Ferric Chelate Reductase Activity Research Articles

Molecular-based characterization and bioengineering of Sorghum bicolor to enhance iron deficiency tolerance in iron-limiting calcareous soils.

Plant biomass can significantly contribute to alternative energy sources. Sorghum bicolor is a promising plant for producing energy, but is susceptible to iron deficiency, which inhibits its cultivation in iron-limiting calcareous soils. The molecular basis for the susceptibility of sorghum to iron deficiency remains unclear. Here, we explored the sorghum genome to identify genes involved in iron uptake and translocation. Iron deficiency-responsive gene expression was comparable to that in other graminaceous plants. A nicotianamine synthase gene, SbNAS1, was induced in response to iron deficiency, and SbNAS1 showed enzyme activity. Sorghum secreted 2'-deoxymugineic acid and other phytosiderophores under iron deficiency, but their levels were relatively low. Intercropping of sorghum with barley or rice rescued iron deficiency symptoms of sorghum. To produce bioengineered sorghum with enhanced tolerance to iron deficiency, we introduced four cassettes into sorghum: 35S promoter-OsIRO2 for activation of iron acquisition-related gene expression, SbIRT1 promoter-Refre1/372 for enhanced ferric-chelate reductase activity, and barley IDS3, and HvNAS1 genomic fragments for enhanced production of phytosiderophores and nicotianamine. The resultant single sorghum line exhibited enhanced secretion of phytosiderophores, increased ferric-chelate reductase activity, and improved iron uptake and leaf greenness compared with non-transformants under iron-limiting conditions. Similar traits were also conferred to rice by introducing the four cassettes. Moreover, these rice lines showed similar or better tolerance in calcareous soils and increased grain iron accumulation compared with previous rice lines carrying two or three comparable cassettes. These results provide a molecular basis for the bioengineering of sorghum tolerant of low iron availability in calcareous soils.

Biochar-based nanoparticles mitigated arsenic toxicity and improved physiological performance of basil via enhancing cation exchange capacity and ferric chelate reductase activity

The modified biochars have positive effects in reducing heavy metal toxicity for plants. However, the mechanism and extent of these effects on mitigating arsenic toxicity and plant performance are not clear. Thus, a pot experiment was conducted as factorial to evaluate the potential of fresh and enriched biochars with potassium and magnesium nano-sulfates [potassium-enriched biochar (K-BC), magnesium-enriched biochar (Mg-BC) in individual and combined forms] on reducing arsenic toxicity (non-contamination, 50, and 100 mg NaAsO2 kg−1 soil) in basil plants. Biochar-related treatments reduced plant arsenic absorption rate (up to 24%), arsenic content of root (up to 38%) and shoot (up to 21%) and root tonoplast H+-ATPase activity (up to 30%). The fresh and particularly enriched biochars improved soil properties (pH, CEC, and available iron content), ferric chelate reductase activity, iron, potassium and magnesium contents of plant tissues, chlorophyll content index, photochemical efficiency of photosystem II, relative electron transport rate, leaf area, and basil growth (shoot and root dry weight). These results revealed that enriched biochars are useful soil amendments for improving physiological performance of plants via reducing heavy metal toxicity and enhancing cation exchange capacity, nutrient availability and ferric chelate reductase activity. Therefore, soil amendment by enriched biochars could be a sustainable solution for enhancing plant productivity in contaminated soils via mitigating environmental impacts. This is an environmentally friendly method for using the natural wastes to overcome the adverse effects of soil pollutants on medicinal plants.

CRISPR/Cas9 edited StbHLH47 lines exhibit altered expression profiling of iron regulating genes and increased iron content in Solanum tuberosum

