A rice FRD3-like (OsFRDL1) gene is expressed in the cells involved in long-distance transport

We identified four putative AtFRD3-like genes (OsFRDL) in the rice genome that exhibited 39.1 to 56.7% amino acid sequence similarities to Arabidopsis FRD3. Of these, we cloned three OsFRDL genes from a cDNA library prepared from iron-deficient rice roots: OsFRDL1, OsFRDL2, and OsFRDL3. OsFRDL1 was expressed weakly in Fe-sufficient roots, and slight expression was induced in the roots of Fe-deficient plants. OsFRDL2 was expressed constitutively in both roots and leaves, and Fe deficiency reduced its expression in leaves. OsFRDL3 was expressed in leaves, but not in roots; Fe deficiency induced slight expression in leaves. An OsFRDL1-sGFP fusion protein was localized in the plasma membrane in onion epidermal cells. The promoter GUS analysis showed that OsFRDL1 was localized in the cells involved in long-distance transport, in both Fe-sufficient and Fe-deficient plants. Furthermore, OsFRDL1 expression was observed during the reproductive stage. These results suggest that OsFRDL1 is a transporter that resides in the plasma membrane of cells involved in long-distant transport.     

EFFECT OF CADMIUM ON PHYSIOLOGICAL RESPONSES OF WHEAT AND CORN TO IRON DEFICIENCY

This work evaluated the effect of cadmium (Cd) on the physiological responses of corn (Zea Mays L.) and wheat (Triticum aestivum L.) to iron (Fe) deficiency. For this purpose, seedlings of corn and wheat were cultivated under controlled conditions, plants were grown in different strength Hoagland's solutions for one month. In the fifth week, some seedlings were still in full strength Hoagland's solution (+Fe) and others were in full strength Hoagland's solutions without iron (?Fe). The plants were exposed to different cadmium (Cd) concentrations for four days. The plant chlorophyll content of young leaves, Fe and Cd content in shoots and roots, biomass production, and phytosiderophores (PS) release by roots were assessed. Results showed that Cd decreased the chlorophyll content of young leaves, accompanied by a significant shoot and root biomass reduction for Fe-deficient and Fe-sufficient wheat and corn across all Cd treatments. However, chlorophyll content and shoot and root biomass of Fe-deficient wheat and corn were lower than Fe-sufficient plants at different Cd concentrations. Iron-deficiency induced Cd accumulation compared to Fe-sufficient in wheat and corn; however, a depressive effect of Cd on iron acquisition in shoots and roots of Fe-deficient and Fe-sufficient wheat and corn across all Cd treatments was observed. Cadmium also inhibited PS release in Fe-deficient and Fe-sufficient wheat and corn. Iron-deficient PS release was higher than Fe-sufficient corn and wheat across all Cd treatments. These results suggested that Cd might reduce capacity of plants to acquire iron from solution by inhibiting PS release.     

Genetic Differences in Resistance to Iron Deficiency Chlorosis in Peanut

Iron (Fe) deficiency has been a widespread problem in peanut (Arachis hypogaea L.) grown on calcareous soils of northern China and has resulted in significant yield losses. Field observations showed considerable variability in visual chlorosis symptoms among peanut cultivars in the same soil. The objective of this study was to confirm the genetic differences in resistance to Fe-deficiency chlorosis in peanut and to identify feasible indicators for screening Fe-efficient genotypes. Resistance to Fe chlorosis of sixteen peanut cultivars grown on calcareous soil was evaluated in the field and physiological responses to Fe-deficiency stress were studied in nutrient solution. There were significant differences in resistance to Fe-deficiency chlorosis among the sixteen peanut cultivars tested, which was identified with SPAD readings, active Fe concentrations in young leaves in the early growth stages, and the pod yield. For Fe-resistant peanut cultivars, Fe-reduction capacity and quality of releasing hydrogen ions from roots increased under Fe-deficiency stress. Highly correlated relationships were observed between the summation of root Fe reduction and field chlorosis scores for sixteen cultivars (r² = 0.79). It was concluded that Fe-reduction capacity was a better physiological indicator for screening Fe-efficient peanut genotypes of the mechanisms measured.     

The effects of Fe deficiency on low molecular weight organic acid exudation and cadmium uptake by Solanum nigrum L.

