黑麦草生长过程中有机酸对镉毒性的影响

研究了低分子量有机酸草酸、柠檬酸、乙酸及高分子量有机酸胡敏酸对黑麦草(Lolium Loinn)生长过程中Cd毒性的影响.结果表明,随着低分子量有机酸浓度增加,Cd毒性有所增强,致使黑麦草中的叶绿素含量降低及黑麦草的生物量降低,递降顺序是草酸＜乙酸＜柠檬酸.而施入胡敏酸后,Cd毒性逐渐减弱,黑麦草中的叶绿素含量及黑麦草生物量逐渐增加.对低分子量有机酸而言,无论迁移到黑麦草茎叶中,还是迁移到黑麦草根系中的Cd,随着施入的有机酸浓度增加,增加顺序为柠檬酸＞乙酸＞草酸.对胡敏酸而言,迁移到黑麦草茎叶和根系中的Cd,随着施入的胡敏酸浓度增加,Cd含量减少,说明其具有降低Cd毒性的作用.另外,根系中Cd含量明显高于茎叶中Cd含量,由此得知,黑麦草根系对Cd有较强的富集作用,并阻止Cd向茎叶中迁移.     

Detection and quantification of ligands involved in nickel detoxification in a herbaceous Ni hyperaccumulator Stackhousia tryonii Bailey

Field-collected, young plants of Ni hyperaccumulator Stackhousia tryonii, grown in a glasshouse for 20 weeks, were exposed to low- (available Ni concentration in the native serpentine soil, i.e. 60 microg g(-1) dry soil) and high- (external application of 1000 ppm) Ni concentrations in the substrate. Nickel concentration in the freeze-dried leaf tissues increased from 3700 microg g(-1) to 13 700 microg g(-1) with soil Ni supplementation, of which >60% was extracted with dilute acid (0.025 M HCl). Nickel supplementation also elicited a 575%, 211%, and 37% increase in the final concentrations of oxalic, citric, and malic acids, respectively, in leaf tissues. Malic acid was the dominant organic acid, followed by citric and oxalic acids. The molar ratio of Ni to malic acid was 1.0, consistent with a role for malate as a ligand for Ni in hyperaccumulating plants, supporting detoxification/transport and storage of this heavy metal in S. tryonii. The total amino acid concentrations in the xylem sap did not change with Ni supplementation (21.7+/-3.7 mM and 17.9+/-5 mM, respectively, for low- and high-nickel-treated plants). Glutamine was the major amino acid in both the low- and high-Ni-treated plants. The concentration of glutamine decreased by >60%, with a corresponding increase in alanine, aspartic acid, and glutamic acid, on exposure to high Ni. A role of amino acids in Ni complexation and transport in S. tryonii is not immediately apparent.     

Roles of Organic Acids and Nitrate in the Long-Distance Transport of Cobalt in Xylem Saps of <Emphasis Type="Italic">Alyssum murale</Emphasis> and <Emphasis Type="Italic">Trifolium subterraneum</Emphasis>

Roles of organic acids and nitrate in the long-distance transport of cobalt (Co) in xylem saps of hyperaccumulator Alyssum murale and non-hyperaccumulator Trifolium subterraneum were studied under hydroponic conditions. Organic acids (oxalic, malic, malonic, citric, and fumaric) and nitrate in xylem sap samples were separated and determined simultaneously by reversed-phase high performance liquid chromatography after solid-phase extraction with nanosized hydroxyapatite. Results indicated that Co treatment significantly increased the concentrations of xylem oxalic and malic acids for the hyperaccumulator A. murale compared to the control but significantly decreased the concentrations of xylem nitrate and malonic acid; concentrations of citric acid in xylem sap samples did not show significant difference between the control and Co treatments. By analyzing the relationship between the concentrations of organic acids, nitrate, and concentrations of Co in xylem saps, it could be concluded that oxalic and malic acids in xylem saps seemed to participate in the long-distance Co translocation process, and citric acid did not relate to the xylem Co transport of A. murale and T. subterraneum. Our work might be very useful for understanding the mechanism of long-distance transport of heavy metals in hyperaccumulator.     

