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.

Integrated multi-omic approach reveals the effect of a Graminaceae-derived biostimulant and its lighter fraction on salt-stressed lettuce plants.

Plant biostimulants are widely applied in agriculture for their ability to improve plant fitness. In the present work, the impact of Graminaceae-derived protein hydrolysate (P) and its lighter molecular fraction F3 (< 1 kDa) on lettuce plants, subjected to either no salt or high salt conditions, was investigated through the combination of metabolomics and transcriptomics. The results showed that both treatments significantly modulated the transcriptome and metabolome of plants under salinity stress, highlighting an induction of the hormonal response. Nevertheless, P and F3 also displayed several peculiarities. F3 specifically modulated the response to ethylene and MAPK signaling pathway, whereas P treatment induced a down-accumulation of secondary metabolites, albeit genes controlling the biosynthesis of osmoprotectants and antioxidants were up-regulated. Moreover, according with the auxin response modulation, P promoted cell wall biogenesis and plasticity in salt-stressed plants. Notably, our data also outlined an epigenetic control of gene expression induced by P treatment. Contrarily, experimental data are just partially in agreement when not stressed plants, treated with P or F3, were considered. Indeed, the reduced accumulation of secondary metabolites and the analyses of hormone pathways modulation would suggest a preferential allocation of resources towards growth, that is not coherent with the down-regulation of the photosynthetic machinery, the CO2 assimilation rate and leaves biomass. In conclusion, our data demonstrate that, although they might activate different mechanisms, both the P and F3 can result in similar benefits, as far as the accumulation of protective osmolytes and the enhanced tolerance to oxidative stress are concerned. Notably, the F3 fraction exhibits slightly greater growth promotion effects under high salt conditions. Most importantly, this research further corroborates that biostimulants' mode of action is dependent on plants' physiological status and their composition, underscoring the importance of investigating the bioactivity of the different molecular components to design tailored applications for the agricultural practice.

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.

Unraveling plant adaptation to single and combined nutrient deficiencies in a dicotyledonous and a monocotyledonous plant species.

Nutrient deficiencies considerably limit agricultural production worldwide. However, while single deficiencies are widely studied, combined deficiencies are poorly addressed. Hence, the aim of this paper was to study single and combined deficiencies of iron (Fe) and phosphorus (P) in barley (Hordeum vulgare) and tomato (Solanum lycopersicum). Plants were grown in hydroponics and root exudation was measured over the growing period. At harvest, root morphology and root and shoot ionome was assessed. Shoot-to-root-ratio decreased in both species and in all nutrient deficiencies, besides in -Fe tomato. Barley root growth was enhanced in plants subjected to double deficiency behaving similarly to -P, while tomato reduced root morphology parameters in all treatments. To cope with the nutrient deficiency barley exuded mostly chelants, while tomato relied on organic acids. Moreover, tomato exhibited a slight exudation increase over time not detected in barley. Overall, in none of the species the double deficiency caused a substantial increase in root exudation. Multivariate statistics emphasized that all the treatments were significantly different from each other in tomato, while in barley only -Fe was statistically different from the other treatments. Our findings highlight that the response of the studied plants in double deficiencies is not additive but plant specific.

Combining micro- and portable-XRF as a tool for fast identification of virus infections in plants: The case study of ASa-Virus in Fraxinus ornus L.

Plant viruses can affect micro- and macro-nutrients homeostasis in woody plants, with fluctuation in the concentration of some elements at the leaf level due to the pathogen activity and/or the plant physiological response to the infection.Leaves of Fraxinus ornus L. (flowering ash) were sampled for three consecutive years in the city of Hamburg (Germany), from both trees showing the typical symptoms of the ash shoestring associated virus infection (ASaV+) and healthy trees (ASaV-). Such leaves were analyzed by µ-XRF, using both laboratory and synchrotron X-ray sources, and large differences between symptomatic and not symptomatic leaves were observed: ASaV+ samples showed uneven element distribution and regions of the lamina with severe depletions of P, S, and Ca. Differently, K appeared more concentrated. Thus, 139 leaflets sampled from various healthy and infected ash trees over the three-year period were analyzed for K and Ca concentration with a portable XRF instrument. We found that the K:Ca concentration ratio was always significantly higher in ASaV+ samples, and this trend was verified for all the samplings over the tree years. We conclude that the K:Ca ratio parameter has potential in the frame of trendsetting diagnostics and could be used, together with visual symptoms, for a rapid, non-destructive, on-site and cheap indirect ASaV detection.

The Application of High-Resolution Melting Analysis to trnL (UAA) Intron Allowed a Qualitative Identification of Apple Juice Adulterations.

