Application of PLA-Based Films to Preserve Strawberries' Bioactive Compounds.

Poly-(Lactic Acid) (PLA) is regarded as one of the most promising bio-based polymers due to its biocompatibility, biodegradability, non-toxicity, and processability. The investigation of the potential of PLA films in preserving the quality of strawberries is fully in line with the current directives on the sustainability of food packaging. The study aims to investigate the effects of PLA films on strawberries' physical and chemical properties, thereby determining whether they can be used as a post-harvest solution to control antioxidant loss, reduce mold growth, and extend the shelf-life of strawberries. Well-designed PLA films with different-sized holes obtained by laser perforation (PLA0, PLA16 and PLA23) were tested against a conventional packaging polypropylene (PP) tray for up to 20 days of storage. Weight loss and mold growth were significantly slower in strawberries packed in PLA films. At the same time, PLA-based films effectively preserved the deterioration of vitamin C content, polyphenols and antioxidant activity compared to the control. Furthermore, among all, the micro-perforated PLA film (PLA23) showed better preservation in the different parameters evaluated. These results could effectively inhibit the deterioration of fruit quality, showing promising expectations as an effective strategy to extend the shelf-life of strawberries.

Ancient Grain Flours with Different Degrees of Sifting: Advances in Knowledge of Nutritional, Technological, and Microbiological Aspects.

Ancient grains have gained considerable attention in recent years, as some research suggests they may be healthier than modern wheat. The present study aims to evaluate the chemical, rheological, and microbiological features of three Southern Italian cultivated ancient wheat varieties: Risciola, Carosella, and Saragolla. ATR-FTIR analyses were performed on the finely ground grain samples of the three varieties. The selected grains were ground with a stone mill, and different sifting degrees (whole-100%, type 1-80%, and type 0-72%) were evaluated. The flours showed a good nutritional profile, a higher amylose/amylopectin ratio, and a lower glycemic index than the literature. The gluten index of the samples was in the range 2.6-28.9%, and the flours can be classified as weak, having a value <30%. The farinographic test showed a short development time, low dough stability, a high softening degree, and water absorption, which increased with the degree of sifting. Microbiological analyses performed on flours from ancient grains at different degrees of sifting show their safety, according to their microbiological parameters, which fall within the legal microbiological requirements established by the European Commission Regulation (EC).

A Comparative Study between Microwave Hydrodiffusion and Gravity (MHG) and Ultrasound-Assisted Extraction (UAE): Chemical and Biological Characterization of Polyphenol-Enriched Extracts from Aglianico Grape Pomace.

The wine industry produces large amounts of grape pomace (GP), a waste that needs to be disposed of properly. Bioactive compounds with high added value can be recovered from GP as an interesting strategy to reduce the environmental impact. Here, two different technologies were employed to recover polyphenol compounds from GP: microwave hydrodiffusion and gravity (MHG) and ultrasound-assisted extraction (UAE). The further purification of UAE and MHG extracts was carried out through solid-phase extraction (SPE) to obtain three fractions, F1, F2 and F3. ATR-FTIR analysis confirmed the presence of sugar and polysaccharide components in F1, as well as non-anthocyanin and anthocyanin compounds in F2 and F3, respectively. Also, the chemical profile was determined by HPLC-UV-DAD, identifying the presence of catechin in F2, and malvidin-3-O-glucoside chloride and cyanidin chloride derivative as the main anthocyanin compounds in F3. The fractions and their parental extracts were characterized for total phenolic content (TPC) and scavenger activity by in vitro assays. We found that F2-MHG and F3-MHG contained phenol contents 6.5 and 8.5 times higher than those of the parental non-fractionated extracts. Finally, F3-MHG (100 µg/mL, w/v) was shown to reduce the proliferation of HT-29 cells.

Mediterranean Diet as a Shield against Male Infertility and Cancer Risk Induced by Environmental Pollutants: A Focus on Flavonoids.

The role of environmental factors in influencing health status is well documented. Heavy metals, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls, dioxins, pesticides, ultrafine particles, produced by human activities put a strain on the body's entire defense system. Therefore, together with public health measures, evidence-based individual resilience measures are necessary to mitigate cancer risk under environmental stress and to prevent reproductive dysfunction and non-communicable diseases; this is especially relevant for workers occupationally exposed to pollutants and/or populations residing in highly polluted areas. The Mediterranean diet is characterized by a high intake of fruits and vegetables rich in flavonoids, that can promote the elimination of pollutants in tissues and fluids and/or mitigate their effects through different mechanisms. In this review, we collected evidence from pre-clinical and clinical studies showing that the impairment of male fertility and gonadal development, as well as cancers of reproductive system, due to the exposure of organic and inorganic pollutants, may be counteracted by flavonoids.

