The impact of iron plaque on La and Nd uptake and translocation in rice (Oryza sativa L.)

The effect of root surface iron plaque formation on the uptake, transfer and accumulation of La and Nd in the rice root system was evaluated by using solution cultures. The results showed that La and Nd pollution stress inhibit formation of rice root surface iron plaques. The amount of La and Nd absorbed by the rice root surface iron plaque rose with the increase of La and Nd solution concentrations. Iron plaque formation on the rice root surface significantly decreases the La and Nd concentrations in rice roots and shoots. At growth solution La concentrations of 0.1, 0.5, and 1.0 mmol.L^? 1, concentrations of La in rice roots with induced iron plaques decreased by 17.1%, 37.4%, and 31.2%, respectively, and concentrations of La in rice shoots decreased by 43.9%, 60.6%, and 27.0%, respectively, when compared to plants with non-induced iron plaques. Also, with Nd solution concentrations of 0.1, 0.5, and 1.0 mmol.L^? 1, the Nd concentrations in rice roots and shoots of plants with induced iron plaques decreased by 21.0–31.7% and 22.7–47.5%, respectively when compared to plants with non-induced iron plaques. Iron plaque formation on the rice root surface affects the accumulation and transfer of La and Nd in rice roots. Accumulation of La and Nd was greater in rice roots than in rice shoots regardless of whether the plants had induced or non-induced iron plaques. Transfer coefficients of iron plague on rice root surface and root system under La treatments were both higher than those under Nd treatment. For rice roots and iron plaques on the root surface, the enrichment coefficient in the La treatment group was less than that in the Nd treatment group, while for rice shoots, the enrichment coefficient in the La treatment group was greater than that in the Nd treatment group. Clearly, the mechanisms governing the effect of iron plaque on La and Nd uptake and transfer in the rice root system are rather complicated.     

Evidence for chemical changes on the root surface of tall fescue in response to infection with the fungal endophyte Neotyphodium coenophialum

Endophyte-infected (E+) tall fescue (Festuca arundinacea Schreb.) plants grown in phosphorus (P) deficient soils accumulate more P in roots and shoots than noninfected isolines. In a growth chamber experiment, four tall fescue genotypes DN2, DN4, DN7, and DN11, infected with their naturally occurring strains of Neotyphodium coenophialum (Morgan-Jones & Gams) Glenn, Bacon & Hanlin, and their noninfected isolines (E-), were cultivated in nutrient solution at two P levels: 31 ppm (P+) and 0 ppm (P-) for 4 wk. The Fe³⁺ reducing activity of extracellular reductants and intact root tissues, and total phenolic concentration in roots and shoots were measured. Endophyte infection significantly increased Fe³⁺ reducing activity rate of extracellular reductants (9.6 × 10^-3 mol Fe³⁺ h-1 g-1 root FW) when compared to E- plants (3.9 × 10^-3) and Fe³⁺ reduction rate of intact root tissues (6.16 and 4.48 mol Fe³⁺ h-1 g-1 root FW, respectively for E+ and E- plants). In response to P deficiency, Fe³⁺ reduction rate of intact root tissues increased in E+ plants by 375% when compared to E- plants, whereas no significant differences were observed when P was provided. Total phenolic concentration was 20% greater in shoots of E+ plants than in E- plants. In response to P deficiency, total phenolic concentration significantly increased in roots of E+ plants by 7%, and decreased in roots of E- plants by 10%. The most active Fe³⁺ reducing zones were located along branching of secondary and tertiary roots. The Fe³⁺ reducing activity on the root surface and total phenolic concentration in roots and shoots increased dramatically in response to endophyte infection, especially under P limiting conditions.Visiting Scientist sponsored by the Fulbright Program No. 21133     

A comparison of ferric-chelate reductase and chlorophyll and growth ratios as indices of selection of quince,pear and olive genotypes under iron deficiency stress

