Metabolome and transcriptome reprogramming underlying tomato drought resistance triggered by a Pseudomonas strain.

Although amelioration of drought stress by Plant Growth Promoting Rhizobacteria (PGPR) is a well-documented phenomenon, the combined molecular and metabolic mechanisms governing this process remain unclear. In these lines, the present study aimed to provide new insights in the underlying drought attenuating mechanisms of tomato plants inoculated with a PGP Pseudomonas putida strain, by using a combination of metabolomic and transcriptomic approaches. Following Differentially Expressed Gene analysis, it became evident that inoculation resulted in a less disturbed plant transcriptome upon drought stress. Untargeted metabolomics highlighted the differential metabolite accumulation upon inoculation, as well as the less metabolic reprograming and the lower accumulation of stress-related metabolites for inoculated stressed plants. These findings were in line with morpho-physiological evidence of drought stress mitigation in the inoculated plants. The redox state modulation, the more efficient nitrogen assimilation, as well as the differential changes in amino acid metabolism, and the induction of the phenylpropanoid biosynthesis pathway, were the main drought-attenuating mechanisms in the SAESo11-inoculated plants. Shifts in pathways related to hormonal signaling were also evident upon inoculation at a transcript level and in conjunction with carbon metabolism regulation, possibly contributed to a drought-attenuation preconditioning. The identified signatory molecules of SAESo11-mediated priming against drought included aspartate, myo-inositol, glutamate, along with key genes related to trehalose, tryptophan and cysteine synthesis. Taken together, SAESo11-inoculation provides systemic effects encompassing both metabolic and regulatory functions, supporting both seedling growth and drought stress amelioration.

Above- and below-ground microbiome in the annual developmental cycle of two olive tree varieties.

The olive tree is a hallmark crop in the Mediterranean region. Its cultivation is characterized by an enormous variability in existing genotypes and geographical areas. As regards the associated microbial communities of the olive tree, despite progress, we still lack comprehensive knowledge in the description of these key determinants of plant health and productivity. Here, we determined the prokaryotic, fungal and arbuscular mycorrhizal fungal (AMF) microbiome in below- (rhizospheric soil, roots) and above-ground (phyllosphere and carposphere) plant compartments of two olive varieties 'Koroneiki' and 'Chondrolia Chalkidikis' grown in Southern and Northern Greece respectively, in five developmental stages along a full fruit-bearing season. Distinct microbial communities were supported in above- and below-ground plant parts; while the former tended to be similar between the two varieties/locations, the latter were location specific. In both varieties/locations, a seasonally stable root microbiome was observed over time; in contrast the plant microbiome in the other compartments were prone to changes over time, which may be related to seasonal environmental change and/or to plant developmental stage. We noted that olive roots exhibited an AMF-specific filtering effect (not observed for bacteria and general fungi) onto the rhizosphere AMF communities of the two olive varieties/locations/, leading to the assemblage of homogenous intraradical AMF communities. Finally, shared microbiome members between the two olive varieties/locations include bacterial and fungal taxa with putative functional attributes that may contribute to olive tree tolerance to abiotic and biotic stress.

Chemical Profiling, Antiproliferative and Antimigratory Capacity of Haberlea rhodopensis Extracts in an In Vitro Platform of Various Human Cancer Cell Lines.

