Ferric Chelate Reductase Research Articles

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 pmol 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.

Interactions between aluminium, iron and silicon in Cucumber sativus L. grown under acidic conditions

Aluminium (Al) is one of the major stressors for plants in acidic soils, negatively affecting plant growth and nutrient balances. Significant efforts have been undertaken to understand mechanisms of Al tolerance in plants. However, little is known of the relevance of iron (Fe) and silicon (Si) nutrition under Al stress conditions. The objectives of this study were to determine whether effects induced by Fe and Si are of importance for limitation of Al moving via xylem in plants (Cucumis sativus L.). Cucumber plants (cv. Phoenix and Solovei) were grown (i) hydroponically in a complete nutrient solution at pH 4.0, either with (+Fe) or in Fe-free (−Fe) nutrient solution, without (−Si) or with (+Si) supply of Si, without (−Al) or with (+Al) exposure of Al and (ii) in soil. Xylem sap concentrations of Al, Fe and Si were measured. To characterise the pattern of xylem sap transport of Al and Fe, metabolomic changes of root tissues were investigated. Although the growth of cucumber plants was not significantly affected by Al3+ (Al-tolerant), Al exposure decreased xylem sap Fe (+Fe plants) and increased ferric chelate reductase (FC-R) activity of roots (−Fe plants). On the other hand, Fe supply greatly mitigated the Al-induced increase in xylem sap Al. The ameliorative effect of Fe depended on plant genotypes and was more pronounced in the more Fe-efficient cultivar Phoenix, which presented the highest level of xylem sap Fe. Xylem sap Fe was positively correlated with root serine, succinic and fumaric acids, suggesting that a probable underlying mechanism of Al tolerance might involve the chelation of Fe by biosynthesis of these chelating compounds. The Si-modulated root succinate increase appears to be of great importance for facilitating long-distance transport of Fe, thereby hindering Al transport from roots to shoots. The results highlight for the first time the importance of both Fe and Si supply in plant exclusion of Al under acidic conditions.

Response of soybean plants to the application of synthetic and biodegradable Fe chelates and Fe complexes

The growing concern over the environmental risk of synthetic chelate application promotes the search for alternatives in Fe fertilization, such as biodegradable chelating agents and natural complexing agents. In this work, plant responses to the application of several Fe treatments (chelates and complexes) was analyzed to study their potential use in Fe fertilization under calcareous conditions. Thus, the root ferric chelate reductase (FCR) activity of soybean (Glycine max cv. Klaxon) plants was determined, and the effectiveness of the Fe chelates and complexes assessed in a pot experiment, by SPAD and fluorescence induction measurements, and the determination of Fe distribution in plant and soil. Additionally, 57Fe Mössbauer spectroscopy was conducted to identify the Fe forms present in the soybean roots. The highest FCR activity was observed for the chelates EDDS/Fe3+ and IDHA/Fe3+; while no activity was observed when using complexes as Fe substrates. In contrast to the FCR data, the pot experiment confirmed that the o,oEDDHA/Fe3+ is the most effective treatment, and the complexes LS/Fe3+ and GA/Fe3+ are able to alleviate Fe chlorosis, also indicated by SPAD data and the maximal quantum efficiency of photosystem II reaction centers as vitality parameters, and the enhanced plant uptake of Fe from natural sources.

Interaction of \u03b3-Fe2O3 nanoparticles with Citrus maxima leaves and the corresponding physiological effects via foliar application

