Unravelling the biostimulant activity of a protein hydrolysate in lettuce plants under optimal and low N availability: a multi-omics approach.

The application of protein hydrolysates (PH) biostimulants is considered a promising approach to promote crop growth and resilience against abiotic stresses. Nevertheless, PHs bioactivity depends on both the raw material used for their preparation and the molecular fraction applied. The present research aimed at investigating the molecular mechanisms triggered by applying a PH and its fractions on plants subjected to nitrogen limitations. To this objective, an integrated transcriptomic-metabolomic approach was used to assess lettuce plants grown under different nitrogen levels and treated with either the commercial PH Vegamin® or its molecular fractions PH1(>10 kDa), PH2 (1-10 kDa) and PH3 (<1 kDa). Regardless of nitrogen provision, biostimulant application enhanced lettuce biomass, likely through a hormone-like activity. This was confirmed by the modulation of genes involved in auxin and cytokinin synthesis, mirrored by an increase in the metabolic levels of these hormones. Consistently, PH and PH3 upregulated genes involved in cell wall growth and plasticity. Furthermore, the accumulation of specific metabolites suggested the activation of a multifaceted antioxidant machinery. Notwithstanding, the modulation of stress-response transcription factors and genes involved in detoxification processes was observed. The coordinated action of these molecular entities might underpin the increased resilience of lettuce plants against nitrogen-limiting conditions. In conclusion, integrating omics techniques allowed the elucidation of mechanistic aspects underlying PH bioactivity in crops. Most importantly, the comparison of PH with its fraction PH3 showed that, except for a few peculiarities, the effects induced were equivalent, suggesting that the highest bioactivity was ascribable to the lightest molecular fraction.

Microplastic influences the ménage à trois among the plant, a fungal pathogen, and a plant growth-promoting fungal species.

Microplastics (MP) can influence a plethora of fungal species within the rhizosphere. Nevertheless, there are few studies on the direct impacts of MPs on soil fungi and their intricate interplay with plants. Here, we investigated the impact of polyethylene microspheres (PEMS) on the ecological interactions between Fusarium solani, a plant pathogenic fungus, and Trichoderma viride, a fungal plant growth promotor, within the rhizosphere of Solanum lycopersicum (tomato). Spores of F. solani and T. viride were pre-incubated with PEMS at two concentrations, 100 and 1000 mg L-1. Mycelium growth, sporulation, spore germination, and elongation were evaluated. Tomato seeds were exposed to fungal spore suspensions treated with PEMS, and plant development was subsequently assessed after 4 days. The results showed that PEMS significantly enhanced the sporulation (106.0 % and 70.1 %) but compromised the spore germination (up to 27.3 % and 32.2 %) and radial growth (up to -5.2% and -21.7 %) of F. solani and T. viride, respectively. Furthermore, the 100 and 1000 mg L-1 concentrations of PEMS significantly (p<0.05) enhanced the mycelium density of T. viride (9.74 % and 22.30 %, respectively), and impaired the germ-tube elongation of F. solani after 4 h (16.16 % and 11.85 %, respectively) and 8 h (4 % and 17.10 %, respectively). In addition, PEMS amplified the pathogenicity of F. solani and boosted the bio-enhancement effect of T. viride on tomato root growth. Further, PEMS enhanced the bio-fungicidal effect of T. viride toward F. solani (p<0.05). In summary, PEMS had varying effects on F. solani and T. viride, impacting their interactions and influencing their relationship with tomato plants. It intensified the beneficial effects of T. viride and increased the aggressiveness of F. solani. This study highlights concerns regarding the effects of MPs on fungal interactions in the rhizosphere, which are essential for crop soil colonization and resource utilization.

An insight into the role of the organic acids produced by Enterobacter sp. strain 15S in solubilizing tricalcium phosphate: in situ study on cucumber.

