Coupling botanical epidemiology and mathematical modeling for the control of Botrytis cinerea in vineyards

Il marciume del grappolo, causato da Botrytis cinerea, è una delle principali malattie che colpiscono la vite. Botrytis cinerea si sviluppa e cresce attivamente come patogeno necrotrofo e saprofita su diversi substrati. In vigneto, B. cinerea può produrre un ampio numero di conidi in un’ampia gamma di condizioni ambientali. Botrytis cinerea ha diversi pathway d’infezione e le infezioni occorrono principalmente in due periodi: dalla fioritura allo sviluppo dei giovani grappoli, e dopo invaiatura alla raccolta. A causa della complessità del ciclo di vita di B. cinerea e all’esistenza di questi diversi pathway d’infezione, i fungicidi vengono normalmente applicati a fine della fioritura (A), pre-chiusura del grappolo (B), invaiatura (C), e prima della raccolta (D). Le applicazioni durante la prima parte della stagione (A e B) hanno come obiettivo: i) la riduzione della germinazione dei conidi e le infezioni fiorali, ii) la prevenzione delle infezioni latenti delle bacche, e iii) la disinfezione dei residui di fioritura. Le applicazioni successive all’invaiatura (C e D) hanno lo scopo non solo di prevenire le infezioni durante la maturazione ma anche di disinfestare i residui di fioritura rimasti all’interno del grappolo. Questa strategia, basata su quattro trattamenti, garantisce una buona protezione contro il marciume del grappolo in vigneto, tuttavia presenta alcuni limiti: i) si tratta di un approccio preventivo che non considera il reale rischio d’infezione, con il conseguente effetto di eseguire trattamenti non necessari; ii) aumenta la probabilità che si sviluppino popolazioni resistenti ai fungicidi; iii) implica potenziali risvolti negativi sulla salute umana e sull’ambiente. Negli ultimi anni, nuove tipologie di prodotti, come gli agenti di biocontrollo (BCAs) e i botanicals, sono state proposte per ridurre l’uso eccessivo di prodotti chimici e aumentare la sostenibilità del controllo del marciume del grappolo in vigneto. Lo scopo di questa tesi di Dottorato è stato: i) comparare diverse strategie di controllo e valutare l’interazione tra fungicidi con diversi timing di applicazione, ii) valutare l’effetto sulla colonizzazione e sporulazione di B. cinerea su residui fiorali di diversi prodotti (fungicidi, agenti di biocontrollo, e botanicals) applicati con diversi timing; iii) valutare un modello meccanicistico sviluppato da González-Domínguez et al. (2015); e iv) includere all’interno di un modello per il biocontrollo, sviluppato da Jeger et al. (2009), l’effetto delle condizioni ambientali sulle interazioni BCA-patogeno.Botrytis bunch rot (BBR), caused by Botrytis cinerea, is one of the main diseases affecting grapevine. Botrytis cinerea develops and actively grows as necrotrophic pathogen and as saprophyte on different substrates. In grapevine, B. cinerea can produce a large number of conidia on bunch and leaf trash and rotted berries under a wide range of environmental conditions. Botrytis cinerea has multiple infection pathways and infection mainly occurs in two periods: from flowering to young cluster development, and after veraison to harvest. Due to the complexity of the B. cinerea life cycle and the existence of these different infection pathways, fungicides are usually applied at the end of flowering (A), pre-bunch closure (B), veraison (C), and before harvest (D). The early season applications (A and B) are aimed at: i) reducing conidial germination and infection of flowers, ii) preventing latent infections of berries, and iii) disinfesting the bunch trash. The later-season applications (i.e., the applications from veraison until harvest; C and D) are aimed not only at preventing berry infection during ripening but also at disinfesting bunch trash in order to reduce the inoculum load. This fungicide schedule may results in unnecessary sprays because the applications are preventive and do not take into account the real risk of BBR infections. In the last decades, new products, like biological control agents (BCAs) and botanicals, have been proposed for reducing the excessive use of chemicals in agriculture and increase the sustainable of BBR control in vineyards. The aim of this Doctoral work was to: i) compare different control strategies and evaluate the interactions among fungicides applied at different timings; ii) investigate the effects of different products (fungicides, biological control agents, and botanicals) applied at different timings on B. cinerea bunch trash colonization and sporulation; iii) evaluate a weather-driven mechanistic model developed by González-Domínguez et al. (2015); and iv) include into a model for biological control, developed by Jeger et al. (2009), the effect of environmental conditions on the pathogen-BCA interactions