BackgroundNutrient-containing nanomaterials have been developed as fertilizers to foster plant growth and agricultural yield through root applications. However, if applied through leaves, how these nanomaterials, e.g. γ-Fe2O3 nanoparticles (NPs), influence the plant growth and health are largely unknown. This study is aimed to assess the effects of foliar-applied γ-Fe2O3 NPs and their ionic counterparts on plant physiology of Citrus maxima and the associated mechanisms.ResultsNo significant changes of chlorophyll content and root activity were observed upon the exposure of 20–100 mg/L γ-Fe2O3 NPs and Fe3+. In C. maxima roots, no oxidative stress occurred under all Fe treatments. In the shoots, 20 and 50 mg/L γ-Fe2O3 NPs did not induce oxidative stress while 100 mg/L γ-Fe2O3 NPs did. Furthermore, there was a positive correlation between the dosages of γ-Fe2O3 NPs and Fe3+ and iron accumulation in shoots. However, the accumulated iron in shoots was not translocated down to roots. We observed down-regulation of ferric-chelate reductase (FRO2) gene expression exposed to γ-Fe2O3 NPs and Fe3+ treatments. The gene expression of a Fe2+ transporter, Nramp3, was down regulated as well under γ-Fe2O3 NPs exposure. Although 100 mg/L γ-Fe2O3 NPs and 20–100 mg/L Fe3+ led to higher wax content, genes associated with wax formation (WIN1) and transport (ABCG12) were downregulated or unchanged compared to the control.ConclusionsOur results showed that both γ-Fe2O3 NPs and Fe3+ exposure via foliar spray had an inconsequential effect on plant growth, but γ-Fe2O3 NPs can reduce nutrient loss due to their the strong adsorption ability. C. maxima plants exposed to γ-Fe2O3 NPs and Fe3+ were in iron-replete status. Moreover, the biosynthesis and transport of wax is a collaborative and multigene controlled process. This study compared the various effects of γ-Fe2O3 NPs, Fe3+ and Fe chelate and exhibited the advantages of NPs as a foliar fertilizer, laying the foundation for the future applications of nutrient-containing nanomaterials in agriculture and horticulture.Graphical abstractγ-Fe2O3 NPs exposed on plants via foliar spray and genes associated with the absorption andtransformation of iron, as well as wax synthesis and secretion in Citrus maxima leaves

Heavy Metals Induce Iron Deficiency Responses at Different Hierarchic and Regulatory Levels.

In plants, the excess of several heavy metals mimics iron (Fe) deficiency-induced chlorosis, indicating a disturbance in Fe homeostasis. To examine the level at which heavy metals interfere with Fe deficiency responses, we carried out an in-depth characterization of Fe-related physiological, regulatory, and morphological responses in Arabidopsis (Arabidopsis thaliana) exposed to heavy metals. Enhanced zinc (Zn) uptake closely mimicked Fe deficiency by leading to low chlorophyll but high ferric-chelate reductase activity and coumarin release. These responses were not caused by Zn-inhibited Fe uptake via IRON-REGULATED TRANSPORTER (IRT1). Instead, Zn simulated the transcriptional response of typical Fe-regulated genes, indicating that Zn affects Fe homeostasis at the level of Fe sensing. Excess supplies of cobalt and nickel altered root traits in a different way from Fe deficiency, inducing only transient Fe deficiency responses, which were characterized by a lack of induction of the ethylene pathway. Cadmium showed a rather inconsistent influence on Fe deficiency responses at multiple levels. By contrast, manganese evoked weak Fe deficiency responses in wild-type plants but strongly exacerbated chlorosis in irt1 plants, indicating that manganese antagonized Fe mainly at the level of transport. These results show that the investigated heavy metals modulate Fe deficiency responses at different hierarchic and regulatory levels and that the interaction of metals with physiological and morphological Fe deficiency responses is uncoupled. Thus, this study not only emphasizes the importance of assessing heavy metal toxicities at multiple levels but also provides a new perspective on how Fe deficiency contributes to the toxic action of individual heavy metals.

Identification and characterization of a grain micronutrient-related OsFRO2 rice gene ortholog from micronutrient-rich little millet (Panicum sumatrense).