Food authenticity plays a pivotal role in the modern age since an increased consumers awareness has led them to pay more attention to food commodities. For this reason, it is important to have reliable and fast techniques able to detect possible adulterations in food, which affect qualitative and economic value. Therefore, the aim of this study was to detect possible adulterations in apple juice from others fruit species (i.e., pear, peach, and kiwi) combining DNA barcoding approach, using trnL (UAA) intron, with high resolution melting analysis (HRMA). A preliminary phylogenetic analysis, using sequences retrieved by the GenBank, confirmed the discriminatory power of trnL (UAA) intron among the four fruit species examined. Moreover, the sequencing of the trnL (UAA) fragments obtained from apple, pear, peach, and kiwi, demonstrated the suitability of an inner shorter sequence, P6 loop, to differentiate the considered species. The HRMA coupled with trnL (UAA) intron allowed discrimination among the four fruits but provided incomplete results for juices. Whereas the HRMA targeting the P6 loop amplicons confirmed the suitability of the technique to qualitatively distinguish fruit juices composed by the combination of apple/pear and apple/peach. However, the impossibility of discriminating apple/kiwi juices from the pure kiwi sample highlighted limitations, most likely related to the DNA extraction process. This hypothesis was further confirmed by analyzing DNA blends obtained by combining nucleic acids extracted from pure matrixes (i.e., apple and kiwi fruits). In this specific case, the application of HRMA allowed both qualitative and quantitative assessment of the samples.

Different vegetal protein hydrolysates distinctively alleviate salinity stress in vegetable crops: A case study on tomato and lettuce.

Plants have evolved diverse plant-species specific tolerance mechanisms to cope with salt stress. However, these adaptive strategies often inefficiently mitigate the stress related to increasing salinity. In this respect, plant-based biostimulants have gained increasing popularity since they can alleviate deleterious effects of salinity. Hence, this study aimed to evaluate the sensitivity of tomato and lettuce plants grown under high salinity and the possible protective effects of four biostimulants based on vegetal protein hydrolysates. Plants were set in a 2 × 5 factorial experimental design completely randomized with two salt conditions, no salt (0 mM) and high salt (120 mM for tomato or 80 mM for lettuce), and five biostimulant treatments (C: Malvaceae-derived, P: Poaceae-derived, D: Legume-derived commercial 'Trainer®', H: Legume-derived commercial 'Vegamin®', and Control: distilled water). Our results showed that both salinity and biostimulant treatments affected the biomass accumulation in the two plant species, albeit to different extents. The salinity stress induced a higher activity of antioxidant enzymes (e.g., catalase, ascorbate peroxidase, guaiacol peroxidase and superoxide dismutase) and the overaccumulation of osmolyte proline in both lettuce and tomato plants. Interestingly, salt-stressed lettuce plants showed a higher accumulation of proline as compared to tomato plants. On the other hand, the treatment with biostimulants in salt-stressed plants caused a differential induction of enzymatic activity depending on the plant and the biostimulant considered. Overall, our results suggest that tomato plants were constitutively more tolerant to salinity than lettuce plants. As a consequence, the effectiveness of biostimulants in alleviating high salt concentrations was more evident in lettuce. Among the four biostimulants tested, P and D showed to be the most promising for the amelioration of salt stress in both the plant species, thereby suggesting their possible application in the agricultural practice.

δ13C as a tool for iron and phosphorus deficiency prediction in crops.

Many studies proposed the use of stable carbon isotope ratio (δ13C) as a predictor of abiotic stresses in plants, considering only drought and nitrogen deficiency without further investigating the impact of other nutrient deficiencies, that is, phosphorus (P) and/or iron (Fe) deficiencies. To fill this knowledge gap, we assessed the δ13C of barley (Hordeum vulgare L.), cucumber (Cucumis sativus L.), maize (Zea mays L.), and tomato (Solanum lycopersicon L.) plants suffering from P, Fe, and combined P/Fe deficiencies during a two-week period using an isotope-ratio mass spectrometer. Simultaneously, plant physiological status was monitored with an infra-red gas analyzer. Results show clear contrasting time-, treatment-, species-, and tissue-specific variations. Furthermore, physiological parameters showed limited correlation with δ13C shifts, highlighting that the plants' δ13C, does not depend solely on photosynthetic carbon isotope fractionation/discrimination (Δ). Hence, the use of δ13C as a predictor is highly discouraged due to its inability to detect and discern different nutrient stresses, especially when combined stresses are present.

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.

