Biocontrol Efficacy of Metschnikowia spp. Yeasts in Organic Vineyards against Major Airborne Diseases of Table Grapes in the Field and in Postharvest.

The aim of this work was to evaluate the efficacy of two antagonistic yeasts, Metschnikowia pulcherrima strain MPR3 and M. fructicola strain NRRL Y-27328 (commercial product NOLI), applied in addition to the "on-farm biological treatments" (BIO) carried out during the production season, for the containment of powdery mildew and grey mould diseases on organic table grapes 'Italia'. The yeast strains were applied in the field three times, and their efficacy was evaluated during the production season and under postharvest conditions. Overall, M. pulcherrima MPR3 combined with BIO treatments reduced disease incidence caused by Erysiphe necator and disease incidence and severity caused by Botrytis cinerea with values between 67.8% and 86.2%, showing higher efficacy than BIO treatments applied alone and in combination with NOLI. Field treatments based on BIO+MPR3 maintained their performance also during fruit storage, protecting grape berries from grey mould development to a greater extent than the other treatments (disease reduction of about 98%). Thus, the presence of M. pulcherrima MPR3 seems to improve disease management both in the field and in postharvest environments, without negative impacts on grape microbial communities. These findings highlight the potential of M. pulcherrima MPR3 as a promising alternative strategy for disease control in organic vineyards and in postharvest, providing sustainable solutions to improve food quality and safety.

XRF analysis searching for fingerprint elemental profile in south-eastern Sicily tomatoes.

The implementation of analytical techniques able to certify food quality and origin in a fast and non-destructive way is becoming a widespread need in the agri-food sector. Among the physical non-destructive techniques, X-ray fluorescence (XRF) spectrometry is often used to analyze the elemental composition of biological samples. In this study, X-ray fluorescence (XRF) elemental profiles were measured on tomato samples belonging to different geographical areas in Sicily (Italy). The purpose of this investigation was aiming to establish a protocol for in-situ measurement and analysis able to provide quality assessment and traceability of PGI agri-food products, specifically sustaining health safety and self qualifying bio-chemical signature. In detail, sampling was performed in one of the most tomato productive area of south-eastern Sicily (Pachino district), characterised by a relative higher amount of Organic Carbon and Cation Exchange Capacity, and compared with samples from other growing areas of Sicily, falling in Ragusa province and Mt. Etna region. Experimental data were analyzed in the framework of multivariate analysis by using principal component analysis and further validated by discriminant analysis. The results show the presence of specific elemental signatures associated to several characterizing elements. This methodology establishes the possibility to disentangle a clear fingerprint pattern associated to the geographical origin of an agri-food product.

Applications of Delayed Luminescence for tomato fruit quality assessment across varied Sicilian cultivation zones.

The food industry places significant emphasis on ensuring quality and traceability as key components of a healthy diet. To cater to consumer demands, researchers have prioritized the development of analytical techniques that can rapidly and non-invasively provide data on quality parameters. In this study, we propose to use the Delayed Luminescence (DL), an ultra-weak and photo-induced emission of optical photons, as a tool for a rapid evaluation of quality profile associated with fruit ripening, in support of traditional analysis methods. Delayed Luminescence measurements have been performed on cherry tomatoes, with and without the PGI "Pomodoro di Pachino" certification, harvested from two different growing areas of south-eastern Sicily (Italy). Then, DL emissions were correlated with soluble solid content and titratable acidity values, which are known to affect the flavor, the commerciality and the maturity degree of tomato fruits. In addition, we evaluated the changes in the DL parameters with respect to the geographical origin of the cherry tomatoes, with the aim of testing the possibility of applying the technique for identification purposes. The signals of Delayed Luminescence appeared to be good indicators of the macromolecular structure of the biological system, revealing structural changes related to the content of total soluble solids present in the juice of tomatoes analyzed, and they appeared unsuitable for authenticating vegetable crops, since the differences in the photon yields emitted by tomato Lots were not related to territory of origin. Thus, our results suggest that DL can be used as a nondestructive indicator of important parameters linked to tomato fruit quality.

Assessing Copper-Alternative Products for the Control of Pre- and Postharvest Citrus Anthracnose.

