Dicarboximides and phenylpyrroles are commonly used fungicides against plant pathogenic ascomycetes. Although their effect on fungal osmosensing systems has been shown in many studies, their ...modes-of-action still remain unclear. Laboratory- or field-mutants of fungi resistant to either or both fungicide categories generally harbour point mutations in the sensor histidine kinase of the osmotic signal transduction cascade.In the present study we compared the mechanisms of resistance to the dicarboximide iprodione and to pyrrolnitrin, a structural analogue of phenylpyrrole fungicides, in Botrytis cinerea. Pyrrolnitrin-induced mutants and iprodione-induced mutants of B. cinerea were produced in vitro. For the pyrrolnitrin-induced mutants, a high level of resistance to pyrrolnitrin was associated with a high level of resistance to iprodione. For the iprodione-induced mutants, the high level of resistance to iprodione generated variable levels of resistance to pyrrolnitrin and phenylpyrroles. All selected mutants showed hypersensitivity to high osmolarity and regardless of their resistance levels to phenylpyrroles, they showed strongly reduced fitness parameters (sporulation, mycelial growth, aggressiveness on plants) compared to the parental phenotypes. Most of the mutants presented modifications in the osmosensing class III histidine kinase affecting the HAMP domains. Site directed mutagenesis of the bos1 gene was applied to validate eight of the identified mutations. Structure modelling of the HAMP domains revealed that the replacements of hydrophobic residues within the HAMP domains generally affected their helical structure, probably abolishing signal transduction. Comparing mutant phenotypes to the HAMP structures, our study suggests that mutations perturbing helical structures of HAMP2-4 abolish signal-transduction leading to loss-of-function phenotype. The mutation of residues E529, M427, and T581, without consequences on HAMP structure, highlighted their involvement in signal transduction. E529 and M427 seem to be principally involved in osmotic signal transduction.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Summary
Understanding the causes of population subdivision is of fundamental importance, as studying barriers to gene flow between populations may reveal key aspects of the process of adaptive ...divergence and, for pathogens, may help forecasting disease emergence and implementing sound management strategies. Here, we investigated population subdivision in the multihost fungus Botrytis cinerea based on comprehensive multiyear sampling on different hosts in three French regions. Analyses revealed a weak association between population structure and geography, but a clear differentiation according to the host plant of origin. This was consistent with adaptation to hosts, but the distribution of inferred genetic clusters and the frequency of admixed individuals indicated a lack of strict host specificity. Differentiation between individuals collected in the greenhouse (on Solanum) and outdoor (on Vitis and Rubus) was stronger than that observed between individuals from the two outdoor hosts, probably reflecting an additional isolating effect associated with the cropping system. Three genetic clusters coexisted on Vitis but did not persist over time. Linkage disequilibrium analysis indicated that outdoor populations were regularly recombining, whereas clonality was predominant in the greenhouse. Our findings open up new perspectives for disease control by managing plant debris in outdoor conditions and reinforcing prophylactic measures indoor.
Aims Nitrogen (N) fertilization is known to modify a plant's susceptibility to necrotrophic diseases. However, the effect of N nutrition on defence is not well known. It was hypothesized that not ...only molecules related to the N metabolism but also main sugars could mediate the effect of plant fertilization on its susceptibility to pathogens. Methods Two necrotrophic fungi, Botrytis cinerea and Sclerotinia sclerotiorum were inoculated on leaves of lettuce plants grown at five nitrate (NO₃⁻) fertilization levels, in three independent experiments. Variations in plant composition at the time of inoculation were linked to the size of lesions observed after 5-6 days. Results Both diseases were favoured by high NO₃⁻ fertilization. However, the highest disease levels were not found in the same experiment for B. cinerea and S. sclerotiorum. Among the components measured, NO₃⁻ and sucrose (SUC) were positively and negatively correlated to the two diseases in the three experiments, but the relationship between SUC and lesion size was more significant for S. sclerotiorum. Water content, N and total carbon (C) were also significantly correlated to the diseases, but the relationships were less straightforward. The ratios of SUC over total sugars and fructose (FRU) over total sugars fitted, very closely for S. sclerotiorum, a negative and positive exponential relationship respectively with lesion size. Absolute or relative glucose levels were not linked to the diseases. Conclusions Plant metabolic modifications induced by variations of N availability conferred the plant variable defence ability, which seemed, at least for S. sclerotiorum, mainly mediated by variations in host SUC and FRU levels. The generalization of these findings to other species would be of interest.