Iron is an essential plant nutrient, and a continuous supply of it is required as it is a key factor in various metabolic processes, including photosynthesis, chlorophyll synthesis, and respiration. Various transcription factors are known to regulate iron homeostasis in plants, and the bHLH transcription factor family is one of them. The StbHLH47 is a homologue of the Arabidopsis POPEYE (PYE), which is known to repress iron homeostasis-related genes in Arabidopsis. Potato is the most consumed vegetable in the world and is low in iron content. We have generated CRISPR/Cas9-edited StbHLH47 lines and performed a detailed analysis of these lines. The analysis revealed that the roots of StbHLH47 edited lines have decreased ferric chelate reductase (FCR) activity compared to the roots of the wild-type (WT) plant. We also observed that CRISPR/Cas9 edited lines have fewer trichomes when compared to the WT plant. The expression of genes associated with iron homeostasis was also measured. Compared to the control, the expression of StbHLH47 was downregulated in the edited lines, while the expression of StNAS4, StOPT3, and StFRO3 was upregulated. This suggests the negative regulation of StbHLH47 in modulating iron. The iron content was also quantified using inductively coupled plasma mass spectrometry (ICP-MS) and found to be increased in the generated transgenic lines when compared to WT plants. Overall, this study reveals that StbHLH47 negatively regulates the expression of iron homeostasis-related genes. StbHLH47 edited lines exhibited decreased FCR activity, changes in phenotype, and increased iron content in the potato plants. Key messageThis study provides novel insight into the role of StbHLH47 in modulating iron content in Solanum tuberosum and controlling the expression of various iron homeostasis-related genes.

The activation of iron deficiency responses of grapevine rootstocks is dependent to the availability of the nitrogen forms

BackgroundIn viticulture, iron (Fe) chlorosis is a common abiotic stress that impairs plant development and leads to yield and quality losses. Under low availability of the metal, the applied N form (nitrate and ammonium) can play a role in promoting or mitigating Fe deficiency stresses. However, the processes involved are not clear in grapevine. Therefore, the aim of this study was to investigate the response of two grapevine rootstocks to the interaction between N forms and Fe uptake. This process was evaluated in a hydroponic experiment using two ungrafted grapevine rootstocks Fercal (Vitis berlandieri x V. vinifera) tolerant to deficiency induced Fe chlorosis and Couderc 3309 (V. riparia x V. rupestris) susceptible to deficiency induced Fe chlorosis.ResultsThe results could differentiate Fe deficiency effects, N-forms effects, and rootstock effects. Interveinal chlorosis of young leaves appeared earlier on 3309 C from the second week of treatment with NO3−/NH4+ (1:0)/-Fe, while Fercal leaves showed less severe symptoms after four weeks of treatment, corresponding to decreased chlorophyll concentrations lowered by 75% in 3309 C and 57% in Fercal. Ferric chelate reductase (FCR) activity was by trend enhanced under Fe deficiency in Fercal with both N combinations, whereas 3309 C showed an increase in FCR activity under Fe deficiency only with NO3−/NH4+ (1:1) treatment. With the transcriptome analysis, Gene Ontology (GO) revealed multiple biological processes and molecular functions that were significantly regulated in grapevine rootstocks under Fe-deficient conditions, with more genes regulated in Fercal responses, especially when both forms of N were supplied. Furthermore, the expression of genes involved in the auxin and abscisic acid metabolic pathways was markedly increased by the equal supply of both forms of N under Fe deficiency conditions. In addition, changes in the expression of genes related to Fe uptake, regulation, and transport reflected the different responses of the two grapevine rootstocks to different N forms.ConclusionsResults show a clear contribution of N forms to the response of the two grapevine rootstocks under Fe deficiency, highlighting the importance of providing both N forms (nitrate and ammonium) in an appropriate ratio in order to ease the rootstock responses to Fe deficiency.

Barley preferentially activates strategy-II iron uptake mechanism under iron deficiency

Plants utilize two main strategies for iron (Fe) uptake from the rhizosphere. Strategy-I is based on the reduction of ferric (Fe3+) to ferrous (Fe2+) iron by ferric chelate reductase (FCR) and is mainly observed in dicots. Strategy-II utilizes the complexation of Fe3+ with phytosiderophores secreted from the plant roots and mainly evolved in Gramineous species, including barley (Hordeum vulgare). Recent studies suggest that some species use a combination of both strategies for more efficient Fe uptake. However, the preference of barley for these strategies is not well understood. This study investigated the physiological and biochemical responses of barley under iron deficiency and examined the expression levels of the genes involved in Strategy-I and Strategy-II mechanisms in the roots. Fe deficiency led to decreased root and shoot lengths, fresh and dry weights, and Fe accumulation in the roots. Parallel to the chlorosis observed in the leaves, FCR activity and rhizosphere acidification were also significantly reduced in the roots, while the release of phytosiderophores increased. Furthermore, Strategy-II genes expressed higher than the Strategy-I genes in the roots under Fe deficiency. These findings demonstrate that Strategy-II is more activated than Strategy-I for Fe uptake in barley roots under Fe-deficient conditions.