BACKGROUND: The release of organic acids (OAs) is considered the main mechanism used by phosphate-solubilizing bacteria (PSB) to dissolve inorganic phosphate in soil. Nevertheless, little is known about the effect of individual OAs produced by a particular PSB in a soil-plant system. For these reasons, the present work aimed at investigating the effect of Enterobacter sp. strain 15S and the exogenous application of its OAs on (i) the solubilization of tricalcium phosphate (TCP), (ii) plant growth and (iii) P nutrition of cucumber. To this purpose two independent experiments have been performed. RESULTS: In the first experiment, carried out in vitro, the phosphate solubilizing activity of Enterobacter 15S was associated with the release of citric, fumaric, ketoglutaric, malic, and oxalic acids. In the second experiment, cucumber plants were grown in a Leonard jar system consisting of a nutrient solution supplemented with the OAs previously identified in Enterobacter 15S (jar's base) and a substrate supplemented with the insoluble TCP where cucumber plants were grown (jar's top). The use of Enterobacter 15S and its secreted OAs proved to be efficient in the in situ TCP solubilization. In particular, the enhancement of the morpho-physiological traits of P-starved cucumber plants was evident when treated with Enterobacter 15S, oxalate, or citrate. The highest accumulation of P in roots and shoots induced by such treatments further corroborated this hypothesis. CONCLUSION: In our study, the results presented suggest that organic acids released by Enterobacter 15S as well as the bacterium itself can enhance the P-acquisition by cucumber plants.

A Matter of Metals: Copper but Not Cadmium Affects the Microbial Alpha-Diversity of Soils and Sediments - a Meta-analysis.

Heavy metal (HM) accumulation in soil affects plants and soil fauna, yet the effect on microbial alpha-diversity remains unclear, mainly due to the absence of dedicated research synthesis (e.g. meta-analysis). Here, we report the first meta-analysis of the response of soil microbial alpha-diversity to the experimental addition of cadmium (Cd) and copper (Cu). We considered studies conducted between 2013 and 2022 using DNA metabarcoding of bacterial and fungal communities to overcome limitations of other cultivation- and electrophoresis-based techniques. Fungi were discarded due to the limited study number (i.e. 6 studies). Bacterial studies resulted in 66 independent experiments reported in 32 primary papers from four continents. We found a negative dose-dependent response for Cu but not for Cd for bacterial alpha-diversity in the environments, only for Cu additions exceeding 29.6 mg kg-1 (first loss of - 0.06% at 30 mg kg-1). The maximal loss of bacterial alpha-diversity registered was 13.89% at 3837 mg kg-1. Our results first highlight that bacterial communities behave differently to soil pollution depending on the metal. Secondly, our study suggests that even extreme doses of Cu do not cause a dramatic loss in alpha-diversity, highlighting how the behaviour of bacterial communities diverges from soil macro-organisms.

Interaction between Sulfate and Selenate in Tetraploid Wheat (Triticum turgidum L.) Genotypes.

Selenium (Se) is an essential micronutrient of fundamental importance to human health and the main Se source is from plant-derived foods. Plants mainly take up Se as selenate (SeO42-), through the root sulfate transport system, because of their chemical similarity. The aims of this study were (1) to characterize the interaction between Se and S during the root uptake process, by measuring the expression of genes coding for high-affinity sulfate transporters and (2) to explore the possibility of increasing plant capability to take up Se by modulating S availability in the growth medium. We selected different tetraploid wheat genotypes as model plants, including a modern genotype, Svevo (Triticum turgidum ssp. durum), and three ancient Khorasan wheats, Kamut, Turanicum 21, and Etrusco (Triticum turgidum ssp. turanicum). The plants were cultivated hydroponically for 20 days in the presence of two sulfate levels, adequate (S = 1.2 mM) and limiting (L = 0.06 mM), and three selenate levels (0, 10, 50 µM). Our findings clearly showed the differential expression of genes encoding the two high-affinity transporters (TdSultr1.1 and TdSultr1.3), which are involved in the primary uptake of sulfate from the rhizosphere. Interestingly, Se accumulation in shoots was higher when S was limited in the nutrient solution.

Integrated Metabolomics and Morpho-Biochemical Analyses Reveal a Better Performance of Azospirillum brasilense over Plant-Derived Biostimulants in Counteracting Salt Stress in Tomato.

