Biochemical Responses to True and Bicarbonate-Induced Iron Deficiency in Grapevine Genotypes

ABSTRACT

Biochemical responses to direct or bicarbonate-induced iron (Fe) deficiency were compared in two Tunisian native grapevine varieties, Khamri (tolerant) and Balta4 (sensitive), and a tolerant rootstock, 140Ru. Woody cuttings of each genotype were irrigated with a nutrient solution containing one of the following: 20 μM Fe (control), 1 μM Fe (direct Fe-deficiency), or 20 μM Fe + 10 mM HCO₃ ^? (indirect bicarbonate-induced Fe-deficiency). Under direct Fe-deficient conditions, lower leaf chlorosis score and higher chlorophyll and leaf Fe contents were found in Khamri and 140Ru compared with Balta4. Moreover, indirect Fe deficiency caused similar effects on these parameters, which were more pronounced in Balta4. Both tolerant genotypes, Khamri and 140Ru, showed higher roots-acidification capacity and phenol release under the direct Fe deficiency compared with the bicarbonate-induced condition. In the sensitive variety, Balta4, no significant changes were found between the control and Fe-deficient plants. Root Fe(III)-reductase activity was strongly stimulated by both types of Fe deficiency in Khamri and 140Ru, and displayed no significant changes in Balta4. In the three genotypes, root and leaf activities of two Fe-containing enzymes, catalase and guaiacol peroxidase, were significantly affected under Fe deficiency (either direct or induced), though to a greater extent in the sensitive variety, Balta4. The latter also displayed higher leaf malonyldialdehyde (MDA) content, traducing an important membrane lipid peroxidation.     

Differences in Response to Iron Deficiency Among Some Lines of Common Bean

Abstract

Five dry bean cultivars (Coco blanc, Striker, ARA14, SVM29‐21, and BAT477) were evaluated for their resistance to iron deficiency on the basis of chlorosis symptoms, plant growth, capacity to acidify the external medium and the root‐associated Fe³⁺‐reduction activity. Plants were grown in nutrient solution supplied or not with iron, 45 µM Fe(III)EDTA. For all cultivars, plants subjected to iron starvation exhibited Fe‐chlorosis. These symptoms were more severe and more precocious in BAT477 and Coco blanc than in the others cultivars. An important acidification of the culture medium was observed between the 4th and the 8th days of iron starvation in Striker, SVM29‐21 and, particularly, ARA14 plants. However, all Fe‐sufficient plants increased the nutrient solution pH. This capacity of acidification appeared more clearly when protons extrusion was measured in 10 mM KCl + 1 mM CaCl₂. The above genotypic differences were maintained: ARA14 showed the higher acidification followed by Coco blanc and BAT477. Iron deficiency led also to an increase of the root‐associated Fe(III)‐reductase activity in all lines. However, genotypic differences were observed: Striker shows the highest capacity of iron reduction under Fe deficiency condition.     

Activities of Iron‐Containing Enzymes in Leaves of Two Tomato Genotypes Differing in Their Resistance to Fe Chlorosis

Abstract

Two tomato (Lycopersicon esculentum Mill., cvs. Pakmor and Target) genotypes differing in resistance to iron (Fe) deficiency were grown in nutrient solution under controlled environmental conditions over 50 days to study the relationships between severity of leaf chlorosis, total concentration of Fe, and activities of Fe‐containing enzymes in leaves. The activities of Fe‐containing enzymes ascorbate peroxidase, catalase, and guaiacol peroxidase, and additionaly the activity of glutathione reductase, an enzyme that does not contain Fe, were measured. Plants were supplied with 2 × 10^?7 M (Fe deficient) and 10^?4 M (Fe sufficient) FeEDTA, respectively. Leaf chlorosis appeared more rapidly and severely in Target (Fe deficiency senstive genotype) than Pakmor (Fe deficiency resistant genotype). On day 50, Pakmor had 2‐fold more chlorophyll than Target under Fe deficiency, while at adequate supply of Fe the two genotypes were very similar in chlorophyll concentration. Despite distinct differences in development of leaf chlorosis and chlorophyll concentrations, Pakmor and Target were very similar in concentrations of total Fe under Fe deficiency. In contrast to Fe concentration, activities of Fe‐containing enzymes were closely related to the severity of leaf chlorosis. The Fe‐containing enzymes studied, especially catalase, showed a close relationship with the concentration of chlorophyll and thus differential sensitivity of tomato genotypes to Fe deficiency. Glutathione reductase did not show relationship between Fe deficiency chlorosis and enzyme activity. The results confirm that measurement of Fe‐containing enzymes in leaves is more reliable than the total concentration of Fe for characterization of Fe nutritional status of plants and for assessing genotypical differences in resistance to Fe deficiency. It appears that Fe deficiency‐resistant genotype contains more physiologically available Fe in tissues than the genotype with high sensitivity to Fe deficiency.     

