RESUMO
Fusarium verticillioides causes multiple diseases of Zea mays (maize) including ear and seedling rots, contaminates seeds and seed products worldwide with toxic chemicals called fumonisins. The role of fumonisins in disease is unclear because, although they are not required for ear rot, they are required for seedling diseases. Disease symptoms may be due to the ability of fumonisins to inhibit ceramide synthase activity, the expected cause of lipids (fatty acids, oxylipins, and sphingolipids) alteration in infected plants. In this study, we explored the impact of fumonisins on fatty acid, oxylipin, and sphingolipid levels in planta and how these changes affect F. verticillioides growth in maize. The identity and levels of principal fatty acids, oxylipins, and over 50 sphingolipids were evaluated by chromatography followed by mass spectrometry in maize infected with an F. verticillioides fumonisin-producing wild-type strain and a fumonisin-deficient mutant, after different periods of growth. Plant hormones associated with defense responses, i.e., salicylic and jasmonic acid, were also evaluated. We suggest that fumonisins produced by F. verticillioides alter maize lipid metabolism, which help switch fungal growth from a relatively harmless endophyte to a destructive necrotroph.
Assuntos
Fumonisinas/toxicidade , Fusarium/química , Germinação , Metabolismo dos Lipídeos/efeitos dos fármacos , Micoses/metabolismo , Doenças das Plantas/microbiologia , Zea mays/efeitos dos fármacos , Ciclopentanos/análise , Ciclopentanos/metabolismo , Ácidos Graxos/análise , Ácidos Graxos/metabolismo , Fumonisinas/farmacologia , Micotoxinas/toxicidade , Oxilipinas/análise , Oxilipinas/metabolismo , Ácido Salicílico/análise , Ácido Salicílico/metabolismo , Esfingolipídeos/análise , Esfingolipídeos/metabolismo , Zea mays/química , Zea mays/crescimento & desenvolvimento , Zea mays/metabolismoRESUMO
Aspergillus flavus is a saprophytic cosmopolitan fungus, capable of infecting crops both pre- and post-harvest and exploiting different secondary metabolites, including aflatoxins. Aflatoxins are known carcinogens to animals and humans, but display no clear effect in host plants such as maize. In a previous study, we mined the genome of A. flavus to identify secondary metabolite clusters putatively involving the pathogenesis process in maize. We now focus on cluster 32, encoding for fungal effectors such as salicylate hydroxylase (SalOH), and necrosis- and ethylene-inducing proteins (npp1 domain protein) whose expression is triggered upon kernel contact. In order to understand the role of this genetic cluster in maize kernel infection, mutants of A. flavus, impaired or enhanced in specific functions (e.g., cluster 32 overexpression), were studied for their ability to cause disease. Within this frame, we conducted histological and histochemical experiments to verify the expression of specific genes within the cluster (e.g., SalOH, npp1), the production of salicylate, and the presence of its dehydroxylated form. Results suggest that the initial phase of fungal infection (2 days) of the living tissues of maize kernels (e.g., aleuron) coincides with a significant increase of fungal effectors such as SalOH and Npp1 that appear to be instrumental in eluding host defences and colonising the starch-enriched tissues, and therefore suggest a role of cluster 32 to the onset of infection.
Assuntos
Aspergillus flavus/patogenicidade , Redes e Vias Metabólicas/genética , Família Multigênica , Zea mays/microbiologia , Aflatoxinas/genética , Aflatoxinas/metabolismo , Aspergilose/genética , Aspergilose/metabolismo , Aspergillus flavus/genética , Aspergillus flavus/fisiologia , Catecóis/metabolismo , Produtos Agrícolas/genética , Produtos Agrícolas/metabolismo , Produtos Agrícolas/microbiologia , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Oxigenases de Função Mista/genética , Oxigenases de Função Mista/metabolismo , Organismos Geneticamente Modificados , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Quercetina/metabolismo , Ácido Salicílico/metabolismo , Sementes , Zea mays/genética , Zea mays/metabolismoRESUMO
Fusarium langsethiae is amongst the most recently discovered pathogens of small grains cereals. F. langsethiae is the main producer, in Europe, of T2 and HT-toxins in small grain cereals, albeit often asymptomatic; this makes its control challenging. The European Union (EU) is pushing hard on the use of biocontrol agents to minimize the use of fungicides and pesticides, which are detrimental to the environment and responsible for serious pollution of the soil and superficial water. In line with EU directives (e.g., 128/2009), here we report the use of protein fractions, purified from the culture filtrate of the basidiomycete Trametes versicolor, for controlling F. langsethiae. T. versicolor, a so-called medicinal mushroom which is applied as a co-adjuvant in oncology and other pathologies as a producer of biological response modifiers. In this study, the exo-proteome of T. versicolor proved highly efficient in inhibiting the growth of F. langsethiae and the biosynthesis of the T2 toxin. Results are promising for its future use as a sustainable product to control F. langsethiae infection in cereals under field conditions.
