Nutraceutical Content and Biological Properties of Lipophilic and Hydrophilic Fractions of the Phytocomplex from Pistacia atlantica Desf. Buds, Roots, and Fruits.

The aim of the present investigation was to obtain 12 aqueous extracts and 1 oil from Pistacia atlantica Desf. subsp. atlantica specimens. The samples differed for processed plant organs (i.e., roots, buds, and fruits), gender and geographical station of the collected trees. Total phenols, flavonoids, and condensed tannins were determined, revealing that bud extracts exhibited the highest phenolic content (386.785 ± 16.227 mg GAE/g DM), followed by fruit and root preparations. Similar results were detected for flavonoids and tannins, whose quantitation ranged from 0.014 ± 0.005 to 74.780 ± 9.724 mg CE/g DM and from 0.037 ± 0.003 to 14.793 ± 0.821 mg CE/g DM, respectively. The biochemical profile of the extracts was further characterized by HPLC-DAD, in terms of specific phenolics. This analysis identified gallic acid as a typical metabolite for ripe fruit, while hydroxytyrosol for female roots and male buds. In parallel, P. atlantica fruit oil was profiled by GC-MS analysis, which detected 37 lipophilic components, including palmitic acid (the major component, ~55%), anacardol, tetradecanol, arachidic acid, squalene, and some terpenes. The samples revealed interesting antioxidant activity, with EC50 values ranging from 0.073 ± 0.001 to 193.594 ± 28.942 mg/mL and from 0.029 ± 0.001 to 103.086 ± 20.540 mg/mL, in that order, for DPPH and reducing power assays. Concerning the total antioxidant capacity, the results ranged from 0.053 ± 0.008 to 51.648 ± 1.659 mg AAE/g DM. Finally, the antimicrobial potential of the plant extracts was estimated against 7 bacterial species and 2 fungal strains, known to be human pathogens, demonstrating a good antibiotic effect for the bud extracts. All these findings strongly suggest that P. atlantica would represent a natural reservoir for novel additives to be used in therapeutic, food, and cosmetic products.

Microparticles from dental calculus disclose paleoenvironmental and palaeoecological records.

Plants have always represented a key element in landscape delineation. Indeed, plant diversity, whose distribution is influenced by geographic/climatic variability, has affected both environmental and human ecology. The present contribution represents a multi-proxy study focused on the detection of starch, pollen and non-pollen palynomorphs in ancient dental calculus collected from pre-historical individuals buried at La Sassa and Pila archaeological sites (Central Italy). The collected record suggested the potential use of plant taxa by the people living in Central Italy during the Copper-Middle Bronze Age and expanded the body of evidence reported by previous palynological and palaeoecological studies. The application of a microscopic approach provided information about domesticated crops and/or gathered wild plants and inferred considerations on ancient environments, water sources, and past health and diseases. Moreover, the research supplied data to define the natural resources (e.g., C4-plant intake) and the social use of the space during that period. Another important aspect was the finding of plant clues referable to woody habitats, characterised by broad-leaved deciduous taxa and generally indicative of a warm-temperate climate and grassy vegetation. Other unusual records (e.g., diatoms, brachysclereids) participated in defining the prehistoric ecological framework. Thus, this work provides an overview on the potential of the human dental calculus analysis to delineate some features of the ancient plant ecology and biodiversity.

Sustainability in Aquaponics: Industrial Spirulina Waste as a Biofertilizer for Lactuca sativa L. Plants.

Aquaponics represents an alternative to traditional soil cultivation. To solve the problem of nutrient depletion that occurs in this biotechnological system, the application of a spirulina-based biofertilizer was assessed. The microalgal waste used in this study came from industrial processing. Four different dilutions of the supernatant portion of this waste were sprayed on lettuce plants cultivated in an aquaponics system installed at the Botanical Gardens of the Tor Vergata University of Rome. The biofertilizer was characterized to evaluate its amount of macro- and micronutrients. The analysis conducted on the plants involved both morpho-biometric aspects and qualitative-quantitative measurements. The experiments showed that the spirulina extract had a positive effect on the growth and nutraceutical content of the lettuce plants; the obtained results highlighted that a dilution of 75% was the best for treatment. The use of the proposed organic and recycled fertilizer could increase the sustainability of crop cultivation and promote the functioning of aquaponics systems.