Quantification of Botrytis cinerea in Grapevine Bunch Trash by Real-Time PCR

Quantification of colonization of grape bunch trash by Botrytis cinerea is crucial for Botrytis bunch rot (BBR) control. A previously developed quantitative polymerase chain reaction (qPCR) method was adapted to quantify B. cinerea DNA in grape bunch trash, and a colonization coefficient (CC) was calculated as the ratio between the DNA concentrations of B. cinerea and of Vitis vinifera. CC values increased linearly with the number of conidia of B. cinerea or the quantity of mycelium of B. cinerea added to the bunch trash increased. CC values also increased linearly in bunch trash samples containing increasing percentages of B. cinerea-colonized bunch trash; in the latter samples, CC values were correlated with subsequent assessments of B. cinerea colonization of trash (as determined by plating on agar) and sporulation on the trash (as determined by spore counts after incubation in humid chambers). The qPCR assay was also validated using trash collected from bunches treated or not treated with fungicides in three vineyards in two seasons. CC values reflected the reduction in sporulation and in latent infections of mature berries caused by fungicide application. The qPCR assay enables rapid, specific, sensitive, and reliable quantification of the degree of colonization of bunch trash by B. cinerea, which makes it a useful tool for studies of the epidemiology and management of BBR

Modelling Biocontrol Agents as Plant Protection Tools

In recent years, researchers have increasingly explored sustainable tools for plant protection against pathogens, including the use of biological control agents (BCAs), which have the potential to complement or replace chemical fungicides. However, global reliance on their use remains relatively insignificant and the factors influencing their efficacy remain unclear. The complex interactions among a target pathogen, a host plant, and the BCA population in a changing environment can be studied by process-based, weather-driven mathematical models, able to interpret the combined effects on BCA efficacy of: (i) BCA mechanism of action, (ii) timing of BCA application with respect to timing of pathogen infection (preventative vs. curative), (iii) temperature and moisture requirements for both pathogen and BCA growth, and (iv) BCA survival capability. When the model was used under three contrasting weather conditions for the control of Botrytis bunch rot in grapevine, BCA efficacy was mostly influenced by environmental conditions, accounting for > 90% of the variance in simulated biocontrol efficacy. These findings indicate that the environmental responses of BCAs should be considered during their selection, BCA survival capability should be considered during both selection and formulation, and weather conditions and forecasts should be considered at the time of BCA application in the field. Different commercial BCAs for the control of Botrytis cinerea showed different environmental requirements and adaptation capabilities; therefore, the most suitable BCA to be used for a specific field application may consider weather conditions and forecasts at the time of intervention

Evaluation of atmospheric indicators in the Adriatic coastal areas: a multi-hazards approach for a better awareness of the current and future climate

