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1.
Brain Connect ; 2024 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-38874973

RESUMO

BACKGROUND AND AIMS: Previous research has focused on static functional connectivity in gait disorders caused by cerebral small vessel disease (CSVD), neglecting dynamic functional connections and network attribution. This study aims to investigate alterations in dynamic functional network connectivity (dFNC) and topological organization variance in CSVD-related gait disorders. METHODS: A total of 85 patients with CSVD, including 41 CSVD patients with gait disorders (CSVD-GD), 44 CSVD patients with non-gait disorders (CSVD-NGD), and 32 health controls (HC) were enrolled in this study. Five networks composed of 10 independent components were selected using independent component analysis. Sliding time window and k-means clustering methods were used for dFNC analysis. The relationship between alterations in the dFNC properties and gait metrics was further assessed. RESULTS: Three reproducible dFNC states were determined (State 1: sparsely connected, State 2: intermediate pattern, State 3: strongly connected). CSVD-GD showed significantly higher fractional windows (FW) and mean dwell time (MDT) in State 1 compared to CSVD-NGD. Higher local efficiency variance was observed in the CSVD-GD group compared to HC, but no differences were found in the global efficiency comparison. Both the FW and MDT in State 1 were negatively correlated with gait speed and step length, and the relationship between MDT of State 1 and gait speed was mediated by overall cognition, information processing speed and executive function. CONCLUSIONS: Our study uncovered abnormal dFNC indicators and variations in topological organization in CSVD-GD, offering potential early prediction indicators and freshening insights into the underlying pathogenesis of gait disturbances in CSVD.

2.
Food Chem ; 448: 139141, 2024 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-38574716

RESUMO

Hydrothermal carbonization (HTC) is an effective strategy for high-value utilization of tea residue (TR), and it was noticed the aqueous phase (AP) has not been extensively studied. This study aimed to investigate the chemical components and characteristics of the AP, and applied it in active food packaging films. The results showed that the total phenolic content of AP was 1.86 mg GAE/mL, and the main compounds in AP were organic acids, alcohols, and amino acids. The AP showed excellent antibacterial activity and antioxidant capacity. The active films were prepared using the casting method. The 4:7-AP/PVA film showed outstanding mechanical properties (tensile strength = 34.18 MPa, elongation at break = 458.67%), antioxidant ability (DPPH scavenging capacity 92.01%), antibacterial activity, water resistance and biocompatibility. The banana preservation test showed the AP/PVA films could successfully prolong the shelf-life of bananas and have the potential to be food packaging films.

3.
J Agric Food Chem ; 72(17): 9637-9646, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38642053

RESUMO

Nucleoside diphosphate kinases (NDPKs) are nucleotide metabolism enzymes that play different physiological functions in different species. However, the roles of NDPK in phytopathogen and mycotoxin production are not well understood. In this study, we showed that Fusarium graminearum FgNdpk is important for vegetative growth, conidiation, sexual development, and pathogenicity. Furthermore, FgNdpk is required for deoxynivalenol (DON) production; deletion of FgNDPK downregulates the expression of DON biosynthesis genes and disrupts the formation of FgTri4-GFP-labeled toxisomes, while overexpression of FgNDPK significantly increases DON production. Interestingly, FgNdpk colocalizes with the DON biosynthesis proteins FgTri1 and FgTri4 in the toxisome, and coimmunoprecipitation (Co-IP) assays show that FgNdpk associates with FgTri1 and FgTri4 in vivo and regulates their localizations and expressions, respectively. Taken together, these data demonstrate that FgNdpk is important for vegetative growth, conidiation, and pathogenicity and acts as a key protein that regulates toxisome formation and DON biosynthesis in F. graminearum.


