FePO4 nanoparticles as a source of nutrients for plants: synthesis and evaluation of their effects on hydroponically grown cucumber and maize seedlings

The nutrient use efficiency (NUE) of crops is typically low, in particular referring to the uptake of nutrients applied through fertilizers. A strategy to improve the NUE could be the development of new and more efficient fertilizers. A promising field in order to achieve this goal could be the use of nanotechnology. Nanomaterials are widely used in medical and pharmaceutical fields, but their application in agriculture and in particular in plant nutrition is at its infancy. A continuous method of FePO4 nanoparticles (FePO4 NPs) synthesis based on the extremely fine and rapid mixing of a FeCl3 solution with a K2HPO4 solution in a mixing chamber was tested for its effectiveness with a laboratory-made system. The proof-of-concept could produce FePO4 particles smaller than 100 nm, reaching the threshold of 50% of particles smaller of 100 nm, a value that is recommended by the European Union for the definition of nanomaterial. A pilot plant for the continuous FePO4 NPs synthesis was set up, using two dosing pumps for solutions pumping, and an HPLC mixing tee as mixing chamber. The system could produce 15 L\u2022h-1 of raw FePO4 NPs suspension. Purification through dyalisis was optimized, together with a stabilization method of FePO4 NPs, called citrate capping, based on the adding of tribasic potassium citrate and thorough vortexing, in order to reduce aggregation and sedimentation of particles on long time periods. FePO4 NPs were then tested for their effectiveness as source of P and Fe on two hydroponically grown crop species, cucumber (Cucumis sativus) and maize (Zea mays). The experiments were designed in order to evaluate the effect of FePO4 NPs as source of both nutrients, or source of sole P and Fe. For this reason, as negative controls were used plants grown without P (-P), without Fe (-Fe), or without both nutrients (-P-Fe). In addition, in order to analyze if the size of FePO4 particles could cause different effects on plants, we included in the experiment a treatment with non-nanometric FePO4 (bulk FePO4). The results showed that nano-sized FePO4 improved the availability of P and Fe, if compared to the non-nano counterpart, as demonstrated by SPAD indexes of leaves and the determination of nutrients concentrations in tissues. Transmission Electron Microscopy (TEM) observations on cucumber roots treated with FePO4 NPs revealed that these particles did not enter into the plant, suggesting as mechanism of delivery of nutrients the dissolution in the apoplast. Gene expression analysis of homologs of AtPHR1, a key regulator of the response to P starvation in Arabidopsis, revealed in cucumber an upregulation of Csa3M608690 in plants grown with FePO4 NPs. The transcriptional behavior of Csa1M024210, homologs of AtBTS, suggested that plants grown with both forms of FePO4 are, with respect of Fe, in good nutritional conditions thus confirming physiological parameters. For maize, the negative modulation of ZmFER-Like gene in response to all treatments suggested a minor role of this gene in the regulation of Fe homeostasis in this plant species, while the upregulation of ZmIRO2 in plants grown with both forms of FePO4 confirmed the sub-optimal nutritional state of the plants

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

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

Water-extractable humic substances speed up transcriptional response of maize roots to nitrate

Humic substances are known to positively influence plant growth and nutrition. In particular, the water-extractable fraction of humic substances (WEHS) has been shown to enhance nitrate acquisition, increasing the activity of high affinity nitrate uptake system. However, molecular bases of this physiological response are not clarified so far. Thus, in the present work, the physiological effect of WEHS on nitrate acquisition in maize roots was correlated with changes in the root transcriptomic profile. Results confirmed that WEHS caused a faster induction of a higher capacity to take up nitrate in maize roots. Comparing the root transcriptomic profile of Nitrate- and Nitrate + WEHS-treated plants with Control (-N) ones, more than 2000 transcripts appeared to be modulated only in the presence of WEHS. Among these, genes involved in nitrate transport and assimilation (NRT1s, NRT2s, NAR2.1, NR, GS, GOGAT, CNX, UPM) were strongly modulated by WEHS. Furthermore, also some genes known to be linked to the nitrogen limitation responses were affected by WEHS, as transcripts coding for transcription factors (as LBD37, NIN-like protein, NFY-A, GRF5) and enzymes of hormones\u2019 metabolism. The modulation of these transcripts might play a crucial role in coordinating the induction to nitrate, favouring its uptake and assimilation in WEHS-treated plants. The overexpression of nitrogen assimilatory genes by WEHS might led to an early feedback regulation of the high affinity nitrate transport system, as being operated by N-metabolites. Results of the present work shed further light on the contribution of the organic soil component to the nitrogen use efficiency in crops. \ua9 2017 Elsevier B.V

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

Abstract 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

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. H(2)bpcd/Fe3+ complex features distinctive chemical characteristics, including moderate stability of the Fe(bpcd)(+) species (log beta(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 H(2)bpcd/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 mu M H(2)bpcd/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 mu M concentration of Fe-57 showed that cucumber roots absorbed H(2)bpcd/Fe-57(3+) at a slower rate than EDTA/Fe-57(3+). The high kinetic inertness of H(2)bpcd/Fe3+ may explain such behavior

Ticagrelor improves endothelial function by decreasing circulating Epidermal Growth Factor (EGF)

Ticagrelor is one of the most powerful P2Y12inhibitor. We have recently reported that, in patients with concomitant Stable Coronary Artery Disease (SCAD) and Chronic Obstructive Pulmonary Disease (COPD) undergoing percutaneous coronary intervention (PCI), treatment with ticagrelor, as compared to clopidogrel, is associated with an improvement of the endothelial function (Clinical Trial NCT02519608). In the present study, we showed that, in the same population, after 1 month treatment with ticagrelor, but not with clopidogrel, there is a decrease of the circulating levels of epidermal growth factor (EGF) and that these changes in circulating levels of EGF correlate with on-treatment platelet reactivity. Furthermore, in human umbilical vein endothelial cells (HUVEC) incubated with sera of the patients treated with ticagrelor, but not with clopidogrel there is an increase of p-eNOS levels. Finally, analyzing the changes in EGF and p-eNOS levels after treatment, we observed an inverse correlation between p-eNOS and EGF changes only in the ticagrelor group. Causality between EGF and eNOS activation was assessed in vitro in HUVEC where we showed that EGF decreases eNOS activity in a dose dependent manner. Taken together our data indicate that ticagrelor improves endothelial function by lowering circulating EGF that results in the activation of eNOS in the vascular endothelium