Metal Interactions in the Ni Hyperaccumulating Population of Noccaea caerulescens Monte Prinzera.

Hyperaccumulation is a fascinating trait displayed by a few plant species able to accumulate large amounts of metal ions in above-ground tissues without symptoms of toxicity. Noccaea caerulescens is a recognized model system to study metal hyperaccumulation and hypertolerance. A N. caerulescens population naturally growing on a serpentine soil in the Italian Apennine Mountains, Monte Prinzera, was chosen for the study here reported. Plants were grown hydroponically and treated with different metals, in excess or limiting concentrations. Accumulated metals were quantified in shoots and roots by means of ICP-MS. By real-time PCR analysis, the expression of metal transporters and Fe deficiency-regulated genes was compared in the shoots and roots of treated plants. N. caerulescens Monte Prinzera confirmed its ability to hypertolerate and hyperaccumulate Ni but not Zn. Moreover, excess Ni does not induce Fe deficiency as in Ni-sensitive species and instead competes with Fe translocation rather than its uptake.

A mechanistic mathematical model for describing and predicting the dynamics of high-affinity nitrate intake into roots of maize and other plant species.

A fully mechanistic dynamical model for plant nitrate uptake is presented. Based on physiological and regulatory pathways and based on physical laws, we form a dynamic system mathematically described by seven differential equations. The model evidences the presence of a short-term positive feedback on the high-affinity nitrate uptake, triggered by the presence of nitrate around the roots, which induces its intaking. In the long run, this positive feedback is overridden by two long-term negative feedback loops which drastically reduces the nitrate uptake capacity. These two negative feedbacks are due to the generation of ammonium and amino acids, respectively, and inhibit the synthesis and the activity of high-affinity nitrate transporters. This model faithfully predicts the typical spiking behavior of the nitrate uptake, in which an initial strong increase of nitrate absorption capacity is followed by a drop, which regulates the absorption down to the initial value. The model outcome was compared with experimental data and they fit quite nicely. The model predicts that after the initial exposure of the roots with nitrate, the absorption of the anion strongly increases and that, on the contrary, the intensity of the absorption is limited in presence of ammonium around the roots.

Upcycling of PHA-producing bacteria for biostimulants production and polyhydroxyalkanoates recovery.

The use of petroleum-based plastic has led to its accumulation in the environment, with negative impacts on the ecosystem and the biota. Polyhydroxyalkanoates (PHAs), biobased and biodegradable plastics produced by microbes, have many commercial applications, however their high production cost means they cannot yet compete with traditional plastics. At the same time, the problem of the growing human population implies that improved crop production is needed to avoid malnutrition. Biostimulants enhance plant growth and have the potential to improve agricultural yields; they can be obtained from biological feedstock, including microbes. Therefore, there is scope for coupling the production of PHAs with that of biostimulants, making the process more cost-efficient and minimizing by-products generation. In this work, low-value agro-zoological residues were processed to obtain PHA-storing bacteria via acidogenic fermentation; PHAs destined for the bioplastic market were extracted, and the protein-rich by-products were turned into protein hydrolysates using different treatment methods, assessing their biostimulant effects in growth trials with tomato and cucumber plants. The results indicate that the best hydrolysis treatment, realizing the highest amount of organic nitrogen (6.8 gN-org/L) while achieving the best PHA recovery (63.2 % gPHA/gTS), is obtained with strong acids. All the protein hydrolysates were effective in improving either roots or leaf development, with various results, depending on the species and the growth method. The acid hydrolysate was the most effective treatment to enhance the development of shoots (21 % increase compared to the control) and roots (16 % increase for the dry weight and 17 % for main root length) of hydroponically-grown cucumber plants, while pot-grown tomatoes, biostimulated via foliar spray, developed bigger shoots (up to 41 %) with the hydrolysate obtained from the alkaline treatment. These preliminary results indicate that simultaneous production of PHAs and biostimulants is feasible, and that commercialization could be achievable given the expected reduction in production costs.

Regreening properties of the soil slow-mobile H2bpcd/Fe3+ complex: Steps forward to the development of a new environmentally friendly Fe fertilizer.