Effects of Low-Molecular-Weight Organic Acids on Gadolinium Accumulation and Transportation in Tomato Plants

Effects of low-molecular-weight organic acids on the accumulation and transportation of gadolinium (Gd) in tomato plants were studied under hydroponic condition. The results indicated that changes of organic acids occurred in the processes of Gd accumulation and transportation in tomato plants which were treated with extraneous Gd solutions. Malic, citric, and succinic acids contributed to both Gd accumulation in roots and transportation in xylem vessels. When Gd was unloaded from the xylem to the leaf cells, formic, lactic, citric, and succinic acids played important roles in Gd accumulation in leaves. When tomato plants were cultured in the uptake solution of Gd-containing malic, citric, or succinic acid for 48 h, the succinic acid in roots and leaves and the malic acid in xylem saps both increased obviously. From the results above, we can conclude that succinic acid had the most important role in Gd accumulation in tomato roots and leaves, while malic acid transported Gd via xylem vessels more effectively.     

Symplastic and apoplastic uptake and root to shoot translocation of nickel in wheat as affected by exogenous amino acids

This study investigated the effect of exogenous amino acids on apoplastic and symplastic uptake and root to shoot translocation of nickel (Ni) in two wheat cultivars. Seedlings of a bread (Triticum aestivum cv. Back Cross) and a durum wheat cultivar (T. durum cv. Durum) were grown in a modified Johnson nutrient solution and exposed to two levels (50 and 100 μM) of histidine, glycine, and glutamine. Application of amino acids resulted in increasing symplastic to apoplastic Ni ratio in roots of both wheat cultivars, although glutamine and glycine were more effective than histidine under our experimental conditions. The amino acid used in the present study generally increased the relative transport of Ni from the roots to shoots in both wheat cultivars. Higher amounts of Ni were translocated to wheat shoots in the presence of histidine than the other amino acids studied, which indicated that histidine was more effective in translocation of Ni from roots to shoots. Amino acids used in the present study largely increased root symplastic Ni, but shoot Ni accumulation was much lower than the total Ni accumulation in roots, indicating a large proportion of Ni was retained or immobilized in wheat roots (either in the apoplastic or symplastic space), with only a very small fraction of Ni being translocated from the root to the shoot. According to the results, glutamine and glycine were more effective than histidine in enhancing the symplastic to apoplastic Ni ratio in the roots, while more Ni was translocated from the roots to the shoots in the presence of histidine.     

Effect of Organic Ligands on Accumulation of Copper in Hyperaccumulator and Nonaccumulator Commelina communis

Better understanding of copper uptake and accumulation regulation in plants is critical to the phytoremediation of copper contaminated soil. This study employed a 30-day pot experiment to assess the relationship between organic ligands and copper accumulation in plants. Hyperaccumulator and nonaccumulator varieties of Commelina communis were used, different organic ligands were applied, and the data of copper accumulation in shoots were collected. The six organic ligands included ethylenediaminetetraacetic acid and organic acids (formic acid, citric acid, malic acid, tartaric acid, and succinic acid). The results showed that organic ligands added to culture increased the copper accumulation both varieties. The results of the copper accumulation in shoots agreed with the study of the root uptake kinetics of copper influx. The addition of organic acids could increase copper accumulation in shoots because the copper influx in roots was increased. The results also indicated that the copper influx of hyperaccumulator roots was higher than that of nonaccumulator roots. This is one of the mechanisms by which a hyperaccumulator could amass large amounts of copper in its shoots. In this accumulation process, little effect on the leaf relative water content was in the hyperaccumulator and nonaccumulator of leaves and normal physiological condition of plants.     

Organic acids are not specifically involved in the nitrate-enhanced Zn hyperaccumulation mechanism in Noccaea caerulescens

