppm) contaminated soils and the potential for Cd phytoextraction. Mo was applied via seed soaking, soil addition and foliar spray. Foliar sprays significantly increased plant biomass, Cd accumulation and bioconcentration. Phenolic concentrations showed significantly positive correlations with Cd accumulation in roots (R ² = 0.793, 0.807 and 0.739) and leaves (R ² = 0.707, 721 and 0.866). Similarly, proline was significantly positively correlated with Cd accumulation in roots (R ² = 0.668, 0.694 and 0.673) and leaves (R ² = 0.831, 0.964 and 0.930). Foliar application was found to be the most effective way to deliver Mo in terms of increase in plant growth, Cd accumulation and production of phenolics and proline.

     

Growth and physiological responses of Pennisetum sp. to cadmium stress under three different soils

Pennisetum sp. was employed as a model species to detect the growth and physiological response to cadmium (Cd) stress at different Cd concentrations (0, 20, 50, and 100 mg kg⁻¹) in three types of soils (yellow brown soil, yellow soil, and red soil). Results showed that the growth of Pennisetum sp. was not significantly influenced by Cd in 20 mg kg⁻¹, but significantly inhibited at higher Cd concentrations in three types of soils. Besides, the higher Cd concentrations, the lower root, stem, and leaf biomass. With Cd concentration of soil increasing, Cd content of root, stem, and leaf increased. Compared with no Cd, high Cd concentrations (50 and 100 mg kg⁻¹) induced the physiological indices (photosynthetic rate, stomatal conductance, transpiration rate) and biochemical indices (nitrate reductase, glutamine synthetase, and glutamate synthase activities) decreasing, but the concentration of NO₃⁻ and NH₄⁺ increasing. The activity of antioxidative enzymes (SOD, POD, and CAT) was disrupted and the content of malondialdehyde (MDA) increasing. Pennisetum sp. could protect cells from damage and maintain normal physiological metabolism via increasing the production of soluble sugar and soluble protein, but soluble proteins and soluble sugars were limited in high concentrations of Cd (50 and 100 mg kg⁻¹). Moreover, the growth and physiological response to Cd are different in the three types of soils. The growth of Pennisetum sp. in yellow brown soil was better than that in other two soils, and the gas exchange rate, antioxidant enzyme activity, and nitrogen metabolism in yellow soil and red soil were more affected by Cd stress than that in yellow brown soil. Overall, Pennisetum sp. had certain tolerance and biosorption ability to Cd in different Cd concentrations and different types of soil. Hence, Pennisetum sp. was a suitable choice for Cd remediation, especially in yellow brown soil.

     

Heavy Metal Uptake by Two Edible Amaranthus Herbs Grown on Soils Contaminated with Lead,Mercury, Cadmium,and Nickel

The uptake of an element by a plant is primarily dependent on the plant species, its inherent controls, and the soil quality. Amaranthus hybridus (green herbs) and Amaranthus dubius (red herbs) were chosen to investigate their response and ability to accumulate and tolerate varying levels of elements in their roots and shoots. Red herbs and green herbs were grown in soil pots contaminated with three mixtures of Cd(II), Ni(II), Pb(II), and Hg(II). Plants in the control treatment were grown in the absence of the heavy metals mixture. The distribution of Cd, Ni, Pb, and Hg in the plants (in roots, stems, and leaves) was determined in two stages. Stage 1, after 5 weeks of plant growth and stage 2, full grown after 10 weeks of growth. In the red herbs the Cd concentration in the leaves at stage 2 was 150 ppm and was present in higher concentrations than Ni, Hg, and Pb. At the highest contamination level, in the green herbs plant, Hg was present in the highest concentration in the root, i.e., 336 ppm at stage 1, while the level in the leaves was 7.12 ppm. Both the green and red herbs species showed an affinity for Ni and Cd with moderate to high levels detected in the leaves, respectively.     