The application of synthetic Fe-chelates stands for the most established agronomical practice to alleviate lime-induced chlorosis, which still constitutes a major agronomic problem. However, the percolation through the soil profile due to the negative charge of the most deployed molecules results in agronomical and environmental problems. H2bpcd/Fe3+ complex features distinctive chemical characteristics, including moderate stability of the Fe(bpcd)+ species (logß ML = 20.86) and a total positive charge, and we studied its behavior in soil and regreening effects on cucumber plants. Soil column experiments have underlined that H2bpcd/Fe3+ is retained in more amounts than EDDHA/Fe3+. The new ligand was not proven to be toxic for the cucumber and maize seedlings. A concentration of 20 µM H2bpcd/Fe3+ attained regreening of Fe-deficient cucumber plants grown in the hydroponic solution supplied with CaCO3, similar to that shown by EDDHA/Fe3+. Experiments with a 2 µM concentration of 57Fe showed that cucumber roots absorbed H2bpcd/57Fe3+ at a slower rate than EDTA/57Fe3+. The high kinetic inertness of H2bpcd/Fe3+ may explain such behavior.

Chemical Characterization of a Collagen-Derived Protein Hydrolysate and Biostimulant Activity Assessment of Its Peptidic Components.

Protein hydrolysates (PHs) are plant biostimulants consisting of oligopeptides and free amino acids exploited in agriculture to increase crop productivity. This work aimed to fractionate a commercial collagen-derived protein hydrolysate (CDPH) according to the molecular mass of the peptides and evaluate the bioactivity of different components. First, the CDPH was dialyzed and/or filtrated and analyzed on maize, showing that smaller compounds were particularly active in stimulating lateral root growth. The CDPH was then fractionated through fast protein liquid chromatography and tested on in vitro grown tomatoes proving that all the fractions were bioactive. Furthermore, these fractions were characterized by liquid chromatography-electrospray ionization-tandem mass spectrometry revealing a consensus sequence shared among the identified peptides. Based on this sequence, a synthetic peptide was produced. We assessed its structural similarity with the CDPH, the collagen, and polyproline type II helix by comparing the respective circular dichroism spectra and for the first time, we proved that a signature peptide was as bioactive as the whole CDPH.

Nodulating white lupins take advantage of the reciprocal interplay between N and P nutritional responses.

The low bioavailability of nutrients, especially nitrogen (N) and phosphorus (P), is one of the most limiting factors for crop production. In this study, under N- and P-free nutrient solution (-N-P), nodulating white lupin plants developed some nodules and analogous cluster root structures characterized by different morphological, physiological, and molecular responses than those observed upon single nutrient deficiency (strong acidification of external media, a better nutritional status than -N+P and +N-P plants). The multi-elemental analysis highlighted that the concentrations of nutrients in white lupin plants were mainly affected by P availability. Gene-expression analyses provided evidence of interconnections between N and P nutritional pathways that are active to promote N and P balance in plants. The root exudome was mainly characterized by N availability in nutrient solution, and, in particular, the absence of N and P in the nutrient solution triggered a high release of phenolic compounds, nucleosides monophosphate and saponines by roots. These morphological, physiological, and molecular responses result from a close interplay between N and P nutritional pathways. They contribute to the good development of nodulating white lupin plants when grown on N- and P-free media. This study provides evidence that limited N and P availability in the nutrient solution can promote white lupin-Bradyrhizobium symbiosis, which is favourable for the sustainability of legume production.

The potential use of Chlamydomonas reinhardtii and Chlorella sorokiniana as biostimulants on maize plants.

Nitrogen deficiency and drought stress are among the major stresses faced by plants with negative consequence on crop production. The use of plant biostimulants is a very promising application in agriculture to improve crop yield, but especially to prevent the effect of abiotic stresses. Algae-derived biostimulants represent an efficient tool to stimulate the root development: while macroalgae have already been widely adopted as a source of biostimulants to improve plants growth and resilience, far less information is available for microalgae. The objective of this work is to investigate the stimulant ability on maize roots of two green algae species, Chlamydomonas reinhardtii and Chlorella sorokiniana, being respectively the model organism for Chlorophyta and one of the most promising species for microalgae cultivation at industrial scale. The results obtained demonstrate that both C. reinhardtii and C. sorokiniana cells promoted the development of maize root system compared to the untreated negative control. C. sorokiniana specifically increased the number of secondary roots, while improved micro-nutrients accumulation on roots and shoots was measured in the case of C. reinhardtii treated plants. When these microalgae-derived biostimulants were applied on plants grown in stress conditions as nitrogen deficiency, improved development of the root system was measured in the case of plants treated with C. sorokiniana biomass. Microalgae cultivation for biostimulant production can thus be considered as a bio-based process providing solutions for improving plant resilience toward stress conditions.

