Is ethylene involved in regulation of root ferric reductase activity of dicotyledonous species under iron deficiency?

Most dicotyledonous species respond to Fe deficiency by developing some mechanisms known as Fe-deficiency responses. The role of ethylene in regulation of root ferric reductase activity of wild-type tomato (Lycopersicon esculentum L.) and its mutant Never ripe (Nr), bean (Phaseolus vulgaris L., cv. Bifeng 80-30), and cucumber (Cucumis sativus L., cv. Xintaimici) plants grown in nutrient solution without Fe supply was studied under controlled condition. The results show that: (i) the tomato mutant Nr, which is insensitive to ethylene, presented rapid increase in root ferric reductase activity after omitting Fe from the nutrient solution; (ii) the initial time for increase in root ferric reductase activity was earlier than that in ethylene production after onset of Fe deficiency in the three species; (iii) like cobalt (3 μM Co²⁺), an inhibitor for ethylene production, high concentration of zinc (50 μM Zn²⁺) and copper (5 μM Cu²⁺) also suppressed the increase in root ferric reductase activity of Fe-starved plants; (iv) under Fe-sufficient conditions, indol-3-butylric acid (IBA) stimulated root ferric reductase activity of cucumber and bean plants, and this stimulating effect could not be suppressed by aminoethoxyvinylglycine (AVG, an inhibitor for ethylene synthesis). These results suggested that ethylene might not be directly involved in the regulation of root ferric reductase activity of Fe-deficient dicotyledonous species.     

Formation of Root Epidermal Transfer Cells in Plantago

The root ultrastructure and transmembrane electron transport activities of two Plantago species have been examined with respect to alterations in response to Fe deficiency, exogenously supplied auxin, and the presence of chromium in the external medium. Both species showed increased ferric reductase activity upon Fe starvation, but they differed in the maximum rates. The addition of chromium to the nutrient solution led to a further enhancement in Fe-ethylenediaminetetraacetate reduction by Fe-deficient plants. In roots of Plantago lanceolata, the enhanced redox activity is associated with the formation of transfer cells in the epidermis. Similar characteristics of rhizodermal cells were observed in Fe-sufficient roots 3 d after application of the auxin analog 2,4-dichlorophenoxy-acetic acid. No structural adaptations occurred in roots of Plantago maritima. A quantitative estimation of the frequencies of transfer cells in root segments of Fe-deficient plants that differ in reduction activity revealed no correlation between the two phenomena. It is concluded that the area of plasmalemma infoldings is not specialized for the enhanced reduction of extracytoplasmatic Fe in response to Fe deficiency. The role of transfer cells in the adaptation to suboptimal Fe availability and the mechanisms triggering their formation are discussed.     

Relationships among iron deficit-induced potato callus growth inhibition,Fe distribution,chlorosis, and oxidative stress amplified by reduced antioxidative enzyme activities

Callus cultures were used to investigate and delineate responses of potato to iron (Fe) deficiency conditions over different culture durations. The morphological responses included chlorotic symptoms, reduced fresh weight and area of callus growth on Fe-deficient medium compared to calli grown under Fe sufficient conditions. Biochemically, potato calli under Fe deficit exhibited decreases in chlorophyll and carotenoid contents, reduction in activities of antioxidant enzymes (peroxidase, catalase and ascorbate peroxidase), as well as an increase in ferric chelate reductase (FCR) activity, lipid peroxidation, phenolic production and hydrogen peroxide (H₂O₂) level. Perls staining revealed sparse Fe distribution in Fe-deficient callus cells whereas Fe was widely distributed and intensely stained among numerous actively dividing cells in Fe-sufficient calli. These responses of calli to Fe deficiency were more pronounced with prolonged exposure to such stress leading to severe chlorosis and/or death of cells in chlorosis-susceptible calli but potential chlorosis-tolerant callus cells maintained their greenness and viability. Over a prolonged period in culture, significantly positive correlations were found among callus fresh weight, chlorophyll and carotenoid contents, antioxidant enzyme activities and lipid peroxidation as Fe supplies to the medium was increased. FCR activity was strongly correlated in a negative manner with Fe deficiency, chlorophyll content and peroxidase activity. The responses of calli to Fe supply can serve as reliable indicators for detecting chlorosis tolerance and/or nutrient deficiency stress.     