Influx,transport, and accumulation of cadmium in plant species grown at different Cd2+ activities

Abstract

Cadmium (Cd) has no known essential biological function, but it is toxic to plants, animals, and humans. A promising approach to prevent Cd from entering the food chain would be to select and/or create Cd‐accumulating plants to remediate contaminated soils or to develop Cd‐excluding plants to reduce Cd flow from soils into foods. The present study was undertaken to examine the differences in Cd influx, transport, and accumulation among five plant species in relation to plant tolerance to Cd toxicity. Ryegrass (Lolium perenne L.) had the least reduction in dry matter which may be due to its lowest Cd transport rate (TR) to shoots at all Cd levels among the plant species tested. White‐clover (Trifolium repens L.) was the most sensitive species to Cd toxicity, likely because of its highest Cd influx rate (IR) and high TR when plants were grown at low Cd²⁺ activity (≤8 μM). The high tolerance of cabbage (Brassica oleracea var. capitata L.) to moderate Cd toxicity (≤14 μM) appeared to be mainly due to the detoxification of Cd inside plant tissue since it recorded the highest TR and relatively high IR for Cd among the tested species. At Cd²⁺ activities up to 28 uM, the Cd uptake ratios of shoot/root for ryegrass were, on average, about 50‐fold and 27‐fold lower than that for cabbage and maize (Zea mays L.), respectively. These results showed that Cd could be easily transported into shoots of cabbage and maize, but was mainly confined to roots of ryegrass. We suggest that influx and transport rates, especially transport rate, could be used as plant physiological parameters for screening Cd‐excluding genotypes among monocotyledonous plants.     

Antioxidative response to Cd in a newly discovered cadmium hyperaccumulator, Arabis paniculata F

A hydroponic experiment was carried out to study the effect of cadmium (Cd) on growth, Cd accumulation, lipid peroxidation, reactive oxygen species (ROS) content and antioxidative enzymes in leaves and roots of Arabis paniculata F., a new Cd hyperaccumuator found in China. The results showed that 22-89 microM Cd in solution enhanced the growth of A. paniculata after three weeks, with 21-27% biomass increase compared to the control. Cd concentrations in shoots and roots increased with increasing Cd supply levels, and reached a maximum of 1662 and 8670 mg kg(-1) Cd dry weight at 178 microM Cd treatment, respectively. In roots, 22-89 microM Cd reduced the content of malondialdehyde (MDA), superoxide (O(2)(-1)) and H(2)O(2) as well as the activities of superoxide dismutase (SOD), guaiacol peroxidase (GPX), ascorbate peroxidase (APX) and glutathione reductase (GR). In leaves, the contents of MDA, O(2)(-1) and H(2)O(2) remained unaffected by 22-89 microM Cd, while 178 microM Cd treatment significantly increased the MDA content, 69.5% higher than that of the control; generally, the activities of SOD, catalase (CAT), GPX and APX showed an increasing pattern with increasing Cd supply levels. Our present work concluded that A. paniculata has a great capability of Cd tolerance and accumulation. Moderate Cd treatment (22-89 microM Cd) alleviated the oxidative stress in roots, while higher level of Cd addition (178 microM) could cause an increasing generation of ROS, which was effectively scavenged by the antioxidative system.     

Remediation mechanisms of mercapto-grafted palygorskite for cadmium pollutant in paddy soil

The immobilization agent was the key factor that determined the success of remediation of heavy metal polluted soil. In this study, mercapto-grafted palygorskite (MP) as a novel and efficient immobilization agent was utilized for the remediation of Cd-polluted paddy soil in pot trials, and the remediation mechanisms were investigated in the aspect of soil chemistry and plant physiology with different rice cultivars as model plants. Mercapto-grafted palygorskite at applied doses of 0.1–0.3% could reduce Cd contents of brown rice and straws of different cultivars significantly. Both reduced DTPA-extractable Cd contents in rhizosphere and non-rhizosphere soil and decreasing Cd contents in iron plaques on rice root surfaces confirmed that MP was an efficient immobilization agent for Cd pollutant in paddy soil. In the aspect of soil chemistry, the pH values of rhizosphere and non-rhizosphere soils had no statistical changes in the MP treatment groups, but their zeta potentials decreased obviously, indicating that MP could enhance the fixation or sorption of Cd on soil compositions. In the aspect of antioxidant system, MP could increase POD activity of rice roots significantly to alleviate the stress of Cd to roots, and resulted in the decrease of T-AOC, SOD, and CAT activities of rice roots of the selected cultivars. MP had no inhabitation or enhancement effects on TSH of rice roots but enhance the contents of MTs and NPT to binding Cd to complete detoxification process. MP as a novel and efficient immobilization agent could complete the remediation effects through soil chemistry and plant physiological mechanisms.

