Summary
Fusarium graminearum is a causal agent of wheat scab and a producer of the trichothecene mycotoxin deoxynivalenol (DON). The expression of trichothecene biosynthesis (TRI) genes and DON ...production are mainly regulated by the cyclic adenosine monophosphate‐protein kinase A (cAMP‐PKA) pathway and two pathway‐specific transcription factors (TRI6 and TRI10). Interestingly, deletion mutants of TRI6 show reduced expression of several components of cAMP signalling, including the FgCAP1 adenylate‐binding protein gene that has not been functionally characterized in F. graminearum. In this study, we show that FgCap1 interacts with Fac1 adenylate cyclase and that deletion of FgCAP1 reduces the intracellular cAMP level and PKA activity. The Fgcap1 deletion mutant is defective in vegetative growth, conidiogenesis and plant infection. It also shows significantly reduced DON production and TRI gene expression, which can be suppressed by exogenous cAMP, indicating a PKA‐dependent regulation of DON biosynthesis by FgCap1. The wild‐type, but not tri6 mutant, shows increased levels of intracellular cAMP and FgCAP1 expression under DON‐producing conditions. Furthermore, the promoter of FgCAP1 contains one putative Tri6‐binding site that is important for its function during DON biosynthesis, but is dispensable for hyphal growth, conidiogenesis and pathogenesis. In addition, FgCap1 shows an actin‐like localization to the cortical patches at the apical region of hyphal tips. Phosphorylation of FgCap1 at S353 was identified by phosphoproteomics analysis. The S353A mutation in FgCAP1 has no effect on its functions during vegetative growth, conidiation and DON production. However, expression of the FgCAP1S353A allele fails to complement the defects of the Fgcap1 mutant in plant infection, indicating the importance of the phosphorylation of FgCap1 at S353 during pathogenesis. Taken together, our results suggest that FgCAP1 is involved in the regulation of DON production via cAMP signalling and subjected to a feedback regulation by TRI6, but the phosphorylation of FgCap1 at S353 is probably unrelated to the cAMP‐PKA pathway because the S353A mutation only affects plant infection.
Summary
Appressorium formation is a key step in the infection cycle of Magnaporthe oryzae. Mst12 is a transcription factor essential for appressorium penetration and invasive growth. In this study we ...used the affinity purification approach to identify proteins that physically associate with Mst12. One of the Mst12‐interacting genes identified was MoMCM1, which encodes a MADS‐box protein orthologous to yeast Mcm1. MoMcm1 interacted with both Mst12 and Mata‐1 in yeast two‐hybrid assays. Deletion of MoMCM1 resulted in the loss of male fertility and microconidium production. The Momcm1 mutant was defective in appressorium penetration and formed narrower invasive hyphae, which may be responsible for its reduced virulence. In transformants expressing MoMCM1–eGFP fusion, GFP signals were observed in the nucleus. We also generated the Momcm1 mst12 double mutant, which was defective in penetration and non‐pathogenic. On hydrophilic surfaces, germ tubes produced by the double mutant were severely curved, and 20% of them formed appressoria. In contrast, the Momcm1 or mst12 mutant did not form appressoria on hydrophilic surfaces. These results suggest that MoMCM1 and MST12 have overlapping functions to suppress appressorium formation under non‐conducive conditions. MoMcm1 may interact with Mst12 and MatA‐1 to regulate germ tube identity and male fertility respectively.