Nitrogen form has been shown to affect Zn uptake, translocation and storage in the Zn-hyperaccumulating plant Noccaea caerulescens but the biochemical processes are not fully understood. Organic acids and amino acids have been implicated in Zn transport and storage. This study aimed to examine the effect of N form on concentrations of organic acids and amino acids and how these metabolites correlated with Zn hyperaccumulation. Plants were grown in nutrient solution with NO₃⁻, NH₄NO₃ or NH₄⁺, supplied with 50 or 300 μM Zn, and buffered at either pH 4.5 or 6.5. The metabolomic profile was determined by gas chromatography mass spectroscopy. The concentration of Zn in shoots, xylem and roots was greatest for the NO₃⁻, pH 6.5 and 300 μM Zn treatments. For all N forms, the lower growth-medium pH raised xylem sap pH but had no influence on Zn concentration or exudation rate of the xylem sap. Nitrate enhanced organic acid production while NH₄⁺ increased amino acid production. Organic acids in the xylem were more responsive to changes in growth-medium pH than N form, and did not correlate with Zn concentration in shoots, roots or xylem. Serine might be directly involved in Zn hyperaccumulation. Phosphoric acid was associated with reduced Zn accumulation in the shoots. Malic acid was not detected in the shoots but responded to cation uptake more than to Zn specifically in the roots. Citric acid responded to cation uptake more than to Zn specifically in the shoots but did not correlate with Zn concentration in the roots or the xylem sap, or any other cations in the roots. In conclusion, organic acids in N. caerulescens are not specifically involved in Zn hyperaccumulation but are involved in regulating pH in the xylem and cation–anion balance in plants.     

外源有机酸对玉米磷吸收及其生长发育的影响

以玉米品种郑单958为材料,通过施加外源有机酸的盆栽试验方法,设置了土壤中不施磷肥和有机酸(CKo)、仅施磷肥(CKP)、施柠檬酸(Tca)、施苹果酸(Tma)和施草酸(Toa)处理,研究外源有机酸对玉米生长过程中的磷素吸收及其生长发育的影响.结果显示:(1)3种外源有机酸均可显著提高土壤中速效磷含量和磷素利用效率,Tca、Tma和Toa处理玉米根际土壤的速效磷含量分别为CKo的2.25、1.96和2.04倍,且各处理间的磷素利用效率依次为:CKp>Tca>Tma>Toa>CKo.(2)在拔节期、抽雄期和成熟期,Tca处理玉米的磷素利用效率依次为CKo的1.16、1.05和1.04倍.(3)3种外源有机酸均可显著提高玉米根系体积和根系活力、单株玉米的氮磷钾的累积量、玉米的生物量及相对籽粒产量.研究表明,外源柠檬酸、苹果酸和草酸均可显著提高玉米根际土壤中固态磷的溶解,促进植株体内营养元素的累积,保证植物生长发育的需要,从而提高玉米的生物量和产量;并以外源柠檬酸处理对玉米磷吸收及其生长发育效果最好.     

Influence of arbuscular mycorrhiza on organic solutes in maize leaves under salt stress

A pot experiment was conducted to examine the effect of the arbuscular mycorrhizal (AM) fungus, Glomus mosseae, on plant biomass and organic solute accumulation in maize leaves. Maize plants were grown in sand and soil mixture with three NaCl levels (0, 0.5, and 1.0 g kg⁻¹ dry substrate) for 55 days, after 15 days of establishment under non-saline conditions. At all salinity levels, mycorrhizal plants had higher biomass and higher accumulation of organic solutes in leaves, which were dominated by soluble sugars, reducing sugars, soluble protein, and organic acids in both mycorrhizal and non-mycorrhizal plants. The relative abundance of free amino acids and proline in total organic solutes was lower in mycorrhizal than in non-mycorrhizal plants, while that of reducing sugars was higher. In addition, the AM symbiosis raised the concentrations of soluble sugars, reducing sugars, soluble protein, total organic acids, oxalic acid, fumaric acid, acetic acid, malic acid, and citric acid and decreased the concentrations of total free amino acids, proline, formic acid, and succinic acid in maize leaves. In mycorrhizal plants, the dominant organic acid was oxalic acid, while in non-mycorrhizal plants, the dominant organic acid was succinic acid. All the results presented here indicate that the accumulation of organic solutes in leaves is a specific physiological response of maize plants to the AM symbiosis, which could mitigate the negative impact of soil salinity on plant productivity.     