Citrus production is worldwide threatened by Colletotrichum spp., causal agents of pre- and postharvest anthracnose. The recent limitation on the use of copper-based antimicrobials, due to its demonstrated noxious effect on the environment, makes the control of this pathogen difficult. Thus, alternative products able to reduce/phase out copper in organic citrus farming are needed. In this study, the efficacy of 11 commercial alternative products were evaluated in vitro, in growth chamber, in open field and in postharvest environments. In vitro, mineral fertilizers, basic substances, essential oils, plant defense stimulators and biological control agents were able to inhibit the mycelial growth with variable efficacy. On artificially infected citrus fruit, almost all tested products significantly reduced disease incidence and severity, but with lower efficacy than copper. The efficacy of mineral fertilizers-based Kiram and Vitibiosap 458 Plus, citrus essential oil-based Prev-Am Plus and chitosan-based Biorend was confirmed in open field trials, in naturally infected citrus fruits. In these trials Biorend was the best alternative product, significantly reducing disease incidence (71% DI reduction) with better results than copper (47.5%). Field treatments reduced the incidence and severity of the disease in postharvest conditions, especially in fruits field-treated three times. Overall, selected products tested in open field can represent a good alternative to copper compounds in the view of future limitation of its use.

Efficacy and mechanism of action of food isolated yeasts in the control of Aspergillus flavus growth on pistachio nuts.

Due to its nutritional characteristics, dried fruit and in particular pistachio is considered an important component in the daily diet. Unfortunately, pistachio nuts can be contaminated during storage with a wide range of pathogenic fungi, including Aspergillus flavus. The present work has evaluated how the use of two yeast strains belonging to the species Wickerhamomyces anomalus and Metschnikowia pulcherrima can inhibit the in vitro and in vivo growth of A. flavus. Both yeast strains demonstrated a good in vitro antifungal activity attributable to a specific mechanism of action, although higher efficacy was evidenced by W. anomalus strain. The production of volatile organic compounds (VOCs) and lytic enzymes was hypothesized as the main mechanisms of action exerted by W. anomalus, while the nutritional competition for iron was assumed as the main biocontrol mechanism for M. pulcherrima. Moreover, from the results of the in vivo test carried out on artificially infected pistachio seeds, it was clear as M. pulcherrima strain showed the same efficacy of W. anomalus in inhibiting the growth and sporulation of A. flavus mold, despite W. anomalus was the most effective strain during in vitro assay. Altogether, these results indicate that both strains could be considered as potential biocontrol agents against A. flavus fungal growth, notwithstanding it is always important considering the tritrophic interaction (yeast-mold- host), which could play a crucial role in determining the final results.

Epiphytic and endophytic microorganisms associated to different cultivar of tomato fruits in greenhouse environment and characterization of beneficial bacterial strains for the control of post-harvest tomato pathogens.

Several bacterial and fungal diseases affect greenhouse-grown tomato crops, causing severe annual yield losses worldwide. The need to reduce chemical compound applications has encouraged the search of alternative approach for the control of tomato diseases, including the use of biological control agents. The presence of total and beneficial microbial populations was investigated on the surface and in the pulp of seven cultivars of tomato fruit coming from eleven greenhouses in the Pachino district (south-east of Sicily), recognized by the European Community with the "Protected Geographical Indication" label. Principal component analysis (PCA) showed that epiphytic and endophytic microbial populations clustered into groups according to the areas of origin. Approximately 240 tomato fruit-associated bacterial isolates were selected and a high percentage of them showed antagonistic activity against Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato, Xanthomonas euvesicatoria pv. perforans, Botrytis cinerea and Alternaria alternata. Analysis of the 16S rRNA gene sequences revealed a predominance of bacteria in Bacillus and Pseudomonas genera, followed by Citrobacter and Enterobacter. The presence of these genera differed according to the geographical areas of tomato samples, whereas their antagonistic capabilities varied according to the five tomato pathogens. The in vitro effectiveness of eight representative bacterial strains belonging to Pseudomonas, Bacillus and Enterobacter genera was confirmed in in vivo assays, carried out on tomato fruit artificially inoculated with A. alternata and B. cinerea. Altogether, these results revealed differences in population density of native microbiota of "Pomodoro di Pachino" fruits and allowed the selection of antagonistic bacterial strains that can be applied in field and in postharvest conditions as alternatives to chemical compounds.