Minor millets are considered as nutrient-rich cereals having significant effect in improving human health. In this study, a rice ortholog of Ferric Chelate Reductase (FRO2) gene involved in plant metal uptake has been identified in iron-rich Little millet (LM) using PCR and next generation sequencing-based strategy. FRO2 gene-specific primers designed from rice genome amplified 2.7Kb fragment in LM genotype RLM-37. Computational genomics analyses of the sequenced amplicon showed high level sequence similarity with rice OsFRO2 gene. The predicted gene structure showed the presence of 6 exons and 5 introns and its protein sequence was found to contain ferric reductase and NOX_Duox_Like_FAD_NADP domains. Further, 3D structure analysis of FCR-LM model protein (494 amino acids) shows that it has 18 helices, 10 beta sheets, 10 strands, 41 beta turn and 5 gamma turn withslight deviation from the FCR-Os structure. Besides, the structures of FCR-LM and FCR-Os were modelled followed by molecular dynamics simulations. The overall study revealed both sequence and structural similarity between the identified gene and OsFRO2. Thus, a putative ferric chelate reductase gene has been identified in LM paving the way for using this approach for identification of orthologs of other metal genes from millets. This also facilitates mining of effective alleles of known genes for improvement of staple crops like rice.

Use of compact and friable callus cultures to study adaptive morphological and biochemical responses of potato (Solanum tuberosum) to iron supply

The efficient utilization, storage and buffering of iron (Fe) is necessary to prevent the development of iron deficiency or toxicity related stresses in plants. It is of practical interest to develop effective in vitro plant tissue culture schemes for generating chlorosis-tolerant and/or Fe-biofortified plants. The objectives of this study were to induce the formation of compact and friable calli from leaf explants of potato and to investigate adaptive responses to Fe nutrition using these in vitro cell culture-based systems. The severity of chlorosis of the callus cultures increased with the withdrawal of Fe supplementation from the growth medium and the culture duration. Ferric chelate reductase (FCR) activity was found to be higher in the compact compared to the friable potato callus cultures. Chlorophyll and carotenoids content in compact calli increased with an increase in Fe supply but in friable calli carotenoids content declined and the chlorophyll level was unaffected. Phenolic content in Fe-deficient compact calli was significantly higher than in friable ones but a reverse trend was observed as the culture period was extended. Peroxidase activity was reduced in both compact and friable calli cultured under Fe-deprived condition compared to those on Fe-sufficient medium. Friable and compact calli displayed differential morphological and biochemical responses to Fe deficiency. These calli also appeared to vary in their sensitivity to Fe-deficiency and the response to Fe nutrition seemed to be related to the culture duration.

Root plant growth promoting rhizobacteria inoculations increase ferric chelate reductase (FC-R) activity and Fe nutrition in pear under calcareous soil conditions

Iron deficiency occurring in calcareous soil is a problem in various plants. It is well known that some soil bacteria can release organic acids that can decrease the pH of the soil rhizosphere. However, there have been no attempts to study the effects of plant growth promoting rhizobacteria (PGPR), including organic acid releasing bacteria, on the organic acid contents of the leaf and FC-R activity in the roots and leaves under calcareous soil conditions. Therefore, pear plants were inoculated with 6 bacterial strains with the aim of acquiring iron under calcareous conditions. Uniform 1-year-old pear cv. Deveci sapling grafted on BA-29 and OHF-333 rootstocks were planted in plastic pots containing 10L of loamy soil at 29.6% CaCO3. All bacteria were applied to the roots as an inoculation before planting. The root and leaf Fe content, FC-R activity, leaf organic acids, and soil Fe content were compared in the Alcaligenes 637Ca, Agrobacterium A18, Staphylococcus MFDCa1, MFDCa2, Bacillus M3 and Pantoea FF1 strains. The study showed that the leaf organic acid content and the Fe content in the soil, root and leaf were significantly affected by the bacterial treatments in pear plants. It was determined that the total and active Fe in the leaf was higher in OHF-333 compared to BA-29 by 7% and 14%, respectively. Furthermore, the leaf FC-R activity of Deveci on OHF-333 was 8% higher than that on BA-29. In the Deveci/BA-29 plants, the 637Ca treatment had the highest root FC-R activity value (107nmolFe+2gr−1FW h−2). The highest leaf FC-R activity value was obtained from the MFDCa1, MFDCa2 and FF1 treatments (58.4, 56.3 and 55.7nmolFe+2gr−1FWh−2, respectively). The bacterial strains used in the present study have an important potential to be used as a biofertilizer to replace the use of iron fertilizers.