Melatonin Promotes Iron Reactivation and Reutilization in Peach Plants under Iron Deficiency.

The yellowing of leaves due to iron deficiency is a prevalent issue in peach production. Although the capacity of exogenous melatonin (MT) to promote iron uptake in peach plants has been demonstrated, its underlying mechanism remains ambiguous. This investigation was carried out to further study the effects of exogenous MT on the iron absorption and transport mechanisms of peach (Prunus persica) plants under iron-deficient conditions through transcriptome sequencing. Under both iron-deficient and iron-supplied conditions, MT increased the content of photosynthetic pigments in peach leaves and decreased the concentrations of pectin, hemicellulose, cell wall iron, pectin iron, and hemicellulose iron in peach plants to a certain extent. These effects stemmed from the inhibitory effect of MT on the polygalacturonase (PG), cellulase (Cx), phenylalanine ammonia-lyase (PAL), and cinnamoyl-coenzyme A reductase (CCR) activities, as well as the promotional effect of MT on the cinnamic acid-4-hydroxylase (C4H) activity, facilitating the reactivation of cell wall component iron. Additionally, MT increased the ferric-chelate reductase (FCR) activity and the contents of total and active iron in various organs of peach plants under iron-deficient and iron-supplied conditions. Transcriptome analysis revealed that the differentially expressed genes (DEGs) linked to iron metabolism in MT-treated peach plants were primarily enriched in the aminoacyl-tRNA biosynthesis pathway under iron-deficient conditions. Furthermore, MT influenced the expression levels of these DEGs, regulating cell wall metabolism, lignin metabolism, and iron translocation within peach plants. Overall, the application of exogenous MT promotes the reactivation and reutilization of iron in peach plants.

Coated Hematite Nanoparticles Alleviate Iron Deficiency in Cucumber in Acidic Nutrient Solution and as Foliar Spray.

Micronutrient iron (Fe) deficiency poses a widespread agricultural challenge with global implications. Fe deficiency affects plant growth and immune function, leading to reduced yields and contributing to the global "hidden hunger." While conventional Fe-based fertilizers are available, their efficacy is limited under certain conditions. Most recently, nanofertilizers have been shown as promising alternatives to conventional fertilizers. In this study, three nanohematite/nanoferrihydrite preparations (NHs) with different coatings were applied through the roots and shoots to Fe-deficient cucumber plants. To enhance Fe mobilization to leaves during foliar treatment, the plants were pre-treated with various acids (citric acid, ascorbic acid, and glycine) at a concentration of 0.5 mM. Multiple physiological parameters were examined, revealing that both root and foliar treatments resulted in improved chlorophyll content, biomass, photosynthetic parameters, and reduced ferric chelate reductase activity. The plants also significantly accumulated Fe in their developing leaves and its distribution after NHs treatment, detected by X-ray fluorescence mapping, implied long-distance mobilization in their veins. These findings suggest that the applied NHs effectively mitigated Fe deficiency in cucumber plants through both modes of application, highlighting their potential as nanofertilizers on a larger scale.

Bicarbonate rather than high pH in growth medium induced Fe-deficiency chlorosis in dwarfing rootstock quince A (Cydonia oblonga Mill.) but did not impair Fe nutrition of vigorous rootstock Pyrus betulifolia.