Assuntos
Agaricales/metabolismo , Antibiose , Grão Comestível/microbiologia , Fusarium/fisiologia , Proteoma , Trametes/metabolismo , Bioensaio , Micotoxinas/biossínteseRESUMO
Plant roots release complex mixtures of bioactive molecules, including compounds that affect the activity and modify the composition of the rhizosphere microbiome. In this work, we investigated the initial phase of the interaction between tomato and an effective biocontrol strain of Trichoderma harzianum (T22). We found that root exudates (RE), obtained from plants grown in a split-root system and exposed to various biotic and abiotic stress factors (wounding, salt, pathogen attack), were able to stimulate the growth and act as chemoattractants of the biocontrol fungus. On the other hand, some of the treatments did not result in an enhanced chemotropism on Fusarium oxysporum f. sp. lycopersici, indicating a mechanism that may be selective for nonpathogenic microbes. The involvement of peroxidases and oxylipins, both known to be released by roots in response to stress, was demonstrated by using RE fractions containing these molecules or their commercial purified analogs, testing the effect of an inhibitor, and characterizing the complex pattern of these metabolites released by tomato roots both locally and systemically.
Assuntos
Fusarium/efeitos dos fármacos , Exsudatos de Plantas/farmacologia , Raízes de Plantas/metabolismo , Solanum lycopersicum/metabolismo , Trichoderma/efeitos dos fármacos , Quimiotaxia , Fusarium/fisiologia , Solanum lycopersicum/microbiologia , Doenças das Plantas/microbiologia , Raízes de Plantas/microbiologia , Microbiologia do Solo , Esporos Fúngicos , Estresse Fisiológico , Trichoderma/fisiologiaRESUMO
Functional properties of cell membranes depend on their composition, particularly on the relative amount of saturated, unsaturated and polyunsaturated fatty acids present in the phospholipids. The aim of this study was to investigate the effect of cell membrane composition on cell fitness, adaptation and stress response in Kluyveromyces lactis. To this purpose, we have deleted the genes FAD2 and FAD3 encoding Δ12 and ω3 desaturases in Kluyveromyces lactis, thus generating mutant strains with altered fatty acid composition of membranes. These strains were viable and able to grow in stressing conditions like hypoxia and low temperature. Deletion of the Δ9 desaturase-encoding gene KlOLE1 resulted in lethality, suggesting that this enzyme has an essential role in this yeast. Transcription of the desaturase genes KlOLE1, FAD2 and FAD3 and cellular localization of the corresponding enzymes, have been studied under hypoxia and cold stress. Our findings indicate that expression of these desaturase genes and membrane composition were modulated by hypoxia and temperature stress, although the changes induced by these and other assayed conditions did not dramatically affect the general cellular fitness.
