Induction of in vivo root ferric chelate reductase activity in fruit tree rootstock

A method has been developed to consistently induce increases in root ferric chelate reductase activity in the fruit tree rootstock GF 677 (Prunus amygdalopersica) grown under iron (Fe) deficiency. Clonal GF 677 plants were grown hydroponically in a growth chamber with 0 or 90 μM Fe(III)‐EDTA. Root ferric chelate reductase activity was measured in vivo using BPDS. Plants grown without Fe developed visible symptoms of chlorosis and had lower root ferric chelate reductase activities than those grown with Fe. Root ferric chelate reductase activities were 0.1–1.9 and 0.6–5.3 nmol of Fe reduced per gram of fresh mass and minute, respectively, in Fe‐deficient and sufficient plants. However, when plants grown without Fe for several days were resupplied with 180 μM of Fe(III)‐EDTA, FC‐R activities increased within 1 day. The FC‐R values after Fe resupply were 20‐fold higher than those found in Fe‐deficient plants and 5‐fold higher than those found in the Fe‐sufficient controls. After three days of the Fe treatments the FC‐R activities had decreased again to the control values. The reduction of Fe was localized at the subapical root zone. In the conditions used we have found no decreases of the nutrient solution pH values, indicating that this type of response is not strong enough to be detected in peach tree rootstocks. Also, no major changes in root morphology have been found in response to Fe deficiency. This ferric chelate reductase induction protocol may be used in screening assays to select rootstock genotypes tolerant to Fe chlorosis.     

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.     

Physiological responses of tomato roots grown in organ culture to iron-deficiency stress

Roots of the Fe-efficient tomato (Lycopersicon esculentum Mill., cultivar Floradel) were cultured in an inorganic medium supplemented with glycine, thiamine, pyridoxine, and nicotinic acid, with sucrose as an energy and carbon source. Iron was supplied as ferric hydroxyethylethylenediaminetriacetic acid (FeHEDTA) and the initial PH was 5.5. Root growth was limited when less than 40 μm FeHEDTA was supplied. Roots grown at lower Fe concentrations decreased the pH of the FCR assay medium to a greater extent than did roots grown at higher Fe concentrations. Cultured roots grown with 10 μm FeHEDTA had increased levels of ferric chelate reductase (FCR) activity compared to roots grown with either lower or higher concentrations of FeHEDTA. Low FCR activity of roots grown at 2.5 or 5 μm FeHEDTA was attributed either to impaired metabolism due to Fe-deficiency or the lack of sufficient Fe for enhanced FCR formation. Roots of hydroponically grown tomato plants exhibited typical increases in FCR activity with Fe-deficiency. Based on these preliminary results, cultured roots were found to exhibit similar Physiological responses to Fe-deficiency stress as intact root systems. Cultured roots should provide a useful system for the investigation of the role of the root in plant Fe-deficiency stress responses as previously suggested by Bienfait et al.(Plant Physiol., 83, 244–247, 1987).     

Iron deficiency stress responses amongst citrus rootstocks

Seedlings of citrus rootstocks differing in lime tolerance were grown in nutrient solution with and without Fe. Proton efflux, release of phenolic compounds and Fe reducing substances and root-mediated reduction of Fe^III in FeEDTA and freshly precipitated Fe(OH)₃ in response to Fe deficiency were determined. Sweet orange, Carrizo citrange and trifoliate orange, the three least tolerant rootstocks used in the study, did not decrease nutrient solution pH in response to Fe deficiency. The more lime tolerant rootstocks, rough lemon, Cleopatra mandarin and sour orange, did decrease nutrient solution pH. But in CaSO₄ solution only sour orange increased H⁺ efflux significantly under Fe deficiency. In response to Fe deficiency, the release of phenolic compounds was increased significantly in rough lemon and Cleopatra mandarin seedlings, while the release of reducing substances was increased significantly in rough lemon, sour orange and trifoliate orange. Rough lemon was the only rootstock to respond to Fe deficiency with an increase in root-mediated reduction of chelated Fe^III at pH 6.5. At pH 8.0, both Fe-deficient rough lemon and Cleopatra mandarin roots reduced higher amounts of Fe^III from freshly precipitated Fe(OH)₃ than Fe-sufficient seedlings. Iron reduction by detached roots of Fe-deficient and Fe-sufficient rough lemon did not follow Michaelis-Menten kinetics at high substrate concentrations. Rates of Fe reduction at low substrate concentrations were inconsistent with the existence of an inducible ferric reductase in response to Fe deficiency.     