Quince (Cydonia oblonga Mill.), pear (Pyrus communis L.) and olive (Olea europaea L.) genotypes were evaluated for their tolerance to iron deficiency stress by growing young plants in three types of aerated nutrient solutions: (1) with iron, (2) without iron or (3) low in iron and with 10 mM bicarbonate. Plants were obtained either from rooted softwood cuttings or from germination of seeds. The degree of tolerance was evaluated with several indices: (1) the chlorophyll content, (2) the root Fe³⁺ reducing capacity and (3) the whole plant relative growth. Fifteen hours before Fe³⁺ reducing capacity determination, iron was applied to the roots of plants with iron-stress, since this method resulted in increasing the reductase activity. All quince and pear genotypes increased the root Fe³⁺ reducing capacity when grown in the treatments for iron-stress, in relation to control plants of the same genotypes. In olive cultivars, the Fe³⁺ reducing capacity was lower in the iron-stress treatments than in the control one. Studying the relationship between relative growth and chlorophyll content for each genotype under iron-stress, in relation to both indices in control plants, a classification of species and genotypes was established. According to that, most olive cultivars and some pear rootstocks and cultivars appear more iron-efficient than quince rootstocks. Our study shows that in some woody species, determining root Fe³⁺ reducing capacity is not the best method to establish tolerance to iron deficiency stress.     

Morphological and physiological responses of root tip cells to Fe<Superscript>2+</Superscript> toxicity in rice

Two genotypes of rice (Oryza sativa L.), Azucena (iron tolerant) and IR64 (iron sensitive), were used to investigate the numbers and survival rates of root border cells (namely, in situ border cells) in plants that were exposed to excess iron (Fe²⁺). Additionally, we examined the changes in the root tip cell morphology and activities of protective enzymes in response to Fe²⁺ toxicity. The results showed that Fe²⁺ toxicity hindered the development of root border cells (RBCs) and that higher Fe²⁺ concentrations caused root cap cell walls to thicken. In the iron-sensitive rice variety, these changes lowered RBC survival rate and lead to programmed cell death. Low concentrations of Fe²⁺ were shown to facilitate the development of RBCs in the iron-tolerant rice variety and that the activities of the protective enzymes superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) were elevated in the iron-tolerant variety, thus suggesting that rice root tips could defend against Fe²⁺ toxicity by producing RBCs, root cap cells, and protective enzymes.     

Redox properties of iron–dithiocarbamates and their nitrosyl derivatives: implications for their use as traps of nitric oxide in biological systems

While the Fe²⁺–dithiocarbamate complexes have been commonly used as NO traps to estimate NO production in biological systems, these complexes can undergo complex redox chemistry. Characterization of this redox chemistry is of critical importance for the use of this method as a quantitative assay of NO generation. We observe that the commonly used Fe²⁺ complexes of N-methyl-D-glucamine dithiocarbamate (MGD) or diethyldithiocarbamate (DETC) are rapidly oxidized under aerobic conditions to form Fe³⁺ complexes. Following exposure to NO, diamagnetic NO–Fe³⁺ complexes are formed as demonstrated by the optical, electron paramagnetic resonance and gamma-resonance spectroscopy, chemiluminescence and electrochemical methods. Under anaerobic conditions the aqueous NO–Fe³⁺–MGD and lipid soluble NO–Fe²⁺–DETC complexes gradually self transform by reductive nitrosylation into paramagnetic NO–Fe²⁺–MGD complexes with yield of up to 50% and the balance is converted to Fe³⁺–MGD and nitrite. In dimethylsulfoxide this process is greatly accelerated. More efficient transformation of NO–Fe³⁺–MGD into NO–Fe²⁺–MGD (60–90% levels) was observed after addition of reducing equivalents such as ascorbate, hydroquinone or cysteine or with addition of excess Fe²⁺–MGD. With isotope labeling of the NO–Fe³⁺–MGD with ⁵⁷Fe, it was shown that these complexes donate NO to Fe²⁺–MGD. NO–Fe³⁺–MGD complexes were also formed by reversible oxidation of NO–Fe²⁺–MGD in air. The stability of NO–Fe³⁺–MGD and NO–Fe²⁺–MGD complexes increased with increasing the ratio of MGD to Fe. Thus, the iron–dithiocarbamate complexes and their NO derivatives exhibit complex redox chemistry that should be considered in their application for detection of NO in biological systems.     

Importance of ferric chelate reductase activity and acidification capacity in root nodules of N2-fixing common bean (Phaseolus vulgaris L.) subjected to iron deficiency

Iron is an essential nutrient for plants, especially in symbiotic N₂-fixing legumes. Although abundant in the soil, iron is generally not available to plants as it is predominantly in an insoluble form (Fe^III) . Mono- and dicotyledonous plants, except Grarnineae, have developed morphological and physiological responses, notably an increase in rhizosphere acidification (H⁺-ATPase) and an enhanced plasma membrane ferric chelate reductase activity (Fe-CR) in the roots. However, studies on the physiological responses of root nodules are lacking. The present study was initiated to investigate the acidification capacity and Fe-CR activity of nodulated roots, and intact nodules, in two contrasting common bean varieties, Coco blanc sensitive to iron deficiency and Flamingo tolerant to iron deficiency. The discovery of an induction of H⁺-ATPase and Fe-CR activities in root nodules of commonbean under iron deficiency, suggests that these organs participate in improving iron availability for the contained bacteroids.     