Iron‐stress response of inoculated and non‐inoculated roots of an iron inefficient soybean cultivar in a split‐root system

The objective of this study was to establish whether the iron‐stress responses observed in T203 soybean (Fe‐inefficient) with active nodules are products of the nodules or of the entire root system. A split‐root system was used in which half the roots of each plant were inoculated and actively fixing nitrogen and the other half were not. Soybean cultivar T203 is normally Fe‐inefficient and does not exhibit the Fe‐stress responses, however an iron‐stress response did occur during active N₂ fixation in earlier studies. This implies that the active nodules influenced the plant's ability to respond to Fe‐deficiency stress. In this study, the Fe‐stress response (H⁺ and reductant release) observed in T203 soybean was limited to the inoculated side of the split‐root system. The severe Fe chlorosis which developed in these plants was overcome in a manner similar to Fe‐efficient cultivars undergoing normal Fe‐stress response and the T203 plants completely regreened. Exudation of H⁺ ions was similar in both the presence and absence of Fe, and was generally limited to inoculated roots. Reductant release was nearly nonexistent from the non‐inoculated roots and was greater for the Fe‐stressed (‐Fe) plants than for non‐stressed (+Fe) plants. Thus, the response observed, which alleviated Fe chlorosis, appeared to be associated with N₂ fixation of the active nodules.     

Mechanisms of exogenous nitric oxide and 24-epibrassinolide alleviating chlorosis of peanut plants under iron deficiency

Iron(Fe) is a crucial transition metal for all living organisms including plants; however, Fe deficiency frequently occurs in plant because only a small portion of Fe is bioavailable in soil in recent years. To cope with Fe deficiency, plants have evolved a wide range of adaptive responses from changes in morphology to altered physiology. To understand the role of nitric oxide(NO) and 24-epibrassinolide(EBR) in alleviating chlorosis induced by Fe deficiency in peanut(Arachis hypogaea L.) plants, we determined the concentration of chlorophylls, the activation, uptake, and translocation of Fe, the activities of key enzymes, such as ferric-chelate reductase(FCR),proton-translocating adenosine triphosphatase(H~+-ATPase), and antioxidant enzymes, and the accumulation of reactive oxygen species(ROS) and malondialdehyde(MDA) of peanut plants under Fe sufficiency(100 μmol L~(-1)ethylenediaminetetraacetic acid(EDTA)-Fe) and Fe deficiency(0 μmol L~(-1)EDTA-Fe). We also investigated the production of NO in peanut plants subjected to Fe deficiency with foliar application of sodium nitroprusside(SNP), a donor of NO, and/or EBR. The results showed that Fe deficiency resulted in severe chlorosis and oxidative stress, significantly decreased the concentration of chlorophylls and active Fe, and significantly increased NO production. Foliar application of NO and/or EBR increased the activity of antioxidant enzymes, superoxide dismutase,peroxidase, and catalase, and decreased the ROS and MDA concentrations, thus enhancing the resistance of plants to oxidative stress.Application of NO also significantly increased Fe translocation from the roots to the shoots and enhanced the transfer of Fe from the cell wall fraction to the cell organelle and soluble fractions. Consequently, the concentrations of available Fe and chlorophylls in the leaves were elevated. Furthermore, the activities of H~+-ATPase and FCR were enhanced in the Fe-deficient plants. Simultaneously,there was a significant increase in NO production, especially in the plants that received NO, regardless of Fe supply. These suggest that NO or EBR, and, especially, their combination are effective in alleviating plant chlorosis induced by Fe deficiency.     