Fire-induced effects on the bioavailability of potentially toxic elements in a polluted agricultural soil: implications for Cr uptake by durum wheat plants.

Fire events can modify the distribution and speciation of potentially toxic elements (PTEs) in soil, especially if they are associated to organic matter (OM). In fact, OM can undergo substantial structural modifications at high temperatures, up to the complete mineralization. The present study aims to investigate the changes of PTEs' bioavailability to durum wheat (Triticum durum Desf.) plants after simulating fire events (up to 300 °C and 500 °C) in an agricultural soil polluted by Cr, Zn, Cu, and Pb. The PTEs' uptake and allocation in plant tissues were assessed using the RHIZOtest system. After the fire simulations, no evident risk of accumulation and translocation in plants was observed for Zn, Pb, and Cu. Conversely, a high accumulation in roots and a significant translocation to shoots were observed for Cr, which reached concentrations of 829 mg kg-1 in roots and 52 mg kg-1 in shoots at 500 °C. Additional experimental evidence suggested that Cr was taken up by plants grown on heated soils as Cr(VI). Once acquired by roots, only a small part of Cr (up to 6%) was translocated to shoots where it was likely present as mobile forms, as evidenced by micro X-ray fluorescence (µ-XRF) analyses. Overall, the results obtained provide evidence that the high temperatures occurring during fire events can increase the mobility and bioavailability of certain PTEs transforming apparently safe environments into potentially dangerous sources of pollution. These processes can ultimately affect the human health through the food chain transfer of PTEs or their migration into surface water and groundwater.

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.

TXRF spectral information enhanced by multivariate analysis: A new strategy for food fingerprint.

The increased costumers' request of safe and high-quality food products makes food traceability a priority for frauds identification and quality certification. Elemental profiling is one of the strategies used for food traceability, and TXRF spectroscopy is widely used in food analysis even if its potentialities have not been fully investigated. In this work, a new method for food traceability using directly TXRF spectra coupled with multivariate analyses, was tested. Twenty-four different beans' genotypes (Phaseolus vulgaris L.) grown onto two different sites have been studied. After the development of the method for beans' analysis, TXRF spectra were collected and processed with PCA combined with SNV and GLSW filter obtaining a perfect clustering of the seeds according to their geographical origin. Finally, using PLS-DA, beans were correctly classified demonstrating that TXRF spectra can be successfully used as fingerprint for food/seed traceability and that elemental quantification procedure is not necessary to this aim.

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.

Plant species-specific impact of polyethylene microspheres on seedling growth and the metabolome.

Microplastics (MPs) are ubiquitous contaminants. In recent decades, the hazardous impacts of MPs on the environment have raised significant concern. However, little attention has been focused on the interaction between MPs and plants in terrestrial agroecosystems. This study aims to investigate the effects of polyethylene microspheres (PE-MS) on the germination, morphology, and metabolism of barley (Hordeum vulgare L.), cucumber (Cucumis sativus L.), and tomato (Solanum lycopersicum L.). Specifically, seeds were soaked in PE-MS solutions at three concentrations (10, 100, and 1000 mg L-1), while control seeds were treated with distilled water. After five days, the morphological parameters of barley (i.e., shoot and root biomass, length, and average diameter) were significantly affected by PE-MS treatment, even at the lowest concentration, without a dose dependency. On the other hand, the effect of PE-MS on the morphological parameters of cucumber and tomato was evident only at the highest concentration (1000 mg L-1). PE-MS also induced metabolomic reprogramming of shoots and roots in all three plant species. There was a downregulation of fatty acids and secondary metabolites (except in tomato shoots). In addition, the response of amino acids and hormones was highly heterogeneous among species and plant parts. In particular, the response of metabolites changed within species among different plant parts. In conclusion, we found a strong influence of MS-PE on the metabolic profile of the three plant species and a positive priming of seedling growth, especially in barley, where all the morphological parameters considered were significantly improved. Further investigations are needed to fully understand the mechanisms underlying MP-plant interactions, especially in the long term.

Copper toxicity compromises root acquisition of nitrate in the high affinity range.