Remedying the iron-deficient maize plants by new synthetic macromolecular chelating agents

The efficacy of Fe³⁺ complexes of polyethers with 8-quinolinol (8QOH) chelating groups attached to the polymer chain at different positions of the aromatic ring or having different chain length for remedying the iron-deficient maize plants was evaluated. The efficacy of chelates of polymers having terminal 8QOH residues was compared with that of complexes of ethylenediaminetetraacetic acid, 8QOH, mixtures of commercial polyethers with isopropylamino end-groups and 8QOH or FeCl₃.6H₂O. It was found that at 30/25 °C (day/night) and photosynthetic photon flux density 1100–1300 μmol m⁻² s⁻¹, the chlorotic maize plants recovered for 4 days of iron re-supply. An increase in the fresh and dry weight, leaf area, net photosynthetic CO₂ uptake of maize leaves, leaf pigment composition and chlorophyll fluorescence was more pronounced in the plants supplied with Fe³⁺ chelates of polymers bearing 8QOH groups attached at 5-position, compared to the other tested Fe³⁺complexes. The importance of the stability of Fe³⁺ complexes, structure of the chelating agent and the necessity of effective ligand exchange between synthetic chelators and free phytosiderophore in iron uptake by strategy II plants was discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Dependency of Iron Reduction on Development of a Unique Root Morphology in Ficus benjamina L

The activity of the Fe³⁺ reductase of excised adventitious roots of Ficus benjamina L., grown in hydroponic culture without iron, was determined by a colorometric assay simplified by the use of a microplate reader. Reductase activity remained the same from pH 4.5 to 6.5 and decreased sharply above pH 6.5. Acetate buffer inhibited reduction. During early stages of root growth, excised roots did not exhibit Fe³⁺ reductase activity. After several weeks and extensive root system development, Fe³⁺ reduction still was not detectable in primary roots, but intermediate and high rates of reduction occurred in lateral and newly formed root clusters, respectively. Clustered roots only developed on plants grown at 0 or very low (<1 micromolar) iron. Microscopic examination revealed the root cluster to be composed of up to 30 lateral roots, usually less than 1 millimeter in diameter and 1 centimeter in length, that were completely covered with root hairs.     

Iron plaque enhances phosphorus uptake by rice (Oryza sativa) growing under varying phosphorus and iron concentrations

Both solution culture and pot experiments were performed to investigate (a) the effects of external Fe (II) concentrations and forms on the formation of iron plaque on the roots of rice (Oryza sativa) and subsequent P adsorption on iron plaque and shoot P concentrations and (b) the effects of soil moisture regimes on the formation of iron plaque and P adsorption on root surfaces and P accumulation in shoots. The results showed that iron plaque was significantly increased with increasing Fe²⁺ concentrations in the solution culture. The amounts of P adsorbed on the iron plaque were increased significantly with external Fe²⁺ concentrations. Although shoot P concentration was not significantly affected by Fe²⁺ treatment after incubation for 2 days, it was significantly increased in the Fe‐treated plants compared with Fe‐deprived ones after incubation for 4 days. Soil culture experiment showed that the formation of iron plaque on root surfaces was promoted by exogenous iron, with greater amount of iron plaque being formed by addition of ferric hydroxide than of ferric oxide. Phosphorus adsorption on iron plaque also increased with the addition of iron oxides, and increasing soil P increased the amounts of P associated with the iron plaque and shoot P concentration. The amounts of iron plaque were almost sixfold higher under flooding condition than under field capacity condition. Plants pretreated under flooding condition generally had higher shoot P concentrations when they were transplanted to solutions with varying P levels, and this was most pronounced in the treatment with highest solution P concentration. The results suggest that iron plaque acts as a nutrient reservoir for phosphorus in the rhizosphere and helps enhance P acquisition by rice.     