Pollen variability in Quercus L. species and relative systematic implications.

This study aims to address one of the challenges related to the complexity of the Quercus L. genus, that is the identification of structural elements favouring the systematic identification of the oak pollen. Thus, in this contribution, we explored the variation of morphometric and chemical parameters in pollen samples collected from 47 different Quercus species and hybrids. Several qualitative (e.g., outline in polar view, class, aperture structures) and quantitative (e.g., diameter, exine and sporoderm thickness, autofluorescence, content in proteins and plant metabolites) features were evaluated by optic microscopy and spectrophotometric assays. Statistical analyses were also carried out to assess significant correlations and clustering effects among the studied taxa, based on phenotypical and biochemical data, to identify the parameters which could be useful for taxonomic discrimination at inter- and intra-specific level. Only few morphological traits showed the potentiality to be diagnostic, such as pollen diameter and outline in polar view. The intensity of pollen autofluorescence varied among the samples but it did not seem to correlate with protein, carotenoid, phenolic and flavonoid content. However, differences in protein and carotenoid levels were detected, suggesting them as possible taxonomic discriminants for oak pollen. Thus, our work represents a step forward in understanding morphology and biochemistry of oak pollen, constitutes an experimental set-up applicable in future systematic studies on other genera, and opens new perspectives for further molecular investigations on Quercus species.

Profile and potential bioactivity of the miRNome and metabolome expressed in Malva sylvestris L. leaf and flower.

Malva sylvestris L. (common mallow) is a plant species widely used in phytotherapy and ethnobotanical practices since time immemorial. Characterizing the components of this herb might promote a better comprehension of its biological effects on the human body but also favour the identification of the molecular processes that occur in the plant tissues. Thus, in the present contribution, the scientific knowledge about the metabolomic profile of the common mallow was expanded. In particular, the phytocomplex of leaves and flowers from this botanical species and the extraction capacity of different concentrations of ethanol (i.e., 95%, 70%, 50%, and 0%; v/v in ddH2O) for it were investigated by spectrophotometric and chromatographic approaches. In detail, 95% ethanol extracts showed the worst capacity in isolating total phenols and flavonoids, while all the hydroalcoholic samples revealed a specific ability in purifying the anthocyanins. HPLC-DAD system detected and quantified 20 phenolic secondary metabolites, whose concentration in the several extracts depended on their own chemical nature and the percentage of ethanol used in the preparation. In addition, the stability of the purified phytochemicals after resuspension in pure ddH2O was also proved, considering a potential employment of them in biological/medical studies which include in vitro and in vivo experiments on mammalian models. Here, for the first time, the expressed miRNome in M. sylvestris was also defined by Next Generation Sequencing, revealing the presence of 33 microRNAs (miRNAs), 10 typical for leaves and 2 for flowers. Then, both plant and human putative mRNA targets for the detected miRNAs were predicted by bioinformatics analyses, with the aim to clarify the possible role of these small nucleic acids in the common mallow plant tissues and to try to understand if they could exert a potential cross-kingdom regulatory activity on the human health. Surprisingly, our investigations revealed that 19 miRNAs out of 33 were putatively able to modulate, in the plant cells, the expression of various chromosome scaffold proteins. In parallel, we found, in the human transcriptome, a total of 383 mRNAs involved in 5 fundamental mammalian cellular processes (i.e., apoptosis, senescence, cell-cycle, oxidative stress, and invasiveness) that theoretically could be bound and regulated by M. sylvestris miRNAs. The evidence collected in this work would suggest that the beneficial properties of the use of M. sylvestris, documented by the folk medicine, are probably linked to their content of miRNAs and not only to the action of phytochemicals (e.g., anthocyanins). This would open new perspectives about the possibility to develop gene therapies based on miRNAs isolated from medicinal plants, including M. sylvestris.

Antioxidant Phytocomplexes Extracted from Pomegranate (Punica granatum L.) Using Hydrodynamic Cavitation Show Potential Anticancer Activity In Vitro.