Increased soil salinity is one of the main concerns in agriculture and food production, and it negatively affects plant growth and crop productivity. In order to mitigate the adverse effects of salinity stress, plant biostimulants (PBs) have been indicated as a promising approach. Indeed, these products have a beneficial effect on plants by acting on primary and secondary metabolism and by inducing the accumulation of protective molecules against oxidative stress. In this context, the present work is aimed at comparatively investigating the effects of microbial (i.e., Azospirillum brasilense) and plant-derived biostimulants in alleviating salt stress in tomato plants by adopting a multidisciplinary approach. To do so, the morphological and biochemical effects were assessed by analyzing the biomass accumulation and root characteristics, the activity of antioxidant enzymes and osmotic stress protection. Furthermore, modifications in the metabolomic profiles of both leaves and root exudates were also investigated by ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC/QTOF-MS). According to the results, biomass accumulation decreased under high salinity. However, the treatment with A. brasilense considerably improved root architecture and increased root biomass by 156% and 118% in non-saline and saline conditions, respectively. The antioxidant enzymes and proline production were enhanced in salinity stress at different levels according to the biostimulant applied. Moreover, the metabolomic analyses pointed out a wide set of processes being affected by salinity and biostimulant interactions. Crucial compounds belonging to secondary metabolism (phenylpropanoids, alkaloids and other N-containing metabolites, and membrane lipids) and phytohormones (brassinosteroids, cytokinins and methylsalicylate) showed the most pronounced modulation. Overall, our results suggest a better performance of A. brasilense in alleviating high salinity than the vegetal-derived protein hydrolysates herein evaluated.

Millimetre-resolution mapping of citrate exuded from soil-grown roots using a novel, low-invasive sampling technique.

The reliable sampling of root exudates in soil-grown plants is experimentally challenging. This study aimed at developing a citrate sampling and mapping technique with millimetre-resolution using DGT (diffusive gradients in thin films) ZrOH-binding gels. Citrate adsorption kinetics, DGT capacity, and stability of ZrOH gels were evaluated. ZrOH gels were applied to generate 2D maps of citrate exuded by white lupin roots grown in a rhizotron in a phosphorus-deficient soil. Citrate was adsorbed quantitatively and rapidly by the ZrOH gels; these gels can be stored after sampling for several weeks prior to analysis. The DGT capacity of the ZrOH gel for citrate depends on the ionic strength and the pH of the soil solution, but was suitable for citrate sampling. We generated for the first time 2D citrate maps of rhizotron-grown plants at a millimetre resolution to measure an illustrated plant response to phosphorus fertilization, demonstrating that DGT-based citrate sampling is suitable for studying root exudation in soil environments, at high spatial resolution. The change of binding material would also allow sampling of other exudate classes and exudation profiles of entire root systems. These aspects are crucial in cultivar breeding and selection.

I Like the Way You Eat It: Lemur (Indri indri) Gut Mycobiome and Geophagy.

Here, we investigated the possible linkages among geophagy, soil characteristics, and gut mycobiome of indri (Indri indri), an endangered lemur species able to survive only in wild conditions. The soil eaten by indri resulted in enriched secondary oxide-hydroxides and clays, together with a high concentration of specific essential micronutrients. This could partially explain the role of the soil in detoxification and as a nutrient supply. Besides, we found that soil subject to geophagy and indris' faeces shared about 8.9% of the fungal OTUs. Also, several genera (e.g. Fusarium, Aspergillus and Penicillium) commonly associated with soil and plant material were found in both geophagic soil and indri samples. On the contrary, some taxa with pathogenic potentials, such as Cryptococcus, were only found in indri samples. Further, many saprotrophs and plant-associated fungal taxa were detected in the indri faeces. These fungal species may be involved in the digestion processes of leaves and could have a beneficial role in their health. In conclusion, we found an intimate connection between gut mycobiome and soil, highlighting, once again, the potential consequent impacts on the wider habitat.

Phytotoxicity of hydrochars obtained by hydrothermal carbonization of manure-based digestate.

