Influence of cadmium on water relations, stomatal resistance, and abscisic Acid content in expanding bean leaves

Ten day old bush bean plants (Phaseolus vulgaris L. cv Contender) were used to analyze the effects of 3 micromolar Cd on the time courses of expansion growth, dry weight, leaf water relations, stomatal resistance, and abscisic acid (ABA) levels in roots and leaves. Control and Cd-treated plants were grown for 144 hours in nutrient solution. Samples were taken at 24 hour intervals. At the 96 and 144 hour harvests, additional measurements were made on excised leaves which were allowed to dry for 2 hours. From the 48 hour harvest, Cd-treated plants showed lower leaf relative water contents and higher stomatal resistances than controls. At the same time, root and leaf expansion growth, but not dry weight, was significantly reduced. The turgor potentials of leaves from Cd-treated plants were nonsignificantly higher than those of control leaves. A significant increase (almost 400%) of the leaf ABA concentration was detected after 120 hours exposure to Cd. But Cd was found to inhibit ABA accumulation during drying of excised leaves. It is concluded that Cd-induced decrease of expansion growth is not due to turgor decrease. The possible mechanisms of Cd-induced stomatal closure are discussed.     

Cadmium distribution and chemical fate in soybean plants

The distribution and chemical behavior of Cd²⁺ in tissues and its chemical form in xylem water of soybean plants (cv. Williams) were investigated. Following root absorption, Cd is strongly retained by roots, with only 2% of the accumulated Cd being transported to leaves; as much as 8% was transported to seeds during seed filling. In vivo xylem exudates contained two anionic Cd complexes in addition to inorganic forms of Cd. Once accumulated in root and leaf tissues, Cd rapidly equilibrated between the insoluble, soluble, and organelle fractions. Of the solubles, which contain 50% of the Cd, >50% was associated with components of >10,000 molecular weight, and <8% was associated with <500 molecular weight components. Cadmium accumulated in soybean seeds was primarily associated with cotyledons. Fractionation of seeds showed the soy proteinate and soy whey to contain 32 and 50% of the accumulated Cd, respectively.     

不同类型镉积累水稻细胞镉化学形态及亚细胞和分子分布

利用水培试验结合亚细胞组分分级分离和凝胶过滤等技术,研究了水稻根和叶中镉的化学结合形态及其亚细胞和分子分布,比较了低镉积累品种“广源占No.3”和高镉积累品种 “珍桂矮”的差异.结果表明：随着营养液中镉浓度的升高,根和叶亚细胞镉含量显著上升,大部分镉积累在细胞壁（F_Ⅰ）和细胞可溶部分（F_Ⅲ）.高镉积累品种“珍桂矮”根和叶中可溶部分镉含量显著高于低镉积累品种“广源占No.3”.根和叶各种形态镉中,以氯化钠提取态占优势,其次是醋酸提取态,盐酸提取态镉含量最低.与“广源占No.3”相比,“珍桂矮”中迁移性较强的去离子水和乙醇提取态镉比例较高.凝胶过滤结果表明,两种类型的水稻可溶部分镉的出峰位置与样品流份中可溶性蛋白的出峰位置大致相同.可溶部分中的镉大多与分子量为3kD的物质结合,属于植物鳌合肽（PCs）或低分子量物质.“广源占No.3” 根系中镉与PCs配合的组分（Cd-PCs）含量远小于“珍桂矮”.“广源占No.3”细胞可溶部分较低的镉含量以及根系中较少的Cd-PCs形成量,降低了镉的移动及其向地上部转运的可能性.     

Coordination and speciation of cadmium in corn seedlings and its effects on macro- and micronutrients uptake

The effect of cadmium (Cd) on both the absorption of important nutrients and the synthesis of low molecular weight thiols (LMWTs) was investigated in corn plants. The inductively coupled plasma-optical emission spectroscopy results demonstrated that the concentration of Cd in tissues (mainly in roots) increased as the concentration in the medium increased. In addition, the concentration of phosphorus increased in roots of Cd treated plants but remained at normal concentration in shoots. On the other hand, the uptake of sulfur (S) followed a similar trend as the Cd uptake. The concentration of S and the production of LMWT were found to increase significantly upon exposure to Cd. The results of the X-ray absorption spectroscopy analyses indicated that Cd within tissues was bound to S ligands with interatomic distances of 2.51–2.52 Å. These results confirm a strong linkage between S uptake and the production of LMWT upon exposure to Cd.     