Haberlea rhodopensis is a Balkan endemic plant that belongs to the Gesneriaceae family, and is believed to have medicinal use and health-promoting properties. This study aimed to (i) prepare aqueous (HAE) and ethanolic (HEE) extracts from the leaves of H. rhodopensis from in vitro propagated plants, (ii) screen for their potential antiproliferative and antimigratory activities, and (iii) chemically characterize both HAE and HEE by identifying compounds which may contribute to their observed bioactivity thereby further supporting their potential use in biomedical applications. The antiproliferative activity of both extracts was assessed against six human cancer cell lines by employing the sulforhodamine-B (SRB) assay. HEE was found to be more potent in inhibiting cancer cell growth as compared to HAE. Therefore, HEE's antimigratory effects were further studied in hepatocellular carcinoma (HepG2) and non-small cell lung adenocarcinoma (A459) cell lines as they were among the most sensitive ones to its antiproliferative activity. HEE was found to exert significant antimigratory concentration-dependent effects in both cell lines assessed with the wound healing assay. Chemical characterization by UPLC-MS/MS analysis identified that HEE contains higher levels of flavonoids, phenolic compounds, pigments (chlorophyll-/-b, lycopene, and ß-carotene), monoterpenoids, and condensed tannins compared to HAE, while HAE, contains higher levels of soluble protein and sugars. Furthermore, HEE demonstrated remarkable antioxidant activity evaluated by the 2,2-diphenyl-1-picrylhydrazyl (DPPH●), 2,2-azinobis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS●+) and ferric reducing/antioxidant power (FRAP) assays. We have obtained comprehensive results highlighting the potential of HEE as a source of bioactive compounds with anticancer properties. Future studies should aim at identifying the chemical constituents responsible for the bioactivities observed, and focus on investigating HEE's effects, in in vivo preclinical cancer models.

Coated Cu-doped ZnO and Cu nanoparticles as control agents against plant pathogenic fungi and nematodes.

In the current study, coated copper nanoparticles with polyethylene glycol 8000 (Cu@PEG NPs) and copper-doped zinc oxide nanoparticles with diethylene glycol (Cu-doped ZnO@DEG NPs) have been synthesized via solvothermal and microwave-assisted process, physicochemical characterized, and studied as nano-fungicides and nano-nematicides. Spheroidal Cu-doped ZnO@DEG NPs and urchin-like Cu@PEG NPs have been isolated with average crystallite sizes of 12 and 21 nm, respectively. The Cu doping (11.3 wt%) in ZnO lattice (88.7 wt%) was investigated by Rietveld refinement analysis and confirmed by X-ray Diffraction (XRD) and X-ray Photoelectron Spectroscopy (XPS). The Cu-doped ZnO@DEG and Cu@PEG NPs revealed a growth inhibition of fungi Botrytis cinerea (B. cinerea) and Sclerotinia sclerotiorum (S. sclerotiorum) and nematode paralysis of Meloidogyne javanica in a dose-dependent manner. Cu-doped ZnO@DEG NPs were more effective against M. javanica (EC50 = 2.60 µg/mL) than the Cu@PEG NPs (EC50 = 25 µg/mL). In contrast, the antifungal activity was approximately similar for both NPs, with EC50 values at 310 and 327 µg/mL against B. cinerea, respectively, and 260 and 278 µg/mL against S. sclerotiorum, respectively. Lettuce (Lactuca sativa) plants were inoculated with S. sclerotiorum or M. javanica and sprayed with either Cu-doped ZnO@DEG NPs or Cu@PEG NPs. The antifungal effect was evaluated based on a disease index (DI), and nematicidal activity was assessed based on the total number of galls and nematode females per root gram. NPs successfully inhibited the growth of both pathogens without causing phytotoxicity on lettuce. The DI were significantly decreased as compared to the positive control (DI = 5.2), estimated equal to 1.7, 2.9 and 2.5 for Cu@PEG NPs, Cu-doped ZnO@DEG NPs and the chemical control (KOCIDE 2000), respectively. The reduction in galling and population of M. javanica ranged from 39.32% to 32.29%, statistically like chemical control. The treatment of lettuce plants with Cu-doped ZnO@DEG NPs increased the leaf net photosynthetic value at 4.60 and 6.66 µmol CO2-2 s-1 in plants inoculated with S. sclerotiorum and M. javanica, respectively, as compared to the control (3.00 µmol CO2-2 s-1). The antioxidant capacity of NPs treated lettuce plants was evaluated as 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity in leaf extracts. Plants inoculated with S. sclerotiorum and sprayed with Cu-doped ZnO@DEG and Cu@PEG NPs, exhibited a 34.22% and 32.70% increase in antioxidant capacity, respectively, higher than the control. Similarly, an increase in antioxidant capacity was measured (39.49 and 37.36%) in lettuce inoculated with M. javanica and treated with Cu-doped ZnO@DEG and Cu@PEG NPs, respectively. Moreover, an increase of phenolic compounds in lettuce leaf tissue treated with NPs was measured as compared to the control. Overall, foliar applied Cu and Cu-doped ZnO NPs could be a promising tool to control phytopathogenic fungi and nematodes contributing to sustainability of agri-food sector.