Assuntos
Resposta ao Choque Frio/fisiologia , Metabolismo Energético/fisiologia , Ácidos Graxos Dessaturases/genética , Kluyveromyces/enzimologia , Anaerobiose , Membrana Celular/metabolismo , Retículo Endoplasmático/enzimologia , Metabolismo Energético/genética , Ácidos Graxos Dessaturases/metabolismo , Fermentação/genética , Fermentação/fisiologia , Deleção de Genes , Kluyveromyces/genética , Estearoil-CoA DessaturaseRESUMO
In the yeast Kluyveromyces lactis, the inactivation of structural or regulatory glycolytic and fermentative genes generates obligate respiratory mutants which can be characterized by sensitivity to the mitochondrial drug antimycin A on glucose medium (Rag(-) phenotype). Rag(-) mutations can occasionally be generated by the inactivation of genes not evidently related to glycolysis or fermentation. One such gene is the hypoxic regulatory gene KlMGA2. In this work, we report a study of the many defects, in addition to the Rag(-) phenotype, generated by KlMGA2 deletion. We analyzed the fermentative and respiratory metabolism, mitochondrial functioning and morphology in the Klmga2Δ strain. We also examined alterations in the regulation of the expression of lipid biosynthetic genes, in particular fatty acids, ergosterol and cardiolipin, under hypoxic and cold stress and the phenotypic suppression by unsaturated fatty acids of the deleted strain. Results indicate that, despite the fact that the deleted mutant strain had a typical glycolytic/fermentative phenotype and KlMGA2 is a hypoxic regulatory gene, the deletion of this gene generated defects linked to mitochondrial functions suggesting new roles of this protein in the general regulation and cellular fitness of K. lactis. Supplementation of unsaturated fatty acids suppressed or modified these defects suggesting that KlMga2 modulates membrane functioning or membrane-associated functions, both cytoplasmic and mitochondrial.
Assuntos
Proteínas de Bactérias/genética , Ácidos Graxos Insaturados/metabolismo , Fermentação/genética , Glucose/metabolismo , Kluyveromyces/metabolismo , Consumo de Oxigênio/genética , Fatores de Transcrição/genética , Antifúngicos/farmacologia , Antimicina A/farmacologia , Cardiolipinas/metabolismo , Hipóxia Celular/fisiologia , Resposta ao Choque Frio/fisiologia , Ergosterol/metabolismo , Regulação Fúngica da Expressão Gênica/genética , Glicólise/genética , Kluyveromyces/efeitos dos fármacos , Kluyveromyces/genética , Proteínas de Membrana/genética , Mitocôndrias/metabolismo , Transcrição Gênica/genéticaRESUMO
Viticulture has been an important economic sector for centuries. In recent decades, global wine production has fluctuated between 250 and almost 300 million hectoliters, and in 2022, the value of wine exports reached EUR 37.6 billion. Climate change and the associated higher temperatures could favor the occurrence of ochratoxin A (OTA) in wine. OTA is a mycotoxin produced by some species of the genera Aspergillus and Penicillium and has nephrotoxic, immunotoxic, teratogenic, hepatotoxic, and carcinogenic effects on animals and humans. The presence of this toxin in wine is related to the type of wine-red wines are more frequently contaminated with OTA-and the geographical location of the vineyard. In Europe, the lower the latitude, the greater the risk of OTA contamination in wine. However, climate change could increase the risk of OTA contamination in wine in other regions. Due to their toxic effects, the development of effective and environmentally friendly methods to prevent, decontaminate, and degrade OTA is essential. This review summarises the available research on biological aspects of OTA prevention, removal, and degradation.
Assuntos
Contaminação de Alimentos , Ocratoxinas , Vinho , Ocratoxinas/análise , Vinho/análise , Contaminação de Alimentos/análise , Contaminação de Alimentos/prevenção & controle , Animais , HumanosRESUMO
In this work, the first specific phytochemical analysis on Odontites vulgaris Moench collected in Central Italy was performed. The aerial parts ethanolic extract was studied and eight compounds were identified: pheophytin a (1), aucubin (2), catalpol (3), shanzhiside methyl ester (4), melampyroside (5), 8-epi-loganin (6), caryoptoside (7) and quinic acid (8). To the best of our knowledge, in this study, compounds (7-8) resulted to be isolated from the genus for the first time. The chemophenetic markers of the family and order were evidenced and several important ecological conclusions could be drawn. The ethanolic extract was also tested for several biological activities showing high effects in the antioxidant, cytoprotective and aflatoxin B1 production inhibitory assays. A brief explanation on these activities under the phytochemical standpoint was also included.