Investigation of responses of some apple (Mallus x domestica Borkh.) cultivars grafted on MM106 and M9 rootstocks to lime-induced chlorosis and oxidative stress

The response of dwarf (M9) and semi dwarf (MM106) clonal apple rootstocks and 5 apple cultivars [Granny Smith (GS), Fuji (F), Royal Gala (RG), Mondial Gala (MG) and Red Chief (RC)] grafted on these rootstocks to lime induced iron chlorosis and iron deficiency induced oxidative stress were investigated. Both rootstocks and cultivars were divided into two groups. One of the group were kept as control group. To the second group, to ensure the lime-induced chlorosis, bicarbonate (HCO3−) was applied. Both groups were grown for two years in a pot experiment. The soil used in the experiment was a clay loam calcareous soil containing 70.9gkg−1 lime. To evaluate the lime-induced chlorosis effects on both rootstocks and cultivars, accumulation of H2O2 and membrane damage (MDA, lipid peroxidation), activities of catalase (CAT, EC 1.11.1.6), superoxide dismutase (SOD, EC 1.15.1.1) and ascorbate peroxidase (APX, EC 1.11.1.11) determined. Bicarbonate treatment reduced active and total Fe and total chlorophyll concentrations, and FCR (leaf ferric chelate reductase) activity while increased the MDA, H2O2, SOD and APX activity of cultivars and rootstocks in both years. These changes and the statistical significances of individual and combined effects of treatment and cultivar or rootstocks showed great differences among the cultivars and rootstocks. So B/C rates helped us to evaluate the tolerance to lime induced chlorosis of the cultivars and rootstocks. B/C referring Fe efficiency ratio calculated as the value for the bicarbonate treatment divided by that for the control for each parameter studied used for tolerance state of cultivars and rootstocks. According to B/C ratios calculated for active and total Fe, FCR, the MG and RG grafted on both rootstocks regarded as tolerant cultivars. In addition to this, the MM106 rootstock was more tolerant than the M9 rootstock under bicarbonate applied condition giving higher B/C ratio for active and total Fe, total chlorophyll, FCR and APX but lower MDA concentrations. It can be concluded that suitable cultivar-rootstock combinations is a good strategy to overcome Fe chlorosis in apple plants grown in calcareous soils.

Silencing of the FRO1 gene and its effects on iron partition in Nicotiana benthamiana

To evaluate the dynamic role of the ferric-chelate reductase enzyme (FCR) and to identify possible pathways of regulation of its activity in different plant organs an investigation was conducted by virus-induced gene silencing (VIGS) using tobacco rattle virus (TRV) to silence the ferric reductase oxidase gene (FRO1) that encodes the FCR enzyme. Half of Nicotiana benthamiana plants received the VIGS vector and the rest remained as control. Four treatments were imposed: two levels of Fe in the nutrient solution (0 or 2.5 μM of Fe), each one with silenced or non-silenced (VIGS-0; VIGS-2.5) plants. Plants grown without iron (0; VIGS-0) developed typical symptoms of iron deficiency in the youngest leaves. To prove that FRO1 silencing had occurred, resupply of Fe (R) was done by adding 2.5 μM of Fe to the nutrient solution in a subset of chlorotic plants (0-R; VIGS-R). Twelve days after resupply, 0-R plants had recovered from Fe deficiency while plants containing the VIGS vector (VIGS-R) remained chlorotic and both FRO1 gene expression and FCR activity were considerably reduced, consequently preventing Fe uptake. With the VIGS technique we were able to silence the FRO1 gene in N. benthamiana and point out its importance in chlorophyll synthesis and Fe partition.