Phenolic Extract from Extra Virgin Olive Oil Induces Different Anti-Proliferative Pathways in Human Bladder Cancer Cell Lines.

Regular consumption of olive oil is associated with protection against chronic-degenerative diseases, such as cancer. Epidemiological evidence indicates an inverse association between olive oil intake and bladder cancer risk. Bladder cancer is among the most common forms of cancer; in particular, the transitional cell carcinoma histotype shows aggressive behavior. We investigated the anti-proliferative effects of a phenolic extract prepared from an extra virgin olive oil (EVOOE) on two human bladder cancer cell lines, namely RT112 and J82, representing the progression from low-grade to high-grade tumors, respectively. In RT112, the EVOOE reduced cell viability (IC50 = 240 µg/mL at 24 h), triggering a non-protective form of autophagy, evidenced by the autophagosome formation and the increase in LC-3 lipidation. In J82, EVOOE induced a strong decrease in cell viability after 24 h of treatment (IC50 = 65.8 µg/mL) through rapid and massive apoptosis, assessed by Annexin V positivity and caspase-3 and -9 activation. Moreover, in both bladder cancer cell lines, EVOOE reduced intracellular reactive oxygen species, but this antioxidant effect was not correlated with its anti-proliferative outcomes. Data obtained suggest that the mixture of phenolic compounds in extra virgin olive oil activates different anti-proliferative pathways.

Nutritional and Chemical-Physical Characterization of Fresh Pasta Gnocchi Prepared with Sea Water as New Active Ingredient.

This study shows the chemical-physical and nutritional results obtained using food-grade sea water for the preparation of fresh pasta Gnocchi with respect to those prepared with tap water. Gnocchi obtained by mixing the flour with seawater (GSW) were compared with traditional Gnocchi made with tap water (GTW). The contents of sodium chloride, macro and micro elements, volatile molecules profile, thermal properties, and morphological analysis were investigated in both Gnocchi types. The analysis of chlorides showed that the samples prepared with sea water had a significantly lower NaCl content after cooking in comparison with those prepared with tap water. These results were also confirmed by the inductively coupled plasma (ICP) analysis for sodium content. The profiles of the volatile molecules acquired by gas chromatography-mass spectrometry (GC-MS) evidenced significant differences between the groups of aromatic molecules of the two typologies of samples. Morphological analysis evidenced that both raw and cooked GSW Gnocchi were structurally tightened whereas GTW Gnocchi showed a labile and weak macromolecular network. In addition, GSW Gnocchi was more thermally stable than GTW Gnocchi, as evidenced by thermogravimetric analysis (TGA).

The Effect of Novel Packaging Technology on Food Safety and Quality.

This Special Issue of Foods, The Effect of Novel Packaging Technology on Food Safety and Quality, contains seven papers that were refereed and selected in accordance with the usual editorial standards of the journal [...].

Active Edible Polysaccharide-Based Coating for Preservation of Fresh Figs (Ficus carica L.).

Fresh figs are very sensitive to microbial spoilage, even in cold storage conditions. Thus, fresh figs are high perishable products during postharvest with microbiological decay that induces an unpleasant taste and smell due to rot, and suitable conservation methods must therefore be applied. The fruit usually is consumed fresh locally, dried, or preserved longer term in other transformed forms. A sustainable approach to extend the shelf-life of figs can be constituted by application of an edible coating able to maintain the quality of the fruit during storage. A comparison between fresh figs in a commercial preservation system, with the figs preserved in an edible coating, and an active edible coating to preserve their quality characteristics was carried out. The coating efficacy was enhanced with the addition of pomegranate peel extract at two different concentrations. The inclusion of a component with high antioxidant activity in an edible coating proved to be an excellent method for preserving the quality of this highly perishable fruit. The application of natural products, obtained from renewable sources, represents a simple and economic strategy, but also a tool capable of preserving the quality of the fruit during the postharvest storage, which is often consumed in production areas due to shelf-life problems.

Phytocompounds vs. Dental Plaque Bacteria: In vitro Effects of Myrtle and Pomegranate Polyphenolic Extracts Against Single-Species and Multispecies Oral Biofilms.