Abstract

The influence of Fe-deficiency on the root exudation of low molecular weight organic acids (LMWOAs), pH alteration and cadmium (Cd) accumulation and translocation were investigated in morel (Solanum nigrum L.) in hydroponic culture experiments. Tartaric, citric, malic and acetic acids were monitored because these acids were abundant and often detected as root exudates. Results showed that Fe-deficient plants excreted large amounts of LMWOAs in comparison with Fe-sufficient plants across all Cd treatments (p <0.05). In both cases the concentrations of the four organic acids were tartaric > citric > malic > acetic. The results showed that the Fe-deficient plants with higher concentrations of LMWOAs accumulated more Cd (p <0.05) and induced a decrease in solution pH compared with the Fe-sufficient plants. Cadmium accumulation in the Fe-deficient and Fe-sufficient plants had significant positive correlations with the exudation of malic and acetic acids (p <0.05 and p<0.01). Cadmium accumulation in the Fe-sufficient plants had a significant (p<0.01) positive correlation with the exudation of tartaric acid, whereas there was a negative correlation (p<0.01) between Cd accumulation and the exudation of tartaric acid in the Fe-deficient plants. No significant correlation between the exudation of citric acid and Cd accumulation was obtained. Our results indicate that Fe-deficiency could induce Cd accumulation and translocation through an increase of LMWOAs exudation and pH alteration, both of which enhance Cd bioavailability.     

Comparison of Iron Availability in Leaves of Barley and Rice

Iron (Fe) is an essential trace element in all eukaryotes. In higher plants, Fe deficiency causes interveinal chlorosis in young leaves. However, in barley and rice, both of which are “Strategy II” plants, the degree and the pattern of Fe-deficiency symptoms differ. In the present study, barley and rice plants were grown in the same container, i.e., by “coculturing,” to compensate for the amount of mugineic acids in rice in the nutrient solution. We examined the differential availability of Fe for distribution and retranslocation in shoots between barley and rice without considering the difference in the iron acquisition ability, which is affected by the differential mugineic acid secretion between barley and rice. Although the Fe concentration of young barley leaves had decreased under the coculture conditions, the SPAD value was similar to that in monocultured barley. In contrast, although there was an increase in the Fe concentration of the young leaves of cocultured rice, the SPAD value decreased, as in the case of monocultured rice. Rice accumulated Fe in old leaves, whereas in barley Fe was efficiently distributed to young leaves. Therefore, the SPAD value of the second leaf in rice remained constantly high. The Fe concentration of the second leaf in barley decreased under Fe-deficient coculture conditions, the SPAD value decreased and the senescence of the second leaf become accelerated. ⁵⁹Fe pulse-labeling experiments suggested that in barley Fe was more efficiently retranslocated from old leaves to young leaves than that in rice. As a result, the level of Fe present in the fraction with a molecular weight lower than the 10,000/water-soluble Fe ratio was higher in the old leaves of barley than in the old leaves of rice under Fe-deficient conditions. Based on the results obtained, we suggest that the distribution and retranslocation characteristics of internal Fe in barley may be well adapted to Fe deficiency.     

Comparison of Iron Availability in Leaves of Barley and Rice

Iron (Fe) is an essential trace element in all eukaryotes. In higher plants, Fe deficiency causes interveinal chlorosis in young leaves. However, in barley and rice, both of which are "Strategy II" plants, the degree and the pattern of Fe-deficiency symptoms differ. In the present study, barley and rice plants were grown in the same container, i.e., by "coculturing," to compensate for the amount of mugineic acids in rice in the nutrient solution. We examined the differential availability of Fe for distribution and retranslocation in shoots between barley and rice without considering the difference in the iron acquisition ability, which is affected by the differential mugineic acid secretion between barley and rice. Although the Fe concentration of young barley leaves had decreased under the coculture conditions, the SPAD value was similar to that in monocultured barley. In contrast, although there was an increase in the Fe concentration of the young leaves of cocultured rice, the SPAD value decreased, as in the case of monocultured rice. Rice accumulated Fe in old leaves, whereas in barley Fe was efficiently distributed to young leaves. Therefore, the SPAD value of the second leaf in rice remained constantly high. The Fe concentration of the second leaf in barley decreased under Fe-deficient coculture conditions, the SPAD value decreased and the senescence of the second leaf become accelerated. ⁵⁹Fe pulse-labeling experiments suggested that in barley Fe was more efficiently retranslocated from old leaves to young leaves than that in rice. As a result, the level of Fe present in the fraction with a molecular weight lower than the 10,000/water-soluble Fe ratio was higher in the old leaves of barley than in the old leaves of rice under Fe-deficient conditions. Based on the results obtained, we suggest that the distribution and retranslocation characteristics of internal Fe in barley may be well adapted to Fe deficiency.     