Nickel in Plants: II. Distribution and Chemical Form in Soybean Plants

The gross tissue distribution, intracellular fate, and chemical behavior of Ni²⁺ in soybean plants (Glycine max cv. Williams) were investigated. Following root absorption, Ni was highly mobile in the plant, with leaves being the major sink in the shoots for Ni during vegetative growth. A senescence >70% of the Ni present in the shoot was remobilized to seeds. Fractionation of root and leaf tissues showed >90% of the Ni to be associated with the soluble fraction of tissues; ultrafiltration of the solubles showed >77% of the Ni to be associated with the 10,000 to 500 molecular weight components of both roots and leaves. Chemical characterization of the soluble components (10,000 to 500 and >500 molecular weight) by thin layer chromatography and electrophoresis resolved a number of Ni-containing organic complexes. Major Ni-containing components formed in the root are transported in the xylem stream, and undergo partial modification on deposition in leaves. Nickel accumulated in seeds is primarily associated with the cotyledons. Chemical fractionation of cotyledon components showed 80% of the Ni to be associated with the soluble whey fraction, while 70% of this fraction was composed of Ni-containing components with molecular weight <10,000.     

Aldehyde oxidase in roots, leaves and seeds of barley (Hordeum vulgare L.)

Aldehyde oxidase (AO, EC 1.2.3.1) proteins in leaves, roots and seeds of barley (Hordeum vulgare L.) plants were studied. Differences in substrate specificity and mobility in native PAGE between AO proteins extracted from roots, leaves and seeds have been observed. Four clear bands of AO reacting proteins were detected in barley plants capable of oxidizing a number of aliphatic and aromatic aldehydes such as indole-3-aldehyde, acetaldehyde, heptaldehyde, and benzaldehyde. Mouse polyclonal antibodies raised against purified maize AO cross-reacted with barley AO proteins extracted from roots, leaves and seeds. At least three different AO proteins were detected in roots on the basis of their mobility during PAGE after native Western blot analysis while in leaves and seeds only one polypeptide cross-reacted with the antibody. SDS-immunoblot analysis showed marked differences in molecular weight between subunits of the AO bands extracted from roots, leaves and seeds. Two distinct subunit bands were observed in roots, with relatively close molecular weights (160 kDa and 145 kDa), while a single subunit with a molecular weight of 150 kDa was observed in leaf and seed extracts.Menadione, a specific and potent inhibitor of animal AO did not affect barley AO proteins. Root and leaf AO differed in their thermostability and susceptibility to exogenous tungstate. The AO proteins in plants may be a group of enzymes with different substrate specificity, tissue distribution and presumably fulfilling different metabolic roles in each plant organ.     

Effects of jasmonate and some other signalling factors on bean and onion growth during the initial phase of cadmium action

Short-time direct and indirect effects of 25 μM Cd on the growth of dicotyledon (Phaseolus coccineus) and monocotyledon (Allium cepa) plants were investigated in the presence of inhibitors of ethylene synthesis, NADPH oxidase, and the octadecanoid pathway. Only 5 min-long action of Cd was enough for inhibition of growth in bean roots, but its recovery time was extended to several days. After 7 h treatment, Cd was significantly accumulated in bean roots, but maximum H₂O₂ accumulation was seen after 1 h. Cd-induced H₂O₂ accumulation decreased especially after addition of ethylene inhibitor silver thiosulphate (STS). Low Cd accumulation and high growth inhibition were observed also in bean leaves and in A. cepa roots. The inhibitors of the octadecanoid pathway greatly weakened the inhibitory effect of Cd in P. coccineus roots, while no significant effect was observed in A. cepa. NADPH oxidase and ethylene blockade reversed (in the case of bean plants and indirectly treated A. cepa plants) or significantly diminished Cd action. Cd-induced growth inhibition of P. coccineus leaves was also alleviated by most inhibitors of the jasmonate pathway and by STS. These results indicate that Cd may have indirect and direct effects on growth processes.     