Numerous environmental pathogens, including Cryptococcus neoformans, Legionella spp., Chlamydophila pneumoniae, Mycobacterium avium, Listeria monocytogenes, Pseudomonas aeruginosa, and Francisella ...tularensis, might have acquired virulence traits via their resistance to predation by amoebae. Genomics and phylogenetics are revealing the evolutionary link between, for example, commensal strains of Escherichia coli and modern pathogens such as enterohaemorrhagic strains of this species (such as O157).\n However, the complexity of the biotic and abiotic environment perceived by obligate biotrophs during colonization of plants (powdery mildews on leaf surfaces inhabited by other microorganisms, for example) or during their dissemination (survival in air or in association with vectors) are also likely to exert selection independent of that due to the host plant genotype per se.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Trans isomers of fatty acids exhibit different health properties. Among them, trans-10,cis-12 conjugated linoleic acid has negative effects on milk fat production and can affect human health. A shift ...from the trans-11 to the trans-10 pathway of biohydrogenation (BH) can occur in the rumen of dairy cows receiving high-concentrate diets, especially when the diet is supplemented with highly unsaturated fat sources. The differences of BH patterns between linoleic acid (LeA) and linolenic acid (LnA) in such ruminal conditions remain unknown; thus, the aim of this work was to investigate in vitro the effects of starch and sunflower oil in the diet of the donor cows and starch level in the incubates on the BH patterns and efficiencies of LeA and LnA. The design was a 4 × 4 Latin square design with 4 cows, 4 periods, and 4 diets with combinations of 21 or 34% starch and 0 or 5% sunflower oil. The rumen content of each cow during each period was incubated with 4 substrates, combining 2 starch levels and either LeA or LnA addition. Capillary electrophoresis single-strand conformation polymorphism of incubates showed that dietary starch decreased the diversity of the bacterial community and the high-starch plus oil diet modified its structure. High-starch diets poorly affected isomerization and first reduction of LeA and LnA, but decreased the efficiencies of trans-11,cis-15-C18:2 and trans C18:1 reduction. Dietary sunflower oil increased the efficiency of LeA isomerization but decreased the efficiency of trans C18:1 reduction. An interaction between dietary starch and dietary oil resulted in the highest trans-10 isomers production in incubates when the donor cow received the high-starch plus oil diet. The partition between trans-10 and trans-11 isomers was also affected by an interaction between starch level and the fatty acid added to the incubates, showing that the trans-10 shift only occurred with LeA, whereas LnA was mainly hydrogenated via the more usual trans-11 pathway, whatever the starch level in the substrate, although the bacterial communities were not different between LeA and LnA incubates. In LeA incubates, trans-10 isomer production was significantly related to the structure of the bacterial community.
Human T-cell leukemia virus type 1 (HTLV-1) infection is associated with adult T-cell leukemia/lymphoma (ATLL), a lymphoproliferative malignancy with a dismal prognosis and limited therapeutic ...options. Recent evidence shows that HTLV-1-transformed cells present defects in both DNA replication and DNA repair, suggesting that these cells might be particularly sensitive to treatment with a small helicase inhibitor. Because the "Werner syndrome ATP-dependent helicase" encoded by the WRN gene plays important roles in both cellular proliferation and DNA repair, we hypothesized that inhibition of WRN activity could be used as a new strategy to target ATLL cells.
Our analysis demonstrates an apoptotic effect induced by the WRN helicase inhibitor in HTLV-1-transformed cells in vitro and ATL-derived cell lines. Inhibition of cellular proliferation and induction of apoptosis were demonstrated with cell cycle analysis, XTT proliferation assay, clonogenic assay, annexin V staining, and measurement of mitochondrial transmembrane potential.
Targeted inhibition of the WRN helicase induced cell cycle arrest and apoptosis in HTLV-1-transformed leukemia cells. Treatment with NSC 19630 (WRN inhibitor) induces S-phase cell cycle arrest, disruption of the mitochondrial membrane potential, and decreased expression of anti-apoptotic factor Bcl-2. These events were associated with activation of caspase-3-dependent apoptosis in ATL cells. We identified some ATL cells, ATL-55T and LMY1, less sensitive to NSC 19630 but sensitive to another WRN inhibitor, NSC 617145.
WRN is essential for survival of ATL cells. Our studies suggest that targeting the WRN helicase with small inhibitors is a novel promising strategy to target HTLV-1-transformed ATL cells.