Effects of Exogenous Salicylic Acid and Nitric Oxide on Peanut Seedlings Growth under Iron Deficiency

ABSTRACTSalicylic acid (SA) and nitric oxide (NO), which are known as important signaling molecules in plants, could be promising compounds for the reduction in stress sensitivity. The aim of the present work was to study the physiological changes in peanut (Arachis hypogaea L.) seedlings grown in growth medium that contained 0.1 mM SA, 0.25 mM sodium nitroprusside (SNP, a NO donor), or in full (SA+SNP) or half [1/2 (SA+SNP)] combined strengths under iron (Fe) deficiency. After 21 days of treatment, Fe deficiency significantly inhibited peanut plant growth, destroyed photosynthetic system, and caused oxidative damages. Addition of SA, SNP, and 1/2 (SA+SNP), especially SA+SNP, alleviated the stress, increased the contents of chlorophylls, and promoted plant growth. They improved Fe uptake, transport, and availability in peanut plants by increasing the activities of H+-ATPase and ferric chelate reductase (FCR), and promoting Fe translocation from cell wall to cell organelle and soluble fraction in leaves. Furthermore, they also effectively mitigated oxidative damages by increasing the activities of antioxidant enzymes in peanut leaves and roots. The results from the present study indicate that application of SA, SNP, or 1/2 (SA+SNP) can overcome the adverse effect of Fe deficiency, but the combined application of SA+SNP is more effective in alleviating Fe deficiency stress.

Nitric oxide signaling is involved in the response to iron deficiency in the woody plant Malus xiaojinensis

To cope with iron (Fe) deficiency, plants have evolved a wide range of adaptive responses from changes in morphology to altered physiological responses. Recent studies have demonstrated that nitric oxide (NO) is involved in the Fe-deficiency response through hormonal signaling pathways. Here, we report that NO plays a significant role in Malus xiaojinensis, an Fe-efficient woody plant. Fe deficiency triggered significant accumulation of NO in the root system, predominantly in the outer cortical and epidermal cells of the elongation zone. The NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt (cPTIO) completely arrested Fe deficiency-induced root hair formation, blocked the increase in root ferric-chelate reductase activity and in root H+ excretion, further reduced the active iron content in young leaves and roots, and prevented the upregulation of the critical Fe-related genes, FIT, MxFRO2-like, and MxIRT1. These conditions were restored under Fe deficiency by treatment with the NO donor, sodium nitroprusside (SNP). Additionally, chlorophyll content and relative expression levels of the genes chlorophyll a deoxygenase (MxCAO) and polyamine oxidase (MxPAO) were not changed significantly following Fe deficiency for 6 d; however, SNP treatment increased MxHEMA gene expression. Interestingly, the Fv/Fm ratio, the maximum quantum yield of photosystem II (PSII), decreased significantly following cPTIO treatment. We observed more severe chlorosis under Fe deficiency with cPTIO treatment for 9 d. These results strongly suggest that NO mediates a range of responses to Fe deficiency in M. xiaojinensis, from morphological changes to the regulation of physiological processes and gene expression.

Ferric-chelate reductase activity is a limiting factor in iron uptake in spinach and kale roots

Iron (Fe) is essential for many vital processes in plants, including chlorophyll biosynthesis, DNA synthesis, nitrogen reduction, and photosynthetic electron transfer. However, free Fe ions also participate in the Fenton reaction, which generates reactive oxygen species that induce oxidative stress, leading to lipid peroxidation, protein oxidation, and DNA mutation. Accordingly, plants have developed strategies to prevent roots from absorbing excess Fe. Here, we investigated Fe homeostasis in hydroponically grown spinach and kale under various Fe concentrations (5-800 µM), as well as the resulting changes in bioactive compound levels. Spinach and kale Fe contents did not increase under Fe concentrations in the nutrient solution of up to 200 µM. When spinach plants (grown in nutrient solution containing 2 µM Fe-EDTA) were transferred to 50 µM Fe-EDTA, ferric-chelate reductase activity in roots rapidly decreased within 24 hours. In kale, ferric-chelate reductase activity also significantly decreased with increasing Fe-EDTA concentration. As Fe did not accumulate in spinach plants, no significant differences in plant growth were observed. However, in kale, root growth was slightly suppressed by high Fe (100-200 µM) levels in the rhizosphere. Comparisons to other studies suggest that variations in phenolic and flavonoid contents are dependent on plant species, but overall, Fe treatment did not result in a significant increase in these compounds in spinach or kale. Our results suggest that in spinach and kale roots, the regulation of Fe contents by ferric-chelate reductase is responsible for maintaining Fe homeostasis.