Modifications in endopeptidase and 20S proteasome expression and activities in cadmium treated tomato (<Emphasis Type="Italic">Solanum lycopersicum</Emphasis> L.) plants

The effects of cadmium (Cd) on cellular proteolytic responses were investigated in the roots and leaves of tomato (Solanum lycopersicum L., var Ibiza) plants. Three-week-old plants were grown for 3 and 10 days in the presence of 0.3–300 μM Cd and compared to control plants grown in the absence of Cd. Roots of Cd treated plants accumulated four to fivefold Cd as much as mature leaves. Although 10 days of culture at high Cd concentrations inhibited plant growth, tomato plants recovered and were still able to grow again after Cd removal. Tomato roots and leaves are not modified in their proteolytic response with low Cd concentrations (≤3 μM) in the incubation medium. At higher Cd concentration, protein oxidation state and protease activities are modified in roots and leaves although in different ways. The soluble protein content of leaves decreased and protein carbonylation level increased indicative of an oxidative stress. Conversely, protein content of roots increased from 30 to 50%, but the amount of oxidized proteins decreased by two to threefold. Proteolysis responded earlier in leaves than in root to Cd stress. Additionally, whereas cysteine- and metallo-endopeptidase activities, as well as proteasome chymotrypsin activity and subunit expression level, increased in roots and leaves, serine-endopeptidase activities increased only in leaves. This contrasted response between roots and leaves may reflect differences in Cd compartmentation and/or complexation, antioxidant responses and metabolic sensitivity to Cd between plant tissues. The up-regulation of the 20S proteasome gene expression and proteolytic activity argues in favor of the involvement of the 20S proteasome in the degradation of oxidized proteins in plants. This paper is dedicated to Nathalie Galtier (1964–2005), who was senior researcher at the INRA Research Center, Villenave d’Ornon, France.     

Differential antioxidative responses to cadmium in roots and leaves of pea (Pisum sativum L. cv. Azad).

Pea (Pisum sativum L. cv. Azad) plants exposed to 4 and 40 microM of Cd for 7 d in hydroponic culture were analysed with reference to the distribution of metal, the accumulation of biomass and the metal's effects on antioxidants and antioxidative enzymes in roots and leaves. Cd-induced a decrease in plant biomass. The maximum accumulation of Cd occurred in roots followed by stems and leaves. An enhanced level of lipid peroxidation and an increased tissue concentration of hydrogen peroxide (H2O2) in both roots and leaves indicated that Cd caused oxidative stress in pea plants. Roots and leaves of pea plants responded differently to Cd with reference to the induction of enhanced activities of most of the enzymes monitored in the present study. These differential responses to Cd were further found to be associated with levels of Cd to which the plants were exposed. Cd-induced enhancement in superoxide dismutase (SOD) activity was more at 40 microM than at 4 microM in leaves. While catalase (CAT) prominently increased in leaves both at 4 and 40 microM Cd, ascorbate peroxidase (APX) showed maximum stimulation at 40 microM Cd in roots. Enhancement in glutathione reductase (GR) activity was also more at 40 microM than at 4 microM Cd in roots. While glutathione peroxidase (GPOX) activity decreased in roots and remained almost unmodified in leaves, glutathione S-transferase (GST) showed pronounced stimulation in both roots and leaves of pea plants exposed to 40 microM Cd. Increased activities of antioxidative enzymes in Cd-treated plants suggest that they have some additive function in the mechanism of metal tolerance in pea plants.     

Cadmium-binding components in soybean plants

Soybean (Glycine max L.) plants exposed to ¹⁰⁹Cd readily absorb the element. Differential centrifugation of leaf, stem, and root homogenates followed by radioassay showed that Cd was associated primarily with the 105,000g supernatant. Separation of this fraction by gel chromatography and subsequent analysis by radioassays revealed that ¹⁰⁹Cd was bound to macromolecules of >50,000, 13,800, and 2,280 molecular weights. The >50,000 and 2,280 molecular weight fractions probably are nonspecific binding of Cd to normal cell components. The 13,800 molecular weight ¹⁰⁹Cd-bound component was found to be inducible by cadmium. It had a high ultraviolet absorbance at 254 nm and a low absorbance at 280 nm at pH 8.6.     