Hydrodynamic cavitation (HC), as an effective, efficient, and scalable extraction technique for natural products, could enable the affordable production of valuable antioxidant extracts from plant resources. For the first time, whole pomegranate (Punica granatum L.) fruits, rich in bioactive phytochemicals endowed with anti-cancer properties, were extracted in water using HC. Aqueous fractions sequentially collected during the process (M1-M5) were lyophilized (L), filtered (A), or used as such, i.e., crude (C), and analyzed for their biochemical profile and in vitro antioxidant power. The fractions M3 and M4 from the L and C series showed the highest antiradical activity and phytochemical content. While the lyophilized form is preferable for application purposes, sample L-M3, which was produced faster and with lower energy consumption than M4, was used to assess the potential antiproliferative effect on human breast cancer line (AU565-PAR) and peripheral blood mononuclear (PBMC) cells from healthy donors. In a pilot study, cell growth, death, and redox state were assessed, showing that L-M3 significantly reduced tumor cell proliferation and intracellular oxygen reactive species. No effect on PBMCs was detected. Thus, the antioxidant phytocomplex extracted from pomegranate quickly (15 min), at room temperature (30 °C), and efficiently showed potential anticancer activity without harming healthy cells.

Looking for Plant microRNAs in Human Blood Samples: Bioinformatics Evidence and Perspectives.

Literature has proposed the existence of a cross kingdom regulation (CRK) between human and plants. In this context, microRNAs present in edible plants would be acquired through diet by the consumer's organism and transported via bloodstream to tissues, where they would modulate gene expression. However, the validity of this phenomenon is strongly debated; indeed, some scholars have discussed both the methodologies and the results obtained in previous works. To date, only one study has performed a bioinformatics analysis on small RNA-sequencing data for checking the presence of plant miRNAs (pmiRNAs) in human plasma. For that investigation, the lack of reliable controls, which led to the misidentification of human RNAs as pmiRNAs, has been deeply criticized. Thus, in the present contribution, we aim to demonstrate the existence of pmiRNAs in human blood, adopting a bioinformatics approach characterized by more stringent conditions and filtering. The information obtained from 380 experiments produced in 5 different next generation sequencing (NGS) projects was examined, revealing the presence of 350 circulating pmiRNAs across the analysed data set. Although one of the NGS projects shows results likely to be attributed to sample contamination, the others appear to provide reliable support for the acquisition of pmiRNAs through diet. To infer the potential biological activity of the identified pmiRNAs, we predicted their putative human mRNA targets, finding with great surprise that they appear to be mainly involved in neurogenesis and nervous system development. Unfortunately, no consensus was identified within the sequences of detected pmiRNAs, in order to justify their stability or capability to be preserved in human plasma. We believe that the issue regarding CKR still needs further clarifications, even if the present findings would offer a solid support that this hypothesis is not impossible.

Leaf-buds of Pistacia atlantica: a novel source of bioactive molecules with high anti-inflammatory, antioxidant, anti-tyrosinase and antimicrobial properties.

In this study, for the first time, the anti-inflammatory, antioxidant, anti-tyrosinase and antimicrobial property of P. atlantica Desf. subsp. atlantica leaf-bud extract have been investigated. The anti-inflammatory activity was determined in vivo by reducing carrageenan-induced hind paw edema in mice, while the antiradical function was evaluated using DPPH, total antioxidant capacity (TAC) and reduction power assays. The extract induced a significant reduction of the edema, from 1 to 6 h in a dose-dependent manner (150, 200 and 300 mg/kg). Histological observations of the inflamed tissues also confirmed this. An effective antioxidant activity of the plant samples was demonstrated, showing an EC50 = 0.183 ± 0.005 mg/mL for the DPPH test, a value of 28.776 ± 2.541 mg AAE/g for the TAC and an EC50 = 0.136 ± 0.003 mg/mL for reducing power. The leaf-bud extract also revealed a good antimicrobial activity against S. aureus and L. monocytogenes (mean diameter of inhibition zones of 13.2 and 17.0 mm, respectively), while a slight antifungal effect was observed. The plant preparation was then documented to inhibit tyrosinase activity, with an EC50 value of 0.098 ± 0.00 mg/mL in a dose-dependent manner. HPLC-DAD analysis revealed that dimethyl-allyl caffeic acid and rutin were the most abundant molecules. The current data documented that P. atlantica leaf-bud extract has strong biological properties and constitutes a potential source of pharmacological molecules.

Influence of environmental conditions on the production of nutraceuticals in Italian edible plant landraces.