Rapid reduction of iron in horse spleen ferritin by thioglycolic acid measured by dispersive X-ray absorption spectroscopy

Summary The release of iron from ferritin is important in the formation of iron proteins and for the management of diseases in both animals and plants associated with abnormal accumulations of ferritin iron. Much more iron can be released experimentally by reduction of the ferric hydrous oxide core than by chelation of Fe³⁺ which has led to the notion that reduction is also the major aspect of iron release in vivo. Variations in the kinetics of reduction of the mineral core of ferritin have been attributed to the redox potential of the reductant, redox properties of the iron core, the structure of the protein coat, the analytical method used to detect Fe²⁺ and reactions at the surface of the mineral. Direct measurements of the oxidation state of the iron during reduction has never been used to analyze the kinetics of reduction, although Mössbauer spectroscopy has been used to confirm the extent of reduction after electrochemical reduction using dispersive X-ray absorption spectroscopy (DXAS). We show that the near edge of X-ray absorption spectra (XANES) can be used to quantify the relative amounts of Fe²⁺ and Fe³⁺ in mixtures of the hydrated ions. Since the nearest neighbors of iron in the ferritin iron core do not change during reduction, XANES can be used to monitor directly the reduction of the ferritin iron core. Previous studies of iron core reduction which measured by Fe²⁺ · bipyridyl formation, or coulometric reduction with different mediators, suggested that rates depended mainly on the redox potential of the electron donor. When DXAS was used to measure the rate of reduction directly, the initial rate was faster than previously measured. Thus, previously measured differences in reduction rates appear to be influenced by the accessibility of Fe²⁺ to the complexing reagent or by the electrochemical mediator. In the later stages of ferritin iron core dissolution, reduction rates drop dramatically whether measured by DXAS or formation of Fe²⁺ complexes. Such results emphasize the heterogeneity of ferritin core structure.     

Inhibition of Fe2+- and Fe3+- induced hydroxyl radical production by the iron-chelating drug deferiprone

Deferiprone (L1) is an effective iron-chelating drug that is widely used for the treatment of iron-overload diseases. It is known that in aqueous solutions Fe²⁺ and Fe³⁺ ions can produce hydroxyl radicals via Fenton and photo-Fenton reactions. Although previous studies with Fe²⁺ have reported ferroxidase activity by L1 followed by the formation of Fe³⁺ chelate complexes and potential inhibition of Fenton reaction, no detailed data are available on the molecular antioxidant mechanisms involved. Similarly, in vitro studies have also shown that L1–Fe³⁺ complexes exhibit intense absorption bands up to 800 nm and might be potential sources of phototoxicity. In this study we have applied an EPR spin trapping technique to answer two questions: (1) does L1 inhibit the Fenton reaction catalyzed by Fe²⁺ and Fe³⁺ ions and (2) does UV–Vis irradiation of the L1–Fe³⁺ complex result in the formation of reactive oxygen species. PBN and TMIO spin traps were used for detection of oxygen free radicals, and TEMP was used to trap singlet oxygen if it was formed via energy transfer from L1 in the triplet excited state. It was demonstrated that irradiation of Fe³⁺ aqua complexes by UV and visible light in the presence of spin traps results in the appearance of an EPR signal of the OH spin adduct (TMIO–OH, a(N)=14.15 G, a(H)=16.25 G; PBN–OH, a(N)=16.0 G, a(H)=2.7 G). The presence of L1 completely inhibited the OH radical production. The mechanism of OH spin adduct formation was confirmed by the detection of methyl radicals in the presence of dimethyl sulfoxide. No formation of singlet oxygen was detected under irradiation of L1 or its iron complexes. Furthermore, the interaction of L1 with Fe²⁺ ions completely inhibited hydroxyl radical production in the presence of hydrogen peroxide. These findings confirm an antioxidant targeting potential of L1 in diseases related to oxidative damage.     

Oncoidal granular iron formation in the Mesoarchaean Pongola Supergroup,southern Africa: Textural and geochemical evidence for biological activity during iron deposition