Changes in sugar beet leaf plasma membrane Fe(III)-chelate reductase activities mediated by Fe-deficiency, assay buffer composition, anaerobiosis and the presence of flavins

Summary Different assay conditions induce changes in the ferric chelate reductase activities of leaf plasma membrane preparations from Fe-deficient and Fe-sufficient sugar beet. With an apoplasttype assay medium the ferric chelate reductase activities did not change significantly when Fe(III)-EDTA was the substrate. However, with ferric citrate as substrate, the effect depended on the citrateto-Fe ratio. When the citrate-to-Fe ratio was 20 1, the effects were practically unappreciable. However, with a lower citrate-to-Fe ratio of 5 1 the activities were significantly lower with the apoplast-type medium than with the standard assay medium. Our data also indicate that anaerobiosis during the assay facilitates the reduction of ferric malate and Fe(III)-EDTA by plasma membrane preparations. Anaerobiosis increased by approximately 50% the plasma membrane ferric chelate reductase activities when Fe(III)-EDTA was the substrate. With ferric malate anaerobiosis increased activities by 70–90% over the values obtained in aerobic conditions. However, with ferric citrate the increase in activity by anaerobiosis was not significant. We have also tested the effect of riboflavin, flavin adenine dinucleotide, and flavin mononucleotide on the plasma membrane ferric chelate reductase activities. The presence of flavins generally increased activities in plasma membrane preparations from control and Fe-deficient plants. Increases in activity were generally moderate (lower than twofold). These increases occurred with Fe(III)-EDTA and Fe(III)-citrate as substrates.Abbreviations BPDS bathophenantroline disulfonate - FC ferric chelate - FC-R ferric chelate reductase - PM plasma membrane     

Proteins under the Control of the Gene for Fe Efficiency in Tomato

Fe-deficient dicotyledons develop Fe-efficiency reactions, such as proton extrusion and ferric chelate reduction activity, which are located in the plasma membranes of the root epidermal cells. The fer mutant of tomato (Lycopersicon esculentum Mill.) cannot develop these reactions. Membranes were isolated from roots of wild-type (FER) and mutant (fer) tomato plants grown on nutrient solution with high and low Fe concentrations. Two proteins were identified which are synthesized under the control of the FER gene.     

The role of nodules in the tolerance of common bean to iron deficiency

Iron is vital for the establishment and function of symbiotic root nodules of legumes. Although abundant in the environment, Fe is often a limiting nutrient for plant growth due to its low solubility and availability in some soils. We have studied the mechanism of iron uptake in the root nodules of common bean to evaluate the role of nodules in physiological responses to iron deficiency. Based on experiments using full or partial submergence of nodulated roots in the nutrient solution, our results show that the nodules were affected only slightly under iron deficiency, especially when the nodules were submerged in nutrient solution in the tolerant cultivar. In addition, fully submerged root nodules showed enhanced acidification of the nutrient solution and showed higher ferric chelate reductase activity than that of partially submerged roots in plants cultivated under Fe deficiency. The main results obtained in this work suggest that in addition to preferential Fe allocation from the root system to the nodules, this symbiotic organ probably develops some mechanisms to respond to iron deficiency. These mechanisms were implied especially in nodule Fe absorption efficiency and in the ability of this organ to take up Fe directly from the medium.     

Iron deficiency enhances the levels of ascorbate,glutathione, and related enzymes in sugar beet roots

Summary.  The effects of Fe deficiency stress on the levels of ascorbate and glutathione, and on the activities of the enzymes ferric chelate reductase, glutathione reductase (EC 1.6.4.2), ascorbate free-radical reductase (EC 1.6.5.4) and ascorbate peroxidase (EC 1.11.1.11), have been investigated in sugar beet (Beta vulgaris L.) roots. Plasma membrane vesicles and cytosolic fractions were isolated from the roots of the plants grown in nutrient solutions in the absence or presence of Fe for two weeks. Plants responded to Fe deficiency not only with a 20-fold increase in root ferric chelate reductase activity, but also with moderately increased levels of the general reductants ascorbate (2-fold) and glutathione (1.6-fold). The enzymes of the ascorbate-glutathione cycle in roots were also affected by Fe deficiency. Glutathione reductase activity was enhanced 1.4-fold with Fe deficiency, associated to an increased ratio of reduced to oxidized glutathione, from 3.1 to 5.2. The plasma membrane fraction from iron-deficient roots showed 1.7-fold higher ascorbate free-radical reductase activity, whereas in the cytosolic fraction the enzyme activity was not affected by Fe deficiency. The activity of the cytosolic hemoprotein ascorbate peroxidase decreased approximately by 50% with Fe deprivation. These results show that sugar beet responds to Fe deficiency with metabolic changes affecting components of the ascorbate-glutathione cycle in root cells. This suggests that the ascorbate-glutathione cycle would play certain roles in the general Fe deficiency stress responses in strategy I plants. Received November 19, 2001; accepted September 30, 2002; published online April 2, 2003 RID="*" ID="*" Correspondence and reprints: Departamento de Nutrición Vegetal, Estación Experimental de Aula Dei, CSIC, Apartado 202, 50080 Zaragoza, Spain.     