Assuntos
Proteínas Fúngicas , Fusarium , Núcleosídeo-Difosfato Quinase , Doenças das Plantas , Esporos Fúngicos , Tricotecenos , Fusarium/genética , Fusarium/enzimologia , Fusarium/metabolismo , Fusarium/crescimento & desenvolvimento , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Tricotecenos/metabolismo , Doenças das Plantas/microbiologia , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/genética , Núcleosídeo-Difosfato Quinase/genética , Núcleosídeo-Difosfato Quinase/metabolismo , Regulação Fúngica da Expressão Gênica , Virulência , Triticum/microbiologia
4.
Mol Plant Pathol ; 25(3): e13443, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38502146

RESUMO

SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) facilitate docking and fusion of vesicles with their target membranes, playing a crucial role in vesicle trafficking and exocytosis. However, the spatial assembly and roles of plasma membrane (PM)-associated SNAREs in phytopathogen development and pathogenicity are not clearly understood. In this study, we analysed the roles and molecular mechanisms of PM-associated SNARE complexes in the banana Fusarium wilt fungus Fusarium oxysporum f. sp. cubense tropical race 4 (FocTR4). Our findings demonstrate that FocSso1 is important for the fungal growth, conidiation, host penetration and colonization. Mechanistically, FocSso1 regulates protein secretion by mediating vesicle docking and fusion with the PM and hyphal apex. Interestingly, a FocSso1-FocSec9-FocSnc1 complex was observed to assemble not only at the fungal PM but also on the growing hyphal apex, facilitating exocytosis. FocSso2, a paralogue of FocSso1, was also found to form a ternary SNARE complex with FocSec9 and FocSnc1, but it mainly localizes to the PM in old hyphae. The functional analysis of this protein demonstrated that it is dispensable for the fungal growth but necessary for host penetration and colonization. The other subunits, FocSec9 and FocSnc1, are involved in the fungal development and facilitate host penetration. Furthermore, FocSso1 and FocSnc1 are functionally interdependent, as loss of FocSso1 leads to mis-sorting and degradation of FocSnc1 in the vacuole and vice versa. Overall, this study provides insight into the formation of two spatially and functionally distinct PM SNARE complexes and their involvement in vesicle exocytosis to regulate development and pathogenicity of FocTR4.


Assuntos
Fusarium , Membrana Celular , Citoplasma , Proteínas SNARE
5.
Int J Biol Macromol ; 261(Pt 2): 129841, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38309401

RESUMO

The transcription factor FgHtf1 is important for conidiogenesis in Fusarium graminearum and it positively regulates the expression of the sporulation-related gene FgCON7. However, the regulatory mechanism underlying its functions is still unclear. The present study intends to uncover the functional mechanism of FgHtf1 in relation to FgCon7 in F. graminearum. We demonstrated that FgCON7 serves as a target gene for FgHtf1. Interestingly, FgCon7 also binds the promoter region of FgHTF1 to negatively regulate its expression, thus forming a negative-feedback loop. We demonstrated that FgHtf1 and FgCon7 have functional redundancy in fungal development. FgCon7 localizes in the nucleus and has transcriptional activation activity. Deletion of FgCON7 significantly reduces conidia production. 4444 genes were regulated by FgCon7 in ChIP-Seq, and RNA-Seq revealed 4430 differentially expressed genes in FgCON7 deletion mutant, with CCAAT serving as a consensus binding motif of FgCon7 to the target genes. FgCon7 directly binds the promoter regions of FgMSN2, FgABAA, FgVEA and FgSMT3 genes and regulates their expression. These genes were found to be important for conidiogenesis. To our knowledge, this is the first study that unveiled the mutual regulatory functions of FgCON7 and FgHTF1 to form a negative-feedback loop, and how the loop mediates sporulation in F. graminearum.


Assuntos
Fusarium , Fatores de Transcrição , Retroalimentação , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Fusarium/fisiologia , Expressão Gênica , Regulação Fúngica da Expressão Gênica , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Doenças das Plantas/microbiologia
6.
Cancer Lett ; 588: 216738, 2024 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-38401887