The application of copper (Cu)-based fungicides for crop protection plans has led to a high accumulation of Cu in soils, especially in vineyards. Copper is indeed an essential micronutrient for plants, but relatively high concentrations in soil or other growth substrates may cause toxicity phenomena, such as alteration of the plant's growth and disturbance in the acquisition of mineral nutrients. This last aspect might be particularly relevant in the case of nitrate ( NO 3 - ) , whose acquisition in plants is finely regulated through the transcriptional regulation of NO 3 - transporters and plasma membrane H+-ATPase in response to the available concentration of the nutrient. In this study, cucumber plants were grown hydroponically and exposed to increasing concentrations of Cu (i.e., 0.2, 5, 20, 30, and 50 µM) to investigate their ability to respond to and acquire NO 3 - . To this end, the kinetics of substrate uptake and the transcriptional modulation of the molecular entities involved in the process have been assessed. Results showed that the inducibility of the high-affinity transport system was significantly affected by increasing Cu concentrations; at Cu levels higher than 20 µM, plants demonstrated either strongly reduced or abolished NO 3 - uptake activity. Nevertheless, the transcriptional modulation of both the nitrate transporter CsNRT2.1 and the accessory protein CsNRT3.1 was not coherent with the hindered NO 3 - uptake activity. On the contrary, CsHA2 was downregulated, thus suggesting that a possible impairment in the generation of the proton gradient across the root PM could be the cause of the abolishment of NO 3 - uptake.

Physiological and Molecular Investigation of Urea Uptake Dynamics in Cucumis sativus L. Plants Fertilized With Urea-Doped Amorphous Calcium Phosphate Nanoparticles.

At present, the quest for innovative and sustainable fertilization approaches aiming to improve agricultural productivity represents one of the major challenges for research. In this context, nanoparticle-based fertilizers can indeed offer an interesting alternative with respect to traditional bulk fertilizers. Several pieces of evidence have already addressed the effectiveness of amorphous calcium phosphate-based nanoparticles as carriers for macronutrients, such as nitrogen (N), demonstrating increase in crop productivity and improvement in quality. Nevertheless, despite N being a fundamental nutrient for crop growth and productivity, very little research has been carried out to understand the physiological and molecular mechanisms underpinning N-based fertilizers supplied to plants via nanocarriers. For these reasons, this study aimed to investigate the responses of Cucumis sativus L. to amorphous calcium phosphate nanoparticles doped with urea (U-ACP). Urea uptake dynamics at root level have been investigated by monitoring both the urea acquisition rates and the modulation of urea transporter CsDUR3, whereas growth parameters, the accumulation of N in both root and shoots, and the general ionomic profile of both tissues have been determined to assess the potentiality of U-ACP as innovative fertilizers. The slow release of urea from nanoparticles and/or their chemical composition contributed to the upregulation of the urea uptake system for a longer period (up to 24 h after treatment) as compared to plants treated with bulk urea. This prolonged activation was mirrored by a higher accumulation of N in nanoparticle-treated plants (approximately threefold increase in the shoot of NP-treated plants compared to controls), even when the concentration of urea conveyed through nanoparticles was halved. In addition, besides impacting N nutrition, U-ACP also enhanced Ca and P concentration in cucumber tissues, thus having possible effects on plant growth and yield, and on the nutritional value of agricultural products.

Identification of an Isoflavonoid Transporter Required for the Nodule Establishment of the Rhizobium-Fabaceae Symbiotic Interaction.

Nitrogen (N) as well as Phosphorus (P) are key nutrients determining crop productivity. Legumes have developed strategies to overcome nutrient limitation by, for example, forming a symbiotic relationship with N-fixing rhizobia and the release of P-mobilizing exudates and are thus able to grow without supply of N or P fertilizers. The legume-rhizobial symbiosis starts with root release of isoflavonoids that act as signaling molecules perceived by compatible bacteria. Subsequently, bacteria release nod factors, which induce signaling cascades allowing the formation of functional N-fixing nodules. We report here the identification and functional characterization of a plasma membrane-localized MATE-type transporter (LaMATE2) involved in the release of genistein from white lupin roots. The LaMATE2 expression in the root is upregulated under N deficiency as well as low phosphate availability, two nutritional deficiencies that induce the release of this isoflavonoid. LaMATE2 silencing reduced genistein efflux and even more the formation of symbiotic nodules, supporting the crucial role of LaMATE2 in isoflavonoid release and nodulation. Furthermore, silencing of LaMATE2 limited the P-solubilization activity of lupin root exudates. Transport assays in yeast vesicles demonstrated that LaMATE2 acts as a proton-driven isoflavonoid transporter.

The hidden effects of agrochemicals on plant metabolism and root-associated microorganisms.