Iron deficiency responses in roots of kiwi

Many dicotyledonous species respond to iron (Fe) deficiency by morphological and physiological changes at root level, which are usually defined as Strategy I. Particularly, these latter modifications include a higher acidification of the external medium and the induction of a high root Fe reductase activity. The aim of this work was to investigate the response of kiwi (Actinidia deliciosa cv. Hayward) plants, which often exhibit Fe chlorosis in the field, to Fe deficiency. Actinidia kept for two weeks in nutrient solution without Fe showed visual deficiency symptoms (leaf chlorosis). Moreover, upon prolonged micronutrient shortage shoot, and to a lesser extent, root dry weight accumulation was greatly impaired. Roots of Fe‐deficient Actinidia showed an increased capacity of net proton extrusion and higher ferric ethylenediaminetetraacetate [Fe(III)EDTA] reductase activity as compared to plants grown in the presence of 10 μM Fe(III)EDTA. Localization of the increased acidification and reductase capacity by means of agar‐technique revealed that these activities are both present in the sub‐apical region of the roots. Re‐supply of Fe after two weeks partially reversed the tendency of the roots to acidify the nutrient solution and to reduce Fe(III)EDTA.     

小金海棠Fe3+-还原酶基因MxFRO在酵母中的表达及其活性检测

为研究苹果属植物抗缺铁的分子生物学机理,从铁高效基因型小金海棠（Malus xiaojinensis）中克隆了Fe3+-还原酶基因MxFRO。MxFRO2的cDNA序列长2283bp,开放阅读框为2166bp,编码722个氨基酸。半定量RT-PCR结果表明MxFRO在小金海棠的根和叶中均受缺铁胁迫诱导。 将MxFRO转入野生型酵母中,结果表明转基因酵母的Fe3+-还原酶活性是对照的2.8倍,因此我们初步推测MxFRO基因编码的蛋白具有Fe3+-还原酶活性。     

Physiological responses of grapevine callus cultures to iron deficiency

Grapevine is considered a ‘Strategy I’ plant because it performs some peculiar biochemical and physiological responses when grown under iron (Fe) deficiency stress conditions. Callus cultures were started from leaf and internode cuts of micropropagated plantlets of two grapevine genotypes well known for their Fe‐chlorosis characteristic: Vitis riparia a very susceptible genotype and Vitis berlandieri a resistant one. Modification of NADH: ferric (Fe³⁺) reductase activity was spectrophotometrically evaluated by following the formation of the complex ferrous (Fe²⁺)‐(BPDS)₃, while the malic and citric acid production were determined in callus cultures grown both in the presence (+Fe) and absence (‐Fe) of Fe. Moreover, a microsomal fraction was isolated from the calli to evaluate the H⁺‐ATPase and the Fe³⁺‐EDTA reductase activities. As expected, calli of the Fe‐efficient genotype (V. berlandieri) was able to enhance Fe³⁺‐EDTA reductase activity when growing under Fe deficiency while the Fe‐chlorosis susceptible V. riparia could not or did it with lower efficiency. Therefore, the H⁺‐ATPase assay showed a higher enzymatic activity in the microsomal fraction isolated from Vitis berlandieri grown without Fe with respect to its control (+Fe). Organic acid determination gave quite contradictory results, specially regarding malic acid which, under our study conditions, seemed not to be linked with the strategies of response to Fe deficiency.     