The management of digestate, the main by-product of the anaerobic digestion (AD) process, is one of the most serious environmental issues. Although digestate is used on arable land as a fertilizer, it can have a negative impact on the environment due to nitrate leaching into the groundwater and ammonia volatilization into the atmosphere, with high economic and environmental disposal costs. Therefore, hydrothermal carbonization (HTC), a thermochemical biomass conversion process, could represent a sustainable and efficient alternative for digestate management. Hydrochar, the solid product of the HTC process, has been recently proposed as a plant growing medium in soilless culture systems (SCS). Here, using cow manure digestate as feedstock, we investigated the influence of the HTC process reaction temperature (180, 220 and 250 °C) and residence time (1 and 3 h) on the physical-chemical properties (pH, electrical conductivity, and mineral element concentrations) of the resulting hydrochars. Furthermore, in order to fully valorize hydrochar as a growing medium, their possible phytotoxic effects and those of their water extracts (prepared at two different concentrations and at different pHs) were tested in germination tests with cress seeds (Lepidium sativum L.). Concentrations of nutrients, heavy metals, organic acids, sugars and furan compounds were determined in the water extracts. Characterization analysis of these hydrochars revealed that they can be distinguished from each other by their physical-chemical properties, which were significantly affected by the two process parameters. Specifically, the HTC temperature had a greater effect on the composition of hydrochars than the residence time. Germination tests found hydrochar water extracts to show significantly lower phytotoxicity than the hydrochars themselves. Notably, the phytotoxic effect of the extracts decreased with increasing extraction ratio and decreasing pH. The chromatographic characterization of extracts identified the presence of potential phytotoxins, such as furan compounds (i.e., hydroxymethylfurfural and furfural). However, before using hydrochars as potential and innovative growing media for plants, their phytotoxicity should be limited, for example through their dilution with other substrates. Overall, AD-HTC coupling could represent a valuable eco-sustainable expedient in the field of biomasses, green economy and waste conversion and, therefore, further investigations in this direction are necessary.

Single and Combined Fe and S Deficiency Differentially Modulate Root Exudate Composition in Tomato: A Double Strategy for Fe Acquisition?

Fe chlorosis is considered as one of the major constraints on crop growth and yield worldwide, being particularly worse when associated with S shortage, due to the tight link between Fe and S. Plant adaptation to inadequate nutrient availabilities often relies on the release of root exudates that enhance nutrients, mobilization from soil colloids and favour their uptake by roots. This work aims at characterizing the exudomic profile of hydroponically grown tomato plants subjected to either single or combined Fe and S deficiency, as well as at shedding light on the regulation mechanisms underlying Fe and S acquisition processes by plants. Root exudates have been analysed by untargeted metabolomics, through liquid chromatography-mass spectrometry as well as gas chromatography-mass spectrometry following derivatization. More than 200 metabolites could be putatively annotated. Venn diagrams show that 23%, 10% and 21% of differential metabolites are distinctively modulated by single Fe deficiency, single S deficiency or combined Fe-S deficiency, respectively. Interestingly, for the first time, a mugineic acid derivative is detected in dicot plants root exudates. The results seem to support the hypothesis of the co-existence of the two Fe acquisition strategies in tomato plants.

Common and specific responses to iron and phosphorus deficiencies in roots of apple tree (Malus × domestica).

Iron and phosphorus are abundant elements in soils but poorly available for plant nutrition. The availability of these two nutrients represents a major constraint for fruit tree cultivation such as apple (Malus × domestica) leading very often to a decrease of fruit productivity and quality worsening. Aim of this study was to characterize common and specific features of plant response to Fe and P deficiencies by ionomic, transcriptomic and exudation profiling of apple roots. Under P deficiency, the root release of oxalate and flavonoids increased. Genes encoding for transcription factors and transporters involved in the synthesis and release of root exudates were upregulated by P-deficient roots, as well as those directly related to P acquisition. In Fe-deficiency, plants showed an over-accumulation of P, Zn, Cu and Mn and induced the transcription of those genes involved in the mechanisms for the release of Fe-chelating compounds and Fe mobilization inside the plants. The intriguing modulation in roots of some transcription factors, might indicate that, in this condition, Fe homeostasis is regulated by a FIT-independent pathway. In the present work common and specific features of apple response to Fe and P deficiency has been reported. In particular, data indicate similar modulation of a. 230 genes, suggesting the occurrence of a crosstalk between the two nutritional responses involving the transcriptional regulation, shikimate pathway, and the root release of exudates.

Azospirillum brasilense inoculation counteracts the induction of nitrate uptake in maize plants.

