Mobilisation of iron and manganese by plant-borne and synthetic metal chelators

Phytosiderophores released by roots of iron-deficient grasses mobilise Fe, Zn, Mn and Cu in calcareous soils. Mobilisation of Fe, Zn and Cu can be explained as the chelation of these metal cations by phytosiderophores. Mobilisation of Mn could not be so explained because phytosiderophores have a much smaller affinity for Mn than for Fe, Cu and Zn. Model experiments have been made with freshly precipitated Fe(OH)₃ and different soils to study the mobilisation of iron and manganese by plant-borne chelating phytosiderophores, the synthetic metal chelators DTPA and the microbial metal chelator sulphonated ferrioxamine B (FOB). Compared with the synthetic chelator DTPA, the plant-borne chelating phytosiderophores mobilised Fe very efficiently, but no change was observed in the Mn mobilisation by phytosiderophores.Different phytosiderophores, as well as the microbial metal chelator FOB, were used to compare the mobilisation of iron and manganese in a calcareous soil.     

Mobilization of iron by phytosiderophores as affected by other micronutrients

It has been shown previously (Treeby et al., 1989) that phytosiderophores, released by roots of iron deficient grasses (Gramineae), mobilize from calcareous soils not only iron (Fe) but also zinc (Zn), manganese (Mn) and copper (Cu). Mobilization of Fe may therefore be impaired by other micronutrient cations. This has been studied in both, model experiments with Fe hydroxide and with a calcareous soil (15% CaCO₃, pH 8.6) amended with micronutrients as sulfate salts.Mobilization of Fe from Fe hydroxide by phytosiderophores (epi-3-hydroxymugineic acid) was not affected by the addition of CaCl₂, MgSO₄ and MnSO₄, slightly inhibited by ZnSO₄ and strongly inhibited by CuSO₄. In a calcareous soil amended with increasing levels of ZnSO₄, MnSO₄ and CuSO₄, mobilization of Fe by phytosiderophores remained uneffected by Zn and Mn amendments but was progressively impaired by increasing levels of Cu amendment, correlated with corresponding enhancement of Cu mobilization.High concentrations of ZnSO₄ and MnSO₄ and relatively high concentrations of CuSO₄ were required for inhibition of Fe mobilization by phytosiderophores. It is therefore concluded that in most calcareous soils phytosiderophores efficiently mobilize Fe, and that phytosiderophores play an important role in Fe acquisition by grasses grown on calcareous soils.     

Light-mediated release of phytosiderophores in wheat and barley under iron or zinc deficiency

The effect of varied light intensity (50 – 600 mol m-2 s-1) on the rate of phytosiderophore release was studied under zinc (Zn) deficiency using a bread (Triticum aestivum L. cv. Aroona) and a durum wheat cultivar (Triticum durum Desf. cv. Durati) differing in zinc (Zn) efficiency and under iron (Fe) deficiency using a barley cultivar (Hordeum vulgare L. Europe). Plants were grown under controlled environmental conditions in nutrient solution for 15 days (wheat plants) or 11 days (barley plants). Phytosiderophore release was determined by measuring capacity of root exudates to mobilize copper (Cu) from a Cu-loaded resin.With increasing light intensity visual Zn deficiency symptoms such as whitish-brown lesions on leaf blade developed rapidly and severely in wheat, particularly in the durum cultivar Durati. In wheat plants supplied well with Zn, increases in light intensity from 100 to 600 mol m-2 s-1 did not clearly affect the rate of phytosiderophore release. However, under Zn deficiency increases in light intensity markedly enhanced release of phytosiderophores in Zn-deficient Aroona, but not in Zn-inefficient Durati. When Fe-deficient barley cultivar Europe was grown first at 220 mol m-2 s-1 and then exposed to 600 mol m-2 s-1 for 24 and 48 h, the rate of release of phytosiderophores was enhanced about 4-fold and 7-fold, respectively. Transfer of Fe-deficient plants from 600 to 50 mol m-2 s-1 for 48 h reduced the rate of release of phytosiderophores by a factor of 7. The effect of light on phytosiderophore release was similar regardless of whether the rate of phytosiderophore release was expressed per plant or per unit dry weight of roots.The results demonstrate a particular role of light intensity in phytosiderophore release from roots under both Zn and Fe deficiency. It is suggested that in the studies concerning the role of phytosiderophore release in expression of Zn or Fe efficiency among and within cereals, a special attention should be given to the light conditions.     