Impact of Geraniol and Geraniol Nanoemulsions on Botrytis cinerea and Effect of Geraniol on Cucumber Plants' Metabolic Profile Analyzed by LC-QTOF-MS.

In the present study, the bioactive substance geraniol was tested in vitro and in planta against B. cinerea on cucumber plants, and the changes in the metabolic profile of cucumber plants inoculated with the pathogen and/or treated with geraniol were monitored by a novel LC-QTOF-MS method employing target and suspect screening. The aforementioned treatments were also studied for their impact on membrane lipid peroxidation calculated as malondialdehyde (MDA) content. Additionally, geraniol-loaded nanoemulsions (GNEs) were synthesized and tested against B. cinerea as an integrated formulation mode of geraniol application. The EC50 values calculated for geraniol and GNEs against B. cinerea were calculated at 235 µg/mL and 105 µg/mL, respectively. The in planta experiment on cucumber plants demonstrated the ability of geraniol and GNEs to significantly inhibit B. cinerea under greenhouse conditions. The LC-QTOF-MS analysis of the metabolic profile of the cucumber plants treated with geraniol demonstrated an increase in the concentration levels of myricetin, chlorogenic acid, and kaempferol rhamnoside, as compared to control plants and the presence of B. cinerea caused an increase in sinapic acid and genistein. These compounds are part of important biosynthetic pathways mostly related to responses against a pathogen attack.

Root Transcriptional and Metabolic Dynamics Induced by the Plant Growth Promoting Rhizobacterium (PGPR) Bacillus subtilis Mbi600 on Cucumber Plants.

Bacillus subtilis MBI600 is a commercialized plant growth-promoting bacterial species used as a biocontrol agent in many crops, controlling various plant pathogens via direct or indirect mechanisms. In the present study, a detailed transcriptomic analysis of cucumber roots upon response to the Bs MBI600 strain is provided. Differentially expressed genes (DEGs) analysis showed altered gene expression in more than 1000 genes at 24 and 48 h post-application of Bs MBI600. Bs MBI600 induces genes involved in ISR and SAR signaling. In addition, genes involved in phytohormone production and nutrient availability showed an upregulation pattern, justifying the plant growth promotion. Biocontrol ability of Bs MBI600 seems also to be related to the activation of defense-related genes, such as peroxidase, endo-1,3(4)-beta-glucanase, PR-4, and thaumatin-like. Moreover, KEGG enriched results showed that differentially expressed genes were classified into biocontrol-related pathways. To further investigate the plant's response to the presence of PGPR, a profile of polar metabolites of cucumber treated with Bs MBI600 was performed and compared to that of untreated plants. The results of the current study gave insights into the mechanisms deployed by this biocontrol agent to promote plant resistance, helping to understand the molecular interactions in this system.

Decoding the potential of a new Pseudomonas putida strain for inducing drought tolerance of tomato (Solanum lycopersicum) plants through seed biopriming.