Root-level inoculation with iron-reducing microorganisms does not enhance iron uptake by southern highbush blueberry plants

The rate-limiting step in the iron uptake mechanism of strategy-I plants is ferric chelate reductase (FCR)-mediated iron reduction. Southern highbush blueberry (SHB, Vaccinium corymbosum L. interspecific hybrids) is a specialty crop that commonly exhibits iron deficiency in agricultural soils due to its low FCR activity. Extracellular iron reduction has been reported in some fungi and peronosporomycetes (subsequently referred to as oomycetes), and this has the potential to increase iron reduction capacity in the rhizosphere and impact plant iron uptake. Here, we isolated and identified fungi and oomycetes from the roots and nutrient solution of hydroponically grown SHB. Then, we built a consortium of iron-reducing microorganisms and used this to study the iron nutrition of inoculated and non-inoculated SHB plants in vitro and ex vitro. We found isolates of nine fungal species and the oomycete Pythium irregulare complex, all of which exhibited detectable levels of extracellular iron reduction. Isolates of Alternaria alternata, Bjerkandera adusta, and Pythium irregulare exhibited extracellular iron reduction that was equal to or higher than that of SHB roots, and they were used to create a microbial consortium for the inoculation experiments. In vitro, inoculation with a consortium of iron-reducing microorganisms did not affect iron reduction capacity in the SHB rhizosphere under iron deficiency conditions, but it increased iron reduction capacity in the SHB rhizosphere under iron abundance conditions. Nevertheless, inoculation treatments generally decreased SHB iron uptake from the medium. Ex vitro, inoculation with a consortium of iron-reducing microorganisms did not affect iron reduction capacity in the SHB rhizosphere. In all experiments, the inoculation treatment did not increase leaf active iron concentration. Altogether, our results suggest that inoculation with iron-reducing microorganisms is not a viable alternative for enhancing SHB iron nutrition.

Alkaline stress and iron deficiency regulate iron uptake and riboflavin synthesis gene expression differently in root and leaf tissue: implications for iron deficiency chlorosis.

Iron (Fe) is an essential mineral that has low solubility in alkaline soils, where its deficiency results in chlorosis. Whether low Fe supply and alkaline pH stress are equivalent is unclear, as they have not been treated as separate variables in molecular physiological studies. Additionally, molecular responses to these stresses have not been studied in leaf and root tissues simultaneously. We tested how plants with the Strategy I Fe uptake system respond to Fe deficiency at mildly acidic and alkaline pH by measuring root ferric chelate reductase (FCR) activity and expression of selected Fe uptake genes and riboflavin synthesis genes. Alkaline pH increased cucumber (Cucumis sativus L.) root FCR activity at full Fe supply, but alkaline stress abolished FCR response to low Fe supply. Alkaline pH or low Fe supply resulted in increased expression of Fe uptake genes, but riboflavin synthesis genes responded to Fe deficiency but not alkalinity. Iron deficiency increased expression of some common genes in roots and leaves, but alkaline stress blocked up-regulation of these genes in Fe-deficient leaves. In roots of the melon (Cucumis melo L.) fefe mutant, in which Fe uptake responses are blocked upstream of Fe uptake genes, alkaline stress or Fe deficiency up-regulation of certain Fe uptake and riboflavin synthesis genes was inhibited, indicating a central role for the FeFe protein. These results suggest a model implicating shoot-to-root signaling of Fe status to induce Fe uptake gene expression in roots.