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.     

Characterization of Phloem iron and its possible role in the regulation of fe-efficiency reactions

`Fe-efficiency reactions' are induced in the roots of dicotyledonous plants as a response to Fe deficiency. The role of phloem Fe in the regulation of these reactions was investigated. Iron travels in the phloem of Ricinus communis L. as a complex with an estimated molecular weight of 2400, as determined by gel exclusion chromatography. The complex is predominantly in the ferric form, but because of the presence of reducing compounds in the phloem sap, there must be a fast turnover in situ between ferric and ferrous (k ≈ 1 min⁻¹). Iron concentrations in R. communis phloem were determined colorimetrically or after addition of ⁵⁹Fe to the nutrient solution. The iron content of the phloem in Fe-deficient plants was lower (7 micromolar) than in Fe-sufficient plants (20 micromolar). Administration of Fe-EDTA to leaves of Phaseolus vulgaris L. increased the iron content of the roots within 2 days, and decreased proton extrusion and ferric chelate reduction. The increase in iron content of the roots was about the same as the difference between iron contents of roots grown on two iron levels with a concomitantly different expression of Fe-efficiency reactions. We conclude that the iron content of the leaves is reflected by the iron content of the phloem sap, and that the capacity of the phloem to carry iron to the roots is sufficient to influence the development of Fe-efficiency reactions. This does not preclude other ways for the shoot to influence these reactions.     

Transfer cell formation in sugar beet roots induced by latent Fe deficiency

Leaves of Fe deficient sugar beets precultured in complete nutrient solution with Fe(III)EDTA remained green during the first 6 days of –Fe treatment when grown in a small nutrient solution volume (0.5 L/plant). After 3 days of –Fe treatment, roots placed in agar showed enhanced H⁺ release and ferric reduction at the tips of young laterals where short root hairs and transfer cells had developed. However, the H⁺ release was too weak to cause a pH decrease of the bulk nutrient solution. Nevertheless, the Fe stress response reactions probably lead to mobilization of Fe from the apoplasmic pool so that chlorosis development was prevented. Slight chlorosis symptoms appeared only after 4 more days of Fe deficiency and the pH of the bulk nutrient solution decreased to pH 4.5 simultaneously with renewed transfer cell formation and subsequent rapid regreening. In the 10 times higher volume of 5 L-Fe solution/plant, laterals with root hairs and transfer cells also showed localized acidification of the agar system. However, the protons released were so diluted that no pH decrease of the bulk solution was measurable. Instead, the leaves showed continuously increasing chlorosis with degenerated chloroplast ultrastructure. It is concluded that root hairs and transfer cells are not only formed under severe chlorosis but, instead, they seem to be an integral part of the adaptive response to latent Fe deficiency.     

Iron-Stress Induced Redox Activity in Tomato (Lycopersicum esculentum Mill.) Is Localized on the Plasma Membrane

Tomato plants (Lycopersicum esculentum Mill.) were grown for 21-days in a complete hydroponic nutrient solution including Fe³⁺-ethylenediamine-di(o-hydroxyphenylacetate) and subsequently switched to nutrient solution withholding Fe for 8 days to induce Fe stress. The roots of Fe-stressed plants reduced chelated Fe at rates sevenfold higher than roots of plants grown under Fe-sufficient conditions. The response in intact Fe-deficient roots was localized to root hairs, which developed on secondary roots during the period of Fe stress. Plasma membranes (PM) isolated by aqueous two-phase partitioning from tomato roots grown under Fe stress exhibited a 94% increase in rates of NADH-dependent Fe³⁺-citrate reduction compared to PM isolated from roots of Fe-sufficient plants. Optimal detection of the reductase activity required the presence of detergent indicating structural latency. In contrast, NADPH-dependent Fe³⁺-citrate reduction was not significantly different in root PM isolated from Fe-deficient versus Fe-sufficient plants and proceeded at substantially lower rates than NADH-dependent reduction. Mg²⁺-ATPase activity was increased 22% in PM from roots of Fe-deficient plants compared to PM isolated from roots of Fe-sufficient plants. The results localized the increase in Fe reductase activity in roots grown under Fe stress to the PM.     