Glutathione and phytochelatin contents in tomato plants exposed to cadmium

The effect of cadmium on growth and contents of glutathione (GSH) and phytochelatins (PCs) were investigated in roots and leaves of tomato plants (Lycopersicon esculentum Mill. cv. 63/5 F1). The accumulation of Cd increased with external Cd concentrations and was considerably higher in roots than in leaves. Dry mass production decreased under Cd treatment especially in leaves. In both roots and leaves, exposure to Cd caused an appreciable decline in GSH contents and increase in PCs synthesis proportional to Cd concentrations in the growth medium. At the same Cd concentration, PCs production was higher in roots than in leaves. The implication of glutathione in PC synthesis was strongly suggested by the use of buthionine sulfoximine (BSO). The major fraction of Cd accumulated by tomato roots was in the form of a Cd-PCs complex.     

Cadmium-induced biochemical responses of Vallisneria spiralis

The following study was carried out to investigate the cadmium (Cd) accumulating potential of Vallisneria. After subjecting plants to different concentrations of Cd, it was observed that plants are able to accumulate ample amount of metal in their roots (5,542 μg g^?1 dw) and leaves (4,368 μg g^?1 dw) in a concentration- and duration-dependent manner. Thus, it is evident that the accumulation in roots was 1.3 times higher than the shoots. It was also noted that with increasing Cd accumulation, roots of the plant appeared darker in color and harder in texture. In response to metal exposure, amount of low molecular weight antioxidants such as cysteine and nonprotein thiols (NP-SH) and activity of enzymes such as APX and GPX were significantly enhanced at lower concentrations of Cd, followed by decline at higher doses. It was also observed that in exposed plants, activity of APX enzyme was higher in roots (ca. 3 times) as compared to leaves. However, chlorophyll and protein content was found to decline significantly in a dose-dependent manner. Results suggested that due to its high accumulation potential, Vallisneria may be effectively grown in water bodies moderately contaminated with Cd.     

Variation in cadmium accumulation potential and tissue distribution of cadmium in tobacco

Variation in Cd accumulation between Nicotiana species but not varieties has been observed in seedlings grown in solution culture with moderate-to-low levels of Cd. Nicotiana tabacum has been characterized as a leaf and root accumulator while Nicotiana rustica is shown to be primarily a root accumulator, having about half the leaf Cd per gram dry weight of N. tabacum. This phenotype is retained in the mature N. rustica plant. To characterize these two species which differ in their modes of Cd accumulation, tissue Cd distribution, partitioning of metal in soluble and insoluble fractions and the contribution of soluble Cd-binding proteins (peptides) to total plant Cd was assessed using mature solution cultured plants. Metal accumulation was highest in the most mature leaves and in young roots. The preponderance of young roots in N. rustica may, in part, account for low leaf/high root Cd accumulation in this species. While Cd-binding peptides appear to be a principal form of Cd in leaves and roots of seedlings and these also occur in mature leaves, Cd is equally distributed between soluble (about 80% as Cd-binding peptide) and uncharacterized insoluble forms in mature plant roots.     

Calcium modifies Cd effect on runner bean plants

The effect of different Ca concentrations in the growth medium on the toxicity of 25 μM CdSO₄ was studied in runner bean plants (var. Pi kny Ja ) at two different growth stages of primary leaves. In young plants growing in a medium with low level of Ca a treatment with Cd for 12 days resulted in Ca accumulation in roots, a strong reduction of the leaf area, a decreased monogalactosyl diacylglycerol/digalactosyl diacylglycerol ratio and efficiency of the photosynthetic apparatus. In leaves of older plants growing under the same conditions, and surviving Cd treatment, a high accumulation of Ca but a low one of Cd, chlorosis of leaves, a decrease of the ratio monogalactosyl diacylglycerol/digalactosyl diacylglycerol and photosynthetic activity were shown. At a high level of Ca in the nutrient medium plant roots showed a remarkably high specificity to accumulate Cd but the toxic effect of the metal on plant growth parameters and content of pigments was decreased. No changes were observed in the level of galactolipids, but changes in fluorescence quenching were recorded. Calcium deficit enhanced the effect of Cd toxicity, including primary photochemistry, whereas excess Ca reduced toxic effects, while it is increasing the nonphotochemical quenching of excitation energy.     