A total of 11 potential plant growth promoting rhizobacteria previously isolated from naturally stressed environments were evaluated for various traits of interest for a beneficial symbiosis with plants, including colonization ability, biofilm formation, motility, exopolysaccharide production and salt tolerance. The vast majority of the strains were found to possess multiple plant growth promoting traits. Nevertheless, the intensity varied among isolates, with those originated from tomato plants being more efficient colonizers. The strain SAESo11, genetically characterized as a Pseudomonas putida member was selected for further investigation of its potential to alleviate drought stress in tomato seedlings. Inoculation with SAESo11 mitigated the negative effects of drought stress as indicated by growth and photosynthetic indices. Furthermore, bacterial inoculation enhanced H2O2 content and malondialdehyde levels in colonized plants. Drought treatment did not further alter the oxidative status of these plants. Similarly, total phenolic content and antioxidant enzyme activity were induced in plant tissues in response to drought stress only at the absence of inoculum. These results indicated that inoculation with the selected strain imposed plants at a priming state, that enabled them to respond more robustly at the exposure to drought stress and efficiently attenuated the drought-induced injury. This state of plant alertness mediated by SAESo11 occurred at no cost to growth, highlighting its role as a potential plant priming agent.

CuZn and ZnO Nanoflowers as Nano-Fungicides against Botrytis cinerea and Sclerotinia sclerotiorum: Phytoprotection, Translocation, and Impact after Foliar Application.

Inorganic nanoparticles (INPs) have dynamically emerged in plant protection. The uptake of INPs by plants mostly depends on the size, chemical composition, morphology, and the type of coating on their surface. Herein, hybrid ensembles of glycol-coated bimetallic CuZn and ZnO nanoparticles (NPs) have been solvothermally synthesized in the presence of DEG and PEG, physicochemically characterized, and tested as nano-fungicides. Particularly, nanoflowers (NFs) of CuZn@DEG and ZnO@PEG have been isolated with crystallite sizes 40 and 15 nm, respectively. Organic coating DEG and PEG (23% and 63%, respectively) was found to protect the NFs formation effectively. The CuZn@DEG and ZnO@PEG NFs revealed a growth inhibition of phytopathogenic fungi Botrytis cinerea and Sclerotinia sclerotiorum in a dose-dependent manner with CuZn@DEG NFs being more efficient against both fungi with EC50 values of 418 and 311 µg/mL respectively. Lettuce (Lactuca sativa) plants inoculated with S. sclerotiorum were treated with the NFs, and their antifungal effect was evaluated based on a disease index. Plants sprayed with ZnO@PEG NFs showed a relatively higher net photosynthetic (4.70 µmol CO2 m-2s-1) and quantum yield rate (0.72) than with CuZn@DEG NFs (3.00 µmol CO2 m-2s-1 and 0.68). Furthermore, the penetration of Alizarin Red S-labeled NFs in plants was investigated. The translocation from leaves to roots through the stem was evident, while ZnO@PEG NFs were mainly trapped on the leaves. In all cases, no phytotoxicity was observed in the lettuce plants after treatment with the NFs.

Monoterpene Enrichments Have Positive Impacts on Soil Bacterial Communities and the Potential of Application in Bioremediation.

We study here how soil bacterial communities of different ecosystems respond to disturbances caused by enrichments with monoterpenes that are common essential oil constituents. We used fenchone, 1,8-cineol and α-pinene, and soils from phrygana, a typical Mediterranean-type ecosystem where aromatic plants abound, and from another five ecosystem types, focusing on culturable bacteria. Patterns of response were common to all ecosystems, but responses themselves were not always as pronounced in phrygana as in the other ecosystems, suggesting that these enrichments are less of a disturbance there. More specifically, soil respiration and abundance of the bacterial communities increased, becoming from below two up to 16 times as high as in control soils (for both attributes) and remained at high levels as long as these compounds were present. Bacteria that can utilize these three compounds as substrates of growth became dominant members of the bacterial communities in the enriched soils. All changes were readily reversible once monoterpene addition stopped. Bacteria with the ability to utilize these monoterpenes as carbon sources were found in soils from all ecosystems, 15 strains in total, suggesting a rather universal presence; of these, six could also utilize the organic pollutants toluene or p-xylene. These results suggest also potential novel applications of monoterpenes in combating soil pollution.