Nitrate and Carbohydrate Effects on Nodulation and Nitrogen Fixation (Acetylene Reduction) Activity of Lentil (Lens esculenta Moench)

Lentils (Lens esculenta Moench, cv. Tekoas) grown in a nutrient solution containing 15 millimolar nitrate had 84% fewer nodules than lentils grown in nitrate-free nutrient solution. Nodules from the nitrate-grown plants weighed 71% less than nodules from the nitrate-free plants. Nitrate-grown plants also fixed much less nitrogen (measured by acetylene reduction) than the nitrate-free plants. When lentils were grown in a solution containing 15 millimolar nitrate and 75 millimolar fructose, glucose, or sucrose, however, the nitrogen fixation activity of their nodules was similar to that of nodules from nitrate-free plants. Leaves of lentils grown in the nitrate-sugar solutions had only about 7% as much nitrate reductase activity and accumulated only 10% as much nitrate as leaves from lentils grown in the nitrate solution alone. Roots of lentils grown in the nitrate-sugar solutions had similar nitrate reductase activity but accumulated only 17 to 25% as much nitrate as roots from lentils grown in the nitrate solution. The results indicate that the added sugars alleviated the inhibitory effects of nitrate on symbiotic nitrogen fixation not only by increasing the carbohydrate supply so lentils could support both nitrogen fixation and nitrate reduction but also by inhibiting the accumulation of nitrate and, hence, lowering nitrate reductase activity in the leaves.     

Iron limitation results in induction of ferricyanide reductase and ferric chelate reductase activities in Chlamydomonas reinhardtii

The green alga Chlamydomonas reinhardtii Dangeard CW-15 exhibited very low rates of plasma-membrane Fe(III) reductase activity when grown under Fe-sufficient conditions. After switching the medium to an Fe-free formulation, both ferricyanide reductase and ferric chelate reductase activities rapidly increased, reaching a maximum after 3 d under iron-free conditions. Both of the Fe(III) reductase activities increased in parallel over time, they exhibited similar K _m values (approximately 10 μM) with respect to Fe(III), displayed the same pH profile of activity, and both exhibited the same degree of light stimulation which could be inhibited by 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea (DCMU). Furthermore, ferricyanide competitively inhibited ferric chelate reduction by iron-limited cells. These results indicate that both Fe(III) reductase activities were mediated by the same iron-limitation-induced plasma-membrane reductase. No evidence was found for the presence of Fe(III)-reducing substances in the culture medium, or for the involvement of active oxygen species in the process of Fe(III) reduction. Chlamydomonas reinhardtii appears to respond to iron limitation in a manner similar to Strategy I higher plants. Received: 24 June 1997 / Accepted: 2 August 1997     

Physiological responses and differential gene expression in Prunus rootstocks under iron deficiency conditions

Two Prunus rootstocks, the Myrobalan plum P 2175 and the interspecific peach-almond hybrid, Felinem, were studied to characterize their biochemical and molecular responses induced under iron-Deficient conditions. Plants of both genotypes were submitted to different treatments using a hydroponic system that permitted removal of Fe from the nutrient solution. Control plants were grown in 90 μM Fe (III)-EDTA, Deficient plants were grown in an iron free solution, and plants submitted to an Inductor treatment were resupplied with 180 μM Fe (III)-EDTA over 1 and 2 days after a period of 4 or 15 days of growth on an iron-free solution. Felinem increased the activity of the iron chelate reductase (FC-R) in the Inductor treatment after 4 days of iron deprivation. In contrast, P 2175 did not show any response after at least 15 days without iron. The induction of the FC-R activity in this genotype was coincident in time with the medium acidification. These results suggest two different mechanisms of iron chlorosis tolerance in both Strategy I genotypes. Felinem would use the iron reduction as the main mechanism to capture the iron from the soil, and in P 2175, the mechanism of response would be slower and start with the acidification of the medium synchronized with the gradual loss of chlorophyll in leaves. To better understand the control of these responses at the molecular level, the differential expression of PFRO2, PIRT1 and PAHA2 genes involved in the reductase activity, the iron transport in roots, and the proton release, respectively, were analyzed. The expression of these genes, estimated by quantitative real-time PCR, was different between genotypes and among treatments. The results were in agreement with the physiological responses observed.     