Autochthonous plant varieties, also referred to as landraces, represent an important genetic resource, being well-adapted to the environment in which they have been selected. Landraces usually show profiles rich in nutraceuticals, making them an effective and valuable alternative to commercial agri-products, as well as potential candidates for crop improvement programs. Basilicata region is recognized as an Italian hotspot for agrobiodiversity, due to its complex orography. Thus, this work aimed to characterize and monitor, for two successive years, the content of secondary metabolites and related antioxidant properties of seven different species, four officinal (i.e., wild fennel - Feoniculum vulgare Mill.; oregano - Origanum vulgare L.; thyme - Thymus vulgaris L.; valerian - Valeriana officinalis L.) and three fruit species (i.e., fig - Ficus carica L. cv. Dottato; sweet cherry Prunus avium L. cv. Majatica; plum - Prunus domestica L. cv. Cascavella Gialla), collected in three different sites of this region. In detail, spectrophotometric tests were performed to assess the concentration of phenolic compounds, flavonoids, and - for officinal plants - also terpenoids, together with the antiradical activity (FRAP assays). In addition, to better typify the phytocomplexes of these landraces, HPLC-DAD and GC-MS analyses were carried out. In general, officinal plants showed higher values of nutraceutical compounds and related bioactivity with respect to fruit species. The data showed how different accessions of the same species had different phytochemical profiles, according to the sampling area and the year of collection, suggesting a role for both genetic and environmental factors in determining the observed results. Therefore, the final goal of this research was also to find a possible correlation between environmental factors and nutraceutics. The greatest correlation was found in valerian, where a lower water intake seemed to lead to a higher accumulation of antioxidants, and in plum, where the flavonoid content correlated positively with high temperatures. All these outcomes contribute at valorising Basilicata landraces for their aptitude to be high-quality foods and, at the same time, promoting the preservation of the agrobiodiversity for this region.

AKT-driven epithelial-mesenchymal transition is affected by copper bioavailability in HER2 negative breast cancer cells via a LOXL2-independent mechanism.

BACKGROUND: The main mechanism underlying cancer dissemination is the epithelial to mesenchymal transition (EMT). This process is orchestrated by cytokines like TGFß, involving "non-canonical" AKT- or STAT3-driven pathways. Recently, the alteration of copper homeostasis seems involved in the onset and progression of cancer. METHODS: We expose different breast cancer cell lines, including two triple negative (TNBC) ones, an HER2 enriched and one cell line representative of the Luminal A molecular subtype, to short- or long-term copper-chelation by triethylenetetramine (TRIEN). We analyse changes in the expression of EMT markers (E-cadherin, fibronectin, vimentin and αSMA), in the levels and activity of extracellular matrix components (LOXL2, fibronectin and MMP2/9) and of copper homeostasis markers by Western blot analyses, immunofluorescence, enzyme activity assays and RT-qPCR. Boyden Chamber and wound healing assays revealed the impact of copper chelation on cell migration. Additionally, we explored whether perturbation of copper homeostasis affects EMT prompted by TGFß. Metabolomic and lipidomic analyses were applied to search the effects of copper chelation on the metabolism of breast cancer cells. Finally, bioinformatics analysis of data on breast cancer patients obtained from different databases was employed to correlate changes in kinases and copper markers with patients' survival. RESULTS: Remarkably, only HER2 negative breast cancer cells differently responded to short- or long-term exposure to TRIEN, initially becoming more aggressive but, upon prolonged exposure, retrieving epithelial features, reducing their invasiveness. This phenomenon may be related to the different impact of the short and prolonged activation of the AKT kinase and to the repression of STAT3 signalling. Bioinformatics analyses confirmed the positive correlation of breast cancer patients' survival with AKT activation and up-regulation of CCS. Eventually, metabolomics studies demonstrate a prevalence of glycolysis over mitochondrial energetic metabolism and of lipidome changes in TNBC cells upon TRIEN treatment. CONCLUSIONS: We provide evidence of a pivotal role of copper in AKT-driven EMT activation, acting independently of HER2 in TNBC cells and via a profound change in their metabolism. Our results support the use of copper-chelators as an adjuvant therapeutic strategy for TNBC.

Exogenous miRNAs from Moringa oleifera Lam. recover a dysregulated lipid metabolism.