Effect of iron plaque outside roots on nutrient uptake by rice (Oryza sativa L.). Zinc uptake by Fe-deficient rice

This solution culture study examined the effect of the deposition of iron plaque on zinc uptake by Fe-deficient rice plants. Different amounts of iron plaque were induced by adding Fe(OH)₃ at 0, 10, 20, 30, and 50 mg Fe/L in the nutrient solution. After 24 h of growth, the amount of iron plaque was correlated positively with the Fe(OH)₃ addition to the nutrient solution. Increasing iron plaque up to 12.1 g/kg root dry weight increased zinc concentration in shoots by 42% compared to that at 0.16 g/kg root dry weight. Increasing the amount of iron plaque further decreased zinc concentration. When the amounts of iron plaque reached 24.9 g/kg root dry weight, zinc concentration in shoots was lower than that in shoots without iron plaque, implying that the plaque became a barrier for zinc uptake. While rice plants were pre-cultured in –Fe and +Fe nutrient solution in order to produce the Fe-deficient and Fe-sufficient plants and then Fe(OH)₃ was added at 20, 30, and 50 mg Fe/L in nutrient solution, zinc concentrations in shoots of Fe-deficient plants were 54, 48, and 43 mg/kg, respectively, in contrast to 32, 35, and 40 mg/kg zinc in shoots of Fe-sufficient rice plants. Furthermore, Fe(OH)₃ addition at 20 mg Fe/L and increasing zinc concentration from 0.065 to 0.65 mg Zn/L in nutrient solution increased zinc uptake more in Fe-deficient plants than in Fe-sufficient plant. The results suggested that root exudates of Fe-deficient plants, especially phytosiderophores, could enhance zinc uptake by rice plants with iron plaque up to a particular amount of Fe.     

Mobilization of cadmium and other metals from two soils by root exudates of Zea mays L., Nicotiana tabacum L. and Nicotiana rustica L.

Soluble root exudates were collected from three plants (Nicotiana tabacum L., Nicotiana rustica L. and Zea mays L.), grown under axenic and hydroponic conditions, in order to study their metal-solubilizing ability for Cd and other cations (Cu, Fe, Mn, Ni, Zn). Nicotiana spp. and Zea mays L. root exudates differed markedly in C/N ratio, sugars vs. amino acids ratio and organic acids content. Metals from two soils were extracted with either root exudate solutions, containing equal amounts of organic carbon, or distilled water as control. In the presence or absence of root exudates, the solubility of Fe and Mn was much higher than of the four other metals tested. Root exudates increased the solubilities of Mn and Cu, whereas those of Ni and Zn were not affected. Root exudates of Nicotiana spp. enhanced the solubility of Cd. The extent of Cd extraction by root exudates (N. tabacum L. N. rustica L. Zea mays L.) was similar to the order of Cd bioavailability to these three plants when grown on soil. An increase in Cd solubility in the rhizosphere of apical root zones due to root exudates is likely to be an important cause of the relatively high Cd accumulation in Nicotiana spp.     

Effect of Trichoderma asperellum strain T34 on iron, copper, manganese, and zinc uptake by wheat grown on a calcareous medium

Rhizosphere microbes may enhance nutrient uptake by plants. Here we studied the effect of Trichoderma asperellum inoculation on the uptake of Fe, Cu, Mn, and Zn by wheat (Triticum aestivum L) grown in a calcareous medium. To this end, an experiment involving two factors, namely Fe enrichment (ferrihydrite enrichment and non-enrichment of the growing medium), and inoculation/non-inoculation with Trichoderma asperellum strain T34, was performed twice under the same conditions. The increase in Fe availability as a result of ferrihydrite enrichment did not enhance plant dry matter production. The effect of T34 on the concentration of Fe, Cu, Mn and Zn, and the total amount of Cu, Mn, and Zn in the aerial parts differed depending on the degree of ferrihydrite enrichment. Inoculation with T34 increased Fe concentration in Fe-deficient media, thus revealing a positive effect of this microorganism on Fe nutrition in wheat. However, T34 significantly decreased the concentration and total amount of Cu, Mn, and Zn in the aerial parts, but only in ferrihydrite-enriched medium. This adverse effect of T34 on Cu, Mn, and Zn uptake by wheat plants may have been related to conditions of restricted availability where potential competition for nutrients between microorganisms and plants can be more marked.     

