Claudia Zadra

General and species-specific transcriptional responses to downy mildew infection in a susceptible (Vitis vinifera) and a resistant (V. riparia) grapevine species.

BackgroundDowny mildew is a destructive grapevine disease caused by Plasmopara viticola (Berk. and Curt.) Berl. and de Toni, which can only be controlled by intensive fungicide treatments. Natural sources of resistance from wild grapevine (Vitis) species are used in conventional breeding approaches, but the signals and effectors involved in resistance in this important crop species are not well understood.ResultsEarly transcriptional changes associated with P. viticola infection in susceptible V. vinifera and resistant V. riparia plants were analyzed using the Combimatrix microarray platform. Transcript levels were measured 12 and 24 h post-inoculation, reflecting the time points immediately preceding the onset of resistance in V. riparia, as determined by microscopic analysis. Our data indicate that resistance in V. riparia is induced after infection, and is not based on differences in basal gene expression between the two species. The strong and rapid transcriptional reprogramming involves the induction of pathogenesis-related proteins and enzymes required for the synthesis of phenylpropanoid-derived compounds, many of which are also induced, albeit to a lesser extent, in V. vinifera. More interestingly, resistance in V. riparia also involves the specific modulation of numerous transcripts encoding components of signal transduction cascades, hypersensitive reaction markers and genes involved in jasmonate biosynthesis. The limited transcriptional modulation in V. vinifera represents a weak attempted defense response rather than the activation of compatibility-specific pathways.ConclusionsSeveral candidate resistance genes were identified that could be exploited in future biotechnological approaches to increase disease resistance in susceptible grapevine species. Measurements of jasmonic acid and methyl jasmonate in infected leaves suggest that this hormone may also be involved in V. riparia resistance to P. viticola.

Hybrid solid anaerobic digestion batch: biomethane production and mass recovery from the organic fraction of solid waste

An experimental apparatus was constructed to perform hybrid solid anaerobic digestion batch processing of the organic fraction of municipal solid waste. The preliminary process was carried out with a high total solids concentration of about 33% w w−1 and with an initial organic load of about 340 kg VS kg−1. The fresh organic fraction to inoculum ratio used to enhance the anaerobic process start-up was 0.910 kg VS kg VS−1. The process was conducted by spreading the percolate on top of the mixture. The percolate was stored in a separate section of the apparatus with a mean hydraulic retention time of about 1 day. During the process, acetate, butyrate and propionate in the percolate reached concentrations ranging from 3000 to 11 000 mg L−1. In spite of these high concentrations, the biomethane produced from both the solid and the percolate was quite high, at about 210 NL kg VS−1. The digestate obtained at the end of the run showed rather good features for being classified as an organic fertilizer according to Italian law. However, a residual phytotoxicity level was detected by a standardized test showing a germination index of about 50%.

Effect of pH on potassium metabisulphite biocidic activity against yeast and human cell cultures

Potassium metabisulphite (PMB) is a common antimicrobial additive in the food industry. In aqueous solutions, PMB leads to complex equilibria according to its concentration, pH and temperature, and different chemical species can be present. In winemaking, PMB is used at low pH, suggesting that the biocidic activity is exerted by sulphur dioxide while, in other applications, it is employed at higher pH values with little if any dissociation. This observation leads to the question of which chemical form is biologically active. For this reason, Saccharomyces cerevisiae cells were subjected to PMB solutions at different pH values and analysed with a Fourier transform infrared spectroscopy (FTIR)-based bioassay, to assess the entity and the type of stress. Cell viability was determined and compared to the metabolomics (FTIR) stress indices, which revealed that the metabolomics fingerprint was an effective description of the cell health state. GC-MS metabolite profiles were obtained to describe (in detail) the changes caused by PMB in the fatty acids region. Human dermal fibroblasts (HDF) were also subjected to PMB stress at pH 7.0 and analysed with the FTIR protocol, in order to compare the response spectra of yeast and human cell cultures.