Nickel-induced changes in carbon metabolism in wheat shoots

In this study, we analyzed the toxic effect of Ni during the development of wheat shoots. Typical developmental alterations in carbon metabolism-related parameters reflecting changes associated with the transition of the seedlings from heterotrophic to autotrophic metabolism were observed in the control shoots between the 1st and the 4th days. Adverse effects of 50 and 100 μM Ni became evident starting from the 4th day of growth on the metal-containing media. We found that Ni-induced stimulation of phosphoenolpyruvate carboxylase (PEPC) activity coincided with decrease in the ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) level and with declines in net photosynthetic rate (P_N) and stomatal conductance (g_s). Application of Ni resulted in increased activities of several dehydrogenases: glucose-6-phosphate dehydrogenase (G6PDH), 6-phosphogluconate dehydrogenase (6PGDH), isocitrate dehydrogenase (NADP-ICDH) and malate dehydrogenase (NADH-MDH). In contrast, the activities of malic enzymes (NADP-ME and NAD-ME) decreased due to Ni stress. Treatment with Ni led to accumulation of glucose and declined concentration of sucrose as well as considerable increases in concentrations of malic and citric acids. Our results indicate that Ni stress redirects the carbon metabolism of developing wheat shoots to provide carbon skeletons for synthesis of amino acids and organic acids as well as to supply reducing power to sustain normal metabolic processes and to support defense mechanisms against oxidative stress.     

Nickel speciation in the xylem sap of the hyperaccumulator Alyssum serpyllifolium ssp. lusitanicum growing on serpentine soils of northeast Portugal

Nickel speciation was studied in the xylem sap of Alyssum serpyllifolium ssp. lusitanicum, a Ni-hyperaccumulator endemic to the serpentine soils of northeast Portugal. The xylem sap was collected from plants growing in its native habitat and characterized in terms of carboxylic and amino acids content. The speciation of nickel was studied in model and real solutions of xylem sap by voltammetric titrations using Square Wave Voltammetry (SWV). The results showed that Ni transport in the xylem sap occurs mainly as a free hydrated cation (about 70%) and complexed with carboxylic acids, mainly citric acid (18%). Altogether, oxalic acid, malic acid, malonic acid and aspartic acid complexed less than 13% of total Ni. A negligible amount bounded to the amino acids, like glutamic acid and glutamine (<1%). Histidine did not play a role in Ni translocation in the xylem sap of A. serpyllifolium under field conditions. Amino acids are one of the main forms of N transport in the xylem sap, and under field conditions, N is usually a limited nutrient. We hypothesize that the translocation of Ni in the xylem sap as a free ion or chelated with carboxylic acids is ‘cheaper’ in terms of N resources.     

Organic acids reduce aluminum toxicity in maize root membranes

Application of 10–50 μ M Al ions to a plasma membrane-enriched microsomal fraction, isolated from roots of maize ( Zea mays L.), resulted in decreased Mg²-ATPase activity. This was probably caused by changes in membrane structure as detected by the use of spin probes. Both enzymatic activity and membrane structure could in part be protected from Al injury when organic acids, similar to those found in maize root tissue, were administered prior to the metal. When stressed by application of Al ions, the Al-tolerant maize hybrid, W64, maintained higher concentrations of organic acids, especially malic and trans -aconitic, than the Al-sensitive maize hybrid, A632. We hypothesize that citric and malic acid, because of their high stability constants with Al and/or the acid's concentration reduce Al toxicity in maize root tissue, especially in the Al-tolerant line.     

Genotypic variability in phosphorus solubilizing activity of root exudates by pigeonpea grown in low-nutrient environments

A pot experiment confirmed that pigeonpea could efficiently utilize various sources of phosphorus (P) (aluminium phosphate, iron phosphate and apatite), irrespective of genotype. A qualitative assay method for iron (Fe)-P solubilizing activity showed that root exudates collected from P-deficient pigeonpea contained Fe-P solubilizing substances and that they were released mainly from root tips. Citric, malic, malonic, succinic and piscidic acids were identified in root exudates. Citric and piscidic acids release from roots was increased by low-P treatment in all the genotypes tested. The release rates of citric and piscidic acids were affected by the P concentration of shoots rather than that of roots. The pigeonpea roots released approximately 5–100 times more piscidic acid than citric acid depending on P stress status, plant age and genotype. When organic acids were added to Alfisols, citric acid was most capable of mobilizing P from the soil, followed by piscidic acid and malic acid. No correlation was found between genotypic variability in the release rates of citric and piscidic acids from the roots under low-P treatment at hydroponic culture and in the growth and P uptake of plants on Alfisols. Although citric and piscidic acids released from pigeonpea roots may play a partial role in solubilizing unavailable insoluble P in soils, the releases were thought to be an unsatisfactory strategy for explaining genotypic variation in low P availability of pigeonpea.     