Influences of ultra‐violet (UV)‐blue light radiation on the growth of cotton. I. Effect on iron nutrition and iron stress response

Cool white fluorescent (CWF) light reduces Fe³⁺ to Fe²⁺ while low pressure sodium (LPS) light does not. Cotton plants grown under CWF light are green, while those yrown under LPS light develop a chlorosis very similar to the chlorosis that develops when the plants are deficient in iron (Fe). It could be that CWF light (which has ultra violet) makes iron more available for plant use by maintaining more Fe²⁺ in the plant. Two of the factors commonly induced by Fe‐stress in dicotyledonous plants‐‐hydroyen ions and reductants released by the roots‐‐were measured as indicators of the Fe‐deficiency stress response mechanism in M8 cotton.

The plants were grown under LPS and CWF light in nutrient solutions containing either NO₃‐N or NH₄‐N as the source of nitrogen, and also in a fertilized alkaline soil. Leaf chlorophyll concentration varied significantly in plants grown under the two light sources as follows: CWF+Fe > LPS+Fe > CWF‐Fe ≥ LPS‐Fe. The leaf nitrate and root Fe concentrations were significantly greater and leaf Fe was generally lower in plants grown under LPS than CWF light. Hydrogen ions were extruded by Fe‐deficiency stressed roots grown under either LPS or CWF light, but “reductants”; were extruded only by the plants grown under CWF light. In tests demonstrating the ability of light to reduce Fe³⁺ to Fe²⁺ in solutions, enough ultra violet penetrated the chlorotic leaf of LPS yrown plants to reduce some Fe³⁺ in a beaker below, but no reduction was evident through a yreen CWF grown leaf.

The chlorosis that developed in these cotton plants appeared to be induced by a response to the source of liyht and not by the fertilizer added. It seems possible that ultra violet liyht could affect the reduction of Fe³⁺ to Fe²⁺ in leaves and thus control the availability of this iron to biological systems requiring iron in the plant.     

Iron inefficient and efficient oat cultivars. II. Characterization of phytosiderophore released in response to Fe deficiency stress

Abstract

Iron‐inefficient TAM 0–312 and Fe‐efficient Coker 227 oats (Strategy II plants) differ in their release of phytosiderophore in response to iron‐deficiency stress—the Fe‐efficient Coker 227 releases a phytosiderophore whereas the Fe‐inefficient TAM 0–312 does not. The phytosiderophore released by Coker 227 oats in response to Fe‐deficiency stress does not appear to transport Fe into the plant as Fe phytosiderophore. When the Fe‐inefficient TAM 0–312 and Fe‐efficient Coker 227 oats were subjected to Fe supplied as Fe²⁺(BPDS)₃, Fe³⁺HEDTA, as Fe³⁺EDDHA, Coker 227 utilized the Fe more efficiently than TAM 0–312 in every case. Both cultivars reduced Fe³⁺ as FeCl₃ to form Fe²⁺(BPOS)₃ and responded better to this form of Fe than Fe supplied as the ferric chelate. Reduction of Fe³⁺ at the root appears to be a factor that facilitates iron uptake by Coker 227 oats and the release of a phytosiderophore appears to make more Fe available at the root that can be reduced and transported to plant tops.     

Uptake of Iron (59Fe) Complexed to Water‐Extractable Humic Substances by Sunflower Leaves

Abstract

A research was carried out to evaluate the leaves' ability to utilize Fe supplied as a complex with water‐extractable humic substances (WEHS) and the long‐distance transport of ⁵⁹Fe applied to sections of fully expanded leaves of intact sunflower (Helianthus annuus L.) plants. Plants were grown in a nutrient solution containing 10 µM Fe(III)‐EDDHA (Fe‐sufficient plants), with the addition of 10 mM NaHCO₃ to induce iron chlorosis (Fe‐deficient plants). Fe(III)‐WEHS could be reduced by sunflower leaf discs at levels comparable to those observed using Fe(III)‐EDTA, regardless of the Fe status. On the other hand, ⁵⁹Fe uptake rate by leaf discs of green and chlorotic plants was significantly lower in Fe‐WEHS‐treated plants, possibly suggesting the effect of light on photochemical reduction of Fe‐EDTA. In the experiments with intact plants, ⁵⁹Fe‐labeled Fe‐WEHS or Fe‐EDTA were applied onto a section of fully expanded leaves. Irrespective of Fe nutritional status, ⁵⁹Fe uptake was significantly higher when the treatment was carried out with Fe‐EDTA. A significant difference was found in the amount of ⁵⁹Fe translocated from treated leaf area between green and chlorotic plants. However, irrespective of the Fe nutritional status, no significant difference was observed in the absolute amount of ⁵⁹Fe translocated to other plant parts when the micronutrient was supplied either as Fe‐EDTA or Fe‐WEHS. Results show that the utilization of Fe complexed to WEHS by sunflower leaves involves an Fe(III) reduction step in the apoplast prior to its uptake by the symplast of leaf cells and that Fe taken up from the Fe‐WEHS complexes can be translocated from fully expanded leaves towards the roots and other parts of the shoot.     