RESUMO

The gastrointestinal tract can be affected by a number of diseases that pancreatic cancer (PC) is a malignant manifestation of them. The prognosis of PC patients is unfavorable and because of their diagnosis at advanced stage, the treatment of this tumor is problematic. Owing to low survival rate, there is much interest towards understanding the molecular profile of PC in an attempt in developing more effective therapeutics. The conventional therapeutics for PC include surgery, chemotherapy and radiotherapy as well as emerging immunotherapy. However, PC is still incurable and more effort should be performed. The molecular landscape of PC is an underlying factor involved in increase in progression of tumor cells. In the presence review, the newest advances in understanding the molecular and biological events in PC are discussed. The dysregulation of molecular pathways including AMPK, MAPK, STAT3, Wnt/ß-catenin and non-coding RNA transcripts has been suggested as a factor in development of tumorigenesis in PC. Moreover, cell death mechanisms such as apoptosis, autophagy, ferroptosis and necroptosis demonstrate abnormal levels. The EMT and glycolysis in PC cells enhance to ensure their metastasis and proliferation. Furthermore, such abnormal changes have been used to develop corresponding pharmacological and nanotechnological therapeutics for PC.


Assuntos
Neoplasias Pancreáticas , Humanos , Linhagem Celular Tumoral , Proliferação de Células/genética , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/genética , Apoptose , Prognóstico
7.
Stress Biol ; 3(1): 35, 2023 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-37676627

RESUMO

Vesicle trafficking is an essential cellular process upon which many physiological processes of eukaryotic cells rely. It is usually the 'language' of communication among the components of the endomembrane system within a cell, between cells and between a cell and its external environment. Generally, cells have the potential to internalize membrane-bound vesicles from external sources by endocytosis. Plants constantly interact with both mutualistic and pathogenic microbes. A large part of this interaction involves the exchange of transport vesicles between the plant cells and the microbes. Usually, in a pathogenic interaction, the pathogen releases vesicles containing bioactive molecules that can modulate the host immunity when absorbed by the host cells. In response to this attack, the host cells similarly mobilize some vesicles containing pathogenesis-related compounds to the pathogen infection site to destroy the pathogen, prevent it from penetrating the host cell or annul its influence. In fact, vesicle trafficking is involved in nearly all the strategies of phytopathogen attack subsequent plant immune responses. However, this field of plant-pathogen interaction is still at its infancy when narrowed down to plant-fungal pathogen interaction in relation to exchange of transport vesicles. Herein, we summarized some recent and novel findings unveiling the involvement of transport vesicles as a crosstalk in plant-fungal phytopathogen interaction, discussed their significance and identified some knowledge gaps to direct future research in the field. The roles of vesicles trafficking in the development of both organisms are also established.

8.
Obesity (Silver Spring) ; 31(10): 2557-2567, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37724054

RESUMO

OBJECTIVE: This study aimed to explore the relationship between white matter hyperintensities (WMHs) and cognitive impairment related to metabolic syndrome (MetS) and the underlying neural network mechanisms. METHODS: This cross-sectional study included 50 participants with MetS and WMHs (MetS-WMHs), 45 with MetS without WMHs, and 50 control participants. All participants underwent resting-state functional magnetic resonance imaging and a detailed cognitive evaluation. A graph theory analysis based on resting-state functional magnetic resonance imaging was conducted to calculate functional network properties. A mediation analysis was conducted to determine the relationship between WMHs and MetS-related cognitive impairment. RESULTS: Compared with the control group, the participants in the MetS-WMHs group displayed lower global efficiency, local efficiency, and nodal efficiency, mainly located in the regions of the salience network. Furthermore, a significant correlation was observed between functional network efficiency and cognitive performance. Mediation analysis indicated that WMHs served as a mediating variable between MetS and cognitive decline, affecting attention/executive function, language, and global cognitive function. CONCLUSIONS: WMHs mediated the association between MetS and cognitive function, with a decline in the efficiency of functional brain networks being a probable neural mechanism.