We document the discovery of the first granular iron formation (GIF) of Archaean age and present textural and geochemical results that suggest these formed through microbial iron oxidation. The GIF occurs in the Nconga Formation of the ca. 3.0–2.8 Ga Pongola Supergroup in South Africa and Swaziland. It is interbedded with oxide and silicate facies micritic iron formation (MIF). There is a strong textural control on iron mineralization in the GIF not observed in the associated MIF. The GIF is marked by oncoids with chert cores surrounded by magnetite and calcite rims. These rims show laminated domal textures, similar in appearance to microstromatolites. The GIF is enriched in silica and depleted in Fe relative to the interbedded MIF. Very low Al and trace element contents in the GIF indicate that chemically precipitated chert was reworked above wave base into granules in an environment devoid of siliciclastic input. Microbially mediated iron precipitation resulted in the formation of irregular, domal rims around the chert granules. During storm surges, oncoids were transported and deposited in deeper water environments. Textural features, along with positive δ⁵⁶Fe values in magnetite, suggest that iron precipitation occurred through incomplete oxidation of hydrothermal Fe²⁺ by iron‐oxidizing bacteria. The initial Fe³⁺‐oxyhydroxide precipitates were then post‐depositionally transformed to magnetite. Comparison of the Fe isotope compositions of the oncoidal GIF with those reported for the interbedded deeper water iron formation (IF) illustrates that the Fe²⁺ pathways and sources for these units were distinct. It is suggested that the deeper water IF was deposited from the evolved margin of a buoyant Fe²⁺_aq‐rich hydrothermal plume distal to its source. In contrast, oncolitic magnetite rims of chert granules were sourced from ambient Fe²⁺_aq‐depleted shallow ocean water beyond the plume.     

Effect of pH and oxalic acid on the reduction of Fe3+ by a biomimetic chelator and on Fe3+ desorption/adsorption onto wood: Implications for brown-rot decay

Fenton reaction is thought to play an important role in wood degradation by brown-rot fungi. In this context, the effect of oxalic acid and pH on iron reduction by a biomimetic fungal chelator and on the adsorption/desorption of iron to/from wood was investigated. The results presented in this work indicate that at pH 2.0 and 4.5 and in the presence of oxalic acid, the phenolate chelator 2,3-dihydroxybenzoic acid (2,3-DHBA) is capable of reducing ferric iron only when the iron is complexed with oxalate to form Fe³⁺-mono-oxalate (Fe(C₂O₄)⁺). Within the pH range tested in this work, this complex formation occurs when the oxalate:Fe³⁺ molar ratio is less than 20 (pH 2.0) or less than 10 (pH 4.5). When aqueous ferric iron was passed through a column packed with milled red spruce (Picea rubens) wood equilibrated at pH 2.0 and 4.5, it was observed that ferric iron binds to wood at pH 4.5 but not at pH 2.0, and the bound iron could then be released by application of oxalic acid at pH 4.5. The release of bound iron was dependent on the amount of oxalic acid applied in the column. When the amount of oxalate was at least 20-fold greater than the amount of iron bound to the wood, all bound iron was released. When Fe–oxalate complexes were applied to the milled wood column equilibrated in the pH range of 2–4.5, iron from Fe–oxalate complexes was bound to the wood only when the pH was 3.6 or higher and the oxalate:Fe³⁺ molar ratio was less than 10. When 2,3-DHBA was evaluated for its ability to release iron bound to the milled wood, it was found that 2,3-DHBA possessed a greater affinity for ferric iron than the wood as 2,3-DHBA was capable of releasing the ferric iron bound to the wood in the pH range 3.6–5.5. These results further the understanding of the mechanisms employed by brown-rot fungi in wood biodegradation processes.     

Protein- and Metal-dependent Interactions of a Prominent Protein in Mussel Adhesive Plaques

The adhesive plaques of Mytilus byssus are investigated increasingly to determine the molecular requirements for wet adhesion. Mfp-2 is the most abundant protein in the plaques, but little is known about its function. Analysis of Mfp-2 films using the surface forces apparatus detected no interaction between films or between a film and bare mica; however, addition of Ca²⁺ and Fe³⁺ induced significant reversible bridging (work of adhesion W_ad ≈ 0.3 mJ/m² to 2.2 mJ/m²) between two films at 0.35 m salinity. The strongest observed Fe³⁺-mediated bridging approaches the adhesion of oriented avidin-biotin complexes. Raman microscopy of plaque sections supports the co-localization of Mfp-2 and iron, which interact by forming bis- or tris-DOPA-iron complexes. Mfp-2 adhered strongly to Mfp-5, a DOPA-rich interfacial adhesive protein, but not to another interfacial protein, Mfp-3, which may in fact displace Mfp-2 from mica. In the presence of metal ions or Mfp-5, Mfp-2 adhesion was fully reversible. These results suggest that plaque cohesiveness depends on Mfp-2 complexation of metal ions, particularly Fe³⁺ and also by Mfp-2 interaction with Mfp-5 at the plaque-substratum interface.     