Physico-chemical and sensory acceptability of no added sugar chocolate spreads fortified with multiple micronutrients.

Vitamin D and magnesium-calcium carbonate nanoparticles were used to fortify a newly developed healthy chocolate spread formulated with inulin and maltitol as sugar replacers and alternative to palm oil to reduce the concentration of saturated fatty acid. These samples were compared with well-known commercially available chocolate spreads in terms of rheology, polyphenols content and in vitro digestion, sensory attributes and willingness to buy. The fortified chocolate spreads showed comparable if not better acceptability than the current products on the market and over 80% of the participants were inclined to buy and 66% prepared to spend 10 to 15% more money on the product enriched with the three micronutrients. The results also demonstrate that the incorporation of nanoparticles could affect the rheological and physio-chemical properties of the formulations and an appropriate ratio between the fat phase and particles seems an important factor to consider.

Evaluation of the Potential Use of a Collagen-Based Protein Hydrolysate as a Plant Multi-Stress Protectant.

Protein hydrolysates (PHs) are a class of plant biostimulants used in the agricultural practice to improve crop performance. In this study, we have assessed the capacity of a commercial PH derived from bovine collagen to mitigate drought, hypoxic, and Fe deficiency stress in Zea mays. As for the drought and hypoxic stresses, hydroponically grown plants treated with the PH exhibited an increased growth and absorption area of the roots compared with those treated with inorganic nitrogen. In the case of Fe deficiency, plants supplied with the PH mixed with FeCl3 showed a faster recovery from deficiency compared to plants supplied with FeCl3 alone or with FeEDTA, resulting in higher SPAD values, a greater concentration of Fe in the leaves and modulation in the expression of genes related to Fe. Moreover, through the analysis of circular dichroism spectra, we assessed that the PH interacts with Fe in a dose-dependent manner. Various hypothesis about the mechanisms of action of the collagen-based PH as stress protectant particularly in Fe-deficiency, are discussed.

Changes in physiological activities and root exudation profile of two grapevine rootstocks reveal common and specific strategies for Fe acquisition.

In several cultivation areas, grapevine can suffer from Fe chlorosis due to the calcareous and alkaline nature of soils. This plant species has been described to cope with Fe deficiency by activating Strategy I mechanisms, hence increasing root H+ extrusion and ferric-chelate reductase activity. The degree of tolerance exhibited by the rootstocks has been reported to depend on both reactions, but to date, little emphasis has been given to the role played by root exudate extrusion. We studied the behaviour of two hydroponically-grown, tolerant grapevine rootstocks (Ramsey and 140R) in response to Fe deficiency. Under these experimental conditions, the two varieties displayed differences in their ability to modulate morpho-physiological parameters, root acidification and ferric chelate reductase activity. The metabolic profiling of root exudates revealed common strategies for Fe acquisition, including ones targeted at reducing microbial competition for this micronutrient by limiting the exudation of amino acids and sugars and increasing instead that of Fe(III)-reducing compounds. Other modifications in exudate composition hint that the two rootstocks cope with Fe shortage via specific adjustments of their exudation patterns. Furthermore, the presence of 3-hydroxymugenic acid in these compounds suggests that the responses of grapevine to Fe availability are rather diverse and much more complex than those usually described for Strategy I plants.

FePO4 NPs Are an Efficient Nutritional Source for Plants: Combination of Nano-Material Properties and Metabolic Responses to Nutritional Deficiencies.

Phosphorous and iron are a macro- and micronutrient, respectively, whose low bioavailability can negatively affect crop productivity. There is ample evidence that the use of conventional P and Fe fertilizers has several environmental and economical disadvantages, but even though great expectations surround nanotechnology and its applications in the field of plant nutrition, little is known about the mechanisms underlying the uptake and use of these sub-micron particles (nanoparticles, NPs) by crop species. This work shows that cucumber and maize plants both use the nutrients borne by FePO4 NPs more efficiently than those supplied as bulk. However, morpho-physiological parameters and nutrient content analyses reveal that while cucumber plants (a Strategy I species with regard to Fe acquisition) mainly use these NPs as a source of P, maize (a Strategy II species) uses them preferentially for Fe. TEM analyses of cucumber root specimens revealed no cell internalization of the NPs. On the other hand, electron-dense nanometric structures were evident in proximity of the root epidermal cell walls of the NP-treated plants, which after ESEM/EDAX analyses can be reasonably identified as iron-oxyhydroxide. It appears that the nutritional interaction between roots and NPs is strongly influenced by species-specific metabolic responses.