Iron and zinc accumulation in winter wheat regulated by NICOTIANAMINE SYNTHASE responded to increasing nitrogen levels

Nitrogen (N) is critical for micronutrient biofortification in wheat grain and is essential for a series of nitrogenous compounds biosynthesis. This study aims to assess the role of improved N supply in iron (Fe) and zinc (Zn) enrichment and expression of genes related to Zn and Fe chelation and transport in winter wheat. Potting and hydroponic culture experiments were conducted to study the effect of increasing N application on Zn and Fe uptake and translocation from roots to leaves and the temporal and spatial gene expression profiles of the NICOTIANAMINE SYNTHASE (NAS) genes in wheat. Plants were grown with low, medium and high N supply levels. The results showed that higher N application increased Fe and Zn content in leaves, and decreased Fe and Zn content in root compared with the lower N supply. High N application also increased the distribution of Fe and Zn from roots to leaves. Expression analysis showed that increased N application resulted in up-regulation of two wheat NAS genes, TaNAS1 and TaNAS2. Highly positive response between NAS genes and increasing N application indicated that abundance nicotianamine (NA) resulted from highly expressed NAS genes might involve in the chelation of Fe and Zn in the phloem and favor Fe and Zn uptake and accumulation in wheat leaves.     

IRON EFFICIENCY IN KENTUCKY BLUEGRASS NOT RELATED TO PHYTOSIDEROPHORE RELEASE

Some Kentucky bluegrass (KBG; Poa pratensis L.) is susceptible to iron (Fe)-deficiency chlorosis. Under Fe-deficiency stress, phytosiderophore is produced and released by the roots of many grasses to solubilize soil Fe and enhance uptake. In other species, quantifying phytosiderophore screens for Fe-deficiency resistant cultivars. A hydroponic study was conducted at 1 and 10 μM solution Fe to variously stress ‘Baron’, ‘Award’, ‘Limousine’, and ‘Rugby II’ KBG cultivars. One μM Fe solution produced more Fe-deficiency stress in all cultivars compared to 10 μM, resulting in greater chlorosis and phytosiderophore release but reduced shoot and root Fe concentrations and shoot weight. Of the four cultivars, Baron was the most susceptible to Fe deficiency and exhibited severe Fe chlorosis and low shoot Fe but, surprisingly, produced the most phytosiderophore. These results imply that Fe-deficiency susceptibility in KBG may be less related to phytosiderophore release and more related to inefficient uptake or utilization mechanisms.     

Effect of iron deficiency on the growth,development and grain yield of some selected upland rice genotypes in the rainforest

Abstract

Iron deficiency is a major production constraint of upland rice in the tropics despite is abundance in the soil. This investigation aimed to explicate the effect of iron deficiency on the growth, development, grain yield and its attributes of some selected upland rice in the rainforest. Field experiments were established at Africa Rice sub-Station, Ibadan, Nigeria. The treatments consisted of 35 upland rice genotypes and availability of iron in the soil (Fe-sufficient and Fe-deficient). The treatments were arranged in alpha lattice design with three replications. It was observed that upland rice sown in iron (Fe) deficient soils had significantly lower growth (plant height, number of tillers and seedling vigor), flowered later, with significantly lower yield attributes (1000 grain weight, filled grain) and grain yield than those sown in Fe-sufficient soils. Conversely, the number of unfilled grains were significantly higher in upland rice sown in Fe-deficient than those in sufficient soils. Percentage yield loss was in the range 98.00% to 22.95% for China best and Faro 65 respectively. Genotypes were identified to be tolerant (Faro 65, NERICA 3 and IRAT 109) and susceptible (Ofada 2, NERICA 5 and China Best) to Fe-deficiency based on their percentage grain yield loss. These evidences suggested that despite the increased phenology of upland rice sown in Fe-deficient soils their reproductive growth was suppressed through increased number of unfilled grains as witnessed in China Best and Faro 64.     

