Biochemical and pharmaceutical traits of Marrubium vulgare L. plants treated with plant growth-promoting bacteria and elevated CO2.

The present research aimed to understand the influence of plant growth-promoting bacteria (PGPB) on various biochemical, nutritional, and pharmaceutical characteristics of Marrubium vulgare plants grown under elevated carbon dioxide (eCO2). To achieve this objective, a pot experiment was carried out, consisting of two treatments, namely: (i) biofertilization (Bf) by a PGPB strain (Micromonospora sp.) and (ii) two different air CO2 levels, including ambient CO2 (aCO2) and eCO2 concentrations (410 and 710 µmol CO2 mol-1, respectively). The improvement in the photosynthesis rate of eCO2 and Bf-treated plants can explain the increase in the production of carbohydrate. This is evidenced by a substantial rise, reaching up to + 75% and 25% in the total sugar and starch content in plants subjected to eCO2 treatment, respectively. Additionally, eCO2-treated plants exhibited a remarkable 102% increase in soluble sugar synthesis, while plants subjected to Bf treatment showed a notable increase of 66%. Such modifications could be the main factor affecting plants carbon and nitrogen metabolism. Although the level of certain amino acids (such as glycine, tyrosine, and phenylalanine) in plants exhibited significant increases in response to eCO2 and Bf, the levels of other amino acids demonstrated enhancements in plants grown under eCO2 (e.g., histidine) or under treatments containing Bf (e.g., alanine and ornithine). Improvements in primary metabolites led to more benefits in plants treated with Bf and CO2 by boosting secondary metabolites accumulation, including phenolics (+ 27-100%), flavonoids (+ 30-92%), and essential oils (up to + 296%), as well as improved antioxidant capacity (FRAP). This remarkable effectiveness was evident in the significant increase in the biomass production, highlighting the synergistic impact of the treatments. Therefore, the interaction of Bf and eCO2 not only induced plant biomass accumulation but also improved the nutritional and pharmaceutical value of M. vulgare plants.

Chromium(VI) Toxicity and Active Tolerance Mechanisms of Wheat Plant Treated with Plant Growth-Promoting Actinobacteria and Olive Solid Waste.

The present study aimed to assess the potential of plant growth-promoting Actinobacteria and olive solid waste (OSW) in ameliorating some biochemical and molecular parameters of wheat (Triticum aestivum) plants under the toxicity of high chromium levels in the soil. With this aim, a pot experiment was conducted, where the wheat plants were treated with a consortium of four Actinobacterium sp. (Bf treatment) and/or OSW (4% w/w) under two levels of nonstress and chromium stress [400 mg Cr(VI) per kg of soil] to estimate the photosynthetic traits, antioxidant protection machine, and detoxification activity. Both Bf and OSW treatments improved the levels of chlorophyll a (+47-98%), carotenoid (+324-566%), stomatal conductance (+17-18%), chlorophyll fluorescence (+12-28%), and photorespiratory metabolism (including +44-72% in glycolate oxidase activity, +6-72% in hydroxypyruvate reductase activity, and +5-44% in a glycine to serine ratio) in leaves of stressed plants as compared to those in the stressed control, which resulted in higher photosynthesis capacity (+18-40%) in chromium-stressed plants. These results were associated with an enhancement in the content of antioxidant metabolites (+10-117%), of direct reactive oxygen species-detoxifying enzymes (+49-94%), and of enzymatic (+40-261%) and nonenzymatic (+17-175%) components of the ascorbate-glutathione cycle in Bf- and OSW-treated plants under stress. Moreover, increments in the content of phytochelatins (+38-74%) and metallothioneins (+29-41%), as markers of detoxification activity, were recorded in the plants treated with Bf and OSW under chromium toxicity. In conclusion, this study revealed that the application of beneficial Actinobacteria and OSW as biofertilization/supplementation could represent a worthwhile consequence in improving dry matter production and enhancing plant tolerance and adaptability to chromium toxicity.

Synergistic Impacts of Plant-Growth-Promoting Bacteria and Selenium Nanoparticles on Improving the Nutritional Value and Biological Activities of Three Cultivars of Brassica Sprouts.