RESPONSES OF “NEWHALL” ORANGE TREES TO IRON DEFICIENCY IN HYDROPONICS: EFFECTS ON LEAF CHLOROPHYLL,PHOTOSYNTHETIC EFFICIENCY,AND ROOT FERRIC CHELATE REDUCTASE ACTIVITY

Orange (Citrus sinensis L. Osb. cv. ‘Newhall’) plants grafted on Citrange troyer rootstock were grown in nutrient solution with 0, 5, 10, or 20 μM iron (Fe), with and without calcium carbonate. Calcium carbonate was added in order to mimic the natural conditions in calcareous soils. Leaf chlorophyll concentration was estimated every 3–4 days using the portable instrument SPAD-502 meter. Chlorophyll fluorescence parameters, photosynthetic capacity estimated from oxygen evolution, leaf Fe concentrations, and root tip ferric chelate reductase activity were measured at the end of the experiment. Plants from the 0 and 5 μM Fe treatments showed leaf chlorosis and had decreased leaf chlorophyll concentrations. Leaves of plants grown in the absence of Fe in the solution had smaller rates of oxygen evolution both in the presence and absence of calcium carbonate, compared with plants grown in the presence of 10 μM Fe. In the absence of calcium carbonate the photosystem II efficiency, estimated from fluorescence parameters, was similar in all treatments. A slight decrease in photosystem II efficiency was observed in plants grown without Fe and in the presence of calcium carbonate. A 2.5-fold increase in root tip ferric chelate reductase activity over the control values was found only when plants were grown with low levels of Fe and in the presence of calcium carbonate.     

Iron uptake in a bicarbonate-resistant cell line of tobacco

Abstract

In alkaline soils, plant growth is impaired mainly by high pH and high concentration of bicarbonates. The bicarbonate concentration increases the pH value, and causes deficiency of iron. A bicarbonate-resistant cell line (BR line) of tobacco (Nicotiana tabacum L. cv. Burley21) was selected by adding excess bicarbonate ions (20 mmol L^?1) to the culture medium. The pH of the medium was buffered 8.0 to 8.3. Under these conditions, about 80% of iron in the medium became insoluble. However, under such conditions, the BR line grew well. In this report, we examined some characteristics of the growth and iron uptake in the BR line under iron-deficient (i.e. high pH or no-iron) condition. At pH 5.8, the Fe³⁺ reduction activity was not significantly different between the non-selected line and the BR line. At pH 8.0, however, the Fe³⁺ reduction activity of the BR line was higher than that of the non-selected line. In no-iron condition, the growth of the non-selected line was markedly reduced after 2 weeks, while the BR line was not affected. The content of malic acid in both lines increased with the medium pH, and the content in the BR line was higher than that in the non-selected line. The BR line was able to adapt to the conditions, which restricted iron uptake, such as high bicarbonate concentration, high pH, and low iron conditions. The high ability of Fe³⁺ reduction was maintained at even high pH conditions. Further, the BR line may be able to improve the utilization of iron in the cells.     

Interactions of ferric ions with olive oil phenolic compounds

The ferric complexing capacity of four phenolic compounds, occurring in olives and virgin olive oil, namely, oleuropein, hydroxytyrosol, 3,4-dihydroxyphenylethanol-elenolic acid (3,4-DHPEA-EA), and 3,4-dihydroxyphenylethanol-elenolic acid dialdehyde (3,4-DHPEA-EDA), and their stability in the presence of ferric ions were studied. At pH 3.5, all compounds formed a reversible 1:1 complex with ferric ions, but hydroxytyrosol could also form complexes containing >1 ferric ion per phenol molecule. At pH 5.5, the complexes between ferric ions and 3,4-DHPEA-EA or 3,4-DHPEA-EDA were relatively stable, indicating that the antioxidant activity of 3,4-DHPEA-EA or 3,4-DHPEA-EDA at pH 5.5 is partly due to their metal-chelating activity. At pH 7.4, a complex containing >1 ferric ion per phenol molecule was formed with hydroxytyrosol. Oleuropein, 3,4-DHPEA-EA, and 3,4-DHPEA-EDA also formed insoluble complexes at this pH. There was no evidence for chelation of Fe(II) by hydroxytyrosol or its derivatives. At all pH values tested, hydroxytyrosol was the most stable compound in the absence of Fe(III) but the most sensitive to the presence of Fe(III).     