COS-OGA Applications in Organic Vineyard Manage Major Airborne Diseases and Maintain Postharvest Quality of Wine Grapes.

In most wine-growing countries of the world the interest for organic viticulture and eco-friendly grape production processes increased significantly in the last decade. Organic viticulture is currently dependent on the availability of Cu and S compounds, but their massive use over time has led to negative effects on environment health. Consequently, the purpose of this study was to evaluate the effectiveness of alternative and sustainable treatments against powdery mildew, gray mold and sour rot under the field conditions on Nero d'Avola and Inzolia Sicilian cultivars. In detail, the efficacy of COS-OGA, composed by a complex of oligochitosans and oligopectates, and its effects in combination with arbuscular mycorrhizal fungi (AMF) were evaluated to reduce airborne disease infections of grape. COS-OGA combined with AMF induced a significant reduction in powdery mildew severity both on Nero d'Avola and Inzolia with a mean percentage decrease of about 15% and 33%, respectively. Moreover, COS-OGA alone and combined with AMF gave a good protection against gray mold and sour rot with results similar to the Cu-S complex (performance in disease reduction ranging from 65 to 100%) on tested cultivars. Similarly, the COS-OGA and AMF integration provided good performances in enhancing average yield and did not negatively impact quality and microbial communities of wine grape. Overall, COS-OGA alone and in combination could be proposed as a valid and safer option for the sustainable management of the main grapevine pathogens in organic agroecosystems.

Bioprospecting of Beneficial Bacteria Traits Associated With Tomato Root in Greenhouse Environment Reveals That Sampling Sites Impact More Than the Root Compartment.

Tomato is subject to several diseases that affect both field- and greenhouse-grown crops. To select cost-effective potential biocontrol agents, we used laboratory throughput screening to identify bacterial strains with versatile characteristics suitable for multipurpose uses. The natural diversity of tomato root-associated bacterial communities was bioprospected under a real-world environment represented by an intensive tomato cultivation area characterized by extraseasonal productions in the greenhouse. Approximately 400 tomato root-associated bacterial isolates, in majority Gram-negative bacteria, were isolated from three compartments: the soil close to the root surface (rhizosphere, R), the root surface (rhizoplane, RP), and the root interior (endorhizosphere, E). A total of 33% of the isolates produced siderophores and were able to solubilize phosphates and grow on NA with 8% NaCl. A total of 30% of the root-associated bacteria showed antagonistic activity against all the tomato pathogens tested, i.e., Clavibacter michiganesis pv. michiganensis, Pseudomonas syringae pv. tomato, Pseudomonas corrugata and Xanthomonas euvesicatoria pv. perforans, and Fusarium oxysporum f. sp. lycopersici. We found that the sampling site rather than the root compartment of isolation influenced bacterial composition in terms of analyzed phenotype. This was demonstrated through a diversity analysis including general characteristics and PGPR traits, as well as biocontrol activity in vitro. Analysis of 16S rRNA gene (rDNA) sequencing of 77 culturable endophytic bacteria that shared multiple beneficial activity revealed a predominance of bacteria in Bacillales, Enterobacteriales, and Pseudomonadales. Their in vitro antagonistic activity showed that Bacillus species were significantly more active than the isolates in the other taxonomic group. In planta activity against phytopathogenic bacteria of a subset of Bacillus and Pseudomonas isolates was also assessed.

Combined application of antagonistic Wickerhamomyces anomalus BS91 strain and Cynara cardunculus L. leaf extracts for the control of postharvest decay of citrus fruit.