Due to the growing world population and increasing environmental stress, improving the production, nutritional quality, and pharmaceutical applications of plants have become an urgent need. Therefore, current research was designed to investigate the impact of seed priming using plant-growth-promoting bacteria (PGPB) along with selenium nanoparticles (SeNPs) treatment on chemical and biological properties of three Brassica oleracea cultivars [Southern star (VA1), Prominence (VA2), Monotop (VA3)]. With this aim, one out of five morphologically different strains of bacteria, namely, JM18, which was further identified via 16S rRNA gene sequencing as a Nocardiopsis species with strong plant-growth-promoting traits, isolated from soil, was used. To explore the growth-promoting potential of Nocardiopsis species, seeds of three varieties of B. oleracea were primed with JM18 individually or in combination with SeNP treatment. Seed treatments increased sprout growth (fresh and dry weights) and glucosinolate accumulation. The activity of myrosinase was significantly increased through brassica sprouts and consequently enhanced the amino-acid-derived glucosinolate induction. Notably, a reduction in effective sulforaphane nitrile was detected, being positively correlated with a decrease in epithiospecifier protein (EP). Consequently, the antioxidant activities of VA2 and VA3, determined by the ferric reducing antioxidant power (FRAP) assay, were increased by 74 and 79%, respectively. Additionally, the antibacterial activities of JM18-treated cultivars were improved. However, a decrease was observed in SeNP- and JM18 + SeNP-treated VA2 and VA3 against Serratia marcescens and Candida glabrata and VA1 against S. marcescens. In conclusion, seed priming with the JM18 extract is a promising method to enhance the health-promoting activities of B. oleracea sprouts.

Earthworm-Driven Changes in Soil Chemico-Physical Properties, Soil Bacterial Microbiota, Tree/Tea Litter Decomposition, and Plant Growth in a Mesocosm Experiment with Two Plant Species.

Earthworms and soil microorganisms contribute to soil health, quality, and fertility, but their importance in agricultural soils is often underestimated. This study aims at examining whether and to what extent the presence of earthworms (Eisenia sp.) affected the (a) soil bacterial community composition, (b) litter decomposition, and (c) plant growth (Brassica oleracea L., broccoli; Vicia faba L., faba bean). We performed a mesocosm experiment in which plants were grown outdoors for four months with or without earthworms. Soil bacterial community structure was evaluated by a 16S rRNA-based metabarcoding approach. Litter decomposition rates were determined by using the tea bag index (TBI) and litter bags (olive residues). Earthworm numbers almost doubled throughout the experimental period. Independently of the plant species, earthworm presence had a significant impact on the structure of soil bacterial community, in terms of enhanced α- and ß-diversity (especially that of Proteobacteria, Bacteroidota, Myxococcota, and Verrucomicrobia) and increased 16S rRNA gene abundance (+89% in broccoli and +223% in faba bean). Microbial decomposition (TBI) was enhanced in the treatments with earthworms, and showed a significantly higher decomposition rate constant (kTBI) and a lower stabilization factor (STBI), whereas decomposition in the litter bags (dlitter) increased by about 6% in broccoli and 5% in faba bean. Earthworms significantly enhanced root growth (in terms of total length and fresh weight) of both plant species. Our results show the strong influence of earthworms and crop identity in shaping soil chemico-physical properties, soil bacterial community, litter decomposition and plant growth. These findings could be used for developing nature-based solutions that ensure the long-term biological sustainability of soil agro- and natural ecosystems.

Understanding the Active Mechanisms of Plant (Sesuvium portulacastrum L.) against Heavy Metal Toxicity.