Nitrogen Starvation Differentially Influences Transcriptional and Uptake Rate Profiles in Roots of Two Maize Inbred Lines with Different NUE.

Nitrogen use efficiency (NUE) of crops is estimated to be less than 50%, with a strong impact on environment and economy. Genotype-dependent ability to cope with N shortage has been only partially explored in maize and, in this context, the comparison of molecular responses of lines with different NUE is of particular interest in order to dissect the key elements underlying NUE. Changes in root transcriptome and NH4+/NO3- uptake rates during growth (after 1 and 4 days) without N were studied in high (Lo5) and low (T250) NUE maize inbred lines. Results suggests that only a small set of transcripts were commonly modulated in both lines in response to N starvation. However, in both lines, transcripts linked to anthocyanin biosynthesis and lateral root formation were positively affected. On the contrary, those involved in root elongation were downregulated. The main differences between the two lines reside in the ability to modulate the transcripts involved in the transport, distribution and assimilation of mineral nutrients. With regard to N mineral forms, only the Lo5 line responded to N starvation by increasing the NH4+ fluxes as supported by the upregulation of a transcript putatively involved in its transport.

FePO4 nanoparticles produced by an industrially scalable continuous-flow method are an available form of P and Fe for cucumber and maize plants.

Nanomaterials are widely used in medical and pharmaceutical fields, but their application in plant nutrition is at its infancy. Phosphorous (P) and iron (Fe) are essential mineral nutrients limiting in a wide range of conditions the yield of crops. Phosphate and Fe fertilizers to-date on the market display low efficiency (P fertilizers) and low persistence in soil (Fe fertilizers) and negatively affect the environment. In the tentative to overcome these problems, we developed a continuous industrially scalable method to produce FePO4 NPs based on the rapid mixing of salt solutions in a mixing chamber. The process, that included the addition of citrate as capping agent allowed to obtain a stable suspension of NPs over the time. The NPs were tested for their effectiveness as P and Fe sources on two hydroponically grown crop species (cucumber and maize) comparing their effects to those exerted by non-nanometric FePO4 (bulk FePO4). The results showed that FePO4 NPs improved the availability of P and Fe, if compared to the non-nano counterpart, as demonstrated by leaf SPAD indexes, fresh biomasses and P and Fe contents in tissues. The results open a new avenue in the application of nanosized material in the field of plant nutrition and fertilization.

The different tolerance to magnesium deficiency of two grapevine rootstocks relies on the ability to cope with oxidative stress.

BACKGROUND: Magnesium (Mg) deficiency causes physiological and molecular responses, already dissected in several plant species. The study of these responses among genotypes showing a different tolerance to the Mg shortage can allow identifying the mechanisms underlying the resistance to this nutritional disorder. To this aim, we compared the physiological and molecular responses (e.g. changes in root metabolome and transcriptome) of two grapevine rootstocks exhibiting, in field, different behaviors with respect to Mg shortage (1103P, tolerant and SO4 susceptible). RESULTS: The two grapevine rootstocks confirmed, in a controlled growing system, their behavior in relation to the tolerance to Mg deficiency. Differences in metabolite and transcriptional profiles between the roots of the two genotypes were mainly linked to antioxidative compounds and the cell wall constituents. In addition, differences in secondary metabolism, in term of both metabolites (e.g. alkaloids, terpenoids and phenylpropanoids) and transcripts, assessed between 1103P and SO4 suggest a different behavior in relation to stress responses particularly at early stages of Mg deficiency. CONCLUSIONS: Our results suggested that the higher ability of 1103P to tolerate Mg shortage is mainly linked to its capability of coping, faster and more efficiently, with the oxidative stress condition caused by the nutritional disorder.

Prospect on Ionomic Signatures for the Classification of Grapevine Berries According to Their Geographical Origin.