     

Characterization of cadmium uptake,translocation, and distribution in young seedlings of two hot pepper cultivars that differ in fruit cadmium concentration

The reasons why some cultivars of hot pepper (Capsicum annuum L.) accumulate low levels of Cd are poorly understood. We aimed to compare the characteristics of Cd uptake and translocation in low-Cd and high-Cd hot pepper cultivars by determining the subcellular locations and chemical forms of Cd, and its distribution among different plant organs. We conducted a hydroponic experiment to investigate the subcellular distribution and chemical forms of Cd in roots, stems, and leaves of a low-Cd (Yeshengchaotianjiao, YCT) and a high-Cd cultivar (Jinfuzaohuangjiao, JFZ). The results showed that the concentrations of Cd in almost all subcellular fractions of roots, and in all chemical forms in roots, were higher in YCT than in JFZ. Compared with YCT, JFZ had higher Cd concentrations in almost all subcellular fractions of stems and leaves, and higher Cd concentrations in almost all chemical forms in stems and leaves. Additionally, YCT had significantly higher total Cd accumulation but a lower Cd translocation rate compared with JFZ. In general, the results presented in this study revealed that root-to-shoot Cd translocation via the xylem is the key physiological processes determining the Cd accumulation level in stems and leaves of hot pepper plants. Immobilization of Cd by the cell walls of different organs is important in Cd detoxification and limiting the symplastic movement of Cd.     

Heavy metal uptake by spinach leaves grown on contaminated soils with lead, mercury, cadmium, and nickel

Spinach plants were grown in soil pots contaminated with increasing mixtures of lead, mercury, cadmium, and nickel salts. Plants in the control soil were grown in the absence of the heavy metals mixture. The elemental distribution of Cd, Ni, Pb, and Hg in the roots and leaves of Spinach (Spinacia Oleracea) was determined in two stages, Stage 1, after five weeks of plant growth and Stage 2, after 10 weeks with full growth. Under the influence of contamination of soil with the heavy metal mixtures, Hg was the most accumulated element in the root of the spinach plant with a concentration of 283 ppm recorded in the highest contaminated soil, followed by Cd at 148 ppm.     

Differential physiological,ultramorphological and metabolic responses of cotton cultivars under cadmium stress

Cadmium (Cd) stress may cause serious physiological, ultramorphological and biochemical anomalies in plants. Cd-induced physiological, subcellular and metabolic alterations in two transgenic cotton cultivars (BR001, GK30) and their parent line (Coker 312) were evaluated using 10, 100 and 1000 μM Cd. Germination, fresh biomass of roots, stems and leaves were significantly inhibited at 1000 μM Cd. Root volume tolerance index significantly increased (124.16%) in Coker 312 at 1000 μM Cd. In non-Cd stressed conditions, electron micrographs showed well-configured root meristem and leaf mesophyll cells. At 1000 μM Cd, greater ultramorphological alterations were observed in BR001 followed by GK30 and Coker 312. These changes were observed in nucleus, vacuoles, mitochondria and chloroplast. Dense precipitates, probably Cd, were seen in vacuoles, which were also attached to the cell walls. A considerable increase in number of nuclei, vacuoles, starch granules and plastoglobuli was observed in the electron micrographs of both roots and leaves at 1000 μM Cd. MDA contents were higher in roots of BR001 at 1000 μM Cd. Mean values of SOD activity in leaves of both BR001 and GK30 at 1000 μM Cd significantly increased as compared to the controls. POD activity in roots of BR001 and Coker 312 was greater at all Cd (10, 100, 1000 μM) levels over the control. Regarding APX, highest percent increase (71.64%) in roots of GK30 at 1000 μM Cd was found. Non-significant differences in CAT activity were observed at all levels of Cd stress in leaves of BR001 and GK30. Both transgenic cotton cultivars and their parental line invariably responded towards Cd stress. However, Coker 312 showed Cd-resistant behavior as compared to its progeny lines (BR001 and GK30).     