Through metabolic analysis, the present research seeks to reveal the defense mechanisms activated by a heavy metals-resistant plant, Sesuvium portulacastrum L. In this regard, shifting metabolisms in this plant were investigated in different heavy metals-contaminated experimental sites, which were 50, 100, 500, 1000, and 5000 m away from a man-fabricated sewage dumping lake, with a wide range of pollutant concentrations. Heavy metals contaminations in contaminated soil and their impact on mineral composition and microbial population were also investigated. The significant findings to emerge from this research were the modifications of nitrogen and carbon metabolisms in plant tissues to cope with heavy metal toxicity. Increased plant amylase enzymes activity in contaminated soils increased starch degradation to soluble sugars as a mechanism to mitigate stress impact. Furthermore, increased activity of sucrose phosphate synthase in contaminated plants led to more accumulation of sucrose. Moreover, no change in the content of sucrose hydrolyzing enzymes (vacuolar invertase and cytosolic invertase) in the contaminated sites can suggest the translocation of sucrose from shoot to root under stress. Similarly, although this study demonstrated a high level of malate in plants exposed to stress, caution must be applied in suggesting a strong link between organic acids and the activation of defense mechanisms in plants, since other key organic acids were not affected by stress. Therefore, activation of other defense mechanisms, especially antioxidant defense molecules including alpha and beta tocopherols, showed a greater role in protecting plants from heavy metals stress. Moreover, the increment in the content of some amino acids (e.g., glycine, alanine, glutamate, arginine, and ornithine) in plants under metal toxicity can be attributed to a high level of stress tolerance. Moreover, strategies in the excitation of the synthesis of the unsaturated fatty acids (oleic and palmitoleic) were involved in enhancing stress tolerance, which was unexpectedly associated with an increase in the accumulation of palmitic and stearic (saturated fatty acids). Taken together, it can be concluded that these multiple mechanisms were involved in the response to stress which may be cooperative and complementary with each other in inducing resistance to the plants.

Biofertilisation with a consortium of growth-promoting bacterial strains improves the nutritional status of wheat grain under control, drought, and salinity stress conditions.

We investigated the effect of plant growth-promoting bacterial strains (PGPB) as biofertilisers on the grain metabolic composition of durum wheat (Triticum durum Desf.). To this aim, we conducted a greenhouse experiment where we grew durum wheat plants supplied with a biofertiliser consortium of four PGPB and/or chemical fertiliser (containing nitrogen, phosphorus, potassium, and zinc), under non-stress, drought (at 40% field capacity), or salinity (150 mM NaCl) conditions. Nutrient accumulations in the grain were increased in plants treated with the biofertiliser consortium, alone or with a half dose of chemical fertilisers, compared to those in no fertilisation treatment. A clear benefit of biofertiliser application in the improvement of protein, soluble sugar, starch, and lipid contents in the grains was observed in comparison with untreated controls, especially under stress conditions. The most striking observation was the absence of significant differences between biofertiliser and chemical fertiliser treatments for most parameters. Moreover, the overall response to the biofertiliser consortium was accompanied by greater changes in amino acids, organic acids, and fatty acid profiles. In conclusion, PGPB improved the metabolic and nutrient status of durum wheat grains to a similar extent as chemical fertilisers, particularly under stress conditions, demonstrating the value of PGPB as a sustainable fertilisation treatment.

Shifts in the Rhizosphere and Endosphere Colonizing Bacterial Communities Under Drought and Salinity Stress as Affected by a Biofertilizer Consortium.

The present research asks how plant growth-promoting bacterial (PGPB) inoculants and chemical fertilizers change rhizosphere and root endophytic bacterial communities in durum wheat, and its dependence on environmental stress. A greenhouse experiment was carried out under drought (at 40% field capacity), or salinity (150 mM NaCl) conditions to investigate the effects of a chemical fertilizer (containing nitrogen, phosphorus, potassium and zinc) or a biofertilizer (a bacterial consortium of four PGPBs). High-throughput amplicon sequencing of the 16S rRNA of the rhizosphere, non-sterilized, or surface-sterilized roots, showed shifts in bacterial communities in response to stress treatments, which were greater for salinity than for drought and tended to show increased oligotrophs relative abundances compared to non-stress controls. The results also showed that Proteobacteria, Acidobacteria, Bacteroidetes, Gemmatimonadetes, Thaumarchaeota, Firmicutes, and Verrucomicrobia had a higher relative abundance in the rhizosphere, while Actinobacteria were more abundant on roots, while Candidatus_Saccharibacteria and Planctomycetes inside roots. The results indicated that the root endophytic bacterial communities were more affected by (bio-) fertilization treatments than those in the rhizosphere, particularly as affected by PGPB inoculation. This greater susceptibility of endophytes to (bio-) fertilizers was associated with increased abundance of the 16S rRNA and acdS genes in plant roots, especially under stress conditions. These changes in root endophytes, which coincided with an improvement in grain yield and photosynthetic capacity of plants, may be considered as one of the mechanisms by which PGPB affect plants.