In the last decades, resistant microbial infection rate has dramatically increased, especially infections due to biofilm-producing strains that require increasingly complex treatments and are responsible for the increased mortality percentages compared with other infectious diseases. Considering that biofilms represent a key factor for a wide range of chronic infections with high drug tolerance, the treatment of biofilm-causing bacterial infections represents a great challenge for the future. Among new alternative strategies to conventional antimicrobial agents, the scientific interest has shifted to the study of biologically active compounds from plant-related extracts with known antimicrobial properties, in order to also evaluate their antibiofilm activity. In this regard, the aim of this study has been to assess the antibiofilm activity of polyphenolic extracts from myrtle leaf and pomegranate peel against oral pathogens of dental plaque, an excellent polymicrobial biofilm model. In particular, the in vitro antibiofilm properties of myrtle and pomegranate extracts, also in binary combination, were highlighted. In addition to inhibiting the biofilm formation, the tested polyphenolic extracts have been proven to destroy both preformed single-species and multispecies biofilms formed by Streptococcus mutans, Streptococcus oralis, Streptococcus mitis, and Rothia dentocariosa oral isolates, suggesting that the new natural sources are rich in promising compounds able to counteract biofilm-related infections.

In vitro Synergy of Polyphenolic Extracts From Honey, Myrtle and Pomegranate Against Oral Pathogens, S. mutans and R. dentocariosa.

The increasing incidence rate of oral diseases, the wide spread of antimicrobial resistance, and the adverse effects of conventional antibiotics mean alternative prevention and treatment options are needed to counteract oral pathogens. In this regard, our study aims to evaluate the antibacterial activity of polyphenolic extracts prepared from acacia honey, myrtle leaves, and pomegranate peel against cariogenic bacteria, such as Streptococcus mutans and Rothia dentocariosa. The chemical-physical parameters of acacia honey and the RP-HPLC polyphenolic profile of pomegranate peel extract have been previously described in our studies, while the characterization of myrtle extract, performed by HPLC analysis, is reported here. All the extracts were used singly and in binary combinations to highlight any synergistic effects. Moreover, the extracts were tested in association with amoxicillin to evaluate their ability to reduce the effective dose of this drug in vitro. The values of minimal inhibitory concentrations and minimal bactericidal concentrations have been used to quantitatively measure the antibacterial activity of the single extracts, while the fractional inhibitory concentration index has been considered as predictor of in vitro anticariogenic synergistic effects. Finally, a time-kill curve method allowed for the evaluation of the bactericidal efficacy of the combined extracts. The microbiological tests suggest that acacia honey, myrtle, and pomegranate extracts are able to inhibit the cariogenic bacteria, also with synergistic effects. This study provides useful and encouraging results for the use of natural extract combinations alone or in association with antibiotics (adjuvant therapy) as a valid alternative for the prevention and treatment of oral infectious diseases.

Phytochemical Characterization and Effects on Cell Proliferation of Lentisk (Pistacia lentiscus) Berry Oil: a Revalued Source of Phenolics.

The ethno-pharmaceutical use of the edible fixed oil produced from lentisk (Pistacia lentiscus) berries covers a long tradition in several Mediterranean regions. Many of the health-promoting properties of lentisk berry oil (LBO) have been associated with the content of polar (poly)phenolic compounds. However, the polar fraction (methanol 80%, v/v) of LBO (LBO-pf) remains poorly and inadequately characterized. We assessed the phytochemical composition (fatty acids, phytosterols and polyphenols) of cold-pressed LBO produced in Cilento (Campania region, Italy) over four years of production (2015-2018). Main phenolic compounds present in LBO-pf were identified and semi-quantified combining ultra-high performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS) and HPLC with diode array detection. Phenolic compounds, also responsible for oil stability and antioxidant properties, are relatively abundant in LBO, compared to other edible oils. LBO-pf induced clear dose-dependent effects on the growth of HT-29 cell line derived from human colorectal adenocarcinoma, as evidenced by the cell cycle arrest. Our data support the health-promoting properties of cold-pressed LBO, which is obtained with good yield from spontaneous plants growing in semiarid regions.

Fast non-destructive assessment of heavy metal presence by ATR-FTIR analysis of crayfish exoskeleton.