The determination of food geographical origin has been an important subject of study over the past decade, with an increasing number of analytical techniques being developed to determine the provenance of agricultural products. Agricultural soils can differ for the composition and the relative quantities of mineral nutrients and trace elements whose bioavailability depends on soil properties. Therefore, the ionome of fruits, vegetables and derived products can reflect the mineral composition of the growth substrate. Multi-elemental analysis has been successfully applied to trace the provenance of wines from different countries or different wine-producing regions. However, winemaking process and environmental and cultural conditions may affect a geographical fingerprint. In this article, we discuss the possibility of applying ionomics in wines classification on a local scale and also by exploiting grape berry analyses. In this regard, we present the ionomic profile of grapevine berries grown within an area of approximately 300 km2 and the subsequent application of chemometric methods for the assignment of their geographical origin. The best discrimination was obtained by using a dataset composed only of rare earth elements. Considering the experiences reported in the literature and our results, we concluded that sample representativeness and the application of a preliminary Principal Component Analysis, as pattern recognition techniques, might represent two necessary starting points for the geographical determination of the geographical origin of grape berries; therefore, on the basis of these observations we also include some recommendations to be considered for future application of these techniques for grape and wines classification.

Growth Stimulatory Effects and Genome-Wide Transcriptional Changes Produced by Protein Hydrolysates in Maize Seedlings.

Protein hydrolysates are an emerging class of crop management products utilized for improving nutrient assimilation and mitigating crop stress. They generally consist of a mixture of peptides and free amino acids derived from the hydrolysis of plant or animal sources. The present work was aimed at studying the effects and the action mechanisms of a protein hydrolysate derived from animal residues on maize root growth and physiology in comparison with the effects induced by either free amino acids or inorganic N supply. The application of the protein hydrolysate caused a remarkable enhancement of root growth. In particular, in the protein hydrolysate-treated plants the length and surface area of lateral roots were about 7 and 1.5 times higher than in plants treated with inorganic N or free amino acids, respectively. The root growth promoting effect of the protein hydrolysate was associated with an increased root accumulation of K, Zn, Cu, and Mn when compared with inorganic N and amino acids treatments. A microarray analysis allowed to dissect the transcriptional changes induced by the different treatments demonstrating treatment-specific effects principally on cell wall organization, transport processes, stress responses and hormone metabolism.

Transcriptional and physiological analyses of Fe deficiency response in maize reveal the presence of Strategy I components and Fe/P interactions.

BACKGROUND: Under limited iron (Fe) availability maize, a Strategy II plant, improves Fe acquisition through the release of phytosiderophores (PS) into the rhizosphere and the subsequent uptake of Fe-PS complexes into root cells. Occurrence of Strategy-I-like components and interactions with phosphorous (P) nutrition has been hypothesized based on molecular and physiological studies in grasses. RESULTS: In this report transcriptomic analysis (NimbleGen microarray) of Fe deficiency response revealed that maize roots modulated the expression levels of 724 genes (508 up- and 216 down-regulated, respectively). As expected, roots of Fe-deficient maize plants overexpressed genes involved in the synthesis and release of 2'-deoxymugineic acid (the main PS released by maize roots). A strong modulation of genes involved in regulatory aspects, Fe translocation, root morphological modification, primary metabolic pathways and hormonal metabolism was induced by the nutritional stress. Genes encoding transporters for Fe2+ (ZmNRAMP1) and P (ZmPHT1;7 and ZmPHO1) were also up-regulated under Fe deficiency. Fe-deficient maize plants accumulated higher amounts of P than the Fe-sufficient ones, both in roots and shoots. The supply of 1 µM 59Fe, as soluble (Fe-Citrate and Fe-PS) or sparingly soluble (Ferrihydrite) sources to deficient plants, caused a rapid down-regulation of genes coding for PS and Fe(III)-PS transport, as well as of ZmNRAMP1 and ZmPHT1;7. Levels of 32P absorption essentially followed the rates of 59Fe uptake in Fe-deficient plants during Fe resupply, suggesting that P accumulation might be regulated by Fe uptake in maize plants. CONCLUSIONS: The transcriptional response to Fe-deficiency in maize roots confirmed the modulation of known genes involved in the Strategy II and revealed the presence of Strategy I components usually described in dicots. Moreover, data here presented provide evidence of a close relationship between two essential nutrients for plants, Fe and P, and highlight a key role played by Fe and P transporters to preserve the homeostasis of these two nutrients in maize plants.

Time-Resolved Investigation of Molecular Components Involved in the Induction of [Formula: see text] High Affinity Transport System in Maize Roots.