Combined Effect of Laboratory-Simulated Fire and Chromium Pollution on Microbial Communities in an Agricultural Soil.

Fire events in agricultural soils can modify not only soil properties but also the structure of soil microbial communities, especially in soils containing high concentrations of potentially toxic elements (PTEs). The recolonization of burned soils can in fact favor the proliferation of certain microorganisms, more adaptable to post-fire soil conditions and higher PTE availability, over others. In this study, we simulated with laboratory experiments the microbial recolonization of an agricultural soil containing high Cr concentrations after heating at 500 °C for 30 min, to mimic the burning of crop residues. Changes in soil properties and Cr speciation were assessed, as well as soil microbial structure by means of 16S rRNA gene sequencing. Both altered soil conditions and increased Cr availability, especially Cr(VI), appeared to be responsible for the reduction in species diversity in heated soils and the proliferation of Firmicutes. Indeed, already after 3 days from the heat treatment, Firmicutes increased from 14% to 60% relative abundance. In particular, Paenibacillus was the most abundant genus identified after the simulation, with an average relative abundance of 40%. These bacteria are known to be good fire-responders and Cr-tolerant. These results could be useful to identify bacterial strains to be used as bioindicators of altered environments and for the recovery of fire-impacted polluted sites.

Fire effects on the distribution and bioavailability of potentially toxic elements (PTEs) in agricultural soils.

In the last years, uncontrolled fires are frequently occurring in forest and agricultural areas as an indirect effect of the rising aridity and global warming or caused by intentional illegal burnings. In addition, controlled burning is still largely used by farmers as an agricultural practice in many parts of the world. During fire events, soil can reach very high temperatures at the soil surface, causing dramatic changes of soil properties and elements biogeochemistry. Among soil elements, also potentially toxic elements (PTEs) can be affected by fires, becoming more or less mobile and bioavailable, depending on fire severity and soil characteristics. Such transformations could be particularly relevant in agricultural soils used for crop productions since fire events could modify PTEs speciation and uptake by plants and associated (micro)organisms thus endangering the whole food-chain. In this review, after describing the effects of fire on soil minerals and organic matter, the impact of fires on PTEs distribution and speciation in soils is presented, as well as their influence on soil microorganisms and plants uptake. The most common experimental methods used to simulate fires at the laboratory and field scale are briefly illustrated, and finally the impact that traditional and innovative agricultural practices can have on PTEs availability in burned agricultural soils is discussed in a future research perspective.

Isolation of Bacteria with Potential Plant-Promoting Traits and Optimization of Their Growth Conditions.

This research aimed at investigating the isolation and identification of bacterial strains with biological nitrogen-fixing capability and phosphate, potassium, and zinc solubilization activities from a durum wheat field under two different tillage practices including 10 years of conventional tillage (CT) and no-tillage (NT) practices. Attempts were also extended to estimate their relative abundances in the soil as well as to develop accurate mathematical models in determining the effect of different temperatures, NaCl concentrations and pH on the growth, and activity of selected isolates. Twelve effective bacterial strains, including Pseudomonas, Acinetobacter, and Comamonas genera, were identified with a great potential to solubilize the insoluble forms of phosphate (from 11.1 to 115.5 mg l-1 at pH 8), potassium (from 32.2 to 35.6 mg l-1 at pH 7), and zinc (from 1.11 to 389.90 mg l-1 at pH 9) as well as to fix N2 gas (from 19.9 to 25.2 mg l-1). To our knowledge, this is the first report of the ability of Comamonas testosteroni and Acinetobacter pittii to fix nitrogen and to solubilize insoluble potassium compound, respectively. Three families, Moraxellaceae, Pseudomonadaceae, and Comamonadaceae, showed a higher percentage of abundance in the NT samples as compared to the CT, but only significant difference was observed in the relative abundance of Pseudomonadaceae (P < 0.01). These strains could be definitively recommended as inoculants to promote plant growth in the wide ranges of pH, salinity levels (with maximum growth and complete inhibition of growth from 0.67-0.92% to 3.5-9.3% NaCl, respectively), and temperatures (2.1-45.1 °C).

Comparative Genomics and In Vitro Plant Growth Promotion and Biocontrol Traits of Lactic Acid Bacteria from the Wheat Rhizosphere.