Eukaryotic cell cycle involves a number of protein kinases important for the onset and progression through mitosis, most of which are well characterized in the budding and fission yeasts and ...conserved in other fungi. However, unlike the model yeast and filamentous fungi that have a single Cdc2 essential for cell cycle progression, the wheat scab fungus Fusarium graminearum contains two CDC2 orthologs. The cdc2A and cdc2B mutants had no obvious defects in growth rate and conidiation but deletion of both of them is lethal, indicating that these two CDC2 orthologs have redundant functions during vegetative growth and asexual reproduction. However, whereas the cdc2B mutant was normal, the cdc2A mutant was significantly reduced in virulence and rarely produced ascospores. Although deletion of CDC2A had no obvious effect on the formation of penetration branches or hyphopodia, the cdc2A mutant was limited in the differentiation and growth of infectious growth in wheat tissues. Therefore, CDC2A plays stage-specific roles in cell cycle regulation during infectious growth and sexual reproduction. Both CDC2A and CDC2B are constitutively expressed but only CDC2A was up-regulated during plant infection and ascosporogenesis. Localization of Cdc2A- GFP to the nucleus but not Cdc2B-GFP was observed in vegetative hyphae, ascospores, and infectious hyphae. Complementation assays with chimeric fusion constructs showed that both the N- and C-terminal regions of Cdc2A are important for its functions in pathogenesis and ascosporogenesis but only the N-terminal region is important for its subcellular localization. Among the Sordariomycetes, only three Fusarium species closely related to F. graminearum have two CDC2 genes. Furthermore, F. graminearum uniquely has two Aurora kinase genes and one additional putative cyclin gene, and its orthologs of CAK1 and other four essential mitotic kinases in the budding yeast are dispensable for viability. Overall, our data indicate that cell cycle regulation is different between vegetative and infectious hyphae in F. graminearum and Cdc2A, possibly by interacting with a stage-specific cyclin, plays a more important role than Cdc2B during ascosporogenesis and plant infection.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The transient response of a turbocharger has an important influence on the performance of engine variable conditions. An accurate estimation of the transient response process time is necessary for ...variable condition control to improve the engine response characteristics. In this study, the effect of the key factors on the turbocharger response speed, such as the turbine inlet flow rate, temperature and rotor inertia is investigated. In addition, the relationships between the turbocharger response time with respect to the response process of the turbine inlet initial state flow rate, turbine inlet end state flow rate, turbine inlet temperature and rotor inertia show correlations. The predicted response time values obtained using the relationships in comparison to the simulated values within the calculation range of this study are within a 10% error.
Ustilaginoidins are a kind of mycotoxins with 9,9'-linked bis-naphtho-γ-pyrones structures produced by the rice false smut pathogen Villosiclava virens. These metabolites displayed a wide range of ...bioactivities, such as teratogenic, cytotoxic, phytotoxic, and antibacterial activities. So far 26 ustilaginoidins have been isolated from V. virens, among which 18 compounds contained stereogenic center(s), however, most of them were unknown for the absolute configurations, except that of ustilaginoidin D. In this study, the absolute structures of these ustilaginoidins were constructed for the first time by analysis of the biosynthetic monomers obtained from a gene knockout mutant (ΔUV_2091) of V. virens. The gene UV_2091 was predicted to encode an enzyme that dimerized the monomeric naphtho-γ-pyrones in V. virens. Knockout of this gene led to the accumulation of three monomers, namely hemiustilaginoidin F (1), epihemiustilaginoidin D (2), and hemiustilaginoidin D (3), but the production of ustilaginoidins was completely blocked. The structures of the monomers were deduced by spectroscopic analysis, in combination with TDDFT ECD calculations for determining the absolute configurations. These compounds were tested for their phytotoxic, cytotoxic, antibacterial, and antifungal activities. Compounds 1 and 3 showed inhibition against the radicle and plumule elongation of rice and lettuce seeds at the tested concentrations. Compound 1 was active against the tested five human cancer cells, with half maximal inhibitory concentrations (IC
s) of 13.2~37.3 μM. Compounds 1~3 inhibited the growth of the tested pathogenic bacteria with minimum inhibitory concentrations of 8~32 µg/mL, while compound 3 exhibited antifungal activity against Magnaporthe oryzae (IC
, 5.21 µg/mL). A comparison of these data with those of the ustilaginoidins provided insights into the structure-bioactivity relationships.
Ascospores are the primary inoculum in the wheat scab fungus Fusarium graminearum that was recently shown to have sexual stage-specific A-to-I RNA editing. One of the genes with premature-stop-codons ...requiring A-to-I editing to encode full-length functional proteins is AMD1 that encodes a protein with a major facilitator superfamily (MFS) domain. Here, we characterized the functions of AMD1 and its UAG to UGG editing event. The amd1 deletion mutant was normal in growth and conidiation but defective in ascospore discharge due to the premature breakdown of its ascus wall in older perithecia, which is consistent with the specific expression of AMD1 at later stages of sexual development. Expression of the wild-type or edited allele of AMD1 but not un-editable allele rescued the defects of amd1 in ascospore discharge. Furthermore, Amd1-GFP localized to the ascus membrane and Amd1 orthologs are only present in ascocarp-forming fungi that physically discharge ascospores. Interestingly, deletion of AMD1 results in the up-regulation of a number of genes related to transporter activity and membrane functions. Overall, these results indicated that Amd1 may play a critical role in maintaining ascus wall integrity during ascus maturation, and A-to-I editing of its transcripts is important for ascospore discharge in F. graminearum.