Assuntos
Antioxidantes , Compostos Fitoquímicos , Componentes Aéreos da Planta , Extratos Vegetais , Componentes Aéreos da Planta/química , Compostos Fitoquímicos/farmacologia , Compostos Fitoquímicos/isolamento & purificação , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Antioxidantes/farmacologia , Antioxidantes/isolamento & purificação , Estrutura Molecular , Itália , HumanosRESUMO
Introduction: The evaluation of biological degradation of waterlogged archeological wood is crucial to choose the conservative and protective treatments to be applied to the wooden material. The waterlogged environmental conditions are characterized by oxygen scarcity, only allowing the growth of adapted microbes capable to degrade the organic wooden material, mainly erosion bacteria and soft-rot fungi. In this work, we characterized and evaluated the biodegradation state and the microbial communities of wooden fragments preserved in storage tanks. These were preserved by waterlogging within the Neolithic village "La Marmotta," currently found under the Bracciano Lake (Lazio, Italy). Methods: The waterlogged wood samples were first identified taxonomically with an optical microscope, also allowing an evaluation of their preservation state. The microbial community was then evaluated through the sequencing of Internal Transcribed Spacer sequences for fungi and 16S for bacteria with the Oxford Nanopore Technologies (ONT) MinION platform. Results: The identified microbial community appears to be consistent with the waterlogged samples, as many bacteria attributable to the erosion of wood and ligninolytic fungi have been sequenced. Discussion: The reported results highlight the first use of targeted metabarcoding by ONT applied to study the biodeterioration of waterlogged archeological wood.
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Pantoea stewartii subsp. stewartii (Pss) causes Stewart's vascular wilt of maize, and it is responsible for serious crop losses. Pss is indigenous to North America and spreads with maize seeds. The presence of Pss has been notified in Italy since 2015. The risk assessment of the entry of Pss in the EU from the United States through seed trade is in the order of magnitude of hundred introductions per year. Several molecular or serological tests were developed for the detection of Pss and used as official analysis for the certification of commercial seeds. However, some of these tests lack adequate specificity, not allowing to correctly discriminate Pss from P. stewartii subsp. indologenes (Psi). Psi is occasionally present in maize seeds and is avirulent for maize. In this study, several Italian isolates of Pss recovered in 2015 and 2018 have been characterized by molecular, biochemical, and pathogenicity tests; moreover, their genomes have been assembled through MinION and Illumina-sequencing procedures. Genomic analysis reveals multiple introgression events. Exploiting these results, a new primer combination has been defined and verified by real-time PCR, allowing the development of a specific molecular test able to detect the presence of Pss down to the concentration of 103 CFU/ml in spiked samples of maize seed extracts. Due to the high analytical sensitivity and specificity achieved with this test, the detection of Pss has been improved disentangling the inconclusive results in Pss maize seed diagnosis, overcoming its misidentification in place of Psi. Altogether, this test addresses the critical issue associated with maize seeds imported from regions where Stewart's disease is endemic.
RESUMO
The nutritional quality of gluten-free (GF) products is usually improved by using flours derived from alternative grains (e.g., pseudocereals and legumes), additives and hydrolysates, leading to long ingredient lists in the labels, that conflict with current customer expectations. In this work, chestnut, carob, and hemp flours were used as mixed ingredients for making a gluten-free type-II sourdough. Three exopolysaccharides-producer lactic acid bacteria, belonging to Leuconostoc mesenteroides, Weissella cibaria, and Leuconostoc pseudomesenteroides, were used, and the fermentation processes (6 log10 cfu/g, 25 °C, 16 h) optimize to maximize the EPS synthesis (15.70 ± 2.1 mg/kg). The chestnut-hemp (70:30) type-II sourdough was included in a rice/corn gluten-free bread recipe also containing psyllium flour as structuring agent. Although the fortification with unfermented flours already led the achievement of 6 g/100 g of fiber (high fiber, Regulation EC n. 1924/2006) and content of magnesium higher than the daily reference intakes, the use of type-II sourdoughs led to a further structural, sensory, and nutritional improvements (e.g., decreasing the main anti-nutritional factor phytic acid). This work demonstrated that the use of ad-hoc selected ingredients and optimized protocol can be used to produce a GF and "clean label" bread with optimal nutritional features and appreciable sensory and structural properties.