This study aimed to isolate lactic acid bacteria (LAB) from wheat rhizosphere, to characterize their in vitro plant growth promoting activities and to differentiate plant-associated LAB from those associated with foods or human disease through comparative genomic analysis. Lactococcus lactis subsp. lactis and Enterococcus faecium were isolated using de Man-Rogosa-Sharpe (MRS) and Glucose Yeast Peptone (GYP) as enrichment culture media. Comparative genomic analyses showed that plant-associated LAB strains were enriched in genes coding for bacteriocin production when compared to strains from other ecosystems. Isolates of L. lactis and E. faecium did not produce physiologically relevant concentrations of the phyto-hormone indolacetic acid. All isolates solubilized high amount of phosphate and 12 of 16 strains solubilized potassium. E. faecium LB5, L. lactis LB6, LB7, and LB9 inhibited the plant pathogenic Fusarium graminearum to the same extent as two strains of Bacillus sp. However, the antifungal activity of the abovementioned LAB strains depended on the medium of cultivation and a low pH while antifungal activity of Bacillus spp. was independent of the growth medium and likely relates to antifungal lipopeptides. This study showed the potential of rhizospheric LAB for future application as biofertilizers in agriculture.

Evaluation of the possible persistence of potential human pathogenic bacteria in olive orchards irrigated with treated urban wastewater.

Under suitable conditions, low-quality, treated urban wastewater (TWW) is an additional water resource for irrigation in water-scarce environments but its use in agriculture requires a careful monitoring of a range of hygiene parameters, including human pathogenic bacteria (HPB). DNA-based microbiological analyses on soil, xylem sap, and leaves surface (phyllosphere) were carried out in an olive (Olea europaea L.) grove located in Southern Italy (Basilicata region). The experimental grove has been managed in two plots for 18 years. The experimental plot (WWtr) was drip irrigated daily with TWW (2800 m3 ha-1 year-1), while the control plot (RFtr) was rainfed. The results of the 16S-rRNA-based metagenomic analysis demonstrated that the phyllosphere had the lowest number of potential HPB (6), compared to soil (22) and xylem (26) compartments. Gammaproteobacteria, including potential HPB, like Pseudomonas and Acinetobacter spp., were significantly higher in WWtr soil and xylem sap, compared to RFtr. A similar trend was observed for Burkholderia spp. (Betaproteobacteria) and Mycobacterium spp. (Actinobacteria). The Firmicutes Enterococcus, Staphylococcus and Streptococcus spp. were more abundant in WWtr xylem sap. The pathogenic Clostridium perfringens was found higher on WWtr leaves (relative abundance 7.17 in WWtr and 1.33 in RFtr) and Enterococcus faecalis in WWtr xylem sap (93.22 in WWtr and 7.08 in RFtr). On the basis of the results obtained, the irrigation with TWW can be considered a realistic and safe agronomic practice in Mediterranean orchards, and an opportunity for farmers and consumers.

Solubilization of insoluble zinc compounds by zinc solubilizing bacteria (ZSB) and optimization of their growth conditions.

In this research, the optimum growth conditions for two zinc solubilizing bacteria (ZSB) have been studied for their potential application as bioinoculants to overcome Zn unavailability in soils. For this purpose, a laboratory-scale experiment was carried out to evaluate the zinc solubilizing ability of 80 plant growth promoting bacteria (PGPB) strains isolated from the rhizosphere of barley and tomato plants. To select effective ZSB, isolates were evaluated on Tris-mineral medium supplemented separately with zinc oxide, zinc carbonate, and zinc phosphate at a concentration of 0.1%. Two strains (Agrobacterium tumefaciens and Rhizobium sp.) were selected, based on a clear halo zone around their colonies in the solid medium supplemented with zinc oxide after 10 days of incubation at 29 °C. Results of solubilization at different pH values showed that these strains had solubilization activity in the range of pH 8-10 while no solubilization was observed at pH 6 and 7. The maximum Zn solubilization values were noted at pH 9: 51.4 mg L-1 (Agrobacterium tumefaciens) and 72.1 mg L-1 (Rhizobium sp). According to findings, bacterial growth was affected by different NaCl concentrations under in vitro condition. The salt concentration required for 50% inhibition of absorbance was 2.11 and 2.27% NaCl for Agrobacterium tumefaciens and Rhizobium sp., respectively. The maximum bacterial growth was observed at about 0.8% NaCl concentration.