Freshwater crayfish are bioindicators of environmental pollution, often used for the assessment of heavy metal (HM) presence in the tissues, a time-consuming and expensive task. In this study, we propose the use of the vibrational spectroscopy to detect in a fast, non-destructive and sensitive way the presence of HM in the cephalothorax exoskeleton of the freshwater crayfish. Incorporation of HM into the cephalothorax exoskeleton was investigated under controlled laboratory conditions. In particular, the cephalothorax exoskeleton of five crayfish species (Astacus leptodactylus, Procambarus clarkii, Austropotamobius pallipes, Faxonius limosus, and Pacifastacus leniusculus) was analyzed by attenuated total reflection-Fourier transformed infrared (ATR-FTIR) spectroscopy in the presence or absence of cadmium (Cd), chromium (Cr), lead (Pb), nickel (Ni), and zinc (Zn) up to 4 weeks at various concentrations (0.01, 0.1, 1, 10, ppm). The ATR-FTIR profile of the crayfish cephalothorax exoskeleton was compatible with the presence of amorphous calcium carbonate, chitin, and proteins. The incubation with the HM revealed two main modifications: the shift of the peak from 859 to 872 cm-1 and the appearance of a peak at 712 cm-1. Both are ascribable to the HM interaction with calcium carbonate. The absorbance of both peaks increased along with the time of incubation, and the HM concentration. We conclude that ATR-FTIR analysis can be a useful, quick, and cost-sensitive tool to detect HM presence in the crayfish cephalothorax exoskeleton. However, it has to be regarded as a non-specific analytical technique for assessing HM contamination, since it is unable to discriminate between different HM.

Autotrophic and Heterotrophic Growth Conditions Modify Biomolecole Production in the Microalga Galdieria sulphuraria (Cyanidiophyceae, Rhodophyta).

Algae have multiple similarities with fungi, with both belonging to the Thallophyte, a polyphyletic group of non-mobile organisms grouped together on the basis of similar characteristics, but not sharing a common ancestor. The main difference between algae and fungi is noted in their metabolism. In fact, although algae have chlorophyll-bearing thalloids and are autotrophic organisms, fungi lack chlorophyll and are heterotrophic, not able to synthesize their own nutrients. However, our studies have shown that the extremophilic microalga Galderia sulphuraria (GS) can also grow very well in heterotrophic conditions like fungi. This study was carried out using several approaches such as scanning electron microscope (SEM), gas chromatography/mass spectrometry (GC/MS), and infrared spectrophotometry (ATR-FTIR). Results showed that the GS, strain ACUF 064, cultured in autotrophic (AGS) and heterotrophic (HGS) conditions, produced different biomolecules. In particular, when grown in HGS, the algae (i) was 30% larger, with an increase in carbon mass that was 20% greater than AGS; (ii) produced higher quantities of stearic acid, oleic acid, monounsaturated fatty acids (MUFAs), and ergosterol; (iii) produced lower quantities of fatty acid methyl esters (FAMEs) such as methyl palmytate, and methyl linoleate, saturated fatty acids (SFAs), and poyliunsaturated fatty acids (PUFAs). ATR-FTIR and principal component analysis (PCA) statistical analysis confirmed that the macromolecular content of HGS was significantly different from AGS. The ability to produce different macromolecules by changing the trophic conditions may represent an interesting strategy to induce microalgae to produce different biomolecules that can find applications in several fields such as food, feed, nutraceutical, or energy production.

Antiproliferative and antioxidant effect of polar hemp extracts (Cannabis sativa L., Fedora cv.) in human colorectal cell lines.

Total phenolic content and antioxidant activity of polar extracts of edible resources from Fedora hemp cultivar (Cannabis sativa L.), namely seed, flour and oil, were evaluated. The main components in the polar extracts were identified using HPLC-DAD and HPLC-ESI-MS/MS. As expected, the molecular profile of components from seeds and flour was strictly similar, dominated by N-trans-caffeoyltyramine. The profile of oil polar extracts contained hydroxycinnamic acid derivatives and cannabinoids at lower extent. While the extracts from hemp seed and flour did not interfere with growth of Caco-2 and HT-29 cell, the one from oil (150 µg/mL) significantly reduced cell viability after 24 h of treatment. This effect was associated with the activation of apoptotic cell death and was independent from the antioxidant capacity of the oil polar extract. Notably, HT-29 cells differentiated with sodium butyrate were not sensitive to the cytotoxic effect of the oil extract.

UHPLC-HR-MS/MS-Guided Recovery of Bioactive Flavonol Compounds from Greco di Tufo Vine Leaves.