Association of soil properties and soil and plant iron to iron deficiency response in rice (Oryza Sativa L.)

Abstract

Problems are invariably encountered when attempts are made to explain the variability in Bray percent yields or plant response in terms of soil or plant iron (Fe). To resolve this inconsistency, the present investigation was initiated to identify a combination of soil extractable Fe, soil properties and form of plant Fe that may be used as a measure of Fe deficiency. The study involved 16 diverse soils, using upland rice (Oryza sativa L.) as the test crop and Fe‐EDDHA [ferric ethylenediamine di (o‐hydroxyl‐phenyl acetic acid)] as source of Fe. The results showed that Bray percent yields were neither related to DTPA (diethylenetriamine pentaacetic acid) or EDTA (ethylenediamine tetraacetic acid) extractable Fe nor with total plant Fe. Even the inclusion of pH, lime, organic carbon and clay data in the regression equations was of no value. However, Bray percent yields were significantly and positively (r = 0.57* ) associated with ferrous Fe (Fe²⁺) in 40‐day‐old rice plants. The explanation concerning variability in Bray percent yields obtained on diverse soils could be increased about one and half 2 times (R²= 0.59*) if the contribution of lime and soil pH was also incorporated in the stepwise regression analysis. The individual contribution to R of lime, pi respectively. Thus, it appears that Fe²⁺ concentration in plants (along with soil pH) may identify Fe deficiency. The critical limit to separate Fe deficient from green rice plants was set at 45 ug Fe²⁺/g in the leaves.     

Does the Sulfur Assimilation Pathway Play a Role in the Response to Fe Deficiency in Maize (Zea mays L.) Plants?

Abstract

The finding that the methionine is the sole precursor of the mugineic acid family phytosiderophores induced us to evaluate whether sulfur assimilation pathway has a role in plant response to Fe deficiency. Maize plants were grown for 10 days in nutrient solution (NS) containing 80 µM Fe in the presence (+S) or absence (?S) of sulfate. After removing the root extraplasmatic iron pool, half of the plants of each treatment (+S and ?S) were transferred to a new Fe deficient NS (0.1 µM final Fe concentration) (?Fe). The remaining plants of each pre‐culture condition (+S and ?S) were transferred to a new NS containing 80 µM Fe (+Fe). Leaves were collected 4 and 24 hours from the beginning of Fe deprivation period and used for chemical analysis and enzyme assays. Results showed that iron content in the leaves was lower in plants grown in S‐deficiency than in those grown in the presence of the macro‐nutrient. Iron deprivation produced an increase in the level of SH compounds in both nutritive conditions (+S and ?S). These observations are suggestive of some relationship between S nutrition and Fe uptake. For this reason, we next investigated the influence of Fe availability on S metabolism through the evaluation of changes in ATPs and OASs activity, the first and the last enzyme of S assimilation pathway respectively. Results showed that S‐starvation increased the activity of both enzymes, but this effect disappeared in plants upon Fe deficiency suggesting that S metabolism is sensitive to Fe availability. Taken together these evidences suggest that S metabolism is sensitive to soil Fe‐availability for plant nutrition and support the hypothesis of S involvement in plant response to Fe deprivation.     

Nutritional Alleviation of Zinc-Induced Iron Deficiency in Indian Mustard and the Effects on Zinc Phytoremediation