Transcriptional reprogramming and phenotypic switching associated with the adaptation of Lactobacillus plantarum C2 to plant niches.

Lactobacillus plantarum has been isolated from a large variety of ecological niches, thus highlighting its remarkable environmental adaptability as a generalist. Plant fermentation conditions markedly affect the functional features of L. plantarum strains. We investigated the plant niche-specific traits of L. plantarum through whole-transcriptome and phenotypic microarray profiles. Carrot (CJ) and pineapple (PJ) juices were chosen as model systems, and MRS broth was used as a control. A set of 3,122 genes was expressed, and 21 to 31% of genes were differentially expressed depending on the plant niche and cell physiological state. L. plantarum C2 seemed to specifically respond to plant media conditions. When L. plantarum was cultured in CJ, useful pathways were activated, which were aimed to sense the environment, save energy and adopt alternative routes for NAD(+) regeneration. In PJ the acidic environment caused a transcriptional switching, which was network-linked to an acid tolerance response involving carbohydrate flow, amino acid and protein metabolism, pH homeostasis and membrane fluidity. The most prominent phenotypic dissimilarities observed in cells grown in CJ and PJ were related to carbon and nitrogen metabolism, respectively. Summarising, a snapshot of a carrot and pineapple sensing and adaptive regulation model for L. plantarum C2 was proposed.

Profile of microbial communities on carbonate stones of the medieval church of San Leonardo di Siponto (Italy) by Illumina-based deep sequencing.

Comprehensive studies of the biodiversity of the microbial epilithic community on monuments may provide critical insights for clarifying factors involved in the colonization processes. We carried out a high-throughput investigation of the communities colonizing the medieval church of San Leonardo di Siponto (Italy) by Illumina-based deep sequencing. The metagenomic analysis of sequences revealed the presence of Archaea, Bacteria, and Eukarya. Bacteria were Actinobacteria, Proteobacteria, Bacteroidetes, Cyanobacteria, Chloroflexi, Firmicutes and Candidatus Saccharibacteria. The predominant phylum was Actinobacteria, with the orders Actynomycetales and Rubrobacteriales, represented by the genera Pseudokineococcus, Sporichthya, Blastococcus, Arthrobacter, Geodermatophilus, Friedmanniella, Modestobacter, and Rubrobacter, respectively. Cyanobacteria sequences showing strong similarity with an uncultured bacterium sequence were identified. The presence of the green algae Oocystaceae and Trebuxiaceae was revealed. The microbial diversity was explored at qualitative and quantitative levels, evaluating the richness (the number of operational taxonomic units (OTUs)) and the abundance of reads associated with each OTU. The rarefaction curves approached saturation, suggesting that the majority of OTUs were recovered. The results highlighted a structured community, showing low diversity, made up of extremophile organisms adapted to desiccation and UV radiation. Notably, the microbiome appeared to be composed not only of microorganisms possibly involved in biodeterioration but also of carbonatogenic bacteria, such as those belonging to the genus Arthrobacter, which could be useful in bioconservation. Our investigation demonstrated that molecular tools, and in particular the easy-to-run next-generation sequencing, are powerful to perform a microbiological diagnosis in order to plan restoration and protection strategies.

The interaction between iron nutrition, plant species and soil type shapes the rhizosphere microbiome.

Plant-associated microorganisms can stimulate plants growth and influence both crops yield and quality by nutrient mobilization and transport. Therefore, rhizosphere microbiome appears to be one of the key determinants of plant health and productivity. The roots of plants have the ability to influence its surrounding microbiology, the rhizosphere microbiome, through the creation of specific chemical niches in the soil mediated by the release of phytochemicals (i.e. root exudates) that depends on several factors, such as plants genotype, soil properties, plant nutritional status, climatic conditions. In the present research, two different crop species, namely barley and tomato, characterized by different strategies for Fe acquisition, have been grown in the RHIZOtest system using either complete or Fe-free nutrient solution to induce Fe starvation. Afterward, plants were cultivated for 6 days on two different calcareous soils. Total DNA was extracted from rhizosphere and bulk soil and 454 pyrosequencing technology was applied to V1-V3 16S rRNA gene region. Approximately 5000 sequences were obtained for each sample. The analysis of the bacterial population confirmed that the two bulk soils showed a different microbial community. The presence of the two plant species, as well as the nutritional status (Fe-deficiency and Fe-sufficiency), could promote a differentiation of the rhizosphere microbiome, as highlighted by non-metric multidimensional scaling (NMDS) analysis. Alphaproteobacteria, Actinobacteria, Chloracidobacteria, Thermoleophilia, Betaproteobacteria, Saprospirae, Gemmatimonadetes, Gammaproteobacteria, Acidobacteria were the most represented classes in all the samples analyzed even though their relative abundance changed as a function of the soil, plant species and nutritional status. To our knowledge, this research demonstrate for the first time that different plants species with a diverse nutritional status can promote the development of a peculiar rhizosphere microbiome, depending on the growth substrate.