Mobilization of Cu and Zn by root exudates of dicotyledonous plants in resin-buffered solutions and in soil

It has been frequently suggested that root exudates play a role in trace metal mobilization and uptake by plants, but there is little in vivo evidence. We studied root exudation of dicotyledonous plants in relation to mobilization and uptake of Cu and Zn in nutrient solutions and in a calcareous soil at varying Cu and Zn supply. Spinach (Spinacia oleracea L.) and tomato (Lycopersicon esculentum L.) were grown on resin-buffered nutrient solutions at varying free ion activities of Cu (pCu 13.0–10.4) and Zn (pZn 10.1–6.6). The Cu and Zn concentrations in the nutrient solution increased with time, except in plant-free controls, indicating that the plant roots released organic ligands that mobilized Cu and Zn from the resin. At same pCu, soluble Cu increased more at low Zn supply, as long as Zn deficiency effects on growth were small. Zinc deficiency was observed in most treatment solutions with pZn ≥ 9.3, but not in nutrient solutions of a smaller volume/plant ratio in which higher Zn concentrations were observed at same pZn. Root exudates of Zn-deficient plants showed higher specific UV absorbance (SUVA, an indicator of aromaticity and metal affinity) than those of non-deficient plants. Measurement of the metal diffusion flux with the DGT technique showed that the Cu and Zn complexes in the nutrient solutions were highly labile. Diffusive transport (through the unstirred layer surrounding the roots) of the free ion only could not explain the observed plant uptake of Cu and of Zn at low Zn²⁺ activity. The Cu and Zn uptake by the plants was well explained if it was assumed that the complexes with root exudates contributed 0.4% (Cu) or 20% (Zn) relative to the free ion. In the soil experiment, metal concentrations and organic C concentrations were larger in the solution of planted soils than in unplanted controls. The SUVA of the soil solution after plant growth was higher for unamended soils, on which the plants were Zn-deficient, than for Zn-amended soils. In conclusion, root exudates of dicotyledonous plants are able to mobilize Cu and Zn, and plants appear to respond to Zn deficiency by exuding root exudates with higher metal affinity.     

The role of phytosiderophores in acquisition of iron and other micronutrients in graminaceous species: An ecological approach

Phytosiderophores (PS) are released in graminaceous species (Gramineae) under iron (Fe) and zinc (Zn) deficiency stress and are of great ecological significance for acquisition of Fe and presumably also of Zn. The potential for release of PS is much higher than reported up to now. Rapid microbial degradation during PS collection from nutrient solution-grown plants is the main cause of this underestimation. Due to spatial separation of PS release and microbial activity in the rhizosphere a much slower degradation of PS can be assumed in soil-grown plants. Concentrations of PS up to molar levels have been calculated under non-sterile conditions in the rhizosphere of Fe-deficient barley plants.Besides Fe, PS mobilize also Zn, Mn and Cu. Despite this unspecific mobilization, PS mobilize appreciable amounts of Fe in calcareous soils and are of significance for chlorosis resistance of graminaceous species. In most species the rate of PS release is high enough to satisfy the Fe demand for optimal growth on calcareous soils.In contrast to the chelates ZnPS and MnPS, FePS are preferentially taken up in comparison with other soluble Fe compounds. In addition, the specific uptake system for FePS (translocator) is regulated exclusively by the Fe nutritional status. Therefore, it seems appropriate to retain the term phytosiderophore instead of phytochelate.     