Leaves of Vitis vinifera cv. Greco di Tufo, a precious waste made in the Campania Region (Italy), after vintage harvest, underwent reduction, lyophilization, and ultrasound-assisted maceration in ethanol. The alcoholic extract, as evidenced by a preliminary UHPLC-HR-MS analysis, showed a high metabolic complexity. Thus, the extract was fractionated, obtaining, among others, a fraction enriched in flavonol glycosides and glycuronides. Myricetin, quercetin, kaempferol, and isorhamnetin derivatives were tentatively identified based on their relative retention time and TOF-MS2 data. As the localization of saccharidic moiety in glycuronide compounds proved to be difficult due to the lack of well-established fragmentation pattern and/or the absence of characteristic key fragments, to obtain useful MS information and to eliminate matrix effect redundancies, the isolation of the most abundant extract's compound was achieved. HR-MS/MS spectra of the compound, quercetin-3-O-glucuronide, allowed us to thoroughly rationalize its fragmentation pattern, and to unravel the main differences between MS/MS behavior of flavonol glycosides and glycuronides. Furthermore, cytotoxicity assessment on the (poly)phenol rich fraction and the pure isolated compound was carried out using central nervous system cell lines. The chemoprotective effect of both the (poly)phenol fraction and quercetin-3-O-glucuronide was evaluated.

Vorinostat Potentiates 5-Fluorouracil/Cisplatin Combination by Inhibiting Chemotherapy-Induced EGFR Nuclear Translocation and Increasing Cisplatin Uptake.

The 5-fluorouracil/cisplatin (5FU/CDDP) combination is one of the most widely used treatment options for several solid tumors. However, despite good anticancer responses, this regimen is often associated with high toxicity and treatment resistance. In our study, we evaluated whether the histone deacetylase inhibitor (HDACi), vorinostat, may induce synergistic antitumor and proapoptotic effects in combination with 5FU/CDDP in squamous cancer cell models. We demonstrated in cancer cell lines, including the intrinsic CDDP-resistant Cal27 cells, that simultaneous exposure to equitoxic doses of vorinostat plus 5FU/CDDP results in strong synergistic antiproliferative and proapoptotic effects related to cell-cycle perturbation and DNA damage induction. These effects were confirmed in vivo in both orthotopic and heterotopic xenograft mouse models of Cal27 cells. Mechanistically, vorinostat reverted 5FU/CDDP-induced EGFR phosphorylation and nuclear translocation, leading to the impairment of nuclear EGFR noncanonical induction of genes such as thymidylate synthase and cyclin D1. These effects were exerted by vorinostat, at least in part, by increasing lysosomal-mediated EGFR protein degradation. Moreover, vorinostat increased platinum uptake and platinated DNA levels by transcriptionally upregulating the CDDP influx channel copper transporter 1 (CTR1). Overall, to our knowledge, this study is the first to demonstrate the ability of vorinostat to inhibit two well-known mechanisms of CDDP resistance, EGFR nuclear translocation and CTR1 overexpression, adding new insight into the mechanism of the synergistic interaction between HDACi- and CDDP-based chemotherapy and providing the rationale to clinically explore this combination to overcome dose-limiting toxicity and chemotherapy resistance.

Rapid Evaluation Methods for Quality of Trout (Oncorhynchus mykiss) Fresh Fillet Preserved in an Active Edible Coating.

In this study different methods were used to evaluate the effectiveness of a carrageenan coating and carrageenan coating incorporating lemon essential oil (ELO) in preserving the physicochemical and olfactory characteristics of trout fillets stored at 4 °C up to 12 days. The fillet morphological structure was analyzed by histological and immunological methods; lipid peroxidation was performed with the peroxide and thiobarbituric acid reactive substances (TBARS) tests. At the same time, two less time-consuming methods, such as Attenuated Total Reflectance-Fourier Transformed Infrared (ATR-FTIR) spectroscopy and the electronic nose, were used. Uncoated trout fillets (UTF) showed a less compact tissue structure than carrageenan-coated threads (CTF) and coated fillets of carrageenan (active) ELO (ACTF), probably due to the degradation of collagen, as indicated by optical microscopy and ATR-FTIR. UTF showed greater lipid oxidation compared to CTF and ACTF, as indicated by the peroxide and TBARS tests and ATR-FTIR spectroscopy. The carrageenan coating containing ELO preserved the olfactory characteristics of the trout fillets better than the carrageenan coating alone, as indicated by the electronic nose analysis. This study confirms that both carrageenan and ELO containing carrageenan coatings slow down the decay of the physicochemical and olfactory characteristics of fresh trout fillets stored at 4 °C, although the latter is more effective.

In vitro intestinal epithelium responses to titanium dioxide nanoparticles.