This study tested the hypothesis that Botyrtis cinerea shows host specialization on tomato and lettuce, using phenotypic and genotypic tools. Strains were isolated from tomato and lettuce grown ...together in the same greenhouse. Forty‐four lettuce strains and 42 tomato strains were investigated for their genetic diversity and their aggressiveness. Both gene diversity and allelic richness were significantly higher in lettuce strains than in tomato strains (P = 0·01). Cluster analysis revealed a clear division of the strains under study into two clusters. However, this structure did not separate the strains according to their host of origin. Tomato strains were significantly more aggressive than lettuce strains when inoculated on tomatoes (P = 0·001), but no significant differences in aggressiveness were observed when the strains were inoculated on lettuce (P = 0·17) or on apple (P = 0·87). The results suggest an absence of clear host specialization of B. cinerea on tomato and lettuce.
Adequate protection of apple trees during the primary contamination period is a cornerstone for management of apple scab. Correct timing of spring treatments is fundamental and thus, much effort has ...been devoted to forecasting ascospore release by Venturia inaequalis. Most models rely on degree‐day accumulation starting from a biofix date established yearly on the basis of biological observations. Here, the potential of using a single calendar date as a biofix and new types of time scales were explored, with the help of numerical optimization with field‐collected data. Using data acquired between 1996 and 2008, the daily rate of development for V. inaequalis primary inoculum was assessed by fitting generic time scale functions, a method that requires the smallest number of assumptions about the effect of temperature on the biological phenomenon. An optimal calendar biofix was established for Provence and use of non‐linear functions relating pseudothecial development rate to temperature for accumulating thermal time was compared with the usual linear response in standard degree‐day models. A model was then constructed using four additional years of data for validation. The predictive value of the model was further improved by adjusting the time scale with ‘accelerating rules’ to take into account the positive influence of rainy days on pseudothecial maturation prior to ascospore release. However, ‘halting rules’ inserted in the time scale to account for dry days during the ascospore release period strongly reduced the predictive value of the model for southern France, suggesting the possible occurrence of strains adapted to dry conditions.
The nutritional status of a plant is known to influence its susceptibility to pathogens. In the case of Botrytis cinerea, the role of nitrogen fertilization of various host plants on disease ...development appears to be variable. This study was carried out to characterize possible variability associated with isolates and inoculum density of B. cinerea in its ability to infect leaf-pruning wounds and to develop stem lesions on tomato plants, as affected by the nitrogen input. Six isolates differing in their aggressiveness to tomato were compared. They all had similar reaction patterns in vitro in response to differential nitrogen levels. In tests on plants grown with contrasted regimes of nitrate fertilization, overall disease severity was lower for all isolates on plants with higher nitrogen inputs, regardless of inoculum concentration. However, differences among isolates were observed in the effect of plant nitrogen nutrition on infection and on lesion expansion. Disease onset was delayed on all plants with higher nitrogen inputs, but the response was greater for isolates with lower aggressiveness on tomato. The highest contrast among isolates was observed with the colonization of stems. The daily rate of stem lesion expansion decreased with increasing nitrogen fertilization levels for the more aggressive isolates, while it increased for the less aggressive isolate. Hypotheses to explain these results are discussed in light of the possible physiological effects of nitrogen fertilization on nutrient availability for the pathogen in the host tissue and of possible production of defence metabolites by the plant.
The question as to why parasites remain generalist or become specialist is a key unresolved question in evolutionary biology. Ampelomyces spp., intracellular mycoparasites of powdery mildew fungi, ...which are themselves plant pathogens, are a useful model for studies of this issue. Ampelomyces is used for the biological control of mildew. Differences in mycohost phenology promote temporal isolation between sympatric Ampelomyces mycoparasites. Apple powdery mildew (APM) causes spring epidemics, whereas other powdery mildew species on plants other than apple cause epidemics later in the season. This has resulted in genetic differentiation between APM and non-APM strains. It is unclear whether there is genetic differentiation between non-APM Ampelomyces lineages due to their specialization on different mycohosts. We used microsatellites to address this question and found no significant differentiation between non-APM Ampelomyces strains from different mycohosts or host plants, but strong differentiation between APM and non-APM strains. A geographical structure was revealed in both groups, with differences between European countries, demonstrating restricted dispersal at the continent scale and a high resolution for our markers. We found footprints of recombination in both groups, possibly more frequent in the APM cluster. Overall, Ampelomyces thus appears to be one of the rare genuine generalist pathogenic fungi able to parasitize multiple hosts in natural populations. It is therefore an excellent model for studying the evolution of pathogens towards a generalist rather than host-specific strategy, particularly in light of the tritrophic interaction between Ampelomyces mycoparasites, their powdery mildew fungal hosts and the mildew host plants.
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Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
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