镉在旱柳中亚细胞分布及存在的化学形态

以2个旱柳无性系幼苗为材料,通过营养液培养并结合差速离心与化学试剂提取法,分析了不同浓度Cd2+胁迫下旱柳叶和根中Cd的亚细胞分布及其存在的化学形态.结果显示,(1)随着培养介质Cd2+浓度升高,旱柳无性系幼苗叶和根中各亚细胞组分Cd含量随之增加.叶片的Cd主要富集于细胞壁、叶绿体和可溶性部分,它们的含量分别占65%～69%、14%～22%、6.8%～7.7%,仅少量Cd发现于膜部分;而根中Cd主要积累于细胞壁和可溶性部分,其中含量分别占59%～66%和14%～25%,Cd在根亚细胞组分中积累量依次为细胞壁>可溶性部分>质体>膜部分.(2)旱柳体内Cd以不同的化学形态存在,大部分为HCl(FHCl)、NaCl(FNaCl)、醋酸(HAC,FHAC)提取态,极少部分为乙醇(EtOH,FEtOH)和水提取态(Fwater),叶和根中5种Cd提取态含量依次为FHCl>FNaCl>FHAC>Fwater>FEtOH,而叶和根中HCl和NaCl提取态Cd占有比例大于30%以上.研究表明,旱柳无性系中Cd主要与蛋白质和有机酸螯合或以金属磷酸盐沉淀的形态存在,其根、叶的细胞壁和液泡在Cd忍耐与解毒中起到重要作用.     

重金属镉在番茄及其害虫西花蓟马中的积累与传递

重金属镉(Cd)是农田土壤中的重要污染源,可在植物和植食性昆虫中积累与传递。本文采用水培法,研究了不同浓度的Cd在番茄Solanum lycopersicum不同组织和在其重要害虫西花蓟马Frankliniella occidentalis体内的积累量。结果表明,随着水培营养液中Cd浓度的增加,番茄植株的根、茎和叶中Cd含量呈增长趋势。根中Cd的积累量远高于茎和叶,当水培溶液中Cd含量为20 mg/L时,根、茎和叶中的积累量分别达19 333.67±233.38、122.67±6.84和147.33±2.96 mg/Kg(干重)。随着Cd浓度的增加,番茄根、茎和叶的鲜重和干重均显著下降。西花蓟马取食Cd处理的番茄叶片后,体内Cd显著积累,最高达1.95±0.36 mg/Kg。同时,Cd积累量的提高进一步影响了以番茄叶片为食的西花蓟马的适合度,降低了其存活率。除对照外,番茄茎-叶的转移系数和叶片对Cd的富集系数均大于1,叶片表现出较强的富集能力。而在所有的试验浓度处理中,西花蓟马对Cd的富集系数和转移系数均小于1,表明Cd未在其体内产生生物放大作用。研究结果明确了Cd在番茄各组织及其害虫中的积累和传递水平,为揭示重金属在农业生态系统食物链中的富集效应提供了基础数据。     

Absorption,distribution and binding of cadmium and zinc in irrigated rice plants

Summary Rice plants are grown in nutrient solutions (Hoagland-Arnon 1) containing different concentrations of cadmium (0.005; 0.010 and 0.100 ppm) and one concentration of zinc (0.070 ppm) traced respectively with¹⁰⁹Cd and⁶⁵Zn. Distribution and localization of both elements are studied at plant and cellular levels. Contrary to zinc, cadmium remains fixed by radicular sites and its translocation index is very low. Centrifugation of roots homogenates showed that % Cd recovered in the residue increases with Cd concentration and emphasize the high capacity of Cd fixation by walls when Cd external concentration is high. The relative distributions of Cd and Zn present at the same concentration in the growing medium are quite identical. In the 100,000×g supernatant fraction of roots, both elements are associated with compounds of low molecular weight, Cd being bound to higher MW ligands than zinc (1000 to 3000 for zinc). The fact that Cd was recovered in fractions in which most of³⁵S-cysteine was present suggests that in plants, like in animals, peptides rich in — SH residues may play a role in the fixation of heavy metals. Contribution no. B 101488 of the Biology-Radiation Protection and Medical Research Programme DG XII CEC.     