Combined use of biocontrol agents and plant extracts can be considered a viable and promising strategy for protecting plant tissues with different synergistic mechanisms of action that improve the antimicrobial activity of the mixtures. Treatments of citrus fruits with Wickerhamomyces anomalus BS91 have been previously reported as effective measures to reduce the incidence of green mold caused by Penicillium digitatum. On the opposite, the knowledge of the antifungal activity of cultivated cardoon (Cynara cardunculus L. var. altilis DC.) leaf extract, vegetable widespread in some Mediterranean areas, is still very limited. In this study, experimental trials were conducted to evaluate the effectiveness in vitro of leaf aqueous, methanolic and ethanolic extracts of C. cardunculus against seven fungal pathogens responsible for considerable food losses in the postharvest stage. In addition, biocontrol yeast W. anomalus BS91 and the three C. cardunculus extracts were tested in vivo both as a single treatment and in mixture, against Penicillium digitatum on 'Tarocco' oranges and 'Femminello' lemons. The combination of W. anomalus BS91 and leaf ethanolic extract reduced with the highest efficacy the incidence and severity of green mold on orange and lemon fruits with respect to the control, and was more effective than treatment with antagonistic yeast or leaf extracts applied alone. Incidence and severity of citrus decay were more consistently reduced when mixtures were applied 24 h before the inoculation of the pathogen, thus suggesting the relevance of preventive treatments. The mixtures of antagonistic W. anomalus BS91 and ethanolic leaf extract were more effective in controlling green mold decay on oranges than on lemons. These results indicate that biocontrol agents and leaf extracts, used in appropriate combination, can provide a stronger protection than when used singularly. However, compatibility between microbial antagonist and antimicrobial extract should be preliminary verified.

Role of biocontrol yeasts Debaryomyces hansenii and Wickerhamomyces anomalus in plants' defence mechanisms against Monilinia fructicola in apple fruits.

The role of killer yeasts of the species Debaryomyces hansenii and Wickerhamomyces anomalus in biocontrol of Monilinia fructicola, and their involvement in plant defence mechanisms against brown rot in apple fruits, were investigated. D. hansenii KI2a and W. anomalus BS91 strains showed the highest in vitro biocontrol activity, inhibiting mycelium growth by 69.53% and 66.08% respectively, as compared to control fungal cultures. Brown rot on apple fruits was significantly reduced by BS91 and two strains of D. hansenii KI2a and AII4b by 92.46%, 85.10% and 70.02%, respectively, in comparison to infected fruits, which did not receive any pre-treatment. In enzymatic tests, the most significant changes in peroxidase (POD) and catalase (CAT) activities were observed in fruits inoculated either with BS91 followed by M. fructicola infection or with AII4b yeast strain alone, where POD activities were significantly higher by 67% and 54%, respectively, and CAT activities were significantly lower by 65% and 68%, respectively, than in untreated control fruits. These results confirmed the ability of killer yeasts to influence host-defence related enzyme activities in apple fruit tissue and may suggest that AII4b killer strain has a potential as biocontrol agent prior to infection by triggering immune response mechanisms in plant tissue, whereas BS91 strain is the most effective during pathogen infection and could be used as biocontrol agent in postharvest disease onset. Accordingly, the antagonistic strains of W. anomalus BS91 and D. hansenii KI2a and AII4b could be proposed as active ingredients for the development of biofungicide against M. fructicola.

Volatile organic compounds (VOCs) produced by biocontrol yeasts.

The Volatile organic compounds (VOCs) produced by biocontrol yeast strains which belong to the Wickerhamomyces anomalus, Metschnikowia pulcherrima, Aureobasidium pullulans and Saccharomyces cerevisiae species were identified by solid phase microextraction (SPME) coupled with Gas Chromatography-Mass Spectrometry (GC-MS). Alcohols (ethyl alcohol, 3-methyl-1-butanol and phenylethyl alcohol) and esters (ethyl acetate and isoamyl acetate) were found to be the main VOCs emitted by the yeast strains, which had different production rate over a 16-day period. In addition, the tested yeast strains showed a remarkable ability to consume oxygen and to produce high percentages of carbon dioxide over a 5 days incubation period in a model system. The yeast strains, which were proven to very efficiently suppress in vivo the growth of postharvest fungal by VOCs, also quickly produced high percentages of ethyl acetate and carbon dioxide. . For all these reasons, we believe that the level of yeast biocontrol efficacy through the production of volatiles could be the result of a synergistic effect between VOCs and carbon dioxide in the packaging environment.

Edible coatings incorporating pomegranate peel extract and biocontrol yeast to reduce Penicillium digitatum postharvest decay of oranges.