ABSTRACT

Indian mustard (Brassica juncea Czern) is a promising species for the phytoextraction of zinc (Zn), but the effectiveness of this plant can be limited by iron (Fe) deficiency under Zn-contaminated conditions. Our objectives were to determine the effects of root-applied Fe and Zn on plant growth, accumulation of Zn in plant tissues, and development of nutrient deficiencies for B. juncea. In the experiment, B. juncea was supplied 6 levels of iron ethylenediamine dihydroxyphenylacetic acid (Fe-EDDHA; 0.625 to 10.0 mg L^?1) and two levels of Zn (2.0 and 4.0 mg L^?1) for 3 weeks in a solution-culture experiment. Nutrient solution pH decreased with decreasing supply of Fe and increasing supply of Zn in solution, indicating that B. juncea may be an Fe-efficient plant. If plants were supplied 2.0 mg Zn L^?1, plant growth was stimulated by increases in Fe supply, but plant growth was not influenced by Fe treatments if plants were supplied 4.0 mg Zn L^?1. Zinc concentration in roots and shoots was suppressed by increasing levels of Fe in solution. Leaf concentrations of Cu, Mn, and P were suppressed also as Fe supply in solutions increased. Iron additions to the nutrient solution were not effective at increasing the Zn-accumulation potential of B. juncea unless plants were supplied the higher level of Zn in solution culture. Even under these conditions, Fe additions were effective only if supplied at low levels in solution culture (1.25 mg Fe L^?1). Results suggest that Fe fertility has limited potential for enhancing Zn phytoextraction by B. juncea, even if plants suffer a suppression in growth from Fe deficiency.     

Nature-inspired soluble iron-rich humic compounds: new look at the structure and properties

Purpose

Humic substances (HS) being natural polyelectrolyte macromolecules with complex and disordered molecular structures are a key component of the terrestrial ecosystem. They have remarkable influence on environmental behavior of iron, the essential nutrient for plants. They might be considered as environmental friendly iron deficiency correctors free of synthetic iron (III) chelates disadvantages. The main goal of this study was to obtain water-soluble iron-rich humic compounds (IRHCs) and to evaluate their efficiency as chlorosis correctors.

Materials and methods

A facile preparation technique of IRHCs based on low-cost and available parent material was developed. The iron-containing precursor (ferrous sulfate) was added dropwisely into alkaline potassium humate solution under vigorous stirring and pH-control. A detailed characterization both of organic and inorganic parts of the compounds was provided, the iron species identification was carried out jointly by EXAFS and Mössbauer spectroscopy. Bioassay experiments were performed using cucumber Cucumis sativus L. as target plants. Plants were grown in modified Hoagland nutrient solution, prepared on deionized water and containing iron in the form of IRHCs. Total iron content in dry plants measured by spectrophotometry after oxidative digestion and the chlorophyll a content determined after acetone extraction from fresh plant were used as parameters illustrating plants functional status under iron deficiency condition.

Results and discussion

The high solubility (up to130 g/l) and iron content (about 11 wt%) of the IRHCs obtained allow considering them to be perspective for practical applications. A set of analytical methods has shown that the main iron species in IRHCs are finely dispersed iron (III) oxide and hydroxide nanoparticles. An application of the precursor solution acidification allows to obtain compounds containing a significant part of total iron (up to 30 %) in the form of partly disordered iron (II–III) hydroxysulphate green rust GR(SO₄ ^2?). Bioavailability of iron from IRHCs was demonstrated using bioassay in cucumber plants grown up on hydroponics under iron deficiency conditions.

Conclusions

The application of iron oxides chemistry for humic substance containing solution was proved to be an effective approach to synthesis of IRHCs. Using bioassay on cucumber plants C. sativus L. under iron deficiency conditions, the efficiency of compounds obtained as chlorosis correctors was demonstrated. Application of water-soluble IRHCs led to significant increase of chlorophyll a content (up to 415 % of the blank) and iron content in plants (up to 364 % of the blank) grown up on hydroponics.     

Iron deficiency‐induced zinc uptake by bread wheat

The present study aimed to test the contribution of the iron (Fe) deficiency‐induced uptake system to zinc (Zn) and copper (Cu) uptake by using bread wheat (Triticum aestivum cv. Bezostaja). For this purpose, two different uptake experiments, long‐term and short‐term, were set up in a nutrient solution culture under controlled growth chamber conditions. For the long‐term experiment, wheat cv. plants were grown with different concentrations of Fe or Zn. Results show that there was an uptake system induced under Fe‐limiting conditions which also contributed to Zn and Cu uptake. However, the Zn deficiency‐induced uptake mechanism affected neither Fe nor Cu uptake by wheat. Short‐term uptake experiments indicate that Fe deficiency‐induced Zn²⁺ uptake was more enhanced than the absorption of Zn‐phytosiderophore (PS) complexes. In addition, the Fe‐deficient plants absorbed more Zn in comparison to those plants supplied with sufficient Fe. Similar tendencies in Zn uptake under Fe deficiency in both short‐ and long‐term experiments suggest that there may be a specific Fe uptake system induced under Fe‐limiting conditions for non‐chelated metals in bread wheat. Moreover, this system also contributes to the transport of inorganic forms of some other metals, such as Zn and Cu. Although evidence is still needed involving the use of molecular biological techniques, it is hypothesized that IRT‐like proteins are responsible for this uptake system. Moreover, the release of Fe deficiency‐induced phytosiderophores and uptake of Fe(III)‐phytosiderophore complexes may not be the only mechanisms involved in the adaptation of wheat to Fe‐limiting conditions.     