Role of Phosphatases,Iron Reducing,and Solubilizing Activity on the Nutrient Acquisition in Mixed Cropped Peanut and Barley

The effect of interspecific complementary and competitive root interactions and rhizosphere effects on primarily phosphorus (P) and iron (Fe) but also nitrogen (N), potassium (K), calcium (Ca), zinc (Zn), and manganese (Mn) nutrition between mixed cropped peanut (Arachis hypogaea L.) and barley (Hordeum vulgare L.). In order to provide more physiological evidence on the mechanisms of interspecific facilitation, phosphatase activities in plant and rhizosphere, root ferric reducing capacity (FR), Fe-solubilizing activity (Fe-SA), and rhizosphere pH were determined. The results of the experiment revealed that biomass yield of peanut and barley was decreased by associated plant species as compared to their monoculture. Rhizosphere chemistry was strongly and differentially modified by the roots of peanut and barley and their mixed culture. In the mixed cropping of peanut/barley, intracellular alkaline and acid phosphatases (AlPase and APase), root secreted acid phosphatases (S-APase), acid phosphatases activity in rhizosphere (RS-APase), and bulk soil (BS-APase) were higher than that of monocultured barley. Regardless of plant species and cropping system, the rhizosphere pH was acidified and concomitantly to this available P and Fe concentrations in the rhizosphere were also increased. The secretion Fe-solubilizing activity (Fe-SA) and ferric reducing (FR) capacity of the roots were generally higher in mixed culture relative to that in monoculture treatments which may improve Fe and Zn nutrition of peanut. Furthermore, mixed cropping improved N and K nutrition of peanut plants, while Ca nutrition was negatively affected by mixed cropping.     

Arbuscular mycorrhizal enhancement of iron concentration by <Emphasis Type="Italic">Poncirus trifoliata</Emphasis> L. Raf and <Emphasis Type="Italic">Citrus reticulata</Emphasis> Blanco grown on sand medium under different pH

The effect of the arbuscular mycorrhizal (AM) fungus (Glomus versiforme) on iron contents by two citrus rootstocks (trifoliate orange [Poncirus trifoliata L. Raf] and red tangerine [Citrus reticulata Blanco]) was studied in sand culture under different pH conditions. Seeds were sown in a mixed substrate (perlite/sand, 1:1 [v/v]) inoculated with or without mycorrhizal inoculum. The experiment was carried out at four pH levels by applying nutrient solution at pH 5.0, 6.0, 7.0, or 8.0 to P. trifoliata and pH 5.2, 6.2, 7.2, or 8.2 to C. reticulata. No AM colonization was found in uninoculated control (NM) and plants, and root colonization in AM plants was depressed under iron deficiency at high pH. Colonization by G. versiforme led to higher dry weights of shoots compared with NM treatments, suggesting that G. versiforme enhanced plant growth. Higher concentration of chlorophyll and active iron, lower ratios of P/Fe and 50(10P+K)/Fe were present in AM plants than NM treatments. Nevertheless, G. versiforme improved root Fe (III) chelate reductase activity of P. trifoliata and C. reticulata. The data indicate that plant uptake and translocation of iron were enhanced and AM fungi may be considered as a potential tool for bioremediation of citrus iron deficiency.     

Response to iron‐deficiency stress of pear and quince genotypes 1

Roots of iron (Fe)‐efficient dicots react to Fe‐deficiency stress by strongly enhancing the ferric (Fe³⁺)‐reductase system and by lowering the rhizo‐sphere pH. In this study, we tested whether such adaptation mechanisms characterize pear and quince genotypes known to have differential tolerance to calcareous and alkaline soils. Two trials were performed using micropagated plants of three quince rootstocks (BA29, CTS212, and MC), three Pyrus communis rootstocks (OHxF51 and two selections obtained at the Bologna University: A28 and B21) and of two pear cultivars (Abbé Fétel and Bartlett, own‐rooted). In the first trial, plants were grown in a nutrient solution with [Fe(+)] and without [Fe(‐)] Fe for 50 days. Their root Fe‐reducing capacity was determined colorimetrically using ferrozine and FeEDTA, and Fe uptake of Fe(+) plants was estimated. In the second trial, the rhizosphere pH of plants grown in an alkaline soil was measured by a micro‐electrode. With the only exception of pears OHxF51 and A28, whose Fe‐reduction rates were similar in Fe(+) and Fe(‐) plants, the Fe‐deficiency stress resulted in a significant decrease in Fe reduction. Among the Fe(‐) plants, the two pear cultivars, OHxF51 and A28, had a higher Fe‐reducing capacity than the quince rootstocks and the cv. Abb6 F. When plants were pre‐treated with Fe, reduction rate was highest in the P. communis rootstocks, intermediate in the own‐rooted cultivars, and lowest in the quinces. Root Fe‐reducing capacity of Fe(+) plants proved to be linearly and positively correlated with Fe uptake and root proton release. Rhizosphere pH was highest in quince MC, intermediate in the other two quinces and in the cv. Abbe F., and lowest in the pear rootstocks and in the cv. Bartlett. Our results indicate that roots of pear and quinces do not increase their ability to reduce the Fe under Fe‐deficiency stress. The genotypical differential tolerance to Fe chlorosis likely reflects differences in the standard reductase system and in the capacity of lowering the pH at the soil/root interface. The determination of the root Fe‐reducing capacity is a promising screening technique for selecting pear root‐stocks efficient in taking up Fe.     