Impacts of Decaying Aromatic Plants on the Soil Microbial Community and on Tomato Seedling Growth and Metabolism: Suppression or Stimulation?

This study provides insight into changes in the features of tomato seedlings growing in soils enriched with spearmint, peppermint, or rosemary leaves and into changes in the microbial communities of these soils used as seedbeds; an organic amendment was also applied as a positive control. While the soil microbial community flourished in the presence of all three aromatic plants, tomato growth was inhibited or stimulated depending on the plant that was used. More specifically, phospholipid fatty acid (PLFA) analysis showed an increase in the total microbial biomass and in the biomass of all the groups examined, except for actinobacteria, and changes in the microbial community structure, with Gram-negative bacteria and fungi being favoured in the mint treatments, in which the microbial biomass was maximized. Seedlings from the rosemary treatment were entirely inhibited; they were at the open-cotyledon stage throughout the experiment. Seedlings from the mint treatments were the heaviest, longest, and had the highest chlorophyll content and photosynthetic yield. Metabolomic analysis showed metabolism enhancement associated with both growth and priming in seedlings from the mint treatments and disruption of metabolic pathways in those from the rosemary treatment. There is a great potential for applying these aromatic plants as soil amendments and as either biostimulants of plant growth or as herbicides.

Comparative Transcriptomics and Metabolomics Reveal an Intricate Priming Mechanism Involved in PGPR-Mediated Salt Tolerance in Tomato.

Plant-associated beneficial strains inhabiting plants grown under harsh ecosystems can help them cope with abiotic stress factors by positively influencing plant physiology, development, and environmental adaptation. Previously, we isolated a potential plant growth promoting strain (AXSa06) identified as Pseudomonas oryzihabitans, possessing 1-aminocyclopropane-1-carboxylate deaminase activity, producing indole-3-acetic acid and siderophores, as well as solubilizing inorganic phosphorus. In this study, we aimed to further evaluate the effects of AXSa06 seed inoculation on the growth of tomato seedlings under excess salt (200 mM NaCl) by deciphering their transcriptomic and metabolomic profiles. Differences in transcript levels and metabolites following AXSa06 inoculation seem likely to have contributed to the observed difference in salt adaptation of inoculated plants. In particular, inoculations exerted a positive effect on plant growth and photosynthetic parameters, imposing plants to a primed state, at which they were able to respond more robustly to salt stress probably by efficiently activating antioxidant metabolism, by dampening stress signals, by detoxifying Na+, as well as by effectively assimilating carbon and nitrogen. The primed state of AXSa06-inoculated plants is supported by the increased leaf lipid peroxidation, ascorbate content, as well as the enhanced activities of antioxidant enzymes, prior to stress treatment. The identified signatory molecules of AXSa06-mediated salt tolerance included the amino acids aspartate, threonine, serine, and glutamate, as well as key genes related to ethylene or abscisic acid homeostasis and perception, and ion antiporters. Our findings represent a promising sustainable solution to improve agricultural production under the forthcoming climate change conditions.

Bacterial Communities in the Rhizosphere and Phyllosphere of Halophytes and Drought-Tolerant Plants in Mediterranean Ecosystems.