Different proportions of cadmium occur as Cd-binding phytochelatin complexes in plants

The aim was to determine cadmium (Cd) speciation in various plants, between buffer-soluble and acid-soluble Cd, and also within the buffer-soluble Cd. A better understanding of Cd speciation shows the relative importance of different biological mechanisms for Cd sequestration. Roots of Pistia stratiodes , Eichhornia crassipes , Agrostis gigantea , Deschampsia caespitosa and wheat Triticum turgidum var. durum were analyzed. Buffer extractions solubilized varying proportions of Cd, ranging from 12% in Eichhornia to 83% in Agrostis . The proportion increased with time of Cd exposure in Pistia . It also increased in wheat roots as the external Cd rose from 0.05 to 0.5 μ M and was lowest in old leaves and highest in roots. The remaining Cd was extractable with acid. Gel filtration resolved buffer-soluble Cd into three peaks distinct from inorganic Cd. Two complexes with phytochelatins and related polythiols were present in all cases, inorganic Cd being prominent only in Eichhornia extracts. The phytochelatin complexes accounted for 2% of the root Cd in Eichhornia to 78% in Agrostis . In wheat, phytochelatins bound 82% of the Cd in roots, 19% in young leaves and 12% in old leaves. The cysteine-rich protein metallothionein from wheat was detected immunologically in the void volume of gel filtrations of old and young leaves, but not of roots, and was distinct from the two phytochelatin-based complexes. Speciation of Cd in the various plants indicated that phytochelatins were not necessarily the major ligands of Cd.     

Analysis of Phaseolus vulgaris Response to Its Association with Trichoderma harzianum (ALL-42) in the Presence or Absence of the Phytopathogenic Fungi Rhizoctonia solani and Fusarium solani

The present study was carried out to evaluate the ability of Trichoderma harzianum (ALL 42-isolated from Brazilian Cerrado soil) to promote common bean growth and to modulate its metabolism and defense response in the presence or absence of the phytopathogenic fungi Rhizoctonia solani and Fusarium solani using a proteomic approach. T. harzianum was able to promote common bean plants growth as shown by the increase in root/foliar areas and by size in comparison to plants grown in its absence. The interaction was shown to modulate the expression of defense-related genes (Glu1, pod3 and lox1) in roots of P. vulgaris. Proteomic maps constructed using roots and leaves of plants challenged or unchallenged by T. harzianum and phytopathogenic fungi showed differences. Reference gels presented differences in spot distribution (absence/presence) and relative volumes of common spots (up or down-regulation). Differential spots were identified by peptide fingerprinting MALDI-TOF mass spectrometry. A total of 48 identified spots (19 for leaves and 29 for roots) were grouped into protein functional classes. For leaves, 33%, 22% and 11% of the identified proteins were categorized as pertaining to the groups: metabolism, defense response and oxidative stress response, respectively. For roots, 17.2%, 24.1% and 10.3% of the identified proteins were categorized as pertaining to the groups: metabolism, defense response and oxidative stress response, respectively.     

Mobility and Translocation of Heavy Metals from Mine Tailings in Three Plant Species after Amendment with Compost and Biosurfactant

An experiment was performed to determine the effects of mine tailings alone mixed with compost or with compost plus crude biosurfactant on the accumulation of heavy metals (Pb, Zn, Cu, Cr, Cd, and Ni) in Acacia retinodes, Nicotiana glauca, and Echinochloa polystachya. The plants were grown in soil, mine tailings, and mine tailings containing compost over a period of seven and five months for shrubs or grass, respectively. The plants Acacia retinodes and Nicotiana glauca grown in mine tailings containing compost showed increases in dry biomass (from 62 to 79%) compared with plants in only tailings. Heavy metals accumulated in the roots and leaves showed high translocation rates of Cr in N. glauca, Cd in A. retinodes, and Ni in E. polystachya. Concentrations of heavy metals in the three plants irrigated with crude biosurfactant were not significantly different from those irrigated with water. Zn and Cd fractions within mine tailings containing compost were bound to carbonate, Pb was bound to residues, and Cu was bound to Fe-oxides. Cd had the highest mobility factor followed in order by Zn, Pb, and Cu. The elevated concentrations of Pb in roots and the low translocation rate for N. glauca and A. retinodes indicate that they are suitable for phytostabilizing Pb and Zn.