Diterpenoids and Triterpenoids from the Resin of Bursera microphylla and Their Cytotoxic Activity

A chemical study of the nonpolar fraction of a methanol-soluble extract of Bursera microphylla resin yielded a variety of di- and triterpenoids. In total, 15 compounds were isolated, of which three are new, namely, malabaricatrienone (1), malabaricatrienol (2), and microphyllanin (3). The antiproliferative activity of the major compounds was evaluated in different murine cancer cell lines (M12.C3.F6 and RAW264.7) and human cancer cells (A549, HeLa, and PC-3). The new compounds (1-3) did not show significant antiproliferative activity. The known compounds ariensin (4), burseran (5), and dihydroclusin diacetate (6) were effective against the RAW264.7 cell line, with IC50 values in the micromolar range.

Role of pathogen-induced volatiles in the Nicotiana tabacum–Golovinomyces cichoracearum interaction

Plant injuries activate signal transduction cascades mediated by the plant hormones, which lead to enhanced expression of defence related genes and/or to changes in the emission of volatile organic compounds that can act as semiochemicals. In this research we demostrated that infection with the biotrophic pathogen Golovinomyces cichoracearum (DC.) V.P. Heluta (ex Erysiphe cichoracearum DC.), the causal agent of powdery mildew, led in the susceptible host Nicotiana tabacum L. cv Havana 425 to an increased emission of volatile compounds including Methyl-jasmonate (MeJA), (E)-2-hexenal and (E)-β-ocimene. Furthermore we investigated the role of these volatiles in the plant-pathogen interaction. Exogenous application of MeJA induced in tobacco an increase in the transcripts level of the defence related genes lipoxygenase, allene oxide cyclase and defensin and a decrease in the severity of the infection. Qualitative and quantitative differences in volatile compounds emission were showed also in MeJA-treated plants, where the emission of (E)-β-ocimene was significantly increased instead (E)-2-hexenal was not detected. Application of (E)-2-hexenal reduced the severity of powdery mildew while application of (E)-β-ocimene did not. Since (E)-2-hexenal did not activate in tobacco the accumulation of the above reported genes transcripts and the plant cell death, the reduction of the infection severity could be attributable to its inhibitory activity on the fungal germ tube growth. Our data highlight the contributions of natural substances that can act, directly or indirectly, against phytopathogens. In the global context of sustainability, food safety and environmental protection, such semiochemicals represent an alternative and promising approach to integrated pest management.

Evaluation of benefits and risks associated with the agricultural use of organic wastes of pharmaceutical origin

Industrial fermentations for the production of pharmaceuticals generate large volumes of wastewater that can be biologically treated to recover plant nutrients through the application of pharmaceutical-derived wastes to the soil. Nevertheless, benefits and risks associated with their recovery are still unexplored. Thus, the aim of the present work was to characterize three potential organic residues (sludge, anaerobic digestate and compost) derived from the wastewater generated by the daptomycin production process. The main parameters evaluated were the physico-chemical properties, potential contaminants (heavy metals, pathogens and daptomycin residues), organic matter stabilization and the potential toxicity towards soil microorganisms and plants. The results showed that all the studied materials were characterized by high concentrations of plant macronutrients (N, P and K), making them suitable for agricultural reuse. Heavy metal contents and pathogens were under the limits established by European and Italian legislations, avoiding the risk of soil contamination. The compost showed the highest organic matter stabilization within the studied materials, whereas the sludge and the anaerobic digestate were characterized by large amounts of labile organic compounds. Although the pharmaceutical-derived fertilizers did not negatively affect the soil microorganisms, as demonstrated by the enzymatic activities, the sludge and the anaerobic digestate caused a moderate and strong phytotoxicity, respectively. The compost showed no toxic effect towards plant development and, moreover, it positively affected the germination and growth in lettuce and barley. The results obtained in the present study demonstrate that the valorization of pharmaceutical-derived materials through composting permits their agricultural reuse and also represents a suitable strategy to move towards a zero-waste production process for daptomycin.