Effects of root medium pH on root water transport and apoplastic pH in red-osier dogwood (Cornus sericea) and paper birch (Betula papyrifera) seedlings.

Soil pH is a major factor affecting plant growth. Plant responses to pH conditions widely vary between different species of plants. However, the exact mechanisms of high pH tolerance of plants are largely unknown. In the present study, we compared the pH responses of paper birch (Betula papyrifera) seedlings, a relatively sensitive species to high soil pH, with red-osier dogwood (Cornus sericea), reported to be relatively tolerant of high pH conditions. We examined the hypotheses that tolerance of plants to high root zone pH is linked to effective control of root apoplastic pH to facilitate nutrient and water transport processes In the study, we exposed paper birch and red-osier dogwood seedlings for six weeks to pH 5, 7 and 9 under controlled-environment conditions in hydroponic culture. Then, we measured biomass, gas exchange, root hydraulic conductivity, ferric chelate reductase (FCR) activity, xylem sap pH and the relative abundance of major elements in leaf protoplasts and apoplasts. The study sheds new light on the rarely studied high pH tolerance mechanisms in plants. We found that compared with paper birch, red-osier dogwood showed greater growth, higher gas exchange, and maintained higher root hydraulic conductivity as well as lower xylem sap pH under high pH conditions. The results suggest that the relatively high pH tolerance of dogwood is associated with greater water uptake ability and maintenance of low apoplastic pH. These traits may have a significant impact on the uptake of Fe and Mn by leaf cells.

Effects of root addition and foliar application of nitric oxide and salicylic acid in alleviating iron deficiency induced chlorosis of peanut seedlings

ABSTRACTNitric oxide (NO) and salicylic acid (SA) are two important signaling molecules, which could alleviate chlorosis of peanut under iron (Fe) deficiency. Here, we further investigated the mechanism of different combinations of sodium nitroprusside (SNP, a nitric oxide donor) and SA supplying in alleviation Fe deficiency symptoms and selected which is the best combination. Thus, peanut was cultivated in hydroponic culture under iron limiting condition with different combinations of SNP and SA application. After 21 days, Fe deficiency significantly inhibited peanut growth, decreased soluble Fe concentration and chlorophyll contents, and disturbed ionic homeostasis. In addition, the content of reactive oxygen species (ROS) and malondialdehyde (MDA) concentration increased, which led the lipid peroxidation. Application of SNP and SA significantly changed Fe trafficking in cells and organs, which increased Fe uptake from nutrient solution, and transport from root to shoot, enhanced the activity of ferric-chelate reductase (FCR), that increased the available Fe in cell organelles, and the active Fe, chlorophyll contents in leaves. Furthermore, ameliorated the inhibition of calcium (Ca), magnesium (Mg) and zinc (Zn) uptake and promoted plant growth in Fe deficiency. At the same time, it increased the activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) to protect the plasmolemma from peroxidation. Results demonstrated that different combinations of SNP and SA application could alleviate the chlorosis of peanut in Fe deficiency by various mechanisms. Such as increased the available Fe and chlorophyll concentrations in leaves, improved the activities of antioxidant enzymes and modulated the mineral elements balance and so on. Foliar application of SNP and SA is the best to protect leaves while directly adding them into nutrient solution is the best to protect roots. These results also indicated that the effects of SNP and SA supplying together to leaves or roots are better than respectively adding to roots and spraying to leaves. The best combination is foliar application of SNP and SA.