Phytosiderophores released by graminaceous species promote 59Fe-uptake in citrus

Chlorosis-susceptible fruit trees growing on calcareous soils have been observed to recover in the presence of grass cover species. However, the physiological mechanisms behind this phenomenon are only scarcely understood. An investigation was carried out to verify whether citrus plants can use ⁵⁹Fe solubilized from a sparingly soluble source by the phytosiderophores (PS) released from graminaceous species. Experiments were performed in hydroponics, using two citrus rootstocks differing in their sensitivity to Fe-deficiency in the field (Poncirus trifoliata × Citrus paradisi, citrumelo “Swingle”, highly susceptible, and Citrus aurantium L., moderately tolerant). Barley (Hordeum vulgare L., cv Europa) was used as a model species for PS-releasing graminaceous plants. Fe-deficient citrus plants increased ⁵⁹Fe-uptake from ⁵⁹Fe-hydroxide supplied inside a dialysis tube, when Fe-deficient barley plants or PS-containing barley root exudates were present in the uptake solution, this effect being particularly evident for the susceptible rootstock. ⁵⁹Fe-uptake from ⁵⁹Fe-hydroxide was also enhanced in Fe-deficient citrumelo “Swingle” in the presence of Fe-deficient Poa pratensis L. and Festuca rubra L., two perennial grasses normally grown in association with fruit trees; no effect was found when Fe-sufficient grasses were employed. The uptake of ⁵⁹Fe by the susceptible citrus rootstock increased in proportion to the amount of 2′-deoxymugineic acid (DMA), the major PS released by Fe-deficient F. rubra, present in the uptake solution. The beneficial effect of F. rubra or P. pratensis was evident from the leaf re-greening observed when Fe-deficient citrumelo “Swingle” plants were grown in association with the grasses in pots filled with a calcareous soil. Leaf re-greening was not observed when citrumelo “Swingle” plants and yellow stripe 3 (ys3) maize (Zea mays L.) mutant plants, unable to release PS, were co-cultivated in pots filled with calcareous soil, unless exogenous PS were added to the soil. Results indicate that graminaceous cover species can improve the Fe-nutrition of fruit trees grown on calcareous soils by enhancing Fe-availability.     

Identification of two novel phytosiderophores secreted by perennial grasses

It has been suggested that some perennial grasses secrete phytosiderophores in response to iron (Fe) deficiency, but the compounds have not been identified. Here, we identified and characterized the phytosiderophores secreted by two perennial grasses, Lolium perenne cv. Tove and Poa pratensis cv. Baron. Root exudates were collected from the roots of Fe-deficient grasses and then purified with various chromatographies. The structure of the purified compounds was determined using both nuclear magnetic resonance and fast atom bombardment mass spectrometry. Both species secreted phytosiderophores in response to Fe deficiency, and the amount of phytosiderophores secreted increased with the development of Fe deficiency. The type of phytosiderophores secreted differed with plant species; L. perenne cv. Tove secreted 3-epihydroxy-2'-deoxymugineic acid (epiHDMA), 2'-deoxymugineic acid (DMA) and an unknown compound, whereas P. pratensis cv. Baron secreted DMA, avenic acid A (AVA) and an unknown compound. Purification and subsequent analysis with nuclear magnetic resonance and mass led to identification of the two novel phytosiderophores; 3-hydroxy-2'-deoxymugineic acid (HDMA) from L. perenne, and 2'-hydroxyavenic acid A (HAVA) from P. pratensis. Both novel phytosiderophores have similar chelating activity to known phytosiderophores.     

Cloning two genes for nicotianamine aminotransferase, a critical enzyme in iron acquisition (Strategy II) in graminaceous plants

Nicotianamine aminotransferase (NAAT), the key enzyme involved in the biosynthesis of mugineic acid family phytosiderophores (MAs), catalyzes the amino transfer of nicotianamine (NA). MAs are found only in graminaceous plants, although NA has been detected in every plant so far investigated. Therefore, this amino transfer reaction is the first step in the unique biosynthesis of MAs that has evolved in graminaceous plants. NAAT activity is dramatically induced by Fe deficiency and suppressed by Fe resupply. Based on the protein sequence of NAAT purified from Fe-deficient barley (Hordeum vulgare) roots, two distinct cDNA clones encoding NAAT, naat-A and naat-B, were identified. Their deduced amino acid sequences were homologous to several aminotransferases, and shared consensus sequences for the pyridoxal phosphate-binding site lysine residue and its surrounding residues. The expression of both naat-A and naat-B is increased in Fe-deficient barley roots, while naat-B has a low level of constitutive expression in Fe-sufficient barley roots. No detectable mRNA from either naat-A or naat-B was present in the leaves of either Fe-deficient or Fe-sufficient barley. One genomic clone with a tandem array of naat-B and naat-A in this order was identified. naat-B and naat-A each have six introns at the same locations. The isolation of NAAT genes will pave the way to understanding the mechanism of the response to Fe in graminaceous plants, and may lead to the development of cultivars tolerant to Fe deficiency that can grow in calcareous soils.     