Heavy metal uptake by two edible Amaranthus herbs grown on soils contaminated with lead, mercury, cadmium, and nickel

The uptake of an element by a plant is primarily dependent on the plant species, its inherent controls, and the soil quality. Amaranthus hybridus (green herbs) and Amaranthus dubius (red herbs) were chosen to investigate their response and ability to accumulate and tolerate varying levels of elements in their roots and shoots. Red herbs and green herbs were grown in soil pots contaminated with three mixtures of Cd(II), Ni(II), Pb(II), and Hg(II). Plants in the control treatment were grown in the absence of the heavy metals mixture. The distribution of Cd, Ni, Pb, and Hg in the plants (in roots, stems, and leaves) was determined in two stages. Stage 1, after 5 weeks of plant growth and stage 2, full grown after 10 weeks of growth. In the red herbs the Cd concentration in the leaves at stage 2 was 150 ppm and was present in higher concentrations than Ni, Hg, and Pb. At the highest contamination level, in the green herbs plant, Hg was present in the highest concentration in the root, i.e., 336 ppm at stage 1, while the level in the leaves was 7.12 ppm. Both the green and red herbs species showed an affinity for Ni and Cd with moderate to high levels detected in the leaves, respectively.     

Analysis and characterization of cultivable heavy metal-resistant bacterial endophytes isolated from Cd-hyperaccumulator Solanum nigrum L. and their potential use for phytoremediation

This study investigates the heavy metal-resistant bacterial endophytes of Cd-hyperaccumulator Solanum nigrum L. grown on a mine tailing by using cultivation-dependent technique. Thirty Cd-tolerant bacterial endophytes were isolated from roots, stems, and leaves of S. nigrum L. and classified by amplified ribosomal DNA-restriction analysis into 18 different types. Phylogenetic analysis based on 16S rDNA sequences showed that these isolates belonged to four groups: Actinobacteria (43%), Proteobacteria (23%), Bacteroidetes (27%) and Firmicutes (7%). All the isolates were then characterized for their plant growth promoting traits as well as their resistances to different heavy metals; and the actual plant growth promotion and colonization ability were also assessed. Four isolates were re-introduced into S. nigrum L. under Cd stress and resulted in Cd phytotoxicity decrease, as dry weights of roots increased from 55% to 143% and dry weights of above-ground from 64% to 100% compared to the uninoculated ones. The total Cd accumulation of inoculated plants increased from 66% to 135% (roots) and from 22% to 64% (above-ground) compared to the uninoculated ones. Our research suggests that bacterial endophytes are a most promising resource and may be the excellent candidates of bio-inoculants for enhancing the phytoremediation efficiency.     

Australian native plant species Carpobrotus rossii (Haw.) Schwantes shows the potential of cadmium phytoremediation

Many polluted sites are typically characterized by contamination with multiple heavy metals, drought, salinity, and nutrient deficiencies. Here, an Australian native succulent halophytic plant species, Carpobrotus rossii (Haw.) Schwantes (Aizoaceae) was investigated to assess its tolerance and phytoextraction potential of Cd, Zn, and the combination of Cd and Zn, when plants were grown in soils spiked with various concentrations of Cd (20–320 mg kg^?1 Cd), Zn (150–2,400 mg kg^?1 Zn) or Cd + Zn (20?+?150, 40?+?300, 80?+?600 mg kg^?1). The concentration of Cd in plant parts followed the order of roots > stems > leaves, resulting in Cd translocation factor (TF, concentration ratio of shoots to roots) less than one. In contrast, the concentration of Zn was in order of leaves > stems > roots, with a Zn TF greater than one. However, the amount of Cd and Zn were distributed more in leaves than in stems or roots, which was attributed to higher biomass of leaves than stems or roots. The critical value that causes 10 % shoot biomass reduction was 115 μg g^?1 for Cd and 1,300 μg g^?1 for Zn. The shoot Cd uptake per plant increased with increasing Cd addition while shoot Zn uptake peaked at 600 mg kg^?1 Zn addition. The combined addition of Cd and Zn reduced biomass production more than Cd or Zn alone and significantly increased Cd concentration, but did not affect Zn concentration in plant parts. The results suggest that C. rossii is able to hyperaccumulate Cd and can be a promising candidate for phytoextraction of Cd from polluted soils.     