Escherichia coli ferredoxin-NADP+ reductase and oxygen-insensitive nitroreductase are capable of functioning as ferric reductase and of driving the Fenton reaction

Two free flavin-independent enzymes were purified by detecting the NAD(P)H oxidation in the presence of Fe(III)-EDTA and t-butyl hydroperoxide from E. coli. The enzyme that requires NADH or NADPH as an electron donor was a 28 kDa protein, and N-terminal sequencing revealed it to be oxygen-insensitive nitroreductase (NfnB). The second enzyme that requires NADPH as an electron donor was a 30 kDa protein, and N-terminal sequencing revealed it to be ferredoxin-NADP⁺ reductase (Fpr). The chemical stoichiometry of the Fenton activities of both NfnB and Fpr in the presence of Fe(III)-EDTA, NAD(P)H and hydrogen peroxide was investigated. Both enzymes showed a one-electron reduction in the reaction forming hydroxyl radical from hydrogen peroxide. Also, the observed Fenton activities of both enzymes in the presence of synthetic chelate iron compounds were higher than their activities in the presence of natural chelate iron compounds. When the Fenton reaction occurs, the ferric iron must be reduced to ferrous iron. The ferric reductase activities of both NfnB and Fpr occurred with synthetic chelate iron compounds. Unlike NfnB, Fpr also showed the ferric reductase activity on an iron storage protein, ferritin, and various natural iron chelate compounds including siderophore. The Fenton and ferric reductase reactions of both NfnB and Fpr occurred in the absence of free flavin. Although the k _cat/K _m value of NfnB for Fe(III)-EDTA was not affected by free flavin, the k _cat/K _m value of Fpr for Fe(III)-EDTA was 12-times greater in the presence of free FAD than in the absence of free FAD.     

The pH Requirement for in Vivo Activity of the Iron-Deficiency-Induced "Turbo" Ferric Chelate Reductase (A Comparison of the Iron-Deficiency-Induced Iron Reductase Activities of Intact Plants and Isolated Plasma Membrane Fractions in Sugar Beet)

The characteristics of the Fe reduction mechanisms induced by Fe deficiency have been studied in intact plants of Beta vulgaris and in purified plasma membrane vesicles from the same plants. In Fe-deficient plants the in vivo Fe(III)-ethylenediaminetetraacetic complex [Fe(III)-EDTA] reductase activity increased over the control values 10 to 20 times when assayed at a pH of 6.0 or below ("turbo" reductase) but increased only 2 to 4 times when assayed at a pH of 6.5 or above. The Fe(III)-EDTA reductase activity of root plasma membrane preparations increased 2 and 3.5 times over the controls, irrespective of the assay pH. The Km for Fe(III)-EDTA of the in vivo ferric chelate reductase in Fe-deficient plants was approximately 510 and 240 [mu]M in the pH ranges 4.5 to 6.0 and 6.5 to 8.0, respectively. The Km for Fe(III)-EDTA of the ferric chelate reductase in intact control plants and in plasma membrane preparations isolated from Fe-deficient and control plants was approximately 200 to 240 [mu]M. Therefore, the turbo ferric chelate reductase activity of Fe-deficient plants at low pH appears to be different from the constitutive ferric chelate reductase.     

Effect of ammonium nutrition on the activity of nitrate reductase in the roots of apple seedlings

Active extracts of nitrate reductase were prepared from theroots of apple seedlings c.v. Granny Smith which were grownin nutrient solution under controlled enviromental conditions.The nutrient solutions contained various ratios of nitrate andammonium ions but all the treatments contained a total of 112ppm nitrogen. Maximum nitrate reductase activity in the roots was obtainedwhen plants were supplied with nitrate as the sole source ofnitrogen. Roots grown in solution containing only ammonium nitrogenhad little or no activity. When plants were supplied with bothforms of nitrogen in the nutrient solution, the presence ofammonium ions markedly lowered the activity of nitrate reductasein the roots. Plants supplied with 98 ppm nitrate nitrogen plus14 ppm ammonium nitrogen had activities only half those of plantsgrown in nitrate alone. Plants supplied with equal amounts ofammonium and nitrate nitrogen had activities less than one sixththose of plants grown in nitrate alone. (Received June 3, 1972; )     