This study investigated the potential use of two edible coatings, chitosan (CH) and locust bean gum (LBG), which incorporated chemically characterized water pomegranate peel extract (WPPE) or methanol pomegranate peel extract (MPPE) and the biocontrol agent (BCA) Wickerhamomyces anomalus, to control the growth of Penicillium digitatum and to reduce the postharvest decay of oranges. CH and LBG including pomegranate peel extracts (PPEs) at different concentrations were tested in vitro against P. digitatum to determine their antifungal efficacy; at the same time, the tolerance of viable cells of W. anomalus to increasing concentrations of WPPE and MPPE extracts was assessed. The potential application of selected bioactive coatings was evaluated in vivo on oranges, which had been artificially inoculated with P. digitatum, causal agent of green mold decay. CH incorporating MPPE or WPPE at all concentrations was able to inhibit in vitro P. digitatum, while LBG was active only at the highest MPPE or WPPE concentrations. W. anomalus BS91 was slightly inhibited only by MPPE-modified coatings, while no inhibition was observed by WPPE, which was therefore selected for the in vivo trials on oranges artificially inoculated with P. digitatum. The experimental results proved that the addition of 0.361 g dry WPPE/mL, both to CH and LBG coatings, significantly reduced disease incidence (DI) by 49 and 28% respectively, with respect to the relative controls. Besides the combination CH or LBG + WPPE, the addition of W. anomalus cells to coatings strengthened the antifungal effect with respect to the relative controls, as demonstrated by the significant reduction of DI (up to 95 and 75% respectively). The findings of the study contribute to the valorization of a value-added industrial byproduct and provide a significant advancement in the development of new food protectant formulations, which benefit from the synergistic effect between biocontrol agents and natural bioactive compounds.

Effects of Sublabeled Rates of Dazomet and Metam-Sodium Applied Under Low-Permeability Films on Calonectria Microsclerotia Survival.

Infested soil is the primary inoculum source for Calonectria spp. for initiating disease in ornamental and forestry crops. The effects of dazomet and metam-sodium on survival of microsclerotia of 28 isolates belonging to 19 Calonectria spp. were evaluated in this study under nursery conditions. Two experiments with exotic Calonectria spp. in plastic containers in a greenhouse and three trials with endemic species in field plots were performed during different seasons. The containers and plots were artificially infested with Calonectria microsclerotia differentiated on carnation leaf tissues. Basamid (dazomet) was applied at 100, 160, 200, 400, and 500 kg/ha, while Divapan (metam-sodium) was applied at 250, 350, 400, 700, and 1,000 liters/ha in both the containers and plots. The fumigants were applied under virtually and totally impermeable films. Fungal survival was evaluated after 21 days using leaf tissues collected from treated soil and plated on potato dextrose agar, and the ability of microsclerotia to cause infection was tested on red clover. The survival of Calonectria inocula and microsclerotia decreased with increasing fumigant rates. In the greenhouse trials, where Basamid was applied at 200, 400, and 500 kg/ha and Divapan at 400, 700, and 1,000 liters/ha, no viable microsclerotia were recovered for 14 exotic Calonectria spp., whereas viable inocula of Calonectria hongkongensis, C. naviculata, and C. sulawesiensis were retrieved from the fumigated plots. Low rates of Basamid (100 and 160 kg/ha) and Divapan (250 and 350 liters/ha) were less effective at reducing Calonectria viability and, for these treatments, the rate of microsclerotia survival was highly variable among the different isolates and species. Furthermore, totally impermeable film significantly enhanced fumigant performance. Relative to endemic Calonectria spp., all of the treatments killed microsclerotia of C. polizzii and C. pauciramosa independent from fumigant, rate, and film. This research demonstrated the possibility of reducing the application rates by up to 160 kg/ha for Basamid and 400 liters/ha for Divapan under low-permeability films (virtually impermeable film or totally impermeable film) for eradicating or reducing the primary inoculum of Calonectria spp. in soil.

Performance evaluation of volatile organic compounds by antagonistic yeasts immobilized on hydrogel spheres against gray, green and blue postharvest decays.