Assuntos
Cannabis , Pão/microbiologia , Fermentação , Dieta Livre de Glúten , Valor Nutritivo , Farinha/microbiologiaRESUMO
Two of the mycotoxins of greatest agroeconomic significance are aflatoxin B1 (AFB1), and ochratoxin A (OTA). It has been reported that extracts from some wood-decaying mushrooms, such as Lentinula edodes and Trametes versicolor showed the ability to inhibit AFB1 or OTA biosynthesis. Therefore, in our study, a wide screening of 42 isolates of different ligninolytic mushrooms was assayed for their ability to inhibit the synthesis of OTA in Aspergillus carbonarius and AFB1 in Aspergillus flavus, in order to find a metabolite that can simultaneously inhibit both mycotoxins. The results showed that four isolates produce metabolites able to inhibit the synthesis of OTA, and 11 isolates produced metabolites that inhibited AFB1 by >50%. Two strains, the Trametes versicolor strain TV117 and the Schizophyllum commune strain S.C. Ailanto, produced metabolites able to significantly inhibit (>90%) the synthesis of both mycotoxins. Preliminary results suggest that the mechanism of efficacy of the S. commune rough and semipurified polysaccharides could be analogous to that found previously for Tramesan®, by enhancing the antioxidant response in the target fungal cells. The overall results indicate that S. commune's polysaccharide(s) could be a potential agent(s) in biological control and/or a useful component of the integrated strategies able to control mycotoxin synthesis.
Assuntos
Agaricales , Micotoxinas , Ocratoxinas , Micotoxinas/metabolismo , Aspergillus flavus/metabolismo , Agaricales/metabolismo , Trametes/metabolismo , Ocratoxinas/metabolismo , Aflatoxina B1/metabolismoRESUMO
This work studies the incidence of Fusarium spp. on wheat kernels about current and future climatic conditions in Italy. Epidemiological analyses were performed from 2007 to 2013 and the resulting dataset was used to find correlations between the disease incidence of five important Fusarium species monitored in Italy (Fusarium graminearum, F. langsethiae, F. sporotrichioides, F. poae and F. avenaceum) and climatic and geographical parameters. Probabilistic-based modelling of the actual distribution of Fusarium spp. was achieved by using the Zero-inflated Poisson regression. The probabilistic geographical distribution of the Fusarium species was assessed by applying future climatic scenarios (RCPs 4.5 and 8.5). The shift from current to future climatic scenarios highlighted changes on a national and regional scale. The tightening of environmental conditions from the RCP4.5 to 8.5 scenarios resulted in a sporadic presence of F. avenaceum only in the northern region of Italy. Fusarium graminearum was plentifully present in the current climate, but the tightening of minimum and maximum temperatures and the decrease of precipitation between May-June in the RCP8.5 no longer represents the optimum conditions for it. Fusarium langsethiae was currently distributed in all of Italy, showing an increase in the probability of detecting it by moving from high to low latitudes and from low to high longitudes in the RCP8.5. Fusarium poae, unlike other Fusarium species, grows and develops in arid climatic conditions. High values of F. poae were recorded at low latitudes and longitudes. Under the RCP scenarios, it showed high incidence probabilities in the southeast and northeast areas of Italy. Fusarium sporotrichioides is scarcely present in Italy, found at high latitudes and in the central areas. Climate change altered this distribution, and the chances of discovering it increased significantly moving to southern Italy. Overall, the study shows that climate change conditions are likely to lead to an increase in the incidence of Fusarium species on wheat kernels in Italy, highlighting the importance of developing strategies to mitigate the effects of climate change on wheat production, quality, and safety.