Plant-microorganism-soil interactions influence the Fe availability in the rhizosphere of cucumber plants.

Iron (Fe) is a very important element for plants, since it is involved in many biochemical processes and, often, for the low solubility of the natural Fe sources in soil, plants suffer from Fe - deficiency, especially when grown on calcareous soils. Among the numerous plant growth-promoting rhizobacteria (PGPR) that colonize the rhizosphere of agronomically important crops, Azospirillum brasilense has been shown to exert strong stimulating activities on plants, by inducing alterations of the root architecture and an improvement of mineral nutrition, which could result from an enhancement of ion uptake mechanisms as well as by increased bioavailability of nutrients. Some studies have also established that A. brasilense can act as biocontrol agent, by preventing the growth and/or virulence of phytopathogens, most likely through the production of microbial siderophores that sequester Fe from the soil. Despite microbial siderophores complexed with Fe could be an easily accessible Fe source for plants, the possible involvement of A. brasilense in improving Fe nutrition in plants suffering from the micronutrient deficiency has not been investigated yet. Within the present research, the characterization of the physiological and biochemical effects induced by Fe starvation and PGPR inoculation in cucumber plants (Cucumis sativus L. cv. Chinese Long) was carried out. The analyses of root exudates released by hydroponically grown plants highlighted that cucumber plants respond differently depending on the nutritional status. In addition, following the cultivation period on calcareous soil, also the root exudates found in the extracts suggested a peculiar behaviour of plants as a function of the treatment. Interestingly, the presence of the inoculum in soil allowed a faster recovery of cucumber plants from Fe-deficiency symptoms, i.e. increase in the chlorophyll content, in the biomass and in the Fe content of leaves. These observations might suggest a feasible application of A. brasilense in alleviating symptoms generated by Fe-limiting growth condition in cucumber plants.

Duodenal and faecal microbiota of celiac children: molecular, phenotype and metabolome characterization.

BACKGROUND: Epidemiology of celiac disease (CD) is increasing. CD mainly presents in early childhood with small intestinal villous atrophy and signs of malabsorption. Compared to healthy individuals, CD patients seemed to be characterized by higher numbers of Gram-negative bacteria and lower numbers Gram-positive bacteria. RESULTS: This study aimed at investigating the microbiota and metabolome of 19 celiac disease children under gluten-free diet (treated celiac disease, T-CD) and 15 non-celiac children (HC). PCR-denaturing gradient gel electrophoresis (DGGE) analyses by universal and group-specific primers were carried out in duodenal biopsies and faecal samples. Based on the number of PCR-DGGE bands, the diversity of Eubacteria was the higher in duodenal biopsies of T-CD than HC children. Bifidobacteria were only found in faecal samples. With a few exceptions, PCR-DGGE profiles of faecal samples for Lactobacillus and Bifidobacteria differed between T-CD and HC. As shown by culture-dependent methods, the levels of Lactobacillus, Enterococcus and Bifidobacteria were confirmed to be significantly higher (P = 0.028; P = 0.019; and P = 0.023, respectively) in fecal samples of HC than in T-CD children. On the contrary, cell counts (CFU/ml) of presumptive Bacteroides, Staphylococcus, Salmonella, Shighella and Klebsiella were significantly higher (P = 0.014) in T-CD compared to HC children. Enterococcus faecium and Lactobacillus plantarum were the species most diffusely identified. This latter species was also found in all duodenal biopsies of T-CD and HC children. Other bacterial species were identified only in T-CD or HC faecal samples. As shown by Randomly Amplified Polymorphic DNA-PCR analysis, the percentage of strains identified as lactobacilli significantly (P = 0.011) differed between T-CD (ca. 26.5%) and HC (ca. 34.6%) groups. The metabolome of T-CD and HC children was studied using faecal and urine samples which were analyzed by gas-chromatography mass spectrometry-solid-phase microextraction and 1H-Nuclear Magnetic Resonance. As shown by Canonical Discriminant Analysis of Principal Coordinates, the levels of volatile organic compounds and free amino acids in faecal and/or urine samples were markedly affected by CD. CONCLUSION: As shown by the parallel microbiology and metabolome approach, the gluten-free diet lasting at least two years did not completely restore the microbiota and, consequently, the metabolome of CD children. Some molecules (e.g., ethyl-acetate and octyl-acetate, some short chain fatty acids and free amino acids, and glutamine) seems to be metabolic signatures of CD.