Increasing climate resilience to global warming is one of the main challenges of the last few decades. Effective local measures have to be adopted to provide concrete solutions to the current and expected impacts of climate change. This is the goal of the AdriaClim Italia-Croatia Interreg Project (https://www.italy-croatia.eu/web/adriaclim), aimed at supporting the development of regional and local climate change adaptation plans for the Adriatic coastal regions. For this purpose, an exhaustive number of atmospheric climate indicators have been identified and evaluated across nine pilot areas to assess the current and expected main climate hazards affecting these regions, considering the worst-case emissions scenario (Representative Concentration Pathway RCP 8.5). The proposed analyses are provided by the results of the regional climate atmospheric model developed within the AdriaClim Project. The selected climate indicators are used to assess the possible evolution of the climate hazard across the pilot areas, covering different hazards, such as thermal discomfort, drought, and hydrological instability. A site-dependent investigation of the atmospheric climate indicators is proposed to emphasize which regions are more affected than others by the investigated climate hazards, thus warranting more attention in defining and proposing new adaptation strategies. The results highlight increasing temperatures (up to +3°C) across the Adriatic coastal regions, with more emphasis on the Northern Adriatic, where the combined effect with the relevant decrease in precipitation (down to −2 mm/day) may lead to severe drought conditions in the coming decades. In contrast, precipitation-related diseases may hit more Central and South Italy than the Northern Adriatic, except for the Emilia-Romagna region, which is found to be highly sensitive to both hazard categories. Finally, it is relevant to emphasize that these analyses have to be carefully considered in supporting adaptation strategies due to the lack of uncertainty estimates representing a fundamental element for decision-makers

A novel direct activator of AMPK inhibits prostate cancer growth by blocking lipogenesis

5′AMP-activated kinase (AMPK) constitutes a hub for cellular metabolic and growth control, thus representing an ideal therapeutic target for prostate cancers (PCas) characterized by increased lipogenesis and activation of mTORC1 pathway. However, whether AMPK activation itself is sufficient to block cancer cell growth remains to be determined. A small molecule screening was performed and identified MT 63–78, a specific and potent direct AMPK activator. Here, we show that direct activation of AMPK inhibits PCa cell growth in androgen sensitive and castration resistant PCa (CRPC) models, induces mitotic arrest, and apoptosis. In vivo, AMPK activation is sufficient to reduce PCa growth, whereas the allelic loss of its catalytic subunits fosters PCa development. Importantly, despite mTORC1 blockade, the suppression of de novo lipogenesis is the underpinning mechanism responsible for AMPK-mediated PCa growth inhibition, suggesting AMPK as a therapeutic target especially for lipogenesis-driven PCas. Finally, we demonstrate that MT 63–78 enhances the growth inhibitory effect of AR signaling inhibitors MDV3100 and abiraterone. This study thus provides a rationale for their combined use in CRPC treatment

Bioaccumulation of Trace Elements in the Muscle of the Blackmouth Catshark Galeus melastomus from Mediterranean Waters

Environmental pollution, particularly in the marine environment, has become a significant concern due to the increasing presence of pollutants and their adverse effects on ecosystems and human health. This study focuses on the bioaccumulation of trace elements in the muscle tissue of the blackmouth catshark (Galeus melastomus) from different areas in the Mediterranean Sea. Trace elements are of interest due to their persistence, toxicity, and potential for bioaccumulation. This research aims to assess the distribution and accumulation of trace elements in the muscle tissue of G. melastomus and investigate their potential impact on the deep-sea environment of the Mediterranean. The focused areas include the Ligurian Sea, the northern and central Tyrrhenian Sea, the southern Tyrrhenian Sea, the Ionian Sea, the Pantelleria Waters, and the Gela Waters. Samples were collected following established protocols, and trace element analysis was conducted using inductively coupled plasma mass spectrometry. The study provides data on the concentrations of 17 trace elements, namely aluminum, arsenic, cadmium, cobalt, copper, manganese, molybdenum, nickel, zinc, selenium, strontium, lead, chromium, iron, barium, bismuth, and uranium. The findings contribute to a better understanding of trace element bioaccumulation patterns in elasmobranch species, specifically G. melastomus, and highlight the potential risks associated with chemical contamination in the Mediterranean Sea. This research emphasizes the importance of studying the impacts of pollutants on marine organisms, particularly those occupying key ecological roles, like sharks, to support effective conservation and management strategies

Alternatives to CU Applications in Viticulture. How R&D Projects Can Provide Applied Solutions, Helping to Establish Legislation Limits