Nitrogen (N) represents one of the limiting factors for crop growth and productivity and to date has been widely supplied via external application of fertilizers. However, the use of plant growth-promoting rhizobacteria (PGPR) might represent a valuable tool to further improve plant nutrition. This study examines the influence of Azospirillum brasilense strain Cd on nitrate uptake in maize (Zea mays) plants, focusing on the high-affinity transport system (HATS). Plants were induced with nitrate (500 µM) and either inoculated or not with Azospirillum. Inoculation decreased the nitrate uptake rate in induced plants, suggesting that Azospirillum may negatively affect HATS in the short term. The expression dynamics of ZmNF-YA and ZmLBD37 suggested that Azospirillum affected the N balance in the plants, most probably by supplying them with reduced N, i.e. NH4+. This was further corroborated by measurements of total N and the expression of ammonium transporter genes. Overall, our data demonstrate that Azospirillum can counteract the plant response to nitrate induction, albeit without compromising N nutrition. This suggests that the agricultural application of microbial inoculants requires fine-tuning of external fertilizer inputs.

The potential of two different Avena sativa L. cultivars to alleviate Cu toxicity.

Agronomic strategies as intercropping might be applied to reduce plant-available copper (Cu) in Cu-contaminated soils. Thus, our aim was to characterize two different oat cultivars, Avena sativa L. cv. Fronteira and cv. Perona for their ability to tolerate and/or phytostabilize Cu. Copper toxicity reduced plant biomass of both cultivars. The exudate analysis revealed the presence of phenolic compounds and phytosiderophores, yet with a different pattern between the cultivars: cv. Fronteira showed a Cu-concentration and time-dependent release of phenolic compounds, while cv. Perona down-regulated this release during the second week of treatment. Copper concentration increased linearly in all the tissues analysed with increasing Cu concentration showing yet a different compartmentalization: cv. Fronteira and cv. Perona preferentially accumulated Cu in the apoplasm and symplast, respectively. This higher accumulation of Cu in the apoplasm strongly reduces the available binding sites, leading to a competitive absorption with other macro-and micronutrients (e.g. Ca, Mn, Zn). Furthermore, in both cultivars Cu toxicity led to a significant reduction of shoot phosphorus content. The ionomic profile and compartmentalization of Cu together with the root activities demonstrate the different tolerance mechanism towards Cu toxicity of the two oat cultivars. In particular, cv. Fronteira seems to adopt an exclusion strategy based on accumulating Cu in the apoplasm and on the exudation of phenolic compounds. Thus, this cultivar could reduce the mobility and the consequent soil bioavailability of Cu playing an important role as phytostabilizer plant in intercropping systems in Cu-contaminated vineyards or orchards.

Growth and chemical changes in the rhizosphere of black oat (Avena strigosa) grown in soils contaminated with copper.

Copper based pesticides are used to protect vineyards from fungal infections. Plants like black oats (Avena strigosa Schreb) can promote chemical changes in the rhizosphere, reducing copper (Cu) bioavailability in contaminated soils. The objective of this study was to evaluate how copper additions would affect growth, morphology and nutrient uptake by black oats and how the plants affect the chemical composition in rhizosphere and bulk soil. The soil was collected in grassland of southern Brazil. The soil was air-dried, adjusted pH and added phosphorus and potassium amendments, and then it was incubated. Three Cu levels were established in the soil with the addition of 0, 40 and 80 mg Cu kg-1. The experimental design consisted of pots containing 8 plants with 10 kg of soil. Rhizosphere (2 kg of soil) and bulk (8 kg of soil) separated by a 30 µm nylon membrane. Black oat plants were grown for 54 days. The soil and solution were chemically characterized throughout cultivation for Cu speciation. At 54 days after emergence, the soil was sampled and proceeded chemical analysis and plants were collected to determine yield dry matter, morphological parameters and nutrient concentration. Black oat plants induce increase of pH and dissolved organic carbon in the rhizosphere. These root-induced processes increase the percentage of complexed chemical species and decrease free Cu+2 in soil solution, decreasing Cu toxicity. However, soil contamination with Cu induces morphological changes and nutritional imbalances. Black oats could thus be planted along with vineyards, for such increasing protect the soil and promote nutrient cycling, as well as reduce the free Cu available fraction due to the root-induced modifications in the rhizosphere.