Quince A (Cydonia oblonga Mill.), a typical dwarfing rootstock in pear cultivation, is susceptible to iron (Fe) deficiency in calcareous soils. The aim of this study was to compare the strategies in Fe uptake and utilization in dwarfing rootstock quince A (low Fe efficiency) versus a typical vigorous rootstock Pyrus betulifolia (PB) with high Fe efficiency. Quince A and PB were grown in nutrient solution (pH 6.3) for 4 weeks followed by three pH treatments: pH6.3, pH8.3a (adjusted with hydroxide) and pH8.3b (adjusted with bicarbonate). The Fe uptake and utilization indicators of the rootstocks were assessed at the onset of chlorosis symptoms (after 58 days of treatments). In contrast to PB, quince A exhibited Fe deficiency chlorosis under bicarbonate (pH8.3b). Bicarbonate stimulated the root proton secretion, inhibited root growth and ferric chelate reductase (FCR) activity in both PB and quince A, whereas high pH without bicarbonate (pH8.3a) stimulated only root proton release. Both species accumulated more Fe in roots under high pH treatments than under pH6.3, resulting in Fe sufficiency in leaves. Both high pH treatments increased the activity of leaf FCR in PB and quince A. However, extractable Fe(II) concentration in leaves was increased by high pH treatments in PB only. This study demonstrated that depressed Fe(III) reduction in leaves caused by bicarbonate rather than high pH explained Fe deficiency in quince A grown in bicarbonate-containing medium.

Silencing of FRO1 gene affects iron homeostasis and nutrient balance in tomato plants

AbstractBackgroundIron chlorosis is an abiotic stress of worldwide importance affecting several agronomic crops. It is important to understand how plants maintain nutrient homeostasis under Fe deficiency and recovery.AimsWe used the virus‐induced gene silencing (VIGS) method to elucidate the role of the FRO1 gene in tomato plants and identify the impact on regulation of the root ferric‐chelate reductase (FCR) activity and nutritional homeostasis.MethodsTomato plantlets cv. “Cherry” were transferred into half‐strength Hoagland's nutrient solution containing 0.5 µM of Fe (Fe0.5). In phase I, two treatments were established: control (Fe0.5) plants and VIGS‐0.5 plants corresponding to plants with the FRO1 gene silenced. In phase II, plants from Fe0.5 and VIGS‐0.5 were transferred to new nutrient solution and then grown for a further 14 days under 0 and 10 µM of Fe (as 0.5 µM would not be enough for the larger plants during phase II). Therefore, four treatments were imposed: Fe0, Fe10, VIGS‐0, and VIGS‐10.ResultsVIGS‐0.5 plants had significantly lower chlorophyll (Chl) and root FCR activity compared to the respective non‐silenced plants and retained more Cu and Zn in the roots at the expense of stems (Cu) or young leaves (Zn). Iron concentration in roots and stems decreased in FRO1 gene‐silenced plants, compared to control plants, but the allocation to different organs was similar in both treatments.ConclusionsThere was a partial recovery of leaf Chl in the VIGS‐10 plants and a higher concentration of Fe in all organs. In contrast, the allocation of Cu to roots decreased in the VIGS‐10 plants.

Enhanced Germination and Growth of Alfalfa with Seed Presoaking and Hydroponic Culture in Fe2O3 Magnetic Nanoparticles

Nanofertilizers can be used as potential contenders to provide macro- and micronutrients to plants. Specifically, iron-oxide-based engineered nanoparticles are promising candidates for next-generation iron-deficiency fertilizers. Aiming to investigate the effect of Fe2O3 magnetic nanoparticles on plant germination and growth, alfalfa seeds were presoaked by a series of concentrations (0, 50, 100, 150, and 200 mg/L) of Fe2O3 NPs in this study. The germination rate, root length, chlorophyll content, and activity of the antioxidant enzymes (superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT)) were detected. Further, hydroponic alfalfa young seedlings were treated with 50 mg/L Fe2O3 NPs, with 50 mg/L Fe-EDTA as the control. The growth index, chlorophyll content, metal (Fe, Cu, Zn, and Mn) content, and ferric-chelate reductase activity were detected. The results indicated that the presoaking of Fe2O3 NPs promoted alfalfa seed germination in all treatments. Short-time of Fe2O3 NPs presoaking has not affected chlorophyll and iron content while increasing antioxidase (SOD, POD, and CAT) activities at 50 and 200 mg/L concentrations. Fe2O3 NPs of 50 mg/L have the most beneficiary impact on alfalfa seed germination by presoaking. Fe2O3 NPs of 50 mg/L enhanced the seedlings’ growth and increased chlorophyll content, Fe content, and Mn content of hydroponic alfalfa compared with the control. Moreover, ferric chelate reductase activity was significantly lower in alfalfa roots exposure to Fe2O3 NPs than in the control. The results showed that 50 mg/L Fe2O3 NPs could be used as an appropriate and effective treatment concentration for seed pretreatment and fertilizer application. The growth-promoting effect of Fe2O3 NPs on alfalfa is related to its nanostructured properties. More in-depth research is still needed to understand the growth-promoting mechanism of Fe2O3 NPs and the impact of Fe2O3 NPs on the field environment.