Copper (Cu) and its based preparations have been used for over 200 years to control fungi and bacterial diseases in cultivated plants. Downy mildew caused by the obligate biotrophic oomycete Plasmopara viticola is one of the most relevant and recurrent diseases of grapevines. Recently, the use of Cu is being limited by some regulations because of its high impact at different levels (health and environmental problems). Due to its accumulation in soil, this metal causes a little controversy with the principles of sustainable production. Therefore, international legislation and initiatives have recently been arisen to start limiting its use, with the main goal to replace it. In this framework, some alternatives have been tested and others are recently being developed to replace, at least partially, the use of Cu in viticulture. Many of them, are being developed and tested under the scope of research and development EU funded projects. To not compromise sustainability targets in viticulture, results from these R&D projects need to be considered to assess the present risks of using Cu in viticulture and to better support establishing limits for its applications, considering soils vulnerability, while no sustainable alternatives are available in the market

Staging of osteonecrosis of the jaw requires computed tomography for accurate definition of the extent of bony disease

Management of osteonecrosis of the jaw associated with antiresorptive agents is challenging, and outcomes are unpredictable. The severity of disease is the main guide to management, and can help to predict prognosis. Most available staging systems for osteonecrosis, including the widely-used American Association of Oral and Maxillofacial Surgeons (AAOMS) system, classify severity on the basis of clinical and radiographic findings. However, clinical inspection and radiography are limited in their ability to identify the extent of necrotic bone disease compared with computed tomography (CT). We have organised a large multicentre retrospective study (known as MISSION) to investigate the agreement between the AAOMS staging system and the extent of osteonecrosis of the jaw (focal compared with diffuse involvement of bone) as detected on CT. We studied 799 patients with detailed clinical phenotyping who had CT images taken. Features of diffuse bone disease were identified on CT within all AAOMS stages (20%, 8%, 48%, and 24% of patients in stages 0, 1, 2, and 3, respectively). Of the patients classified as stage 0, 110/192 (57%) had diffuse disease on CT, and about 1 in 3 with CT evidence of diffuse bone disease was misclassified by the AAOMS system as having stages 0 and 1 osteonecrosis. In addition, more than a third of patients with AAOMS stage 2 (142/405, 35%) had focal bone disease on CT. We conclude that the AAOMS staging system does not correctly identify the extent of bony disease in patients with osteonecrosis of the jaw

Reduction of Botrytis cinerea Colonization of and Sporulation on Bunch Trash.

Botrytis bunch rot (BBR) of grapevine, caused by Botrytis cinerea, is commonly managed by fungicide (FUN) sprays at flowering (A), at prebunch closure (B), at veraison (C), and before harvest. Applications at A, B, and C are recommended to reduce B. cinerea colonization of bunch trash and the production of conidia during berry ripening. The effects of these applications were previously evaluated as reductions in BBR severity at harvest rather than as reductions in bunch trash colonization and sporulation by B. cinerea. This study investigated the effects of FUNs (a commercial mixture of fludioxonil and cyprodonil), biological control agents (BCAs; Aureobasium pullulans and Trichoderma atroviride), and botanicals (BOTs; a commercial mixture of eugenol, geraniol, and thymol) applied at different timings (A, B, C, or ABC) compared with a nontreated control (NT) on B. cinerea bunch trash colonization and sporulation in vineyards. The ability of B. cinerea to colonize the bunch trash (as indicated by B. cinerea DNA content) and sporulate (as indicated by the number of conidia produced under optimal laboratory conditions) was highly variable, and this variability was higher between years (2015 to 2018) than among the three vineyards and three sampling times (i.e., 1 week after applications at A, B, and C). B. cinerea sporulation on bunch trash was significantly lower in plots treated with FUN than in NT in only 3 of 18 cases (3 vineyards × 2 years × 3 sampling times). FUN applications, however, significantly reduced B. cinerea colonization of bunch trash compared with NT; for colonization, BCA efficacy was similar to that of FUN, but BOT efficacy was variable. For all products, colonization reduction was the same with application at A versus ABC, meaning that the effect of an early season application lasted from flowering to 1 week after veraison. These results indicate that the early season control of B. cinerea is important to reduce the saprophytic colonization of bunch trash, especially when the risk of BBR is high