Assuntos
Fusarium , Mudança Climática , Itália , Temperatura , Grão ComestívelRESUMO
For about a decade, olive groves in Apulia (Southern Italy) have been progressively destroyed by Olive Quick Decline Syndrome (OQDS), a disease caused by the bacterium Xylella fastidiosa subsp. pauca (Xfp). Recently, we described an additional wilting syndrome affecting olive trees in that area. The botryosphaeriaceous fungus Neofusicoccum mediterraneum was found associated with the diseased trees, and its high virulence toward olive trees was demonstrated. Given the common features with Branch and Twig Dieback (BTD) of olive tree, occurring in Spain and California, we suggested that the observed syndrome was BTD. During our first survey, we also found a botryosphaeriaceous species other than N. mediterraneum. In the present article, we report the morphological and molecular characterization of this fungal species which we identified as Neofusicoccum stellenboschiana. In the study, we also included for comparison additional N. stellenboschiana isolates obtained from olive trees in Latium and Tuscany region (Central Italy). The occurrence of N. stellenboschiana in olive trees is reported here for the first time in the northern hemisphere. The pathogenicity and virulence were tested in nine inoculation trials, where the Apulian N. stellenboschiana isolate was compared with the isolate from Latium and with the Apulian isolate of N. mediterraneum. Both isolates of N. stellenboschiana proved pathogenic to olive trees. They caused evident bark canker and wood discolouration when inoculated at the base of the stem of two/three-year-old trees and on one-year-old twigs. However, virulence of N. stellenboschiana was significantly lower, though still remarkable, compared with N. mediterraneum in term of necrosis progression in the bark and the wood and capacity of wilting the twigs. Virulence of N. stellenboschiana and N. mediterraneum did not substantially change when inoculations were performed in spring/summer and in autumn, suggesting that these fungal species have the potential to infect and damage olive trees in all seasons. The high thermotolerance of N. stellenboschiana was also revealed with in vitro growth and survival tests. The high virulence of these Botryosphaeriaceae species highlights their contribution in BTD aetiology and the necessity to investigate right away their diffusion and, possibly, the role of additional factors other than Xfp in the general decline of olive groves in Apulia. Hence the importance of assessing the degree of overlap of BTD/Botryosphariaceae with OQDS/Xfp is discussed.
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We investigate laccase-mediated detoxification of aflatoxins, fungal carcinogenic food contaminants. Our experimental comparison between two aflatoxins with similar structures (AFB1 and AFG2) shows significant differences in laccase-mediated detoxification. A multi-scale modeling approach (Docking, Molecular Dynamics, and Density Functional Theory) identifies the highly substrate-specific changes required to improve laccase detoxifying performance. We employ a large-scale density functional theory-based approach, involving more than 7000 atoms, to identify the amino acid residues that determine the affinity of laccase for aflatoxins. From this study we conclude: (1) AFB1 is more challenging to degrade, to the point of complete degradation stalling; (2) AFG2 is easier to degrade by laccase due to its lack of side products and favorable binding dynamics; and (3) ample opportunities to optimize laccase for aflatoxin degradation exist, especially via mutations leading to π-π stacking. This study identifies a way to optimize laccase for aflatoxin bioremediation and, more generally, contributes to the research efforts aimed at rational enzyme optimization.
Assuntos
Aflatoxinas , Aflatoxinas/análise , Aflatoxina B1/química , Lacase/metabolismo , Simulação de Dinâmica Molecular , Contaminação de Alimentos/análiseRESUMO
Among the various factors correlated with toxin production in fungi, oxidative stress is a crucial one. In relation to this, an important role is played by oxidative stress-related receptors. These receptors can transduce the "oxidative message" to the nucleus and promote a transcriptional change targeted at restoring the correct redox balance in the cell. In Aspergillus parasiticus, the knockout of the ApyapA gene, a homologue of the yeast Yap-1, disables the fungus's capacity to restore the correct redox balance in the cell. As a consequence, the onset of secondary metabolism and aflatoxins synthesis is triggered. Some clues as to the involvement of oxidative stress in the regulation of ochratoxin A (OTA) synthesis in Aspergillus ochraceus have already been provided by the disruption of the oxylipin-producer AoloxA gene. In this paper, we add further evidence that oxidative stress is also involved in the regulation of OTA biosynthesis in A. ochraceus. In fact, the use of certain oxidants and, especially, the deletion of the yap1-homologue Aoyap1 further emphasize the role played by this stress in controlling metabolic and morphological changes in A. ochraceus.