The low ferric chelate reductase activity and high apoplastic pH in leaves cause iron deficiency chlorosis in ‘Huangguan’ pears grafted onto quince A grown in calcareous soil

Quince (Cydonia oblonga Mill.) is a dwarfing rootstock often used in pear cultivation. However, the pear cultivar ‘Huangguan’ (Pyrus bretschneideri Rehd. cv) grafted onto quince A (HG-QA) suffered iron (Fe) deficiency chlorosis in the calcareous soils of North China, while ‘Huangguan’ grafted onto Pyrus betulifolia (HG-PB) did not. We hypothesized that the higher leaf apoplastic pH of HG-QA than HG-PB causes Fe deficiency chlorosis by restricting Fe transportation within leaves. Three-year-old rootstocks quince A and Pyrus betulifolia and the combinations HG-QA and HG-PB were grown in the same orchard in a calcareous soil. Newly expanded leaves were analysed for Fe related properties. Micro X-ray fluorescence analysis was used for Fe mapping within the individual leaves. The extractable Fe(II) concentrations in quince A and HG-QA leaves were much lower than those in Pyrus betulifolia and HG-PB, even though the total leaf Fe concentrations were similar. More Fe was located near the leaf midrib in quince A and HG-QA than in Pyrus betulifolia and HG-PB. These were correlated with the lower ferric chelate reductase (FCR) activity in the chlorotic leaves of the quince A rootstock than Pyrus betulifolia and the lower FCR activity and higher apoplastic pH in the chlorotic leaves of the HG-QA rootstock-scion combination than HG-PB. In conclusion, the reduced Fe availability within the leaves caused by the lower FCR activity and higher apoplastic pH compared with those of HG-PB led to HG-QA leaf chlorosis in calcareous soils. This result implies that rootstocks are involved in the regulation of leaf Fe utilization by scions.

Apoplast utilisation of nanohaematite initiates parallel suppression of RIBA1 and FRO1&3 in Cucumis sativus

Nanoscale Fe containing particles can penetrate the root apoplast. Nevertheless, cell wall size exclusion questions that for Fe mobilisation, a close contact between the membrane integrating FERRIC REDUCTASE OXIDASE (FRO) enzymes and Fe containing particles is required. Haematite nanoparticle suspension, size of 10–20 nm, characterized by 57Fe Mössbauer spectroscopy, TEM, ICP and SAED was subjected to Fe utilisation by the flavin secreting model plant cucumber (Cucumis sativus). Alterations in the structure and distribution of the particles were revealed by 57Fe Mössbauer spectroscopy, HRTEM and EDS element mapping. Biological utilisation of Fe resulted in a suppression of Fe deficiency responses (expression of CsFRO 1, 2 &3 and RIBOFLAVIN A1; CsRIBA1 genes and root ferric chelate reductase activity). Haematite nanoparticles were stacked in the middle lamella of the apoplast. Fe mobilisation is evidenced by the reduction in the particle size. Fe release from nanoparticles does not require a contact with the plasma membrane. Parallel suppression in the CsFRO 1&3 and CsRIBA1 transcript amounts support that flavin biosynthesis is an inclusive Fe deficiency response involved in the reduction-based Fe utilisation of Cucumis sativus roots. CsFRO2 is suggested to play a role in the intracellular Fe homeostasis.

Physiological and Transcriptome Analyses Revealed the Mechanism by Which Deferoxamine Promotes Iron Absorption in Cinnamomum camphora.