Agrochemicals are commonly used in agriculture to protect crops and ensure yields. Several of them are mobile within the plant and, being perceived as xenobiotics regardless of their protective/curative roles, they induce a reprogramming of secondary metabolism linked to the detoxification processes even in the absence of phenotype symptoms. Moreover, it is well documented that plants are able to shape the microbial population at the rhizosphere and to significantly affect the processes occurring therein thanks to the root exudation of different metabolites. Here we show that plant metabolic response to foliarly-applied pesticides is much broader than what previously thought and includes diverse and compound-specific hidden processes. Among others, stress-related metabolism and phytohormones profile underwent a considerable reorganization. Moreover, a distinctive microbial rearrangement of the rhizosphere was recorded following foliar application of pesticides. Such effects have unavoidably energetic and metabolic costs for the plant paving the way to both positive and negative aspects. The understanding of these effects is crucial for an increasingly sustainable use of pesticides in agriculture.

Can Inoculation With the Bacterial Biostimulant Enterobacter sp. Strain 15S Be an Approach for the Smarter P Fertilization of Maize and Cucumber Plants?

Phosphorus (P) is an essential nutrient for plants. The use of plant growth-promoting bacteria (PGPB) may also improve plant development and enhance nutrient availability, thus providing a promising alternative or supplement to chemical fertilizers. This study aimed to evaluate the effectiveness of Enterobacter sp. strain 15S in improving the growth and P acquisition of maize (monocot) and cucumber (dicot) plants under P-deficient hydroponic conditions, either by itself or by solubilizing an external source of inorganic phosphate (Pi) [Ca3(PO4)2]. The inoculation with Enterobacter 15S elicited different effects on the root architecture and biomass of cucumber and maize depending on the P supply. Under sufficient P, the bacterium induced a positive effect on the whole root system architecture of both plants. However, under P deficiency, the bacterium in combination with Ca3(PO4)2 induced a more remarkable effect on cucumber, while the bacterium alone was better in improving the root system of maize compared to non-inoculated plants. In P-deficient plants, bacterial inoculation also led to a chlorophyll content [soil-plant analysis development (SPAD) index] like that in P-sufficient plants (p < 0.05). Regarding P nutrition, the ionomic analysis indicated that inoculation with Enterobacter 15S increased the allocation of P in roots (+31%) and shoots (+53%) of cucumber plants grown in a P-free nutrient solution (NS) supplemented with the external insoluble phosphate, whereas maize plants inoculated with the bacterium alone showed a higher content of P only in roots (36%) but not in shoots. Furthermore, in P-deficient cucumber plants, all Pi transporter genes (CsPT1.3, CsPT1.4, CsPT1.9, and Cucsa383630.1) were upregulated by the bacterium inoculation, whereas, in P-deficient maize plants, the expression of ZmPT1 and ZmPT5 was downregulated by the bacterial inoculation. Taken together, these results suggest that, in its interaction with P-deficient cucumber plants, Enterobacter strain 15S might have solubilized the Ca3(PO4)2 to help the plants overcome P deficiency, while the association of maize plants with the bacterium might have triggered a different mechanism affecting plant metabolism. Thus, the mechanisms by which Enterobacter 15S improves plant growth and P nutrition are dependent on crop and nutrient status.

Disentangling the Possible Drivers of Indri indri Microbiome: A Threatened Lemur Species of Madagascar.

Research on the gut microbiome may help with increasing our understanding of primate health with species' ecology, evolution, and behavior. In particular, microbiome-related information has the potential to clarify ecology issues, providing knowledge in support of wild primates conservation and their associated habitats. Indri (Indri indri) is the largest extant living lemur of Madagascar. This species is classified as "critically endangered" by the IUCN Red List of Threatened Species, representing one of the world's 25 most endangered primates. Indris diet is mainly folivorous, but these primates frequently and voluntarily engage in geophagy. Indris have never been successfully bred under human care, suggesting that some behavioral and/or ecological factors are still not considered from the ex situ conservation protocols. Here, we explored gut microbiome composition of 18 indris belonging to 5 different family groups. The most represented phyla were Proteobacteria 40.1 ± 9.5%, Bacteroidetes 28.7 ± 2.8%, Synergistetes 16.7 ± 4.5%, and Firmicutes 11.1 ± 1.9%. Further, our results revealed that bacterial alpha and beta diversity were influenced by indri family group and sex. In addition, we investigated the chemical composition of geophagic soil to explore the possible ecological value of soil as a nutrient supply. The quite acidic pH and high levels of secondary oxide-hydroxides of the soils could play a role in the folivorous diet's gut detoxification activity. In addition, the high contents of iron and manganese found the soils could act as micronutrients in the indris' diet. Nevertheless, the concentration of a few elements (i.e., calcium, sulfur, boron, nickel, sodium, and chromium) was higher in non-geophagic than in geophagic soils. In conclusion, the data presented herein provide a baseline for outlining some possible drivers responsible for the gut microbiome diversity in indris, thus laying the foundations for developing further strategies involved in indris' conservation.