Enhanced cadmium accumulation in maize roots—the impact of organic acids

Low molecular weight organic acids are important components of root exudates and therefore, knowledge regarding the mechanisms of cadmium (Cd) uptake and distribution within plants under the influence of organic acids, is necessary for a better understanding of Cd behavior in the plant–soil system. In this study, acetic and malic acids increased the uptake of Cd by maize (Zea mays L. cv. TY2) roots and enhanced Cd accumulation in shoots under hydroponic conditions. Concentration-dependent net Cd influx in the presence and absence of organic acids could be resolved into linear and saturable components. The saturable component followed Michaelis–Menten kinetics, which indicated that Cd uptake across the plasma membrane was transporter-mediated. While the K _m values were similar, the V _max values in the presence of acetic and malic acids were respectively 6.0 and 3.0 times that of the control. Zinc transporters were the most probable pathways for Cd accumulation. It was hypothesized that Cd(II)–organic acid complexes associated with the root zone, could decompose and liberate Cd²⁺ for subsequent absorption by maize roots; and that in the layer of the roots or within the root free space, depletion of Cd²⁺ was buffered by the presence of Cd(II)–organic acid complexes. Plant response to elevated Cd levels involved overproduction of organic acids in maize roots as a resistance mechanism to alleviate Cd toxicity.     

Enhancement of cr(lll) phytoaccumulation

Chromium (III) accumulation in high biomass agricultural crops, sunflower (Helianthus annum) and Indian mustard (Brassica juncea) was studied using four soils (pH 4.6 to 7.6) contaminated with different rates of CrCl₃.6H₂O in the presence of synthetic chelate and organic acids. Chromium is essential for normal glucose metabolism in humans and animals, but its contamination and recovery from soils is of environmental concern. Adding ethylenediaminetetraacetic acid (EDTA), citric acid, or oxalic acid to Cr(III)‐contaminated soils significantly increased Cr concentration in plant shoots and roots. Adding Cr(III) complexes of EDTA, citric acid, and oxalic acid to soils dramatically increased (>200‐fold) Cr concentration in shoots and roots. Plant growth was severely decreased but was dependent on soil type, chelate rate, form, and time of chelate application. Chelates and organic acids enhanced Cr(III) accumulation, but its toxic effects were not avoided. Chromium(III) complexes were as toxic to plants as Cr(VI). The phytoaccumulation and recovery of Cr(III) from soils were limited and depended on soil type.     

Calcium complexes in the xylem sap of apple shoots

Summary Concentrations of total calcium, ionic calcium, citric and malic acids have been measured in xylem sap extracted from apple shoots. Ionic calcium, as measured by an ion selective electrode, was about 50 per cent of the total calcium. The remainder of the soluble calcium was present as complexes with citric and malic acids. The implication of these findings on the mobility of calcium in the xylem is discussed.     

Carboxylate metabolism in sugar beet plants grown with excess Zn

The effects of Zn excess on carboxylate metabolism were investigated in sugar beet (Beta vulgaris L.) plants grown hydroponically in a growth chamber. Root extracts of plants grown with 50 or 100 μM Zn in the nutrient solution showed increases in several enzymatic activities related to organic acid metabolism, including citrate synthase and phosphoenolpyruvate carboxylase, when compared to activities in control root extracts. Root citric and malic acid concentrations increased in plants grown with 100 μM Zn, but not in plants grown with 50 μM Zn. In the xylem sap, plants grown with 50 and 100 μM Zn showed increases in the concentrations of citrate and malate compared to the controls. Leaves of plants grown with 50 or 100 μM Zn showed increases in the concentrations of citric and malic acid and in the activities of citrate synthase and fumarase. Leaf isocitrate dehydrogenase increased only in plants grown with 50 μM Zn when compared to the controls. In plants grown with 300 μM Zn, the only enzyme showing activity increases in root extracts was citrate synthase, whereas the activities of other enzymes decreased compared to the controls, and root citrate concentrations increased. In the 300 μM Zn-grown plants, the xylem concentrations of citric and malic acids were higher than those of controls, whereas in leaf extracts the activity of fumarase increased markedly, and the leaf citric acid concentration was higher than in the controls. Based on our data, a metabolic model of the carboxylate metabolism in sugar beet plants grown under Zn excess is proposed.     