Diabetic nephropathy (DN) is a major complication of diabetes. Interleukin-1 receptor-associated kinase 2 (IRAK2) has been implicated in various diseases. This study aimed to investigate the role of ...IRAK2 in DN progression and its association with inflammation and the nuclear factor-kappa B (NF-κB) signaling pathway. DN model mice were generated by intraperitoneal injection of streptozotocin. IRAK2 expression was upregulated in the DN model mice. IRAK2 knockdown increased weight and reduced blood glucose levels in DN model mice. In addition, IRAK2 downregulation improved glomerular morphology in DN mice. IRAK2 knockdown reduced the levels of kidney damage biomarkers (24-h urinary protein, urine albumin-creatinine ratio, and plasma creatinine) and inflammatory cytokines (IL-6, tumor necrosis factor TNF-α, TNF-1R, and TNF-2R). Moreover, IRAK2 activated the NF-κB signaling pathway in DN model mice. Overexpression of NF-κB exacerbated DN progression, and IRAK2 knockdown reversed these effects. IRAK2 promoted DN progression and inflammation by activating the NF-κB signaling pathway. These findings suggest that IRAK2 is a potential therapeutic target for DN treatment.
The wheat head blight disease caused by
is a major concern for food security and the health of both humans and animals. As a pathogenic microorganism,
produces virulence factors during infection to ...increase pathogenicity, including various macromolecular and small molecular compounds. Among these virulence factors, secreted proteins and deoxynivalenol (DON) are important weapons for the expansion and colonization of
. Besides the presence of virulence factors, sexual reproduction is also crucial for the infection process of
and is indispensable for the emergence and spread of wheat head blight. Over the last ten years, there have been notable breakthroughs in researching the virulence factors and sexual reproduction of
. This review aims to analyze the research progress of sexual reproduction, secreted proteins, and DON of
, emphasizing the regulation of sexual reproduction and DON synthesis. We also discuss the application of new gene engineering technologies in the prevention and control of wheat head blight.
The natural dispersion of oil depends on the oil types, wave-mixing energy, and the temperature and salinity of water. Laboratory experiments were conducted to investigate the effects of these ...factors on oil dispersion. The results demonstrated that the increase in temperature significantly enhanced the oil dispersion efficiency, particularly for low-viscosity oils. At 30 °C, the dispersion efficiency is 2 times higher than that at 15 °C, while salinity has no significant effect on dispersion efficiency. Nonlinear fitting results revealed an exponential increase in dispersion efficiency with the energy dissipation rate. Furthermore, partial correlation analysis was employed to examine the effects of oil density, viscosity, and surface tension on dispersion efficiency. The results indicated a high correlation between density, viscosity, and dispersion efficiency (r = −0.801, r = −0.812), whereas the correlation coefficient of surface tension was low (r = −0.286). Based on these findings, linear and nonlinear regression models were established between dispersion efficiency and density and viscosity, enabling a rough estimation of oil spill dispersion efficiency under low sea state conditions.
Head blight caused by Fusarium graminearum is an important disease of wheat and barley. Its genome contains chromosomal regions with higher genetic variation and enriched for genes expressed in ...planta, suggesting a role of chromatin modification in the regulation of infection-related genes. In a previous study, the FTL1 gene was characterized as a novel virulence factor in the head blight fungus. FTL1 is homologous to yeast SIF2, which is a component of the Set3 complex. Many members of the yeast Set3 complex, including Hos2 histone deacetylase (HDAC), are conserved in F. graminearum. In this study, we characterized the HDF1 gene that is orthologous to HOS2. HDF1 physically interacted with FTL1 in yeast two-hybrid assays. Deletion of HDF1 resulted in a significant reduction in virulence and deoxynivalenol (DON) production. The Δhdf1 mutant failed to spread from the inoculation site to other parts of wheat heads or corn stalks. It was defective in sexual reproduction and significantly reduced in conidiation. Expression of HDF1 was highest in conidia in comparison with germlings and hyphae. Deletion of HDF1 also resulted in a 60% reduction in HDAC activity. Microarray analysis revealed that 149 and 253 genes were down- and upregulated, respectively, over fivefold in the Δhdf1 mutant. Consistent with upregulation of putative catalase and peroxidase genes, the Δhdf1 mutant was more tolerant to H2O2 than the wild type. Deletion of the other two class II HDAC genes had no obvious effect on vegetative growth and resulted in only a minor reduction in conidiation and virulence in the Δhdf2 mutant. Overall, our results indicate that HDF1 is the major class II HDAC gene in F. graminearum. It may interact with FTL1 and function as a component in a well-conserved HDAC complex in the regulation of conidiation, DON production, and pathogenesis.
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