Assuntos
Aspergillus ochraceus/genética , Aspergillus ochraceus/metabolismo , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Ocratoxinas/biossíntese , Fatores de Transcrição/metabolismo , OxirreduçãoRESUMO
In 2013, Xylella fastidiosa (Xf) was detected for the first time in Apulia and, subsequently, recognized as the causal agent of the olive quick decline syndrome (OQDS). To contain the disease, the olive germplasm was evaluated for resistance to Xf, identifying cultivars with different susceptibility to the pathogen. Regarding this, the resistant cultivar Leccino has generally a lower bacterial titer compared with the susceptible cultivar Ogliarola salentina. Among biomolecules, lipids could have a pivotal role in the interaction of Xf with its host. In the grapevine Pierce's disease, fatty acid molecules, the diffusible signaling factors (DSFs), act as regulators of Xf lifestyle and are crucial for its virulence. Other lipid compounds derived from fatty acid oxidation, namely, oxylipins, can affect, in vitro, biofilm formation in Xf subsp. pauca (Xfp) strain De Donno, that is, the strain causing OQDS. In this study, we combined high-performance liquid chromatography-mass spectrometry-MS-based targeted lipidomics with supervised learning algorithms (random forest, support vector machine, and neural networks) to classify olive tree samples from Salento. The dataset included samples from either OQDS-positive or OQDS-negative olive trees belonging either to cultivar Ogliarola salentina or Leccino treated or not with the zinc-copper-citric acid biocomplex Dentamet®. We built classifiers using the relative differences in lipid species able to discriminate olive tree samples, namely, (1) infected and non-infected, (2) belonging to different cultivars, and (3) treated or untreated with Dentamet®. Lipid entities emerging as predictors of the thesis are free fatty acids (C16:1, C18:1, C18:2, C18:3); the LOX-derived oxylipins 9- and 13-HPOD/TrE; the DOX-derived oxylipin 10-HPOME; and diacylglyceride DAG36:4(18:1/18:3).
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Lipids are central at various stages of host-pathogen interactions in determining virulence and modulating plant defense. Free fatty acids may act as substrates for oxidizing enzymes [e.g., lipoxygenases (LOXs) and dioxygenases (DOXs)] that synthesize oxylipins. Fatty acids and oxylipins function as modulators of several pathways in cell-to-cell communication; their structural similarity among plant, fungal, and bacterial taxa suggests potential in cross-kingdom communication. We provide a prospect of the known role of fatty acids and oxylipins in fungi and bacteria during plant-pathogen interactions. In the pathogens, oxylipin-mediated signaling pathways are crucial both in development and host infection. Here, we report on case studies suggesting that oxylipins derived from oleic, linoleic, and linolenic acids are crucial in modulating the pathogenic lifestyle in the host plant. Intriguingly, overlapping (fungi-plant/bacteria-plant) results suggest that different inter-kingdom pathosystems use similar lipid signals to reshape the lifestyle of the contenders and occasionally determine the outcome of the challenge.
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Aspergillus fumigatus is a disease-causing, opportunistic fungus that can establish infection due to its capacity to respond to a wide range of environmental conditions. Secreted proteins and metabolites, which play a critical role in fungal-host interactions and pathogenesis, are modulated by epigenetic players, such as bromodomain and extraterminal domain (BET) proteins. In this study, we evaluated the in vitro and in vivo capability of the BET inhibitor JQ1 to modulate the extracellular proteins and virulence of A. fumigatus. The abundance of 25 of the 76 extracellular proteins identified through LC-MS/MS proteomic analysis changed following JQ1 treatment. Among them, a ribonuclease, a chitinase, and a superoxide dismutase were dramatically downregulated. Moreover, the proteomic analysis of A. fumigatus intracellular proteins indicated that Abr2, an intracellular laccase involved in the last step of melanin synthesis, was absent in the JQ1-treated group. To investigate at which level this downregulation occurred and considering the ability of JQ1 to modulate gene expression we checked the level of ABR2, Chitinase, and Superoxide dismutase mRNA expression by qRT-PCR. Finally, the capacity of JQ1 to reduce the virulence of A. fumigatus has been proved using Galleria mellonella larvae, which are an in vivo model to evaluate fungal virulence. Overall, the promising activity exhibited by JQ1 suggests that A. fumigatus is sensitive to BET inhibition and BET proteins may be a viable target for developing antifungal agents.
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Biscogniauxia mediterranea is the causal agent of charcoal disease, affecting oak decline under the trigger of various biotic and abiotic factors, including climate change. Here, we report the genome assembly of an Italian B. mediterranea strain obtained using hybrid sequencing technologies combining long and short reads.