Plasma membrane ferric reductase activity of iron-limited algal cells is inhibited by ferric chelators

Iron-limited cells of the green alga Chlorella kesslerii use a reductive mechanism to acquire Fe(III) from the extracellular environment, in which a plasma membrane ferric reductase reduces Fe(III)-chelates to Fe(II), which is subsequently taken up by the cell. Previous work has demonstrated that synthetic chelators both support ferric reductase activity (when supplied as Fe(III)-chelates) and inhibit ferric reductase. In the present set of experiments we extend these observations to naturally-occurring chelators and their analogues (desferrioxamine B mesylate, schizokinen, two forms of dihydroxybenzoic acid) and also two formulations of the commonly-used herbicide N-(phoshonomethyl)glycine (glyphosate). The ferric forms of the larger siderophores (desferrioxamine B mesylate, schizokinen) and Fe(III)-N-(phoshonomethyl)glycine (as the isopropylamine salt) all supported rapid rates of ferric reductase activity, while the iron-free forms inhibited reductase activity. The smaller siderophores/siderophore precursors, 2,3- and 3,4-dihydroxybenzoic acids, did not support high rates of reductase in the ferric form but did inhibit reductase activity in the iron-free form. Bioassays indicated that Fe(III)-chelates that supported high rates of ferric reductase activity also supported a large stimulation in the growth of iron-limited cells, and that an excess of iron-free chelator decreased the growth rate. With respect to N-(phosphonomethyl)glycine, there were differences between the pure compound (free acid form) and the most common commercial formulation (which also contains isopropylamine) in terms of supporting and inhibiting ferric reductase activity and growth. Overall, these results suggest that photosynthetic organisms that use a reductive strategy for iron acquisition both require, and are potentially simultaneously inhibited by, ferric chelators. Furthermore, these results also may provide an explanation for the frequently contradictory results of N-(phosphonomethyl)glycine application to crops: we suggest that low concentrations of this molecule likely solubilize Fe(III), making it available for plant growth, but that higher (but sub-lethal) concentrations decrease iron acquisition by inhibiting ferric reductase activity.     

Role of hormones in the induction of iron deficiency responses in Arabidopsis roots

In "strategy I" plants, several alterations in root physiology and morphology are induced by Fe deficiency, although the mechanisms by which low Fe levels are translated into reactions aimed at alleviating Fe shortage are largely unknown. To prove whether changes in hormone concentration or sensitivity are involved in the adaptation to suboptimal Fe availability, we tested 45 mutants of Arabidopsis defective in hormone metabolism and/or root hair formation for their ability to increase Fe(III) chelate reductase activity and to initiate the formation and enlargement of root hairs. Activity staining for ferric chelate reductase revealed that all mutants were responsive to Fe deficiency, suggesting that hormones are not necessary for the induction. Treatment of wild-type plants with the ethylene precursor 1-aminocyclopropane-1-carboxylic acid caused the development of root hairs in locations normally occupied by non-hair cells, but did not stimulate ferric reductase activity. Ectopic root hairs were also formed in -Fe roots, suggesting a role for ethylene in the morphological responses to Fe deficiency. Ultrastructural analysis of rhizodermal cells indicated that neither Fe deficiency nor 1-aminocyclopropane-1-carboxylic acid treatment caused transfer-cell-like alterations in Arabidopsis roots. Our data indicate that the morphological and physiological components of the Fe stress syndrome are regulated separately.     

Nitrate Reductase Activity in Calcifugous and Calcicolous Tomatoes as Affected by Iron Stress

Calcicolous plants are generally more Fe-efficient than calcifugous plants, because they respond to Fe stress by releasing H-ions and “reductants” from their roots that causes Fe to become available. The objective of our study was to determine if differential response to Fe stress in calcicolous and calcifugous varieties affects nitrate reductase activity. T3238FER (Fe-efficient) and T3238fer (Fe-inefficient) tomato (Lycopersicon esculentum Mill.) cultivars were grown in nutrient solutions supplied with N as NH₄⁺-N plus NO₃^?-N, and as NO₃^?-N only. The chemical reactions induced by Fe stress concomitantly increased nitrate reductase activity in roots and tops of calcicolous, but not in calcifugous tomato. This nitrate reductase activity decreased, however, when Fe was made available to the plants. When Fe stress was eliminated by adding Fe, nitrate reductase activity was comparable in the two cultivars.