Iron-Deficiency Responses in Cereal Seedlings

Acidity release as a characteristic of iron (Fe) deficiency stress responses was investigated during early seedling establishment of maize (Zea mays L.), barley (Hordeum vulgare L.), winter wheat (Triticum aestivum L.), and winter rye (Secale cereale L.). Ferric reduction as another characteristic of Fe deficiency stress responses was also investigated in maize. This has previously been considered a process only occurring in iron-stressed dicotyledonous plants. Measurements were made of the Fe efflux from maize endosperm under Fe-deficient and non-limiting conditions. The scutellum of Fe-stressed cereal seedlings (all species investigated) increased agar acidification by 30–40%, while manganese (Mn) or zinc (Zn) stressed seedlings did not demonstrate this reaction. Among the maize genotypes studied (‘VIR-7569’ and ‘VIR-7882’), the Fe-stressed acid release was more prolonged in relatively less Fe-efficient cv. ‘VIR-7569.’ In maize, the induced enhancement of acid release was accompanied by an increase in Fe efflux from endosperm. Conversely, maize scutellum Fe^{3 +}-reductase activity was not deficiency-dependent on Fe, indicating that enhanced acidity release is a biological phenomenon facilitating Fe supply at early development of graminaceous plants. In cereal seedlings, the coexistence of two marker responses to Fe-deficiency (Strategy I and Strategy II), which are usually typical of different taxonomic groups (dicotyledonous and graminaceous plants) can be regarded as evidence in favour of the hypothesis of common genesis in dicotyledonous and graminaceous plants.     

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.     

Ecophysiology of iron homeostasis in plants

In nature, iron (Fe) occurs in abundance and ranks fourth among all elements on Earth’s surface. Still, its availability to plants is reduced, once this element is in the form of hydrated oxides, which can limit plant productivity and biomass production. On the other hand, in high concentrations, this essential micronutrient for the plants can become a toxic agent, increasing the environmental contamination. Fe is necessary for the maintenance of essential processes like respiration and photosynthesis, participating in the electron transport chain and in the conversion between Fe²⁺ and Fe³⁺, being a key element for carbon dioxide (CO₂) fixation and, therefore, important for crop production of cultivated or natural species. The balance of Fe should be strictly controlled, because both its deficiency and its toxicity affect the physiological process of plants. In aerated soils Fe is present in the form of Fe³⁺, which is the oxidized form and is less available to plants, so these organisms have developed different strategies for absorption, transport and storage of Fe. Deficiency and excess of Fe correlate with local soil conditions and with the care adopted in plant nutrition during the phenological phases and/or in the course of its cultivation. In situations of excessive accumulation of Fe in tissues, an enhancement of hydroxyl radical generation (OH^?) occurs by Fenton reaction. Here, we review the nutritional, genetic and ecophysiological aspects of uptake, translocation and accumulation of Fe ions in plants growing under conditions of deficiency or toxicity of this metal.     