Effect of endophyte-infection on growth parameters and Cd-induced phytotoxicity of Cd-hyperaccumulator Solanum nigrum L

The aim of this work was to evaluate effects of endophytic bacterium inoculation on plant growth and assess the possible mechanism of endophyte in heavy metal phytoremediation. Seeds of Solanum nigrum L. were inoculated with endophyte Serratia nematodiphila LRE07 and were subjected to Cd in the growing medium. Cd produced a significant inhibition on plant growth and a reduction in the content of photosynthetic pigments. The inoculation of endophytic bacterium alleviated the Cd-induced changes, resulting in more biomass production and higher photosynthetic pigments content of leaves compared with non-symbiotic ones. The beneficial effect was more obvious at relatively low Cd concentration (10 μM). Based on the alteration of nutrient uptake and activated oxygen metabolism in infected plants, the possible mechanisms of endophytic bacterium in Cd phytotoxicity reduction can be concluded as uptake enhancement of essential mineral nutrition and improvement in the antioxidative enzymes activities in infected plant.     

重金属污染土壤接种丛枝菌根真菌对蚕豆毒性的影响

采用盆栽实验的方法,研究了重金属(包括Cu、Zn、Pb和Cd)复合污染和接种丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)Glomus mosseae对蚕豆(Vicia faba)生长及DNA损伤的影响.结果表明,虽然接种菌根真菌对蚕豆生物量的影响并不显著,但是却显著影响植物对重金属的吸收,接种菌根真菌对蚕豆吸收4种重金属元素的作用有差异.采用单细胞凝胶电泳(single cell gel electrophoresis,SCGE)法研究接种菌根真菌对蚕豆叶片的DNA损伤的影响,与重金属吸收的结果相吻合.结果表明,接种处理可显著增加蚕豆叶片的DNA损伤程度,这与接种处理可提高植物的重金属吸收相一致.     

Effect of cadmium on nodulation and N2-fixation of soybean in contaminated soils

The effects of cadmium stress on nodulation, N2-fixation capabilities of the root nodule, the change in ultrastructure of the root nodule, soybean growth, and the distribution of cadmium in plants were studied. The results obtained show that the nodulation of soybean roots was greatly inhibited by the addition of Cd, especially at the addition level of 10 and 20 mg kg(-1) soil. The inhibition of plant growth, especially the root growth, increased as the cadmium concentration increased, with deleterious effects observed for the roots. The weight ratio of soybean root/leaf decreased as the Cd concentration increased, which might explain the reason for nodulation decreases. The results also indicate that N2-fixation of root nodule was stimulated to some extent at the low levels of Cd addition, but decreased sharply with further increase of the Cd concentration. High Cd levels were also associated with changes in the ultrastructure of root nodule, in which the effective N2-fixing area was reduced and the N2-fixing cells in the area also reduced. In addition, the results also reveal that the content of Cd in different parts of the plants was as follows: roots > stems > seeds, indicating that the accumulation of Cd by roots is much larger than that by any other part of the soybean plant, and might cause deleterious effects to root systems.     

Effect of EDTA and NTA on cadmium distribution and translocation in Pennisetum purpureum Schum cv. Mott

The primary objective of this research was to investigate the cadmium (Cd) distribution in Pennisetum purpurem (Napier grass) in the presence of 30 mg/L of Cd and different types and concentrations of chelating agents (ethylenediaminetetraacetic acid disodium dihydrate (EDTA), nitrilotriacetic acid (NTA), and EDTA-NTA mixtures). Plant samples were collected every 15 d during a 105-d experimental period. Accumulation of Cd in each part of the plant was determined using atomic absorption spectrometer (AAS), and the distribution of Cd was determined by laser ablation inductively coupled plasma mass spectrometer (LA-ICP-MS) and synchrotron radiation micro X-ray fluorescence (SR-micro-XRF). The highest concentrations of Cd accumulation of 889?±?53 mg kg^?1 in the underground part (roots) and 265?±?26 mg kg^?1 in the aboveground part (stems and leaves) in the presence of 1:1 M ratio of Cd:EDTA after 30 d of exposure were observed. Plants grown in the presence of either NTA or EDTA-NTA mixtures showed significant lower Cd accumulation levels. The LA-ICP-MS analysis showed that Cd was primarily accumulated in the aboveground part (stems and leaves), especially in the xylem and intercalary meristem. In addition, translocation factor was very low. Thus, P. purpurem could be considered as a candidate plant for cadmium phytostabilization.