Application of chelator-buffered nutrient solution technique in studies on zinc nutrition in rice plant (Oryza sativa L.)

It has been difficult to impose different degrees of Zn deficiency on Poaceae species in nutrient solution because most chelators which would control Zn to low activities also bind Fe³⁺ so strongly that Poaceae species cannot obtain adequate Fe. Recently, a method has been developed to provide buffered Fe²⁺ at levels adequate for rice using Ferrozine (FZ), and use of other chelators to buffer the other micronutrient cations. The use of Fe²⁺ buffered with FZ in nutrient solutions in which Zn is buffered with HEDTA or DTPA was evaluated for study of Zn deficiency in rice compared to a conventional nutrient solution technique. The results showed that growth of rice plants in FZ+HEDTA-buffered nutrient solution was similar to that in the conventional nutrient solution. Severe zinc deficiency symptoms were observed in 28-day-old rice seedlings cultured with HEDTA-buffered nutrient solution at Zn²⁺ activities < 10^-10.6 M. With increasing free Zn²⁺ activities, concentrations of Zn, Fe, Cu, and Mn in shoots and roots were quite similar for the FZ+HEDTA-buffered nutrient solution and the conventional nutrient solution techniques. The percentages of water soluble Zn, Fe, Cu and Mn in shoots with HEDTA-buffered nutrient solution were also similar to those with the conventional solution. However, with DTPA-buffered nutrient solution, the rice seedlings suffered severe Fe deficiency; adding more FeFZ₃ corrected the Fe-chlorosis but shifted microelement buffering. Further, much higher total Zn concentrations are required to provide adequate Zn²⁺ in DTPA-buffered solutions, and the contents of Mn and Cu in shoots and roots cultured with DTPA-buffered solutions were much higher than those with the conventional or HEDTA-buffered solutions. In conclusion, DTPA-buffered nutrient solutions are not suitable but the FZ/HEDTA-buffered nutrient solution technique can be used to evaluate genotypic differences in zinc efficiency in rice.     

Trace elements in the hydrologic cycle of a forest ecosystem

Summary Concentrations of Cu, Fe, Mn, and Zn were measured in bulk atmospheric precipitation, throughfall, stemflow, and soil solutions at 10−, 15−, 25−, and 30-cm depths, in aEucalyptus globulus forest in the Berkeley hills, California, during the 1974–75 wet season after each main storm event. Litter and plant samples were analyzed. There was some similarity in the behavior of Cu, Fe, and Zn, but Mn behaved differently. Mn and Zn are largely deposited on the forest canopy by impaction during dry-deposition episodes, whereas most of the Cu and Fe input occurs in rain. For the hydrologic components measured, concentrations of Cu and Fe increase in the order: precipitation<throughfall<stemflow <soil solutions. For Zn the order is: precipitation<stemflow<throughfall<soil solutions. Concentrations of Cu, Zn, and Fe in the soil solution fluctuate with downward movement of wetting fronts and are negatively correlated with pH. Concentrations of Fe in soil solution are about 10 times greater than those of throughfall and stemflow; the corresponding relative differences for Cu and Zn were much less. Plant uptake of Mn exceeds that of Cu, Zn, and Fe. The increases in Mn concentrations from precipitation to throughfall and stemflow are much greater than those for Cu, Zn, and Fe because precipitation has very low Mn concentrations. The concentration series for Mn is: precipitation<soil solutions<throughfall<stemflow. Concentrations of Mn in the soil solution are negatively correlated with pH. During the dry summer Mn accumulates in the soil, but is quickly flushed by early rains of the wet season.     