The aim of the study was to investigate the bacterial community diversity and structure by means of 16S rRNA gene high-throughput amplicon sequencing, in the rhizosphere and phyllosphere of halophytes and drought-tolerant plants in Mediterranean ecosystems with different soil properties. The locations of the sampled plants included alkaline, saline-sodic soils, acidic soils, and the volcanic soils of Santorini Island, differing in soil fertility. Our results showed high bacterial richness overall with Proteobacteria and Actinobacteria dominating in terms of OTUs number and indicated that variable bacterial communities differed depending on the plant's compartment (rhizosphere and phyllosphere), the soil properties and location of sampling. Furthermore, a shared pool of generalist bacterial taxa was detected independently of sampling location, plant species, or plant compartment. We conclude that the rhizosphere and phyllosphere of native plants in stressed Mediterranean ecosystems consist of common bacterial assemblages contributing to the survival of the plant, while at the same time the discrete soil properties and environmental pressures of each habitat drive the development of a complementary bacterial community with a distinct structure for each plant and location. We suggest that this trade-off between generalist and specialist bacterial community is tailored to benefit the symbiosis with the plant.

Plant growth promoting rhizobacteria isolated from halophytes and drought-tolerant plants: genomic characterisation and exploration of phyto-beneficial traits.

Plant growth promoting rhizobacteria (PGPR) are able to provide cross-protection against multiple stress factors and facilitate growth of their plant symbionts in many ways. The aim of this study was to isolate and characterize rhizobacterial strains under natural conditions, associated with naturally occurring representatives of wild plant species and a local tomato cultivar, growing in differently stressed Mediterranean ecosystems. A total of 85 morphologically different rhizospheric strains were isolated; twenty-five exhibited multiple in vitro PGP-associated traits, including phosphate solubilization, indole-3-acetic acid production, and 1-aminocyclopropane-1-carboxylate deaminase activity. Whole genome analysis was applied to eight selected strains for their PGP potential and assigned seven strains to Gammaproteobacteria, and one to Bacteroidetes. The genomes harboured numerous genes involved in plant growth promotion and stress regulation. They also support the notion that the presence of gene clusters with potential PGP functions is affirmative but not necessary for a strain to promote plant growth under abiotic stress conditions. The selected strains were further tested for their ability to stimulate growth under stress. This initial screening led to the identification of some strains as potential PGPR for increasing crop production in a sustainable manner.

Antisense-mediated S-adenosyl-L-methionine decarboxylase silencing affects heat stress responses of tobacco plants.

Understanding the molecular mode(s) of plant tolerance to heat stress (HS) is crucial since HS is a potential threat to sustainable agriculture and global crop production. Polyamines (PAs) seem to exert multifaceted effects in plant growth and development and responses to abiotic and biotic stresses, presumably via their homeostasis, chemical interactions and contribution to hydrogen peroxide (H2O2) cellular 'signatures'. Downregulation of the apoplastic POLYAMINE OXIDASE (PAO) gene improved thermotolerance in tobacco (Nicotiana tabacum L.) transgenics. However, in the present work we show that transgenic tobacco plants with antisense-mediated S-ADENOSYL-L-METHIONINE DECARBOXYLASE silencing (AS-NtSAMDC) exhibited enhanced sensitivity and delayed responses to HS which was accompanied by profound injury upon HS removal (recovery), as assessed by phenological, physiological and biochemical characteristics. In particular, the AS-NtSAMDC transgenics exhibited significantly reduced rate of photosynthesis, as well as enzymatic and non-enzymatic antioxidants. These transgenics suffered irreversible damage, which significantly reduced their growth potential upon return to normal conditions. These data reinforce the contribution of increased PA homeostasis to tolerance, and can move forward our understanding on the PA-mediated mechanism(s) conferring tolerance to HS that might be targeted via traditional or biotechnological breeding for developing HS tolerant plants.

Sweet cherry fruit cracking: follow-up testing methods and cultivar-metabolic screening.

BACKGROUND: Rain-induced fruit cracking is a major physiological problem in most sweet cherry cultivars. For an in vivo cracking assay, the 'Christensen method' (cracking evaluation following fruit immersion in water) is commonly used; however, this test does not adequately simulate environmental conditions. Herein, we have designed and evaluated a cracking protocol, named 'Waterfall method', in which fruits are continuously wetted under controlled conditions. RESULTS: The application of this method alone, or in combination with 'Christensen method, was shown to be a reliable approach to characterize sweet cherry cracking behavior. Seventeen cherry cultivars were tested for their cracking behavior using both protocols, and primary as well as secondary metabolites identification was performed in skin tissue using a combined GC-MS and UPLC-MS/MS platform. Significant variations of some of the detected metabolites were discovered and important cracking index-metabolite correlations were identified. CONCLUSIONS: We have established an alternative/complementary method of cherry cracking characterization alongside to Christiansen assay.