RELATIVE SUSCEPTIBILITY OF SOME PRUNUS ROOTSTOCKS IN HYDROPONICS TO IRON DEFICIENCY

The susceptibility of eleven Prunus rootstocks to iron (Fe) deficiency was studied in hydroponics by growing them with 20 μM Fe, 0 μM Fe or 3 μM Fe+10 mM sodium bicarbonate. Based on the intensity of leaf chlorosis, the peach-almonds PR 204/84, Stylianidis K and KID2, produced at the Pomology Institute of Naoussa (Greece), showed the same or even greater tolerance than GF 677, the Greek peach-almond Retsou x Nemaguard, the plum-almond Myrandier 617 and the peaches GF 305, IDS 37, Greek wild peach seedling the greatest susceptibility whereas the plums St. Julien GF655/2 and Myrobalan 29C intermediate. Rootstocks without Fe presented significantly lower nitrogen and Fe whereas with bicarbonate significantly lower nitrogen, phosphorus, Fe and zinc. Root ferric chelate reductase activity was significantly increased in ?Fe rootstocks but negatively correlated with their tolerance; physiological and morphological changes were observed along a zone of a few centimeters length, 1–2 mm behind the root tip.     

Root and Leaf Ferric Chelate Reductase Activity in Pond Apple and Soursop

Abstract

Root and leaf ferric chelate reductase (FCR) activity in Annona glabra L. (pond apple), native to subtropical wetland habitats and Annona muricata L. (soursop), native to nonwetland tropical habitats, was determined under iron (Fe)-sufficient and Fe-deficient conditions. One-year-old seedlings of each species were grown with 2, 22.5, or 45 µM Fe in a nutrient solution. The degree of tolerance of Fe deficiency was evaluated by determining root and leaf FCR activity, leaf chlorophyll index, Fe concentration in recently mature leaves, and plant growth. Root FCR activity was generally lower in soursop than in pond apple. Eighty days after plants were put in nutrient solutions, leaf FCR activity of each species was lower in plants grown with low Fe concentrations (2 µM) than in plants grown with high (22.5 or 45 µM) Fe concentrations in the nutrient solution. Leaves of pond apple grown without Fe became chlorotic within 6 weeks. The Fe level in the nutrient solution had no effect on fresh and dry weights of soursop. Lack of Fe decreased the leaf chlorophyll index and Fe concentration in recently matured leaves less in soursop than in pond apple. The rapid development of leaf chlorosis in low Fe conditions and low root and leaf FCR activities of pond apple are probably related to its native origin in wetland areas, where there is sufficient soluble Fe for adequate plant growth and development. The higher leaf FCR activity and slower growth rate of soursop compared to pond apple may explain why soursop did not exhibit leaf chlorosis even under low Fe conditions.     

硝酸还原酶基因启动子NRE2元件缺失对烟草氮代谢的影响

为了探究硝酸还原酶基因启动子中硝酸盐响应元件NRE2缺失对烟草植株氮代谢的影响,以烤烟品种K326为材料,采用CRISPR/Cas9基因编辑技术获得了烟草NIA11和NIA2基因启动子中NRE2元件缺失的突变体材料,并于同一氮素营养条件下培养,测定烟草植株氮代谢相关生理指标.结果表明,与野生型相比,单突变体和双突变体叶...     