Iron deficiency causes chlorosis and growth inhibition in Cinnamomum camphora, an important landscaping tree species. Siderophores produced by plant growth-promoting rhizobacteria have been widely reported to play an indispensable role in plant iron nutrition. However, little to date has been determined about how microbial siderophores promote plant iron absorption. In this study, multidisciplinary approaches, including physiological, biochemical and transcriptome methods, were used to investigate the role of deferoxamine (DFO) in regulating Fe availability in C. camphora seedlings. Our results showed that DFO supplementation significantly increased the Fe2+ content, SPAD value and ferric-chelate reductase (FCR) activity in plants, suggesting its beneficial effect under Fe deficiency. This DFO-driven amelioration of Fe deficiency was further supported by the improvement of photosynthesis. Intriguingly, DFO treatment activated the metabolic pathway of glutathione (GSH) synthesis, and exogenous spraying reduced glutathione and also alleviated chlorosis in C. camphora. In addition, the expression of some Fe acquisition and transport-related genes, including CcbHLH, CcFRO6, CcIRT2, CcNramp5, CcOPT3 and CcVIT4, was significantly upregulated by DFO treatment. Collectively, our data demonstrated an effective, economical and feasible organic iron-complexing agent for iron-deficient camphor trees and provided new insights into the mechanism by which siderophores promote iron absorption in plants.

Increase in root branching enhanced ferric-chelate reductase activity under iron stress in potato (Solanum tuberosum)

In response to Fe-deficiency, various dicots increase their root branching to improve ferric-chelate reductase activity. It still remains unclear, whether the response caused by Fe-deficiency ultimately improves the plant's ability to withstand Fe-deficiency. In this experiment conducted at ICAR-Central Potato Research Institute, Regional Station, Shillong during 2020, we demonstrated a substantial increase in the growth of the lateral root of potato genotype (CP 3443), when grown in the iron-stress, in relation to control plants, and the total lateral root number is well linked to ferric-chelate reductase (FCR) activity. These findings showed that FCR is involved in root Fe uptake in potato (Solanum tuberosum L.) and they suggest a role in Fe distribution throughout the plant. In view of these findings, the Fe-deficiency induced increases in the lateral roots suggested that these play a significant role in Fe-deficiency tolerance in potato, which can serve as useful trait for the identification of chlorosis tolerance and/or nutrient-deficiency stress.

Growth and physiological impairments in Fe-starved alfalfa are associated with the downregulation of Fe and S transporters along with redox imbalance

BackgroundIron (Fe) is an essential plant nutrient. Its deficiency is a major constraint in crop production systems, affecting crop yield and quality. It is therefore important to elucidate the responses and adaptive mechanisms underlying Fe-deficiency symptoms in alfalfa.Materials and methodsThe experiment was carried out on 12-day-old alfalfa plants grown in hydroponics under Fe-sufficient and Fe-deficient conditions.ResultsThe Fe-starved alfalfa showed decreased plant biomass, chlorophyll score, PSII efficiency, and photosynthesis performance index in young leaves under low Fe. Further, Fe shortage reduced the Fe, Zn, S and Ca concentration in root and shoot of alfalfa accompanied by the marked decrease of MsIRT1, MsZIP, MsSULTR1;1, MsSULTR1;2 and MsSULTR1;3 transcripts in root and shoot. It indicates that retardation caused by Fe-deficiency was also associated with the status of other elements, especially the reduced Fe and S may be coordinately attributed to the photosynthetic damages in Fe-deficient alfalfa. The ferric chelate reductase activity accompanied by the expression of MsFRO1 in roots showed no substantial changes, indicating the possible involvement of this Strategy I response in Fe-deficient alfalfa. However, the proton extrusion and expression of MsHAI1 were significantly induced following Fe-deficiency. In silico analysis further suggested their subcellular localization in the plasma membrane. Also, the interactome map suggested the partnership of MsFRO1 with plasma membrane H+-ATPase, transcription factor bHLH47, and nitrate reductase genes, while MsHAI1 partners include ferric reductase-like transmembrane component, plasma membrane ATPase, vacuolar-type H-pyrophosphatase, and general regulatory factor 2. In this study, SOD and APX enzymes showed a substantial increase in roots but unable to restore the oxidative damages in Fe-starved alfalfa.ConclusionThese findings promote further studies for the improvement of Fe-starved alfalfa or legumes through breeding or transgenic approaches.Graphic

Effects of Soil Aeration and Fertilization Practices on Alleviating Iron Deficiency Chlorosis in “Huangguan” Pears Grafted onto Quince A in Calcareous Soils