Aluminum resistance in two cultivars of Zea mays L.: Root exudation of organic acids and influence of phosphorus nutrition

Root exudation of organic acids as Al-chelating compounds and P nutrition have been suggested to play a major role in Al-resistance in higher plants. Effects of Al exposure on maize plant growth, and organic acid root content and root exudation under various levels of P nutrition were examined. Sikuani, a Colombian maize cultivar tolerant to acid soils with high Al saturation, and Corso, a Swiss cultivar, were grown in sterile hydroponic conditions for 21 days. Al-caused inhibition of root growth was lower in Sikuani than in Corso. Al effect on plant growth was decreased with increasing P content in roots. Al content in roots increased with increasing P content and was higher in Sikuani than in Corso. When exposed to Al, the contents in root apices as well as the root exudation of citric and malic acids in Corso and citric, malic and succinic acids in Sikuani increased, and were higher in Sikuani than in Corso. Increased PEP carboxylase (PEPC) activity in root apices after Al exposure partially explained the variations of organic acid content in the roots. These Al-induced changes in PEPC activity, organic acid content and exudation were reduced in plants supplied with higher P concentrations during the 21 days prior to treatment. Increased secretion of organic acids after exposure to Al appeared to be specific to Al and was not totally explained by increased root content in organic acids.     

Organic acids promote the uptake of lanthanum by barley roots

Organic acids play an important role in metal uptake by, and accumulation in, plants. However, the relevant mechanisms remain obscure. Acetic, malic and citric acids increased the uptake of lanthanum (La) by barley (Hordeum vulgare) roots and enhanced La content in shoots under hydroponic conditions. Concentration-dependent net La influx in the absence and presence of organic acids yielded nonsaturating kinetic curves that could be resolved into linear and saturable components. The saturable component followed Michaelis-Menten kinetics. The K(m) values were similar; however, the V(max) values in the presence of acetic, malic and citric acids were 4.3, 2.8, 1.5-times that of the control, respectively. Enhanced uptake of La by organic acids was mediated mainly, but not solely, by Ca(2+) channels. X-ray absorption spectroscopic techniques provided evidence of La-oxygen environment and established that La(III) was coordinated to 11 oxygen atoms that are likely to be involved in the binding of La(III) to barley roots via carboxylate groups and hydration of La(III).     

Differential biochemical responses of wheat shoots and roots to nickel stress: antioxidative reactions and proline accumulation

Wheat (Triticum aestivum L. cv. ‘Zyta’) seedlings were treated with 10, 100 and 200 μM Ni. Tissue Ni accumulation, length, relative water content (RWC), proline and H₂O₂ concentrations as well as the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (POD) and glutathione S-transferase (GST) were studied in the shoots and roots after 6 days of Ni exposure. Treatment with Ni, except for its lowest concentration, resulted in a significant reduction in wheat growth. In comparison to the shoots, the roots showed greater inhibition of elongation, which corresponded with higher accumulation of Ni in these organs. Both shoots and roots responded to Ni application with a decrease in RWC and enhancement in proline concentration. Greater dehydration of the shoot tissue was accompanied by more intense accumulation of proline. Treatment of the wheat seedlings with the highest concentration of Ni led to about 60% increase in H₂O₂ concentration in both studied organs. Apart from CAT, constitutive activities of antioxidative enzymes were much higher in the roots than in the shoots. Exposure of the seedlings to Ni resulted in SOD activity decline, which was more marked in the roots. While the shoots showed a substantial decrease (up to 30%) in CAT activity, in the roots the activity of this enzyme remained unchanged. After Ni application APX, POD and GST activities increased several-fold in the shoots, whereas in the roots they were not significantly altered. The results suggest that differential antioxidative responses of the shoots and roots of wheat seedlings to Ni stress might be related to diverse constitutive levels of antioxidant enzyme activities in both organs.