Plant Uptake of Iron Chelated by Humic Acids of Different Molecular Weights

Abstract

Mobilization of iron (Fe) chelated by humic acids (HA) of low (HA_10,000) and high molecular weight (HA_100,000) fractions and its uptake by plants were investigated in growth experiments with sunflower seedlings. The iron chelates (labeled with ⁵⁹Fe) contained in dialysis bags (mw. cutoff=3500) were placed in minus iron Hoagland solutions as the Fe source and at the same time fulvic acid (FA), EDTA, and low and high molecular weight HA fractions were added in the solutions as mobilizators. Characterization of FA, HA_10,000, and HA_100,000 were performed by infrared spectroscopy and chemical analysis, e.g., total acidity, COOH, and phenolic‐OH content. Roots and leaves were harvested, dried, and ground for Fe activity determination. Iron contents and pH in the nutrient solutions were measured before and after treatments. The supply of Fe to the plants was apparently sufficient, because no Fe deficiency has been detected in the test plants but during the whole absorption period, the pH of the nutrient solution was about 4.5. The Fe contents in leaves indicated that part of the Fe was rapidly transported from roots to leaves. Judging from the Fe contents in leaves, it was assumed that the small size HA_10,000 and EDTA were the most efficient in affecting transport of Fe from root to leaf tissue. FA, HA_10,000, and especially HA_100,000 were unable to penetrate the dialysis bags and, hence, were effective in Fe mobilization only after the Fe, dissociated from the Fe‐HA chelate, has passed the dialysis membrane into the nutrient solutions. In contrast, the small size EDTA was expected to have penetrated the dialysis bags, permitting mobilization of chelated Fe by ligand exchange inside the bags, and transporting the Fe to the roots. The results suggested that the humic substances used in this study were able to form with the Fe³⁺ ion complexes that maintained the iron available to the sunflower plants. In the chemical form of Fe.L, where L was FA o HA, the iron within the bags or in solution or in the roots free space, was available for exchange reactions with the natural sunflower plant chelators for its transport to the leaves.     

Efficacy of an artificial microbial siderophore-Fe(III) with high redox potential on correcting Fe chlorosis in rice

ABSTRACT

Microbial siderophore-chelated Fe(III) is suggested to be an important source of Fe for plants, although it is hardly reduced by plant roots. Here, we investigated the efficacy of the easily reducible artificial microbial siderophore tris[2-{(N-acetyl-N-hydroxy)glycylamino}ethyl]amine (TAGE)-Fe(III) as an alternative Fe source to correct Fe deficiency in rice plants, and compared it to that of the natural siderophore deferoxamine B (DFOB)-Fe(III). We also evaluated the absorption of Fe from TAGE-Fe(III) by the Strategy I-like system of gramineous plants using nicotianamine aminotransferase 1 (naat1) mutant rice, which does not synthesize phytosiderophores. Fe(III)-siderophores were synthesized in vitro. Nipponbare rice and its naat1 mutant were reared in soil and gel cultures to determine Fe availability. Hydroponically grown naat1 mutant seedlings were used for reducibility assays to determine the ability of rice roots to reduce Fe(III) chelated by TAGE or DFOB. The expression of a Fe-deficiency inducible gene was also determined, as well as chlorophyll and Fe concentrations. Reduci bility assays on naat1 mutant seedlings revealed that the reduction level of TAGE-Fe(III) was approximately three times higher than that of DFOB-Fe(III). Application of TAGE-Fe(III) to both culture medium and alkaline soil improved Fe chlorosis, growth, and Fe concentration in both naat1 and wild type plants, whereas application of DFOB-Fe(III) only did so in wild type plants. Easily reducible Fe(III)-chelates such as TAGE-Fe(III) can be a better source of Fe for rice plants than most natural microbial siderophores-Fe(III). Our study also demonstrated that rice plants have the ability to utilize microbial siderophores-Fe(III) as the Fe source through the Strategy I-like Fe acquisition system.     

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.     

Effects of exogenous SA supplied with different approaches on growth,chlorophyll content and antioxidant enzymes of peanut growing on calcareous soil

To assess the role of salicylic acid (SA) supplied with 5 approaches in alleviating chlorosis induced by iron (Fe) deficiency in peanut plants growing on calcareous soil, SA was supplied as soil incorporation, making slow-release particles, seed soaking, irrigation and foliar application. SA application, particularly, SA supplied by slow release particles, dramatically increased growth parameters, yield and quality of peanut, and increased Fe concentration in peanut grain. Meanwhile, SA application increased the H⁺-ATPase activity, reduced pH of soil, increased Fe³⁺-Chelate Reductase (FCR) activity in roots, and increased Fe concentration in roots. Furthermore, SA increased active Fe content and increased chlorophyll content. In addition, SA improved enzymes activities containing superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT), and protected Fe deficiency induced oxidative stress. Therefore, SA has a good effect on alleviating chlorosis induced by Fe deficiency on calcareous soil. However, in the 5 SA supplied approaches, foliar application and making slow release particles were more effective.