Effects of Several Metals on Both Fe(III)‐ and Cu(II)‐Reduction by Roots of Fe‐Deficient Cucumber Plants

Abstract

The ferric‐chelate reductase induced by Fe deficiency is also able to reduce other ions such as Cu²⁺. This Cu(II)‐reduction has been less studied and it has been suggested that Cu²⁺ ion rather than Cu²⁺‐chelate serves as the substrate. Ferric‐chelate reductase activity is inhibited by some metals, but the mechanisms implicated are not known. In the present work we use Fe‐deficient cucumber seedlings to study the interactions of Cu²⁺, Ni²⁺, Mn⁴⁺, and Fe³⁺ on both Fe(III)‐reduction and Cu(II)‐reduction activities. The response of Cu(II)‐reduction activity to Cu concentration, in the presence or absence of citrate, was also studied. Results showed that inhibition of the ferric‐chelate reductase activity by Cu²⁺ or Ni²⁺ could be partially reversed by increasing the concentration of Fe‐EDTA. The Cu(II)‐reduction activity was even stimulated by Fe‐EDTA or Ni²⁺; it was inhibited by a high concentration of Cu²⁺ itself; and it was not affected by the absence of citrate. Mn⁴⁺ caused a moderate inhibition of both Fe(III)‐reduction and Cu(II)‐reduction activities. Results agree with the hypothesis that free Cu²⁺ ion is the substrate for Cu(II)‐reduction and suggest that the mechanisms involved in Fe(III)‐reduction and Cu(II)‐reduction could have some differences and be affected by metals in different ways. The mode of action of metals on the reductase activity are discussed, but they are still not well known.     

Role of root-induced changes in the rhizosphere for iron acquisition in higher plants

Plants can mobilize iron (Fe) in the rhizosphere by non-specific and specific (adaptive) mechanisms. Non-specific mechanisms are, for example, rhizosphere acidification related to high cation-anion uptake ratios, or citric acid excretion. The specific mechanisms are root responses to Fe deficiency and can be classified into two different strategies. The Strategy I is typical for dicots and monocots except for grasses (graminaceous species) and is characterized by increased plasma membrane-bound reductase activity, enhanced net excretion of protons and enhanced release of reducing compounds, mainly phenolics. The reductase activity is stimulated by low pH, and with supply of Fe^III chelates, ferric reduction at the plasma membrane takes place prior to uptake. In contrast, in graminaceous species (Strategy II) these root responses are absent, but enhancement of release of Fe^III chelating compounds - phytosiderophores - takes place. These phytosiderophores are very efficient in mobilizing Fe^III from artificially prepared sparingly soluble inorganic compounds (e.g. Fe^III hydroxide) and from calcareous soils. The ferrated phytosiderophores are taken up by grasses at rates 10² to 10³ times higher than Fe supplied either as synthetic chelate or microbial siderophores (e.g. ferrioxamine B), indicating a specific membrane transport system for ferrated phytosiderophores in roots of grasses. In calcareous soils phytosiderophores not only mobilize Fe, but also Zn, Mn, and Cu by chelation. However, only the Fe^III phytosiderophores are taken up preferentially by Fe deficient grasses. The ecological advantages and disadvantages of Strategy I and Strategy II for Fe acquisition from calcareous soils are discussed.     

EFFECT OF IRON DEFICIENCY ON ROOT FERRIC CHELATE REDUCTASE,PROTON EXTRUSION,BIOMASS PRODUCTION AND MINERAL ABSORPTION OF CITRUS ROOT STOCK ORANGE JASMINE (MURRAYA EXOTICA L.)

Three-week iron (Fe) deficiency stress experiments were conducted using two citrus root stocks, Fe-deficiency tolerant Orange Jasmine (OJ, Murraya exotica L.) and the sensitive Flying Dragon [FD, Poncirus trifoliata var. monstrosa (T. Ito) Swingle]. Root ferric chelate reductase activity and proton extrusion increased in OJ between 12 and 18 d of stress, whereas there was no change in FD. Dry weight of OJ roots increased in contrast to FD which decreased. The Mn content in OJ remained the same even under Fe stress. Zn content in OJ roots doubled while that of FD increased 4-folds. The shoot/root Fe accumulation ratio increased in OJ while it decreased in FD. OJ apparently has mechanisms for increasing root biomass, controlling Fe reutilization and regulating manganese (Mn) and zinc (Zn) absorption in response to Fe deficiency. These mechanisms could help maintain homeostasis under heavy metal stress, which would be useful for improved growth of economically important citrus species.