Detection of the regions of phytosiderophore release from barley roots

Abstract

We devised a method to detect regions of phytosiderophore release from barley (Hordeum vulgare cv. Minorimugi) roots. Plants were grown in Fe-sufficient (+Fe) or Fe-deficient (?-Fe) water cultures for 14 d in a phytotron. Intact or excised roots were sampled and put between two sheets of filter papers just after the onset of the light period. The filter papers with roots were wrapped with vinyl film to avoid drying out and covered with aluminum foil to shade. The roots between the filter paper were kept in the phytotron for 4 h and phytosiderophores (PS) released from roots were absorbed by the filter paper. Then, the shape of the roots was preserved by photocopy and PS released on the filter paper were visualized by the method employed for detecting PS on thin layer chromatography (TLC). Gelatinous Fe solubilizing activity was employed for detecting PS on the filter paper. Released PS were detected as white spots on the orange-colored background and the regions of PS release from the roots were visible on the filter paper. It was evident that the apical zones of roots were the main regions of PS release. It was apparent that a distinct primary root newly formed on the basal parts had higher activity to release PS than the other roots of ?Fe plants. It was also shown that apical root zones of +Fe plants released PS.     

Diurnal variations in absorption and translocation of a ferrated phytosiderophore in barley as affected by iron deficiency

The release of phytosiderophore (PS) from roots of Fe-deficient graminaceous plants follows a distinct diurnal rhythm with maximum release rates occurring usually 3 to 4 hours after the onset of light. However, it remains to be determined whether absorption of the PS-Fe³⁺ complex shows a diurnal rhythmicity similar to that of PS release, Barley plants grown with or without 10 µM FeEDTA for 7 days were fed with ferreted PS (10 µM labelled with ⁵⁹Fe) at 4-h intervals to study the diurnal variations in the absorption and transloca tion of ⁵⁹Fe, The absorption of ⁵⁹Fe, irrespective of the Fe nutritional status of the plants, was higher during the day and lower during the night but did not show any peak throughout the day-night cycle. On the other hand, the translocation of ⁵⁹Fe into shoots of Fe-deficient plants was lower than that of Fe-sufficient plants, while the Fe nutritional status of the plants did not affect the absorption of ⁵⁹Fe by roots, The formation of root apoplastic ⁵⁹Fe was lower during the day and higher during the night, regardless of the Fe nutritional status of plants. Our results showed that the absorption of the PS-Fe³⁺ complex by roots did not follow the PS release pattern.     

Construction of artificial promoters highly responsive to iron deficiency

Iron deficiency-responsive element 1 (IDE1) and IDE2 are cis-acting elements that are responsible for Fe-deficiency-inducible and root-specific expression of the barley (Hordeum vulgare L.) gene IDS2 (Fe-deficiency-specific clone no. 2). Using these cis-acting elements, we aimed to construct super-promoters that would induce prominent gene expression in the roots of Fe-deficient rice plants (Oryza sativa L.). Modules containing IDE1 and IDE2 of the IDS2 promoter were used as repeats or were linked to the Fe-deficiency-responsive promoter of barley IDS3, and were connected to known enhancer-like sequences. Five artificial promoters, as well as the native promoters of barley IDS2 or IDS3, were connected individually upstream of β-glucuronidase (GUS) and were introduced into rice. Transgenic rice plants were grown under control or Fe-deficient conditions, and GUS expression was analyzed. The artificial promoter that contained one module of IDE1 and IDE2 conferred strong Fe-deficiency-inducible GUS expression to the roots of rice plants. Each of the five artificial promoters induced a similar level of GUS expression in Fe-deficient roots, which did not exceed the GUS expression driven by the native IDS2 or IDS3 promoter. Artificial and native promoters induced GUS expression in response to Fe-deficiency in leaves, although the level of expression was lower than that in roots. Histochemical observations revealed that GUS expression driven by artificial and native promoters was spatially similar, and expression was dominant within vascular bundles and root exodermis. These findings suggest that there is coordinated expression of the genes that are involved in Fe-deficiency-induced Fe uptake in rice.     

影响菜豆体内铁再利用效率的因素及其机理

本文在人工气候室中，用营养液培养方法，并结合同位素示踪技术研究了铁的供应状况，两种形态氮素（ＮＯ＾－３－Ｎ和ＮＨ＾＋４－Ｎ）及叶处遮光对菜豆体内铁再利用效率的影响，并对其有关机理进行了深入的研究。结果表明，铁的缺乏有利于累积在根和初生叶中的铁身新生组织中转移，铁的再利用效率明显提高。无论有缺铁还供铁条件下，ＮＨ＾＋４－Ｎ的供应使得菜豆新叶中活性铁含量、新叶叶绿素含量及体内铁的再利用效率都明显高于Ｎ     

INHERITANCE OF RESISTANCE TO IRON DEFICIENCY CHLOROSIS IN CHICKPEA (CICER ARIETINUM L.)