     

Enhancing phytoextraction of Cd by combining poplar (clone “I-214”) with Pseudomonas fluorescens and microbial consortia

The plant–microorganism combinations may contribute to the success of phytoextraction of heavy metal-polluted soil. The purpose of this study was to investigate the effects of cadmium (Cd) soil concentration on selected physiological parameters of the poplar clone “I-214” inoculated at root level with a strain (BT4) of Pseudomonas fluorescens and a commercial product based on microbial consortia (Micosat F Fito®). Plants were subjected to Cd treatment of 40 mg kg^?1 in greenhouse. The effects of plant–microbe interactions, plant growth, leaf physiology, and microbial activity were periodically monitored. Metal concentration and translocation factors in plant tissues proved enhanced Cd uptake in roots of plants inoculated with P. fluorescens and transfer to shoots in plants inoculated with Micosat F Fito®, suggesting a promising strategy for using microbes in support of Cd uptake. Plant–microbe integration increased total removal of Cd, without interfering with plant growth, while improving the photosynthetic capacity. Two major mechanisms of metal phytoextraction inducted by microbial inoculation may be suggested: improved Cd accumulation in roots inoculated with P. fluorescens, implying phytostabilization prospective and high Cd transfer to shoots of inoculated plants, outlining enhanced metal translocation.     

Effects of cadmium on the photosynthetic activity in mature and young leaves of soybean plants

Cadmium (Cd) is a widely spread pollutant and can be easily taken up by crop from soil, resulting in a serious health issue for humans. The objective of this study was to comparatively investigate the photosynthetic activity, chlorophyll a fluorescence, chlorophyll contents, and spectral reflectance in mature and young leaves of soybean plants after being treated with different concentrations of Cd for 10 days. The photosynthetic rate, chlorophyll contents, actual photochemical efficiency of PSII, and photochemical quenching in the young leaves decreased more significantly with increasing concentrations of Cd in the nutrient solution, compared with those in the mature leaves, though the young leaves had less Cd concentrations. Thus, there was more excessive excited energy produced in the young leaves than that in the mature leaves. In the young leaves, due to more excessive excited energy, more reactive oxygen species may be generated, which further damaged the photosynthetic apparatus. It was supported by the fact that the decrease of reflectance in near-infrared wavelengths of the young leaves was more noticeable than that of the mature leaves. In addition, the chlorophyll a fluorescence transients of the young leaves was significantly different from that in the mature leaves, indicating that the electron transport of young leaves were inhibited much more severely than that of the mature leaves. These observations imply that the responses of photosynthetic activity of soybean leaves to Cd stress depend on their growth stage, and the Cd-induced inhibition of photosynthetic activity might be attributed to the decrease in chlorophyll contents and the decrease in mesophyll CO₂ assimilation ability cause by the Cd, which further decreased the consumption of ATP and NADPH, leading to accumulation of NADPH on the acceptor sides of the PSI, and then feedback inhibited electron transport in chloroplasts.     

Effects of high dose copper on plant growth and mineral nutrient (Zn,Fe, Mg,K, Ca) uptake in spinach

Loessal soil is one of the main cultivated soils in northwest China. Part of its distribution area was irrigated with industrial wastewater in past three decades. This caused heavy metal contamination in the soil. It had induced toxicity on crops and also threatened local human health for now. Based on a field plot experiment, effects of different Cu concentrations (from 45 to 2000 mg kg^?1) in loessal soil on spinach plant growth and uptake of mineral nutrients (Zn, Fe, Mg, K, and Ca) by spinach were investigated. The Cu addition increased available concentrations of mineral nutrients in loessal soil and concentrations of Cu, Zn, Mg, and Ca in roots. The translocation of mineral nutrients from roots to leaves was inhibited under Cu addition, inducing their decrease in leaves. The EC₁₀ and EC₅₀ of soil Cu in relative dry weights of leaves were 240.33 mg kg^?1 and 1205.04 mg kg^?1, respectively. The PLS-PM analysis showed that available concentrations of nutrients in soil were only affected by Cu in soil positively, nutrients in roots were mainly affected by Cu in soil and Cu in leaves positively, nutrients in leaves were mainly affected by Cu in roots negatively, translocation of nutrients in spinach and plant growth were principally affected by Cu in leaves negatively, and the total effect of Cu in leaves on nutrients in roots and leaves, translocation of nutrients in spinach, and plant growth was the highest. Our results indicated that the phytotoxicity of Cu including spinach growth inhibition and mineral disorder in spinach was mainly affected by the Cu concentrations in leaves.