Complexes of hydroxamates III.: Equilibrium and kinetic studies on the reactions of iron(III) with monohydroxamic acids

Spectral analysis of iron(III) complexes with acetohydroxamate (AX) and histidinehydroxamate (HX) in the UV-visible region revealed that many species may exist in pH range 1.0–7.5. The solution spectra were unstable in pH range ~2.7–4.0. Different species were obtained from fresh solutions and overnight solutions. The difference was rationalized due to hydrolysis and/or polymerization of complexes in solution, especially in pH range 2.7–4.0. The kinetics of the reactions of Fe(III) with AX and HX were accomplished, and mechanisms were suggested for both systems. In both cases, Fe³⁺ and FeOH²⁺ species were found to be the active species in the complex formation of 1:1 complex.     

FORMATION AND MORPHOLOGY OF AN IRON PLAQUE ON THE ROOTS OF TYPHA LATIFOLIA L. GROWN IN SOLUTION CULTURE

Roots of Typha latifolia L. exposed to Fe²⁺ under reduced conditions in solution culture developed visible coatings (plaques) of an oxidized Fe compound that extended as much as 15-17 μm into the rhizosphere. Iron concentrations were significantly less and discoloration was not apparent on the surface of roots exposed to Fe-(BPDS)₃, Fe³⁺, Fe-EDDHA, and Fe-EDTA. The extent of plaque formation increased with the concentration of Fe²⁺ in solution and with pH of the solution in the range of 3.0 to 4.6. Above pH 4.6, oxidation of Fe²⁺ in the culture solution may have reduced precipitation of Fe on the root surface. Plaque development was most extensive approximately 1.0 cm from the root tip, but all root surfaces showed some Fe staining. Scanning electron micrographs of plaqued roots, grown both in solution culture and in the field, provided support for a model of cast formation by oxidation and precipitation of Fe on external cell surfaces.     

Iron toxicity and stress-induced ethylene production in rice leaves

The relationship among iron toxicity, bronzing symptom, and stress-induced ethylene production (SEP) was investigated in detached rice (Oryza sativa L.) leaves during the vegetative-ripening stage and in whole plants during the vegetative stage. When Fe²⁺ (200 mg L^-1) was applied to the detached leaf through a transpiration stream, SEP was higher in the first leaf than in the second and third leaves from the top and maximal around the panicle primordia initiation stage. The genotype difference in SEP was more pronounced in the second and third leaves than in the first leaf. Bronzing intensity increased as SEP increased; iron concentration increase during treatment in the tissue did not correlate with bronzing intensity or with SEP among the 16 genotypes tested. When the roots of an intact plant were exposed to 300 mg L^-1 of Fe²⁺ in culture solution little stress-induced ethylene was produced. By partially or totally derooting the plant, however, stress-induced ethylene was evoked, indicating that roots reduced the Fe²⁺ uptake so that little stress ethylene is produced in the intact plant. Leaf tissue tolerance for Fe²⁺ may contribute to genotype differences in iron toxicity tolerance of rice plants when roots are injured during transplanting or exposed to toxic substances in the soil.     

Investigation of iron pools in cucumber roots by Mössbauer spectroscopy: direct evidence for the Strategy I iron uptake mechanism

Distinct chemical species of iron were investigated by Mössbauer spectroscopy during iron uptake into cucumber roots grown in unbuffered nutrient solution with or without ⁵⁷Fe-citrate. Mössbauer spectra of iron deficient roots supplied with 10–500 μM ⁵⁷Fe-citrate for 30–180 min and 24 h and iron-sufficient ones, were recorded. The roots were analysed for Fe concentration and Fe reductase activity. The Mössbauer parameters in the case of iron-sufficient roots revealed high-spin iron(III) components suggesting the presence of Fe^III-carboxylate complexes, hydrous ferric oxides and sulfate–hydroxide containing species. No Fe^II was detected in these roots. However, iron-deficient roots supplied with 0.5 mM ⁵⁷Fe^III-citrate for 30 min contained significant amount of Fe^II in a hexaaqua complex form. This is a direct evidence for the Strategy I iron uptake mechanism. Correlation was found between the decrease in Fe reductase activity and the ratio of Fe^II–Fe^III components as the time of iron supply was increased. The data may refer to a higher iron reduction rate as compared to its uptake/reoxidation in the cytoplasm in accordance with the increased reduction rate in iron deficient Strategy I plants.