Iron (Fe)-deficiency chlorosis causes considerable yield losses in chickpea (Cicer arietinum L.) when susceptible genotypes are grown in calcareous soils with high pH. The most feasible method for alleviating Fe deficiency is the selection of suitable cultivars resistant to Fe deficiency chlorosis. ICC 6119 (desi type), which is Fe-deficient chlorosis, was crossed with CA 2969 and Sierra (kabuli types), resistant to Fe deficiency chlorosis. Inheritance of resistance to Fe deficiency in chickpea revealed that the resistance was controlled by a single dominant gene in these genotypes crossed. A negative selection for resistance to Fe deficiency chlorosis will be effective after segregating generations.     

Amino acid sequence of proteins induced in Fe-deficient stressed alfalfa (Medicago sativa L.) roots

Alfalfa (Medicago sativa L.) grows well in soils with a moderately high pH and dissolves insoluble iron in the rhizosphere. We have investigated active uptake mechanisms under Fe-deficient nutrient conditions and the effects of Fe-deficiency on plants. Previously, we observed that Fe-deficient alfalfa roots exuded many compounds (Masaoka et al. 1993) such as fiavonoids. We also identified a new compound “alfafuran” which is a phenol compound and is different from organic acids or phytosiderophore-type amino acid derivatives exuded by Fe-deficient plant roots. This compound is also very effective in dissolving ferric phosphate (Noguchi et al. 1994), suggesting that alfalfa may have developed several strategies against Fe-deficient stress including the exudation of organic compounds like alfafuran which accelerate the Fe³⁺-reducing activity on the root cell membrane to dissolve insoluble iron compounds. Suzuki et al. (1995, 1997) observed that in barley several peptide spots obtained by electrophoresis were induced under Fe-deficient stress when mugineic acid-family phytosiderophores were secreted from the roots. They suggested that these peptides control the mugineic acid synthesis and secretion. We examined the peptides induced in Fe-deficient alfalfa roots.     

The central role of microbial activity for iron acquisition in maize and sunflower

 Maize (Zea mays L.) and sunflower (Helianthus annuus L.) grown on a calcareous soil showed poor growth and/or were chlorotic in spite of abundant Fe in the roots. It has been hypothesized that microbial siderophores chelate Fe (III) in the soil, and that in this form Fe is transported towards the root apoplast. On the calcareous soil, total and apoplastic root Fe concentrations were high, probably because of a high apoplastic pH depressing Fe (III)-reductase activity and thus the Fe²⁺ supply to the cytoplasm. On the acidic soil, total and apoplastic root Fe concentrations were low, probably because of a low apoplastic pH favouring Fe (III) reduction, hence plants showed no Fe-deficiency symptoms. The main objective of the present work was to investigate the role of microbial soil activity in plant Fe acquisition. For this purpose, plants were grown under sterile and non-sterile conditions on a loess loam soil. Plants cultivated under non-sterile conditions grew well, showed no Fe-deficiency symptoms and had fairly high Fe concentrations in the roots in contrast to plants grown in the sterile medium. Low root and leaf Fe concentrations in the axenic treatments indicated that the production of microbial siderophores was totally suppressed. Accordingly, sunflowers were severely chlorotic and this was associated with very poor growth, whereas in maize only growth was drastically reduced. In maize under sterile conditions, root apoplastic and total Fe concentrations were not as low as in sunflowers, which may have indicated that phytosiderophores produced in maize partly sustained Fe acquisition, but due to poor growth were not as efficient in supplying Fe as microbial activity under natural conditions. It may be therefore assumed that in natural habitats soil microbial activity is of pivotal importance for plant Fe acquisition. Received: 11 March 1999     

Physiological basis of iron chlorosis tolerance in rice (Oryza sativa) in relation to the root exudation capacity

Micronutrient deficiency in cultivable soil, particularly that of iron (Fe) and zinc (Zn), is a major productivity constraint in the world. Low Fe availability due to the low solubility of the oxidized ferric forms is a challenge. An experiment was, thus, executed to assess the performance of eight genetically diverse rice genotypes on Fe-sufficient (100 µM) and Fe-deficient (1 µM) nutrient solution, and their ability to recover from Fe deficiency was measured. Fe efficiency under Fe deficiency in terms of biomass production showed a significant positive correlation with the root release of phytosiderophore (PS) (R² = 0.62*). This study shows that the Fe deficiency tolerance of Pusa 33 was related to both a high release of PS by the root and an efficient translocation of Fe from the root to the shoot as the Fe–PS complex, which could be useful for improving the Fe nutrition of rice particularly under aerobic conditions.     