The effect of pH,electrolytes and temperature on the rhizosphere geochemistry of phytosiderophores

Background and aims

Graminaceous plants are grown worldwide as staple crops under a variety of climatic and soil conditions. They release phytosiderophores for Fe acquisition (Strategy II). Aim of the present study was to uncover how the rhizosphere pH, background electrolyte and temperature affect the mobilization of Fe and other metals from soil by phytosiderophores.

Methods

For this purpose a series of kinetic batch interaction experiments with the phytosiderophore 2′-deoxymugineic acid (DMA), a calcareous clay soil and a mildly acidic sandy soil were performed. The temperature, electrolyte concentration and applied electrolyte cation were varied. The effect of pH was examined by applying two levels of lime and Cu to the acidic soil.

Results

Fe mobilization by DMA increased by lime application, and was negatively affected by Cu amendment. Mobilization of Fe and other metals decreased with increasing ionic strength, and was lower for divalent than for monovalent electrolyte cations at equal ionic strength, due to higher adsorption of metal-DMA complexes to the soil. Metal mobilization rates increased with increasing temperature leading to a faster onset of competition; Fe was mobilized faster, but also became depleted faster at higher temperature. Temperature also affected biodegradation rates of metal-DMA complexes.

Conclusion

Rhizosphere pH, electrolyte type and concentration and temperature can have a pronounced effect on Strategy II Fe acquisition by affecting the time and concentration ‘window of Fe uptake’ in which plants can benefit from phytosiderophore-mediated Fe uptake.

     

Induction of the Root Cell Plasma Membrane Ferric Reductase (An Exclusive Role for Fe and Cu)

Induction of ferric reductase activity in dicots and nongrass monocots is a well-recognized response to Fe deficiency. Recent evidence has shown that Cu deficiency also induces plasma membrane Fe reduction. In this study we investigated whether other nutrient deficiencies could also induce ferric reductase activity in roots of pea (Pisum sativum L. cv Sparkle) seedlings. Of the nutrient deficiencies tested (K, Mg, Ca, Mn, Zn, Fe, and Cu), only Cu and Fe deficiencies elicited a response. Cu deficiency induced an activity intermediate between Fe-deficient and control plant activities. To ascertain whether the same reductase is induced by Fe and Cu deficiency, concentration- and pH-dependent kinetics of root ferric reduction were compared in plants grown under control, -Fe, and -Cu conditions. Additionally, rhizosphere acidification, another process induced by Fe deficiency, was quantified in pea seedlings grown under the three regimes. Control, Fe-deficient, and Cu-deficient plants exhibited no major differences in pH optima or Km for the kinetics of ferric reduction. However, the Vmax for ferric reduction was dramatically influenced by plant nutrient status, increasing 16- to 38-fold under Fe deficiency and 1.5- to 4-fold under Cu deficiency, compared with that of control plants. These results are consistent with a model in which varying amounts of the same enzyme are deployed on the plasma membrane in response to plant Fe or Cu status. Rhizosphere acidification rates in the Cu-deficient plants were similarly intermediate between those of the control and Fe-deficient plants. These results suggest that Cu deficiency induces the same responses induced by Fe deficiency in peas.     

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.     

Physiological responses of wheat genotypes grown in chelator-buffered nutrient solutions with increasing concentrations of excess HEDTA