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.

Effects of zinc addition to a copper-contaminated vineyard soil on sorption of Zn by soil and plant physiological responses.

The occurrence of high levels of Cu in vineyard soils is often the result of intensive use of fungicides for the preventive control of foliar diseases and can cause toxicity to plants. Nowadays many grape growers in Southern Brazil have replaced Cu-based with Zn-based products. The aim of the study was to evaluate whether the increase in Zn concentration in a soil with high Cu contents can interfere with the dynamics of these elements, and if this increase in Zn may cause toxicity to maize (Zea mays L.). Soil samples were collected in two areas, one in a vineyard with more than 30 years of cultivation and high concentration of Cu and the other on a natural grassland area adjacent to the vineyard. Different doses of Cu and Zn were added to the soil, and the adsorption isotherms were built following the Langmuir's model. In a second experiment, the vineyard soil was spiked with different Zn concentrations (0, 30, 60, 90, 180, and 270mg Zn kg(-1)) in 3kg pots where maize was grown in a greenhouse for 35 days. When Cu and Zn were added together, there was a reduction in the quantities adsorbed, especially for Zn. Zn addition decreased the total plant dry matter and specific leaf mass. Furthermore, with the increase in the activity of catalase, an activation of the antioxidant system was observed. However, the system was not sufficiently effective to reverse the stress levels imposed on soil, especially in plants grown in the highest doses of Zn. At doses higher than 90Znmgkg(-1) in the Cu-contaminated vineyard soil, maize plants were no longer able to activate the protection mechanism and suffered from metal stress, resulting in suppressed dry matter yields due to impaired functioning of the photosynthetic apparatus and changes in the enzymatic activity of plants. Replacement of Cu- by Zn-based fungicides to avoid Cu toxicity has resulted in soil vineyards contaminated with these metals and damaging of plant photosynthetic apparatus and enzyme activity.

Transcriptomic analysis highlights reciprocal interactions of urea and nitrate for nitrogen acquisition by maize roots.

Even though urea and nitrate are the two major nitrogen (N) forms applied as fertilizers in agriculture and occur concomitantly in soils, the reciprocal influence of these two N sources on the mechanisms of their acquisition are poorly understood. Therefore, molecular and physiological aspects of urea and nitrate uptake were investigated in maize (Zea mays), a crop plant consuming high amounts of N. In roots, urea uptake was stimulated by the presence of urea in the external solution, indicating the presence of an inducible transport system. On the other hand, the presence of nitrate depressed the induction of urea uptake and, at the same time, the induction of nitrate uptake was depressed by the presence of urea. The expression of about 60,000 transcripts of maize in roots was monitored by microarray analyses and the transcriptional patterns of those genes involved in nitrogen acquisition were analyzed by real-time reverse transcription-PCR (RT-PCR). In comparison with the treatment without added N, the exposure of maize roots to urea modulated the expression of only very few genes, such as asparagine synthase. On the other hand, the concomitant presence of urea and nitrate enhanced the overexpression of genes involved in nitrate transport (NRT2) and assimilation (nitrate and nitrite reductase, glutamine synthetase 2), and a specific response of 41 transcripts was determined, including glutamine synthetase 1-5, glutamine oxoglutarate aminotransferase, shikimate kinase and arogenate dehydrogenase. Also based on the real-time RT-PCR analysis, the transcriptional modulation induced by both sources might determine an increase in N metabolism promoting a more efficient assimilation of the N that is taken up.

Phosphorus and iron deficiencies induce a metabolic reprogramming and affect the exudation traits of the woody plant Fragaria×ananassa.