In North China, the high-quality pear cultivar “Huangguan” (Pyrus bretschneideri Rehd. cv), which is grafted onto dwarf quince A (Cydonia oblonga Mill. cv) rootstock and grown in calcareous soil, experiences severe iron (Fe) deficiency; this deficiency greatly constrains tree growth as well as fruit yield and quality. Therefore, we evaluated the effects of six practices for alleviating chlorosis caused by Fe deficiency in “Huangguan” grafted onto quince A (HG-QA). The practices included ridging with landscape fabric mulching as a control, flattening with landscape fabric mulching (FM), ridging without landscape fabric mulching (R), flattening without landscape fabric mulching (F), Fe fertilizer application in soil (SFe), foliar Fe application (FFe), and manure application (M). The results showed that the leaf Fe concentration increased by 356% under FFe, compared to that under the control, but the practice failed to alleviate Fe deficiency chlorosis. In contrast, an increase in leaf Fe concentration and chlorosis alleviation were observed under F. F alleviated chlorosis mainly by increasing the root ferric-chelate reductase activity. These results indicate that Fe uptake and utilization in leaves are independent biochemical processes and soil aeration improvement have positive effect on increasing Fe uptake. M improved both the soil active Fe concentration and leaf Fe utilization. Thus, manure application should be the first choice for alleviating Fe deficiency chlorosis in HG-QA grown in calcareous soils. Combining manure application with other practices that increase Fe uptake would likely be an effective way to address the problem of Fe deficiency chlorosis.

Differential response of pea (Pisum sativum L.) genotypes to iron deficiency in relation to the growth, rhizosphere acidification and ferric chelate reductase activities

Calcareous soils are known problematic lands for agricultural systems because of the low availability of nutrients, particularly iron (Fe). The so-called strategy I plant (e. g. Pea, Pisum sativum L.) which groups dicotyledons and monocots other than grasses, developed root membrane activities that contribute to the improvement of Fe availability. Among the functions considered to be a critical phase in iron absorption is rhizosphere acidification by H-ATPase and Fe(III) reduced by Fe(III) chelate reducctase (FeCR). In order to experimentally investigate the importance of root FeCR in Fe nutrition, its relationship with rhizosphere acidification and the genotypic differences in response to iron deficiency in pea (Pisum sativum L.), a glasshouse experiment was conducted hydroponically on four genotypes Merveille de Kelvedon (MK); Lincoln (Lin); Douce de Provence (DP) and Alexandra (Alex). Plants of each genotype were distributed into two plots, the first one received full nutrient solution (+ Fe), the second one received nutrient solution devoid of iron (- Fe). Plant growth, Fe distribution, SPAD index and root acidification and ferric chelate reductase activities were evaluated. Fe deficiency decreased plant growth and SPAD index along with the significant increase of H-ATPase and FeCR activities. Some genotypic differences were observed as follows; Alex showed high tolerance to Fe deprivation as compared to other genotypes. Important H-ATPase and FeCR activities, high Fe use efficiency and adequate membrane efficiency are the main reasons for this tolerance. These physiological parameters could be used as tools of tolerance for further breeding programs

Silicon induces metallochaperone-driven cadmium binding to the cell wall and restores redox status through elevated glutathione in Cd-stressed sugar beet.

Cadmium (Cd) is toxic; however, whether silicon (Si) alleviates Cd toxicity was never studied in sugar beet. The study was conducted on 2-week-old sugar beet cultivated in the presence or absence of Cd (10μM CdSO4 ) and Si (1 mM Na2 SiO3 ) in hydroponic conditions. The morphological impairment and cellular damages observed in sugar beet upon Cd toxicity were entirely reversed due to Si. Si substantially restored the energy-providing ability, absorbed energy flux, and electron transport toward PSII, which might be correlated with the upregulation of BvIRT1 and ferric chelate reductase activity leading to the restoration of Fe status in Cd-stressed sugar beet. Although Si caused a reduction of shoot Cd, the root Cd substantially increased under Cd stress, a significant part of which was retained in the cell wall rather than in the root vacuole. While the concentration of phytochelatin and the expression of BvPCS3 (PHYTOCHELATIN SYNTHASE 3) showed no changes upon Si exposure, Si induced the expression of BvHIPP32 (HEAVY METAL-ASSOCIATED ISOPRENYLATED PLANT PROTEIN 32) in the Cd-exposed root. The BvHIPP32 and AtHIPP32 metallochaperone proteins are localized in the cell wall and they share similar sequence alignment, physiochemical properties, secondary structure, cellular localization, motif locations, domain association, and metal-binding site (cd00371) linked to the metallochaperone-like protein. It suggests that Si reduces the Cd level in shoot by retaining the excess Cd in the cell wall of roots due to the induction of BvHIPP32 gene. Also, Si stimulates glutathione-related antioxidants along with the BvGST23 expression, inferring an ascorbate-glutathione ROS detoxification pathway in Cd-exposed plants.