Proteomic and metabolic analysis reveals novel sweet cherry fruit development regulatory points influenced by girdling.

Despite the application of girdling technique for several centuries, its impact on the metabolic shifts that underly fruit biology remains fragmentary. To characterize the influence of girdling on sweet cherry (Prunus avium L.) fruit development and ripening, second-year-old shoots of the cultivars 'Lapins' and 'Skeena' were girdled before full blossom. Fruit characteristics were evaluated across six developmental stages (S), from green-small fruit (stage S1) to full ripe stage (stage S6). In both cultivars, girdling significantly altered the fruit ripening physiognomy. Time course fruit metabolomic along with proteomic approaches unraveled common and cultivar-specific responses to girdling. Notably, several primary and secondary metabolites, such as soluble sugars (glucose, trehalose), alcohol (mannitol), phenolic compounds (rutin, naringenin-7-O-glucoside), including anthocyanins (cyanidin-3-O-rutinoside, cyanidin-3-O-galactoside, cyanidin-3.5-O-diglucoside) were accumulated by girdling, while various amino acids (glycine, threonine, asparagine) were decreased in both cultivars. Proteins predominantly associated with ribosome, DNA repair and recombination, chromosome, membrane trafficking, RNA transport, oxidative phosphorylation, and redox homeostasis were depressed in fruits of both girdled cultivars. This study provides the first system-wide datasets concerning metabolomic and proteomic changes in girdled fruits, which reveal that shoot girdling may induce long-term changes in sweet cherry metabolism.

Metagenomic Characterization Reveals Pronounced Seasonality in the Diversity and Structure of the Phyllosphere Bacterial Community in a Mediterranean Ecosystem.

We explore how the phyllosphere microbial community responds to a very seasonal environment such as the Mediterranean. For this, we studied the epiphytic bacterial community of a Mediterranean ecosystem in summer and winter, expecting to detect seasonal differences at their maximum. With high-throughput sequencing (HTS), we detected the operational taxonomic units (OTUs) present in the phyllosphere and also in the surrounding air. The epiphytic community is approximately five orders of magnitude denser than the airborne one and is made almost exclusively by habitat specialists. The two communities differ considerably but Proteobacteria and Actinobacteria are dominant in both. Of the five most abundant phyllosphere OTUs, two were closely related to Sphingomonas strains, one to Methylobacterium and the other two to Rhizobiales and Burkholderiales. We found the epiphytic community to become much richer, more distinct, even and diverse, denser and more connected in summer. In contrast, there was no difference in the level of bacterial colonization of the phyllosphere between the two seasons, although there were seasonal differences for individual taxonomic groups: Firmicutes, Gemmatimonadetes and Chlroroflexi had a higher participation in summer, whereas the major Proteobacteria classes presented reverse patterns, with Betaproteobacteria increasing in summer at the expense of the prominent Alphaproteobacteria.

An integrated metabolomic and gene expression analysis identifies heat and calcium metabolic networks underlying postharvest sweet cherry fruit senescence.