A balanced diet is critical for human health, and edible plants play an important role in providing essential micronutrients as well as specific microRNAs (miRNAs) that can regulate human gene expression. Here we present the effects of Moringa oleifera (MO) miRNAs (mol-miRs) on lipid metabolism. Through in silico studies we identified the potential genes involved in lipid metabolism targeted by mol-miRs. To this end, we tested the efficacy of an aqueous extract of MO seeds (MOES), as suggested in traditional African ethnomedicine, or its purified miRNAs. The biological properties of MO preparations were investigated using a human derived hepatoma cell line (HepG2) as a model. MOES treatment decreased intracellular lipid accumulation and induced apoptosis in HepG2. In the same cell line, transfection with mol-miRs showed similar effects to MOES. Moreover, the effect of the mol-miR pool was investigated in a pre-obese mouse model, in which treatment with mol-miRs was able to prevent dysregulation of lipid metabolism.

Neolithic dental calculi provide evidence for environmental proxies and consumption of wild edible fruits and herbs in central Apennines.

Looking for a biological fingerprint relative to new aspects of the relationship between humans and natural environment during prehistoric times is challenging. Although many issues still need to be addressed in terms of authentication and identification, microparticles hidden in ancient dental calculus can provide interesting information for bridging this gap of knowledge. Here, we show evidence about the role of edible plants for the early Neolithic individuals in the central Apennines of the Italian peninsula and relative cultural landscape. Dental calculi from human and animal specimens exhumed at Grotta Mora Cavorso (Lazio), one of the largest prehistoric burial deposits, have returned an archaeobotanical record made up of several types of palaeoecological proxies. The organic fraction of this matrix was investigated by a multidisciplinary approach, whose novelty consisted in the application of next generation sequencing to ancient plant DNA fragments, specifically codifying for maturase K barcode gene. Panicoideae and Triticeae starches, together with genetic indicators of Rosaceae fruits, figs, and Lamiaceae herbs, suggested subsistence practices most likely still based on wild plant resources. On the other hand, pollen, and non-pollen palynomorphs allowed us to outline a general vegetational framework dominated by woodland patches alternated with meadows, where semi-permanent settlements could have been established.

Pollen record of the Late Pleistocene-Holocene stratigraphic sequence and current plant biodiversity from Grotta Mora Cavorso (Simbruini Mountains, Central Italy).

Grotta Mora Cavorso (Jenne, Latium), a complex karstic system in Central Italy, has returned one of the most precious Prehistoric palaeontological and anthropological heritage. Through the analysis of pollen spectra and charcoals from cave stratigraphic levels (Late Pleistocene final phases-Holocene), the overall vegetation trend of the site was pointed out. Although taphonomy and palynology of cave deposits are complex, pollen assemblage represents a reliable source for inferring past vegetation; indeed, climatic, environmental, and cultural interactions determine fossil pollen record. Site formation processes and postdepositional bias should be generally considered in the analysis of stratigraphic sequences used to define paleoenvironmental conditions. The sediment deposits from Grotta Mora Cavorso showed a vegetation pattern point in out a progressive increase in woody plants from lower levels upward. Palynological investigations highlighted a changing environment predominantly characterized by cooler and perhaps more humid conditions than today, with plant subalpine and marsh communities nearby the cave. The ecological requirements of the identified plant taxa supplied useful indications to reconstruct ancient and modern environments of the Simbruini Mounts and the Upper Aniene River valley. This scenario, in accordance with previous faunistic and carpological findings and palynological analyses from Latium, provided a further perspective on the vegetation history, biodiversity, and climate of an important crossroads between the Adriatic and the Tyrrhenian coasts.

Phytochemical and biological investigations on the pollen from industrial hemp male inflorescences.

Cannabis sativa L. belongs to the Cannabaceae family and includes annual, robust, fast-growing and generally dioecious plants. Industrial hemp, and particularly the inflorescences, has been recently recognized as important source of bioactive extracts with antioxidant and antimicrobial effects. The goal of the present study was to explore botanical, phytochemical, and biological properties of water and hydroalcoholic hemp pollen extracts from male inflorescences. The extracts from hemp pollen were found rich in phenolic compounds, such as hydroxytyrosol, coumaric acid, and hesperitin. The phenolic profile was also consistent with the observed scavenging/reducing, enzyme inhibitory, and antimicrobial properties of the extracts. Regarding the antimicrobial effects, Escherichia coli, Trycophyton rubrum, and T. tonsurans were the most sensitive to growth inhibitory effects (MIC values: 9.92-79.37 µg/mL) of the extracts. Whereas, null effects on prostate PC3 and myocyte C2C12 cell viability, in the range 1-1000 µg/mL, are consistent with MIC values and suggest extracts' biocompatibility. The experimental data obtained, which are not reflected in the literature as the topic of hemp pollen is almost completely unexplored, confirm the innovativeness of a product obtained directly from bees, which in the face of greater variability and complexity can reserve promising applications in food, pharmaceutical, and cosmetic sectors.