Effects of a humic acid and its size-fractions on the bacterial community of soil rhizosphere under maize (Zea mays L.).

The effects of a humic acid (HA) and its size-fractions on plants carbon deposition and the structure of microbial communities in the rhizosphere soil of maize (Zea mays L.) plants were studied. Experiments were conducted in rhizobox systems that separate an upper soil-plant compartment from a lower compartment, where roots are excluded from the rhizosphere soil by a nylon membrane. The upper rhizobox compartment received the humic additions, whereas, after roots development, the rhizosphere soil in the lower compartment was sampled and sliced into thin layers. The lux-marked biosensor Pseudomonas fluorescens 10586 pUCD607 biosensor showed a significant increase in the deposition of bioavailable sources of carbon in the rhizosphere of soils when treated with bulk HA, but no response was found for treatments with the separated size-fractions. PCR-DGGE molecular fingerprintings revealed that the structure of rhizosphere microbial communities was changed by all humic treatments and that the smaller and more bioavailable size-fractions were more easily degraded by microbial activity than the bulk HA. On the other hand, highly hydrophobic and strongly associated humic molecules in the bulk HA required additional plant rhizodeposition before their bio-transformation could occur. This work highlights the importance of applying advanced biological and biotechnological methods to notice changes occurring in plant rhizodeposition and rhizosphere microbial activity. Moreover, it suggests correlations between the molecular properties of humic matter and their effects on microbial communities in the rhizosphere as mediated by root exudation.

Different fecal microbiotas and volatile organic compounds in treated and untreated children with celiac disease.

This study aimed at investigating the fecal microbiotas of children with celiac disease (CD) before (U-CD) and after (T-CD) they were fed a gluten-free diet and of healthy children (HC). Brothers or sisters of T-CD were enrolled as HC. Each group consisted of seven children. PCR-denaturing gradient gel electrophoresis (DGGE) analysis with V3 universal primers revealed a unique profile for each fecal sample. PCR-DGGE analysis with group- or genus-specific 16S rRNA gene primers showed that the Lactobacillus community of U-CD changed significantly, while the diversity of the Lactobacillus community of T-CD was quite comparable to that of HC. Compared to HC, the ratio of cultivable lactic acid bacteria and Bifidobacterium to Bacteroides and enterobacteria was lower in T-CD and even lower in U-CD. The percentages of strains identified as lactobacilli differed as follows: HC (ca. 38%) > T-CD (ca. 17%) > U-CD (ca. 10%). Lactobacillus brevis, Lactobacillus rossiae, and Lactobacillus pentosus were identified only in fecal samples from T-CD and HC. Lactobacillus fermentum, Lactobacillus delbrueckii subsp. bulgaricus, and Lactobacillus gasseri were identified only in several fecal samples from HC. Compared to HC, the composition of Bifidobacterium species of T-CD varied, and it varied even more for U-CD. Forty-seven volatile organic compounds (VOCs) belonging to different chemical classes were identified using gas-chromatography mass spectrometry-solid-phase microextraction analysis. The median concentrations varied markedly for HC, T-CD, and U-CD. Overall, the r(2) values for VOC data for brothers and sisters were equal to or lower than those for unrelated HC and T-CD. This study shows the effect of CD pathology on the fecal microbiotas of children.