MAIN CONCLUSION: Ηeat and calcium treatments reprogram sweet cherry fruit metabolism during postharvest senescence as evidenced by changes in respiration, amino acid metabolism, sugars, and secondary metabolites shift. Heat and calcium treatments are used to improve postharvest fruit longevity; however, the exact mechanism remains poorly understood. To characterize the impact of these treatments on sweet cherries metabolism, 'Lapins' fruits were treated with heat or CaCl2 solutions and their combination and subsequently were exposed at room temperature, for up to 4 days, defined as senescence period. Single and combined heat and calcium treatments partially delayed fruit senescence, as evidenced by changes in fruit colour darkening, skin penetration force, and respiration activity. Calcium content was noticeably increased by heat in Ca-treated fruit. Several primary metabolites, including amino acids, organic acids, and alcohols, were decreased in response to both treatments, while many soluble sugars and secondary metabolites were increased within 1 day post-treatment. Changes of several metabolites in heat-treated fruits, especially esculetin, peonidin 3-O-glucoside and peonidin 3-O-galactoside, ribose, pyroglutamate, and isorhamnetin-3-O-rutinoside, were detected. The metabolome of fruit exposed to calcium also displayed substantial modulations, particularly in the levels of galactose, glycerate, aspartate, tryptophan, phospharate rutin, and peonidin 3-O-glucoside. The expression of several genes involved in TCA cycle (MDH1, IDH1, OGDH, SUCLA2, and SDH1-1), pectin degradation (ADPG1) as well as secondary (SK1, 4CL1, HCT, and BAN), amino acids (ALDH18A1, ALDH4A1, GS, GAD, GOT2, OPLAH, HSDH, and SDS), and sugar (PDHA1 and DLAT) metabolism were affected by both treatments. Pathway-specific analysis further revealed the regulation of fruit metabolic programming by heat and calcium. This work provides a comprehensive understanding of metabolic regulation in response to heat and calcium during fruit senescence.

Metabolic features underlying the response of sweet cherry fruit to postharvest UV-C irradiation.

The impact of ultraviolet-C (UV-C) irradiation on sweet cherry fruit was studied. Following harvest, fruits (cv. Sweetheart) were exposed to different doses of UV-C (0, 1.2, 3.0 or 6.0 kJ m-2) and then cold stored (0 °C) for 10 days. Treatments with UV-C delayed most ripening features and reduced pitting symptoms, particularly following prolonged UV-C application. Also, application of the highest UV-C dose inhibited pectin degradation and delayed skin resistance to penetration. An activation of antioxidants capacity and bioactive compounds, such as flavonoids and phenolics was observed. Illumination with UV-C diminished respiration and altered metabolite profile in whole fruit and skin samples. Several amino acids (eg., threonine and aspartate), sugars, (eg., glucose and fructose) and alcohols (e.g., inositol and mannitol) were modulated by long-term UV-C treatment in whole cherry fruit. Various metabolites, including malate, galacturonate, oxoproline and glutamine were also modulated by UV-C skin tissue. These data enhance our understanding of UV-C function in fruit biology.

Metagenomic Characterization of Bacterial Communities on Ready-to-Eat Vegetables and Effects of Household Washing on their Diversity and Composition.

Ready-to-eat (RTE) leafy salad vegetables are considered foods that can be consumed immediately at the point of sale without further treatment. The aim of the study was to investigate the bacterial community composition of RTE salads at the point of consumption and the changes in bacterial diversity and composition associated with different household washing treatments. The bacterial microbiomes of rocket and spinach leaves were examined by means of 16S rRNA gene high-throughput sequencing. Overall, 886 Operational Taxonomic Units (OTUs) were detected in the salads' leaves. Proteobacteria was the most diverse high-level taxonomic group followed by Bacteroidetes and Firmicutes. Although they were processed at the same production facilities, rocket showed different bacterial community composition than spinach salads, mainly attributed to the different contributions of Proteobacteria and Bacteroidetes to the total OTU number. The tested household decontamination treatments proved inefficient in changing the bacterial community composition in both RTE salads. Furthermore, storage duration of the salads at refrigeration temperatures affected the microbiome, by decreasing the bacterial richness and promoting the dominance of psychrotropic bacteria. Finally, both salads were found to be a reservoir of opportunistic human pathogens, while washing methods usually applied at home proved to be inefficient in their removal.