Strawberries are a very popular fruit among berries, for both their commercial and economic importance, but especially for their beneficial effects for human health. However, their bioactive compound content is strictly related to the nutritional status of the plant and might be affected if nutritional disorders (e.g. Fe or P shortage) occur. To overcome nutrient shortages, plants evolved different mechanisms, which often involve the release of root exudates. The biochemical and molecular mechanisms underlying root exudation and its regulation are as yet still poorly known, in particular in woody crop species. The aim of this work was therefore to characterize the pattern of root exudation of strawberry plants grown in either P or Fe deficiency, by investigating metabolomic changes of root tissues and the expression of genes putatively involved in exudate extrusion. Although P and Fe deficiencies differentially affected the total metabolism, some metabolites (e.g. raffinose and galactose) accumulated in roots similarly under both conditions. Moreover, P deficiency specifically affected the content of galactaric acid, malic acid, lysine, proline, and sorbitol-6-phosphate, whereas Fe deficiency specifically affected the content of sucrose, dehydroascorbic acid, galactonate, and ferulic acid. At the same time, the citrate content did not change in roots under both nutrient deficiencies with respect to the control. However, a strong release of citrate was observed, and it increased significantly with time, being +250% and +300% higher in Fe- and P-deficient plants, respectively, compared with the control. Moreover, concomitantly, a significant acidification of the growth medium was observed in both treatments. Gene expression analyses highlighted for the first time that at least two members of the multidrug and toxic compound extrusion (MATE) transporter family and one member of the plasma membrane H(+)-ATPase family are involved in the response to both P and Fe starvation in strawberry plants.

Traceability of different apple varieties by multivariate analysis of isotope ratio mass spectrometry data.

RATIONALE: The awareness of customers of the origin of foods has become an important issue. The growing demand for foods that are healthy, safe and of high quality has increased the need for traceability and clear labelling. Thus, this study investigates the capability of C and N stable isotope ratios to determine the geographical origin of several apple varieties grown in northern Italy. METHODS: Four apple varieties (Cripps Pink, Gala, Golden Delicious, Granny Smith) have been sampled in orchards located in the Districts of Bolzano, Ferrara, Verona and Udine (northern Italy). Carbon (δ(13) C) and nitrogen (δ(15) N) isotope values of the whole apple fruits and three sub-fractions (peel, pulp and seed) have been determined simultaneously by isotope ratio mass spectrometry. RESULTS: The δ(13) C and δ(15) N values of apples and apple sub-fractions, such as peel, seed and pulp, were significantly affected by the geographical origin and the fruit variety. The four varieties could be distinguished to a certain extent only within each district. A 99% correct identification of the samples according to their origin was, however, achieved by cross validation with the 'leave-one-out' method. CONCLUSIONS: This study proves the potential of stable isotopes to discriminate the geographical origin of apples grown in orchards located only a few hundreds of kilometres apart. Stable isotopes were also able to discriminate different apple varieties, although only within small geographical areas.

Iron allocation in leaves of Fe-deficient cucumber plants fed with natural Fe complexes.

Iron (Fe) sources available for plants in the rhizospheric solution are mainly a mixture of complexes between Fe and organic ligands, including phytosiderophores (PS) and water-extractable humic substances (WEHS). In comparison with the other Fe sources, Fe-WEHS are more efficiently used by plants, and experimental evidences show that Fe translocation contributes to this better response. On the other hand, very little is known on the mechanisms involved in Fe allocation in leaves. In this work, physiological and molecular processes involved in Fe distribution in leaves of Fe-deficient Cucumis sativus supplied with Fe-PS or Fe-WEHS up to 5 days were studied combining different techniques, such as radiochemical experiments, synchrotron micro X-ray fluorescence, real-time reverse transcription polymerase chain reaction and in situ hybridization. In Fe-WEHS-fed plants, Fe was rapidly (1 day) allocated into the leaf veins, and after 5 days, Fe was completely transferred into interveinal cells; moreover, the amount of accumulated Fe was much higher than with Fe-PS. This redistribution in Fe-WEHS plants was associated with an upregulation of genes encoding a ferric(III) -chelate reductase (FRO), a Fe(2+) transporter (IRT1) and a natural resistance-associated macrophage protein (NRAMP). The localization of FRO and IRT1 transcripts next to the midveins, beside that of NRAMP in the interveinal area, may suggest a rapid and efficient response induced by the presence of Fe-WEHS in the extra-radical solution for the allocation in leaves of high amounts of Fe. In conclusion, Fe is more efficiently used when chelated to WEHS than PS and seems to involve Fe distribution and gene regulation of Fe acquisition mechanisms operating in leaves.