Wickerhamomyces anomalus, Metschnikowia pulcherrima, Aureobasidium pullulans and Saccharomyces cerevisiae yeasts were tested for their ability to survive and synthesize antifungal volatile organic compounds (VOCs) both in vitro and in vivo conditions when immobilized on commercial hydrogel spheres. The results showed a good survival of all yeasts on hydrogel spheres up to 10 days of incubation. Moreover, VOCs produced in vitro by tested yeasts inhibited Botrytis cinerea, Penicillium digitatum and P. italicum radial growth and conidial germination, with the highest antagonistic activity reported for W. anomalus and A. pullulans strains. Experimental in vivo trials performed on strawberry and mandarin fruits proved the ability of VOCs to reduce significantly postharvest decays on artificially wounded tissues. Comprehensively, the best efficacy was detected for W. anomalus, which totally inhibited gray mold decay on strawberry fruits and significantly reduced green mold infections on mandarin fruits. On the other hand, blue mold decay on mandarin fruits was more effectively managed by A. pullulans VOCs. Accordingly, hydrogel spheres used as a support for VOC-generating yeasts could open a new way for the employment of this polymeric material as a bio-emitter in postharvest packaging.

Potential Role of Exoglucanase Genes (WaEXG1 and WaEXG2) in the Biocontrol Activity of Wickerhamomyces anomalus.

The use of yeasts, including Wickerhamomyces anomalus, as biocontrol agents of fungi responsible for postharvest diseases of fruits and vegetables has been investigated for the past two decades. Among a variety of mechanisms, the production of glucanases coded by the "killer genes" WaEXG1 and WaEXG2 have been reported to play a role in the ability of yeast to inhibit other fungi. The objective of the present study was to determine the expression of these genes by RT-qPCR, utilizing gene-specific primers, when W. anomalus was grown on grape berries and oranges that were either non-inoculated or inoculated with Botrytis cinerea or Penicillium digitatum, or in minimal media supplemented with cell walls of various plant pathogens and different amounts of glucose. Results indicated that WaEXG2 was more responsive than WaEXG1 to the nutritional environment (including the addition of glucose to cell wall-amended media) in vitro and appeared to play a greater role in the cellular metabolism of W. anomalus. WaEXG2 expression also appeared to be more responsive to the presence of cell walls of P. digitatum and B. cinerea than other fungal species, whereas the same level of induction was not seen in vivo when the yeast was grown in wounded/pathogen-inoculated fruits.

Postharvest biocontrol ability of killer yeasts against Monilinia fructigena and Monilinia fructicola on stone fruit.

The antagonistic effects of Debaryomyces hansenii KI2a, D. hansenii MI1a and Wickerhamomyces anomalus BS91 were tested against Monilinia fructigena and Monilinia fructicola in in vitro and in vivo trials. All yeast strains demonstrated antifungal activity at different levels depending on species, strain and pathogen. D hansenii KI2a and W. anomalus BS91 showed the highest biocontrol activity in vitro; the production of hydrolytic enzymes, killer toxins and volatile organic compounds (VOCs) were hypothesized as their main mechanisms of action against pathogens. D hansenii KI2a and W. anomalus BS91 significantly reduced brown rot incidence and severity on peach and plum fruits artificially inoculated with M. fructigena and M. fructicola, especially when applied 24 h before pathogen inoculation. On the opposite, D. hansenii MI1a exhibited weak antagonistic activity towards M. fructigena on peach and plum fruits and was ineffective against M. fructicola. The noticeable ability of W. anomalus BS91 to control brown rot could be also correlated with its high capacity to colonize the wound tissue and to increase its population density. Accordingly, the antagonistic strains of D. hansenii and W. anomalus could be proposed as active ingredients for the development of biofungicides against Monilinia species that are responsible for considerable economic losses in stone fruit crops.

The effect of locust bean gum (LBG)-based edible coatings carrying biocontrol yeasts against Penicillium digitatum and Penicillium italicum causal agents of postharvest decay of mandarin fruit.