The chelator-buffered nutrient solutions containing excess chelator have been used frequently in the micronutrient research, but potential toxicity of the excess chelator has not been ascertained. The present study was conducted to test effects of four concentrations of excess HEDTA [ N-(2-hydroxyethyl)ethylenedinitrilotriacetic acid] and two levels of total Zn on growth, root exudation, and nutrient uptake and transport by Triticum aestivum L. (cv. Aroona) and Triticum turgidum L. conv. durum (Desf.) MacKey (cv. Durati) genotypes differing in tolerance to Zn deficiency. Excess HEDTA at 50 μM reduced root and shoot growth and caused visual toxicity symptoms (necrotic lesions) on leaves; these effects were generally absent at lower concentrations of excess HEDTA. Root exudation of phytosiderophores increased with increasing concentrations of excess HEDTA at deficient and sufficient Zn levels, and was higher in Zn-deficiency-tolerant Aroona than in Zn-deficiency-sensitive Durati wheat. Shoot and root Zn concentrations showed a saturable response to increasing Zn²⁺ activities in solution. Excess HEDTA at 50 μM caused an increase in shoot concentrations of Fe and a decrease in concentrations of Mn and Cu. An average rate of Zn uptake increased with an increase in Zn²⁺ ionic activity in solution, with Zn-deficiency-tolerant Aroona having a higher rate of Zn uptake than Zn-deficiency-sensitive Durati in the deficiency range of Zn²⁺ activities. Average uptake rates of Mn and Cu decreased with an increase in concentration of excess HEDTA. Similar observations were noted for transport of Mn and Cu to shoots, while Zn transport to shoots was proportional to Zn²⁺ activities in solution. It was concluded that excess HEDTA at 50 μM adversely affects wheat growth and physiology, while excess of 25 μM or less does not cause measurable toxicity. This revised version was published online in June 2006 with corrections to the Cover Date.     

Overcoming Fe deficiency by a transgenic approach in rice

Iron (Fe) is an essential microelement for plant growth. Fe availability is particularly limited on calcareous soils, which have high pH. Approximately 30% of the world's soils are considered calcareous with low Fe availability, which results in extensive areas of Fe deficiency in plants. Some graminaceous plants are known to secrete high amounts of mugineic acid family phytosiderophores (MAs) under Fe deficiency. This Fe acquisition system is called the Strategy-II mechanism. Tolerance to Fe deficiency in graminaceous plants is thought to depend on the quantity of MAs secreted by plants under Fe deficiency stress. This system was utilized to enhance the tolerance of rice to low Fe availability. Transgenic rice expressing the barley naat genes, one of the genes for the enzymes on the biosynthetic pathway of MAs, showed tolerance to low Fe availability when grown in a calcareous soil.     

Effect of peanut mixed cropping with gramineous species on micronutrient concentrations and iron chlorosis of peanut plants grown in a calcareous soil

To gain a better understanding of the mechanisms of improvement of iron nutrition of peanut (Arachis hypogaea L.) intercropped with maize (Zea mays L.) in calcareous soil, both greenhouse and field experiments were conducted to investigate the rhizosphere (phytosiderophores) effects from maize, barley, oats and wheat with different phytosiderophores release rates on iron nutrition and other micronutrients in calcareous soil. Six cropping treatments were examined in a greenhouse experiment: peanut grown separately in monoculture, normal peanut/maize intercropping (two genotypes: Danyu13, Zhongdan12), peanut/barley intercropping, peanut/oats intercropping, and peanut/wheat intercropping. Additionally, we investigated in a field experiment the same five cropping systems as the greenhouse experiment (maize/peanut intercropping not including Zhongdan12). Our results show that the chlorophyll and active Fe concentrations in the young leaves of the peanut in the intercropping system with different gramineous species were much higher than those of the peanut in monoculture. In greenhouse conditions, the Fe concentration in the shoots of peanut plants grown in the intercropping systems of two maize genotypes separately were 1.40–1.44, 1.47–1.64 and 1.15–1.42 times higher respectively than those of peanut plants grown in monocropping at 55, 60 and 70 days. In particular, the Fe concentration in shoots of peanut plants grown in the intercropping systems of barley, oats and wheat were not only higher than those in monocropping but also higher than those in peanut intercropped cropping with maize. In the field, the concentration of Fe in shoot of intercropped peanut plants in rows 1–3 from gramineous species were significantly higher than in monocropping at the flowering stage. Simultaneously with iron nutrition variation in peanut, Zn and Cu concentrations of intercropped grown peanut increased significantly compared to those in monocropping in the greenhouse experiment, and different intercropping treatments generally increased the Zn and Cu content in the shoot of peanut in the field. Systemic mechanisms may be involved in adaptation to nutrient stresses at the whole plant level. The study suggests that a reasonable intercropping system of nutrient efficient species should be considered to prevent or mitigate iron and zinc deficiency of plants in agricultural practice.