Molecular characterization of Fe-acquisition genes causing decreased Fe uptake and photosynthetic inefficiency in Fe-deficient sunflower

Iron (Fe) deficiency in plants hinders growth and yield. Thus, this study aims to elucidate the responses and molecular characterization of genes in Fe-deficient sunflower. The study was conducted on 14 days-old sunflower plants cultivated in hydroponic culture under Fe-sufficient and Fe-deficient conditions. The Fe-starved sunflower showed substantial decrease in plant biomass, SPAD score, quantum yield efficiency of PSII (Fv/Fm), photosynthetic performance index (Pi_ABS). Further, Fe shortage reduced Fe and Zn concentrations in roots and shoots, accompanied by a marked decrease of HaNramp1 and HaZIP1 expression in roots, suggesting the association of Zn status contributing to photosynthetic inefficiency in sunflower. The ferric chelate reductase (FCR) activity, along with HaFRO2 and HaIRT1 transcripts, were constitutively expressed, suggesting that sunflower plants can regulate FCR activity, although the lack of bioavailable Fe in the rhizosphere strongly corresponds to the limited Fe uptake in sunflower. The substantial increase of proton extrusion in roots and the localization of Fe-related genes in the plasma membrane are also evident in sunflower as common responses to Fe-deficiency by this Strategy I plant species. Analysis showed that three motifs of Fe-related proteins were linked to the ZIP zinc transporter. The interactome map revealed the close partnership of these Fe-related genes in addition to FRU gene encoding putative transcription factor linked to Fe uptake response. The cis-regulatory analysis of promoter suggested the involvement of auxin, salicylic acid, and methyl jasmonate-responsive elements in the regulatory process in response to Fe deficiency. These findings may be beneficial to develop Fe-efficient sunflower plants through breeding or genome editing approaches.

Adaptive strategies to minimise iron limiting stress under climate change scenario: A study pertinent to iron stress response of two soybean (Glycine max (L.) Merr.) Genotypes

The response of soybean genotypes to iron deficiency stress under elevated CO2 and temperature condition could be guided by three critical rhizospheric processes viz. proton (H+) extrusion, root exudation and ferric chelate reductase (FCR) activity. In the present study, the iron efficient and responsive (FeER) genotype recorded an impressive performance over iron inefficient and responsive (FeIR) genotype in counteracting iron deficiency stress but experienced modest stress caused by the combined interaction between the genotype (G), environment (E) and HCO3- ion (B). The antagonistic interaction between Fe2+ with HCO3- ion resulted in greater iron stress. Plants grown in the absence of bicarbonate have significantly higher total chlorophyll content (1.79±0.04 mg g-1, mean ± SE) than plants grown in presence of bicarbonate (1.48 ± 0.06 mg g-1). To warfare the constraints in Fe availability, especially under more stressed e-[CO2+T] environmental condition in the presence of bicarbonate ion, the root system of iron efficient genotype of soybean (FeER) exuded out significantly higher amount of low molecular weight organic acids. Furthermore, the presence of bicarbonate ion in the nutrient solution exacerbated the iron deficiency stress and consequently resulted in higher proton extrusion (~1.2 fold increase), lower ferric chelate reductase activity (~1.3 fold decrease) and higher organic acid exudation (up to ~1.9 fold increase in malic acid). The intra-specific variability between contrasting genotypes and their response to elevated CO2 and temperature could be exploited to remediate emerging Fe deficiency of soybean plants under climate change scenario.