Titanium dioxide (TiO2) is enclosed in many consumer products including pharmaceuticals, cosmetics, and foods. TiO2 (E171) is daily ingested as mixed nano- and submicron-sized particles since it is approved as a white colorant in Europe in a wide variety of food products, Noteworthy, the relevant risk assessment has never been satisfactorily concluded and growing alarms for human hazards deriving from TiO2 exposure are incrementally reported. The objective of the present study was to establish conceivable mechanisms by which nano-sized TiO2 particles affect physiological function of the intestinal epithelium layer. The well-established Caco-2 cell line differentiated for 21 days on permeable supports was used as a predictive model of the human intestinal mucosa to identify the biological response triggered by TiO2 particles. Exposure to 42 µg/mL TiO2 nanoparticles disrupted the tight junctions-permeability barrier with a prompt effect detectable after 4 h incubation time and wide effects on barrier integrity at 24 h. Transport and ultrastructural localization of TiO2 nanoparticles were determined by ICP-OES, TEM and ESI/EELS analysis, respectively. Nano-sized particles were efficiently internalized and preferentially entrapped by Caco-2 monolayers. Storage of TiO2 nanoparticles inside the cells affected enterocytes viability and triggered the production of pro-inflammatory cytokines, including TNF-α and IL-8. Taken together these data indicate that nano-sized TiO2 particles exert detrimental effects on the intestinal epithelium layer.

Comparative Study of Chemical, Biochemical Characteristic and ATR-FTIR Analysis of Seeds, Oil and Flour of the Edible Fedora Cultivar Hemp (Cannabis sativa L.).

A series of chemical and biochemical parameters of edible hemp resources (seeds, oil, and flour) from the monoecious EU registered hemp genotype Fedora, was determined, including fatty acid profile, phytosterol composition, total phenolics, antioxidant activity, macro- and micro-elements. The fatty acid ω-3/ω-6 approached the nutritionally optimal 3/1 ratio. ß-sitosterol and other phytosterols sterols dominated the unsaponifiable fraction. Hemp seeds, flour, and oil contained 767 ± 41, 744 ± 29, and 21 ± 5 mg GAE kg-1 total polyphenols, respectively. The antioxidant potential of Fedora flour and seeds, evaluated through the DPPH (2,2-Diphenyl-1-picrylhydrazyl) assay, was higher than that of oil. K and Mg were the most abundant macro-elements, particularly in flour, while the concentration of trace elements was Fe > Cu > Ni > Mn. The presence of an array of bioactive compound candidate Fedora products as health-promoting food matrices. The ATR-FTIR spectra of hemp-derived products indicated the proximate composition of macro-nutrients.

The Association With Two Different Arbuscular Mycorrhizal Fungi Differently Affects Water Stress Tolerance in Tomato.

Arbuscular mycorrhizal (AM) fungi are very widespread, forming symbiotic associations with ∼80% of land plant species, including almost all crop plants. These fungi are considered of great interest for their use as biofertilizer in low-input and organic agriculture. In addition to an improvement in plant nutrition, AM fungi have been reported to enhance plant tolerance to important abiotic and biotic environmental conditions, especially to a reduced availability of resources. These features, to be exploited and applied in the field, require a thorough identification of mechanisms involved in nutrient transfer, metabolic pathways induced by single and multiple stresses, physiological and eco-physiological mechanisms resulting in improved tolerance. However, cooperation between host plants and AM fungi is often related to the specificity of symbiotic partners, the environmental conditions and the availability of resources. In this study, the impact of two AM fungal species (Funneliformis mosseae and Rhizophagus intraradices) on the water stress tolerance of a commercial tomato cultivar (San Marzano nano) has been evaluated in pots. Biometric and eco-physiological parameters have been recorded and gene expression analyses in tomato roots have been focused on plant and fungal genes involved in inorganic phosphate (Pi) uptake and transport. R. intraradices, which resulted to be more efficient than F. mosseae to improve physiological performances, was selected to assess the role of AM symbiosis on tomato plants subjected to combined stresses (moderate water stress and aphid infestation) in controlled conditions. A positive effect on the tomato indirect defense toward aphids in terms of enhanced attraction of their natural enemies was observed, in agreement with the characterization of volatile organic compound (VOC) released. In conclusion, our results offer new insights for understanding the molecular and physiological mechanisms involved in the tolerance toward water deficit as mediated by a specific AM fungus. Moreover, they open new perspectives for the exploitation of AM symbiosis to enhance crop tolerance to abiotic and biotic stresses in a scenario of global change.