Forensic Application of Genetic and Toxicological Analyses for the Identification and Characterization of the Opium Poppy (Papaver somniferum L.).

BACKGROUND: A reliable and science-based taxonomic determination of Papaver somniferum L. (opium poppy), the illegal species of the genus Papaver, may have practical and legal implications for law enforcement. P. somniferum is a controlled plant because of its narcotic substances, such as morphine and codeine. As poppy plants have rather similar morphological features, both chemical and genetic analysis are required in order to achieve an accurate characterization of such species. The chemical structures of alkaloids are extremely variable even within the same species, which is why the genetic approach may lead to a more scientific Papaver sp. differentiation. The aim of our study was the taxonomic identification of poppy capsules seized by the Italian Police Forces being considered as potential P. somniferum derivatives. METHODS: The alkaloids detected using gas chromatography/mass-spectrometry (GC/MS) were morphine, codeine, thebaine, noscapine, meconin, hydrocotarnine, and traces of papaverine. Further genetic analysis was carried out simultaneously using three plastid DNA barcoding regions (matK, trnH-psbA, and rbcL) for the samples' identification. RESULTS: The Random Amplification of Polymorphic DNA (RAPD) method showed that the analysed samples were genetically identical. CONCLUSIONS: The morphological, toxicological, and genetic profile of the samples revealed that they belonged to P. somniferum species. Furthermore, the alkaloid content of dried poppy capsules might be used to investigate and track their origin.

Phytochemicals and quality level of food plants grown in an aquaponics system.

BACKGROUND: Beyond nutrition, fruits and vegetables can be considered as natural sources of bioactive molecules, for which beneficial effects on human health are widely recognised. To improve food quality, soilless growing systems could represent a good strategy for promoting a sustainable food production chain, although the nutritional and nutraceutical properties of their products should be investigated in depth. The main quality traits and the volatile and non-volatile secondary metabolites of Solanum lycopersicum L., Petroselinum crispum (Mill.) Fuss and Ocimun basilicum L. grown in an aquaponics system and in organic farming were quantified and compared. RESULTS: On a fresh basis, soil-grown P. crispum and O. basilicum showed significantly higher total phenolics and antioxidant activity compared to aquaponic crops, whereas, on a dry basis, both plants showed opposite results. Soil-grown S. lycopersicum was significantly richer in total phenolics, whereas the aquaponic type showed a higher antioxidant activity. Aquaponics induced the accumulation of resveratrol in P. crispum, rosmarinic acid and myricetin in O. basilicum, and lycopene in S. lycopersicum. Among the volatile compounds, in O. basilicum, linalool was the main constituent in both treatments, whereas τ-cadinol represented the second constituent in aquaponic crops. The volatile profiles of P. crispum did not differ significantly between the two cultivation methods. CONCLUSION: The overall quality of organic and aquaponics cultures appeared to be comparable. The results showed that aquaponic farming method can be an innovative, rapid and sustainable way of producing quality food. © 2021 Society of Chemical Industry.

Exploiting the Potential in Water Cleanup from Metals and Nutrients of Desmodesmus sp. and Ampelodesmos mauritanicus.

Increasing levels of freshwater contaminants, mainly due to anthropogenic activities, have resulted in a great deal of interest in finding new eco-friendly, cost-effective and efficient methods for remediating polluted waters. The aim of this work was to assess the feasibility of using a green microalga Desmodesmus sp., a cyanobacterium Nostoc sp. and a hemicryptophyte Ampelodesmos mauritanicus to bioremediate a water polluted with an excess of nutrients (nitrogen and phosphorus) and heavy metals (copper and nickel). We immediately determined that Nostoc sp. was sensitive to metal toxicity, and thus Desmodesmus sp. was chosen for sequential tests with A. mauritanicus. First, A. mauritanicus plants were grown in the 'polluted' culture medium for seven days and were, then, substituted by Desmodesmus sp. for a further seven days (14 days in total). Heavy metals were shown to negatively affect both the growth rates and nutrient removal capacity. The sequential approach resulted in high metal removal rates in the single metal solutions up to 74% for Cu and 85% for Ni, while, in the bi-metal solutions, the removal rates were lower and showed a bias for Cu uptake. Single species controls showed better outcomes; however, further studies are necessary to investigate the behavior of new species.