Valorization of a pharmaceutical organic sludge through different composting treatments

Nowadays, the agricultural reuse of pharmaceutical sludge is still limited due to environmental and agronomic issues (e.g. low stabilization of the organic matter, phytotoxicity). The aim of the present study was to evaluate the characteristics of a pharmaceutical sludge derived from the daptomycin production and to study the possibility of improving its quality through composting. The pharmaceutical sludge showed high content of macronutrients (e.g. total Kjeldahl N content was 38 g kg-1), but it was also characterized by high salinity (7.9 dS m-1), phytotoxicity (germination index was 36.7%) and a low organic matter stabilization. Two different mixtures were prepared (mixture A: 70% sludge + 30% wood chips w/w, mixture B: 45% sludge + 45% wood chips + 10% cereal straw w/w) and treated through static composting using two different aeration systems: active and passive aeration. The mixtures resulted in the production of two different compost, and the evolution of process management parameters was different. The low total solids and organic matter content of mixture A led to the failure of the process. The addition of cereal straw in mixture B resulted in increased porosity and C/N ratio and, consequently, in an optimal development of the composting process (e.g. the final organic matter loss was 54.1% and 63.1% for the passively and actively aerated treatment, respectively). Both passively and actively aerated composting of mixture B improved the quality of the pharmaceutical sludge, by increasing its organic matter stabilization and removing phytotoxicity.

Correction to: Jasmonate-mediated defence responses, unlike salicylate-mediated responses, are involved in the recovery of grapevine from bois noir disease

CorrectionFollowing publication of the original article [1], it came to the attention of the authors that they had omitted to acknowledge the University of Parma. The Acknowledgement section should read as follows: “The authors kindly acknowledge the University of Parma (Department of Chemistry, Life Sciences and Environmental Sustainability; formerly Department of Life Sciences/Evolutionary and Functional Biology) for the transfer of funds obtained from the Ager project: GIALLUMI DELLA VITE: TECNOLOGIE INNOVATIVE PER LA DIAGNOSI E LO STUDIO DELLE INTERAZIONI PIANTA/PATOGENO, BANDO AGER VITICOLTURA DA VINO”.

Changes in the Composition of Soil Dissolved Organic Matter After Application of Poultry Manure

The aim of this study is to investigate if and how the fertilisation with poultry manure can affect the composition of dissolved organic matter (DOM) in soil solution. Moreover, these effects were studied in relation with different soil depths and time. In this study, the profiles and vertical distribution of selected biomarker compounds were also investigated to provide a fingerprint of manure contribution DOM. The results showed that the application of poultry manure affected the DOM content of soil immediately after the poultry manure application at different soil depths. Moreover, it was demonstrated, by studying the organic compounds in the soil solution, that sterol compounds as cholesterol and coprostanol might be used as biomarkers of animal manure amendment.

In vitro toxicity evaluation of estragole-containing preparations derived from Foeniculum vulgare Mill. (fennel) on HepG2 cells

Estragole, a common component of herbs and spices, is a wellknown genotoxic hepatocarcinogen in rodents, whereas its potential toxic effect in humans is still debated. In the European contest, one of the major sources of human exposure to this phytochemical is Foeniculum vulgare Mill. (fennel). Therefore, the aim of this study was to evaluate the in vitro toxicity of estragole in the context of two complex phytochemical mixtures derived from fennel: fennel seed powder (FSPw) and fennel seed essential oil (FSEO). The estragole-containing preparations were analysed for their ability to cause cytotoxicity, genotoxicity, apoptosis and cell cycle perturbation in the human hepatoma (HepG2) cell line. None of the tested concentrations of FSPw induced DNA damage, nor apoptosis or cell cycle perturbation. Although FSEO did not induce any genetic damage as well, it exerted marked dose-dependent apoptotic effects on HepG2 cells with a concurrent cell cycle arrest in G2/M at the highest tested dose. Although prospective analyses are required to clarify the observed toxic effects of FSEO, our results support the hypothesis that the genotoxicity of estragole may be significantly reduced or null in the context of botanical mixtures.