The induction, i.e., the rapid increase of nitrate ([Formula: see text]) uptake following the exposure of roots to the anion, was studied integrating physiological and molecular levels in maize roots. Responses to [Formula: see text] treatment were characterized in terms of changes in [Formula: see text] uptake rate and plasma membrane (PM) H+-ATPase activity and related to transcriptional and protein profiles of NRT2, NRT3, and PM H+-ATPase gene families. The behavior of transcripts and proteins of ZmNRT2s and ZmNRT3s suggested that the regulation of the activity of inducible high-affinity transport system (iHATS) is mainly based on the transcriptional/translational modulation of the accessory protein ZmNRT3.1A. Furthermore, ZmNRT2.1 and ZmNRT3.1A appear to be associated in a ∼150 kDa oligomer. The expression trend during the induction of the 11 identified PM H+-ATPase transcripts indicates that those mainly involved in the response to [Formula: see text] treatment are ZmHA2 and ZmHA4. Yet, partial correlation between the gene expression, protein levels and enzyme activity suggests an involvement of post-transcriptional and post-translational mechanisms of regulation. A non-denaturing Deriphat-PAGE approach allowed demonstrating for the first time that PM H+-ATPase can occur in vivo as hexameric complex together with the already described monomeric and dimeric forms.

Short-Term Treatment with the Urease Inhibitor N-(n-Butyl) Thiophosphoric Triamide (NBPT) Alters Urea Assimilation and Modulates Transcriptional Profiles of Genes Involved in Primary and Secondary Metabolism in Maize Seedlings.

To limit nitrogen (N) losses from the soil, it has been suggested to provide urea to crops in conjunction with the urease inhibitor N-(n-butyl) thiophosphoric triamide (NBPT). However, recent studies reported that NBPT affects urea uptake and urease activity in plants. To shed light on these latter aspects, the effects of NBPT were studied analysing transcriptomic and metabolic changes occurring in urea-fed maize seedlings after a short-term exposure to the inhibitor. We provide evidence that NBPT treatment led to a wide reprogramming of plant metabolism. NBPT inhibited the activity of endogenous urease limiting the release and assimilation of ureic-ammonium, with a simultaneous accumulation of urea in plant tissues. Furthermore, NBPT determined changes in the glutamine, glutamate, and asparagine contents. Microarray data indicate that NBPT affects ureic-N assimilation and primary metabolism, such as glycolysis, TCA cycle, and electron transport chain, while activates the phenylalanine/tyrosine-derivative pathway. Moreover, the expression of genes relating to the transport and complexation of divalent metals was strongly modulated by NBPT. Data here presented suggest that when NBPT is provided in conjunction with urea an imbalance between C and N compounds might occur in plant cells. Under this condition, root cells also seem to activate a response to maintain the homeostasis of some micronutrients.

Early transcriptomic response to Fe supply in Fe-deficient tomato plants is strongly influenced by the nature of the chelating agent.

BACKGROUND: It is well known that in the rhizosphere soluble Fe sources available for plants are mainly represented by a mixture of complexes between the micronutrient and organic ligands such as carboxylates and phytosiderophores (PS) released by roots, as well as fractions of humified organic matter. The use by roots of these three natural Fe sources (Fe-citrate, Fe-PS and Fe complexed to water-extractable humic substances, Fe-WEHS) have been already studied at physiological level but the knowledge about the transcriptomic aspects is still lacking. RESULTS: The (59)Fe concentration recorded after 24 h in tissues of tomato Fe-deficient plants supplied with (59)Fe complexed to WEHS reached values about 2 times higher than those measured in response to the supply with Fe-citrate and Fe-PS. However, after 1 h no differences among the three Fe-chelates were observed considering the (59)Fe concentration and the root Fe(III) reduction activity. A large-scale transcriptional analysis of root tissue after 1 h of Fe supply showed that Fe-WEHS modulated only two transcripts leaving the transcriptome substantially identical to Fe-deficient plants. On the other hand, Fe-citrate and Fe-PS affected 728 and 408 transcripts, respectively, having 289 a similar transcriptional behaviour in response to both Fe sources. CONCLUSIONS: The root transcriptional response to the Fe supply depends on the nature of chelating agents (WEHS, citrate and PS). The supply of Fe-citrate and Fe-PS showed not only a fast back regulation of molecular mechanisms modulated by Fe deficiency but also specific responses due to the uptake of the chelating molecule. Plants fed with Fe-WEHS did not show relevant changes in the root transcriptome with respect to the Fe-deficient plants, indicating that roots did not sense the restored cellular Fe accumulation.