Investigating the Drought and Salinity Effect on the Redox Components of Sulla Coronaria (L.) Medik.

For the Mediterranean region, climate models predict an acceleration of desertification processes, thus threatening agriculture. The present work aimed to investigate the effect of drought and salinity on Sulla coronaria (L.) Medik., a Mediterranean forage legume, for understanding plant defence systems activated by these stressors. In detail, we focused our attention on the variations on the plant redox status. Plants were subjected to suboptimal watering and irrigation with sodium chloride (NaCl) solutions. The same salt treatment was applied for in vitro tests on seedlings. Water content did not change after treatments. Salt negatively influenced seed germination and seedling development, but it did not affect photosynthesis parameters, contrary to what was observed in adult plants. Proline concentration increased in all samples, while abscisic acid level increased exclusively in seedlings. NaCl caused accumulation of superoxide anion in plants and seedlings and hydrogen peroxide only in seedlings; nevertheless, lipid peroxidation was not detected. Total phenolics, glutathione, expression level, and activity of antioxidant enzymes were assayed, revealing a complex antiradical molecular response, depending on the type of stress and development stage. Our results confirm Sulla as a drought- and salt-tolerant species and highlight its ability to counteract oxidative stress. This evidence suggests a key role for the redox components, as signal transduction messengers, in Sulla acclimation to desertification. Finally, plants and seedlings showed different acclimation capacity to salinity, revealing a greater genomic plasticity for seedlings.

Environmental implications and evidence of natural products from dental calculi of a Neolithic-Chalcolithic community (central Italy).

In this contribution, we investigated the role of plants in the prehistoric community of Casale del Dolce (Anagni, FR, central Italy), through microparticles recovered from dental calculus. The finding of a great amount of pollen types, even in form of compact lumps, could indicate use of natural substances, such as honeybee products and/or conifer resins. This plant-microremain record also suggested environmental implications relative to the Neolithic and Chalcolithic period. Additionally, the stability of the tartar microenvironment had preserved starches and other microparticles, such as one epidermal trichome, a sporangium, and fragments of plant tissue, rarely detected in ancient dental calculus. The detection of secondary metabolites in the ancient matrix confirmed the familiarity of this community with plant resources. All these data supply various interesting food for thought and expand the knowledge about the potential of dental calculus in archaeological and archaeobotanical fields with a special focus on palaeoecology.

Arabidopsis Defense against the Pathogenic Fungus Drechslera gigantea Is Dependent on the Integrity of the Unfolded Protein Response.

Drechslera gigantea Heald & Wolf is a worldwide-spread necrotrophic fungus closely related to the Bipolaris genus, well-known because many member species provoke severe diseases in cereal crops and studied because they produce sesterpenoid phytoxins named ophiobolins which possess interesting biological properties. The unfolded protein response (UPR) is a conserved mechanism protecting eukaryotic cells from the accumulation of unfolded/misfolded proteins in the endoplasmic reticulum (ER). In plants, consolidated evidence supports the role of UPR in the tolerance to abiotic stress, whereas much less information is available concerning the induction of ER stress by pathogen infection and consequent UPR elicitation as part of the defense response. In this study, the infection process of D. gigantea in Arabidopsis thaliana wild type and UPR-defective bzip28 bzip60 double mutant plants was comparatively investigated, with the aim to address the role of UPR in the expression of resistance to the fungal pathogen. The results of confocal microscopy, as well as of qRT-PCR transcript level analysis of UPR genes, proteomics, microRNAs expression profile and HPLC-based hormone analyses demonstrated that ophiobolin produced by the fungus during infection compromised ER integrity and that impairment of the IRE1/bZIP60 pathway of UPR hampered the full expression of resistance, thereby enhancing plant susceptibility to the pathogen.