Effects of nitric oxide and Fe supply on recovery of Fe deficiency induced chlorosis in peanut plants

The effects of nitric oxide (NO) and/or iron (Fe) supplied to Fe deficient plants have been investigated in peanut (Arachis hypogaea L.) grown in Hoagland nutrient solution with or without Fe. Two weeks after Fe deprivation, recovery was induced by addition of 250 μM sodium nitroprusside (SNP, a NO donor) and/or 50 μM Fe (Fe-EDTA) to the Fe deprived (-Fe) nutrient solution. Activities of antioxidant enzymes, leaf chlorophyll (Chl), and active Fe content decreased, whereas activities of H+-ATPase, ferric-chelate reductase (FCR), nitrate reductase, and nitric oxide synthase and NO production increased in Fe deficient plants, consequently an Fe chlorosis symptom appeared obviously. In contrast, these symptoms disappeared gradually after two weeks with NO and/or Fe supply, which caused an increases in leaf Chl and active Fe content, especially following by co-treatment with NO and Fe to values found in Fe sufficient plants. Increased activities of antioxidant enzymes (superoxide dismutase, peroxidase, and catalase) and decreased accumulation of reactive oxygen species (H2O2, O 2 •− ) and malondialdehyde enhanced the ability of resistance to oxidative stress. Supplied NO alone had the obvious effect on increased NO production and on activity of H+-ATPase and FCR, whereas root length and root/shoot ratio were most effectively increased by Fe supplied alone. Co-treatment with NO and Fe did the best effects on recovery peanut chlorosis symptoms by significantly increased Chl and available Fe content and adjusted distribution of Fe and other mineral elements (Ca, Mg, and Zn) in both leaves and roots.

Role of foliar application of 24-epibrassinolide in response of peanut seedlings to iron deficiency

Limited information is available on the role of brassinosteroids (BRs) in response of plants to nutrient deficiency. To understand the functions of BRs in response to iron deficiency, we investigated the effect of 24-epibrassinolide (EBR) on activities of ferric-chelate reductase (FCR), H+-ATPase, Ca2+-ATPase, nitrate reductase (NR), antioxidant enzymes, Fe and other minerals content and distribution, chlorophylls, soluble protein, free proline, reactive oxygen species, and malondialdehyde in peanut (Arachis hypogea L.) plants subjected to Fe deficiency (10−5 M Fe(III)-EDTA) with foliar application of EBR (0, 10−8, 5.0×10−8, 10−7, 5.0×10−7, and10−6 M). Results show that EBR increased Fe translocation from roots to shoots and increased Fe content in cell organelles. Activities of antioxidant enzymes increased and so the ability of resistance to oxidative stress was enhanced. As result of enhancement of H+-ATPase and Ca2+-ATPase activities, the inhibition of Fe, Ca, Mg, and Zn uptake and distribution was ameliorated. Chlorophyll, soluble protein, and free proline content also increased and consequently, chlorosis induced by Fe deficiency was alleviated. The results demonstrate that EBR had a positive role in regulating peanut growth and development under Fe deficiency and an optimal concentration appeared to be 10−7 M.

The memory of iron stress in strawberry plants

To provide information towards optimization of strategies to treat Fe deficiency, experiments were conducted to study the responses of Fe-deficient plants to the resupply of Fe. Strawberry (Fragaria × ananassa Duch.) was used as model plant. Bare-root transplants of strawberry (cv. ‘Diamante’) were grown for 42 days in Hoagland's nutrient solutions without Fe (Fe0) and containing 10 μM of Fe as Fe-EDDHA (control, Fe10). For plants under Fe0 the total chlorophyll concentration of young leaves decreased progressively on time, showing the typical symptoms of iron chlorosis. After 35 days the Fe concentration was 6% of that observed for plants growing under Fe10. Half of plants growing under Fe0 were then Fe-resupplied by adding 10 μM of Fe to the Fe0 nutrient solution (FeR). Full Chlorophyll recovery of young leaves took place within 12 days. Root ferric chelate-reductase activity (FCR) and succinic and citric acid concentrations increased in FeR plants. Fe partition revealed that FeR plants expressively accumulated this nutrient in the crown and flowers. This observation can be due to a passive deactivation mechanism of the FCR activity, associated with continuous synthesis of succinic and citric acids at root level, and consequent greater uptake of Fe.