Strains belonging to Wickerhamomyces anomalus, Metschnikowia pulcherrima and Aureobasidium pullulans species were tested in vitro as biocontrol agents (BCAs) against the post-harvest pathogenic molds Penicillium digitatum and Penicillium italicum. Moreover, studies aimed at screening the antifungal activity of selected yeast strains in vivo conditions against P. digitatum and P. italicum, and investigated the efficacy of a polysaccharidic matrix, locust bean gum (LBG), enriched with the tested BCAs, in controlling postharvest decays in artificially inoculated mandarins. The population dynamics of BCAs on wounds and the magnitude of peroxidase (POD) and superoxide dismutase (SOD) in fruit tissues were also investigated after treatments of mandarins with antagonistic yeasts. W. anomalus BS91, M. pulcherrima MPR3 and A. pullulans PI1 provided excellent control of postharvest decays caused by P. digitatum and P. italicum on mandarins, both when the yeasts were used alone and in combination with LBG, which enhanced the yeast cell viability over time. Finally, the increased activity of POD and lower decrease in SOD activity in response to BCAs application in mandarin fruits confirmed their involvement in the biocontrol mechanism.

Draft genome of a Xanthomonas perforans strain associated with pith necrosis.

Xanthomonas perforans causes bacterial spot of tomato and pepper. A genome draft of an unusual isolate (strain 4P1S2), differing in that it was associated with stem pith necrosis, was assembled from Illumina MiSeq sequencing data using the draft of X. perforans strain 91-118 as a reference. The resulting draft (accession number JRWW00000000) largely overlapped with the reference draft. In addition, the reads not mapping on the reference assembly were selected and used for a further assembly, that revealed a large putative plasmid. The analysis of the predicted proteins showed only few gene features that could be potentially implicated in the switch of a phytopathological behavior.

Biocontrol ability and action mechanism of food-isolated yeast strains against Botrytis cinerea causing post-harvest bunch rot of table grape.

Strains belonging to the species Saccharomyces cerevisiae, Wickerhamomyces anomalus, Metschnikowia pulcherrima and Aureobasidium pullulans, isolated from different food sources, were tested in vitro as biocontrol agents (BCAs) against the post-harvest pathogenic mold Botrytis cinerea. All yeast strains demonstrated antifungal activity at different levels depending on species and medium. Killer strains of W. anomalus and S. cerevisiae showed the highest biocontrol in vitro activity, as demonstrated by largest inhibition halos. The competition for iron and the ability to form biofilm and to colonize fruit wounds were hypothesized as the main action mechanisms for M. pulcherrima. The production of hydrolytic enzymes and the ability to colonize the wounds were the most important mechanisms for biocontrol activity in A. pullulans and W. anomalus, which also showed high ability to form biofilm. The production of volatile organic compounds (VOCs) with in vitro and in vivo inhibitory effect on pathogen growth was observed for the species W. anomalus, S. cerevisiae and M. pulcherrima. Our study clearly indicates that multiple modes of action may explain as M. pulcherrima provide excellent control of postharvest botrytis bunch rot of grape.

Effects of Fungicide Treatments for the Control of Epidemic and Exotic Calonectria Diseases in Italy.

The efficacy of 11 fungicides was evaluated for the control of Calonectria infections on bottlebrush (Callistemon "Masotti") and feijoa (Acca sellowiana), with special emphasis on Calonectria pauciramosa and C. morganii, which are the most destructive species in Italian ornamental nurseries. Three nursery experiments were performed with the selected fungicides in order to determine their ability to prevent leaf spot caused by C. morganii on bottlebrush and leaf spot and crown and root rot caused by C. pauciramosa on bottlebrush and feijoa. All fungicides were effective in reducing disease infections, except for cyproconazole, propamocarb + fosetyl-Al, and K phosphite that were the least effective in reducing C. morganii leaf spot. In contrast, K phosphite proved more effective in reducing crown and root rot caused by C. pauciramosa. Fungicides were also evaluated in growth-cabinet experiments for their ability to reduce incidence and severity of leaf spot on bottlebrush caused by the exotic pathogens C. pseudomexicana, C. tunisiana, C. polizzii, and C. mexicana. Copper hydroxide, fosetyl-Al, prochloraz, prochloraz + cyproconazole, and tebuconazole were always effective in reducing Calonectria leaf spot on bottlebrush. However, some differences in levels of control might be attributable to Calonectria isolate. Overall, this study clearly indicates that new fungicides can be employed for chemical management of Calonectria infections in ornamental nurseries.