缺铁水稻根表铁膜对硒的转运和吸收的影响

Under anaerobic conditions, ferric hydroxide deposits on the surface of rice roots and affects uptake and translocation of certain nutrients. In the present study, rice plants were cultured in Fe-deficient or sufficient solutions and placed in a medium containing selenium （Se） for 2 h. Then, FeSO4 was added at the various concentrations of 0, 10, 40, or 70 mg L-1 to induce varying levels of iron plaque on the root surfaces and subsequent uptake of Se was monitored. The uptake of Se was inhibited by the iron plaque, with the effect proportional to the amount of plaque induced. The activity of cysteine synthase was decreased with increasing amounts of iron plaque on the roots. This may be the important reason for iron plaque inhibition of Se translocation. At each level of iron plaque, Fe-deficient rice had more Se than Fe-sufficient rice. Furthermore, with plaque induced by 20 mg Fe L-1, plants from Fe-deficient media accumulated more Se than those from Fe-sufficient media, as the Se concentration was increased from 10 to 30 or 50 mg L^-1. We found that phytosiderophores, highly effective iron chelating agents, could desorb selenite from ferrihydrite. Root exudates of the Fe-deficient rice, especially phytosiderophores in the exudates, could enhance Se uptake by rice plants with iron plaque.     

Iron Mobilization and Mineralogical Alterations Induced by Iron-Deficient Cucumber Plants (Cucumis sativus L.) in a Calcareous Soil

Dicotyledons cope with ion (Fe) shortage by releasing low-molecular-weight organic compounds into the rhizosphere to mobilize Fe through reduction and complexation mechanisms. The effects induced by these root exudates on soil mineralogy and the connections between Fe mobilization and mineral weathering processes have not been completely clarified. In a batch experiment, we tested two different kinds of organic compounds commonly exuded by Fe-deficient plants, i.e., three organic acids (citrate, malate, and oxalate) and three flavonoids (rutin, quercetin, and genistein), alone or in combination, for their ability to mobilize Fe from a calcareous soil and modify its mineralogy. The effect of root exudates on soil mineralogy was assessed in vivo by cultivating Fe-deficient and Fe-sufficient cucumber plants (Cucumis sativus L.) in a RHIZOtest device. Mineralogical analyses were performed by X-ray powder diffraction. The batch experiment showed that citrate and, particularly, rutin (alone or combined with organic acids or genistein) promoted Fe mobilization from the soil. The combinations of rutin and organic acids modified the soil mineralogy by dissolving the amorphous fractions and promoting the formation of illite. These mineralogical alterations were significantly correlated with the amount of Fe mobilized from the soil. The RHIZOtest experiment revealed a drastic dissolution of amorphous components in the rhizosphere soil of Fe-deficient plants, possibly caused by the intense release of phenolics, amino acids, and organic acids, but without any formation of illite. Both batch and RHIZOtest experiments proved that exudates released by cucumber under Fe deficiency concurred to the rapid modification (on a day-scale) of the mineralogy of a calcareous soil.     

IRON IS REQUIRED FOR THE INDUCTION OF ROOT FERRIC CHELATE REDUCTASE ACTIVITY IN IRON-DEFICIENT TOMATO

Two mutants of tomato and their corresponding wild-type genotypes, Tfer/TFER and chloronerva/Bonner Beste, were grown in nutrient solution under conditions leading to iron (Fe) deficiency. Iron deficiency caused decreases in growth, leaf chlorosis, and changes in the morphology of roots. Ferric chelate reductase activities of whole roots were generally lower in Fe-deficient plants than in control, Fe-sufficient plants. Plants grown for 7 days without Fe, however, had transient increases in whole root ferric chelate reductase activity after the addition of small amounts of Fe (2 μM) to the nutrient solution. Also, adding sequential 0.5 μM Fe pulses to the nutrient solution led to high whole root ferric chelate reductase activities. Similar results were obtained with a protocol using excised root tips instead of whole root systems to measure ferric chelate reductase activities. The protocol using root tips generally gave higher ferric chelate reductase rates than the method using whole roots, due to the localized expression of the enzyme in the distal root zones.