Assuntos
Disfunção Cognitiva , Síndrome Metabólica , Substância Branca , Humanos , Estudos Transversais , Síndrome Metabólica/complicações , Síndrome Metabólica/diagnóstico por imagem , Substância Branca/diagnóstico por imagem , Disfunção Cognitiva/diagnóstico por imagem , Disfunção Cognitiva/etiologia , Cognição
9.
Protein Cell ; 14(9): 653-667, 2023 09 14.
Artigo em Inglês | MEDLINE | ID: mdl-37707322

RESUMO

Lipophagy, the selective engulfment of lipid droplets (LDs) by autophagosomes for lysosomal degradation, is critical to lipid and energy homeostasis. Here we show that the lipid transfer protein ORP8 is located on LDs and mediates the encapsulation of LDs by autophagosomal membranes. This function of ORP8 is independent of its lipid transporter activity and is achieved through direct interaction with phagophore-anchored LC3/GABARAPs. Upon lipophagy induction, ORP8 has increased localization on LDs and is phosphorylated by AMPK, thereby enhancing its affinity for LC3/GABARAPs. Deletion of ORP8 or interruption of ORP8-LC3/GABARAP interaction results in accumulation of LDs and increased intracellular triglyceride. Overexpression of ORP8 alleviates LD and triglyceride deposition in the liver of ob/ob mice, and Osbpl8-/- mice exhibit liver lipid clearance defects. Our results suggest that ORP8 is a lipophagy receptor that plays a key role in cellular lipid metabolism.


Assuntos
Autofagia , Gotículas Lipídicas , Animais , Camundongos , Autofagossomos , Homeostase , Triglicerídeos
10.
Aging (Albany NY) ; 15(16): 8113-8136, 2023 08 17.
Artigo em Inglês | MEDLINE | ID: mdl-37595258

RESUMO

OBJECTIVE: Pyroptosis is a form of programmed cell death that is essential for immunity. Herein, this study was conducted to uncover the implication of pyroptosis in immunomodulation and tumor microenvironment (TME) in gastric cancer. METHODS: Prognostic pyroptosis-related genes were extracted to identify different pyroptosis phenotypes and pyroptosis genomic phenotypes via unsupervised clustering analysis in the gastric cancer meta-cohort cohort (GSE15459, GSE62254, GSE84437, GSE26253 and TCGA-STAD). The activation of hallmark gene sets was quantified by GSVA and immune cell infiltration was estimated via ssGSEA and CIBERSORT. Through PCA algorithm, pyroptosis score was conducted. The predictors of immune response (TMB and IPS) and genetic mutations were evaluated. The efficacy of pyroptosis score in predicting immune response was verified in two anti-PD-1 therapy cohorts. RESULTS: Three different pyroptosis phenotypes with different prognosis, biological pathways and tumor immune microenvironment were established among 1275 gastric cancer patients, corresponding to three immune phenotypes: immune-inflamed, immune-desert, and immune-excluded. According to the pyroptosis score, patients were separated into high and low pyroptosis score groups. Low pyroptosis score indicated favorable survival outcomes, enhanced immune responses, and increased mutation frequency. Moreover, low pyroptosis score patients displayed more clinical benefits from anti-PD-1 and prolonged survival time. CONCLUSION: Our findings uncovered a nonnegligible role of pyroptosis in immunomodulation and TME multiformity and complicacy in gastric cancer. Quantifying the pyroptosis score in individual tumors may tailor more effective immunotherapeutic strategies.


Assuntos
Neoplasias Gástricas , Humanos , Piroptose , Imunoterapia , Imunomodulação , Fenótipo , Microambiente Tumoral
11.
Langmuir ; 39(33): 11851-11863, 2023 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-37556777

RESUMO

E-OLCN photocatalyst was synthesized by oxygen doping of low molecular weight carbon nitride (LCN) with ethanol solvent stripping. The enhanced light absorption, fast electron transport rate, and photogenerated carrier separation efficiency of E-OLCN leads to the excellent photocatalytic degradation performance compared with the original materials. The synergistic effect of oxygen doping and ethanol solvent stripping plays a significant role for the modulation of electronic and structural properties of the prepared catalysts. Methyl orange (MO) and rhodamine B (RhB) are chosen as typical pollutants for the application of photocatalytic degradation. The E-OLCN sample exhibits outstanding photocatalytic degradation performance, where the rate constant k (1 × 10-2 min-1) of E-OLCN (1.68) is 2.9 times than that of O-LCN (0.58) and 8.8 times than that of pristine LCN (0.19) for MO. Moreover, modulated E-OLCN shows good stability after cycling experiments and the activity still achieved 90%. The detailed mechanism for MO degradation was proposed with the technical support of liquid chromatography-mass spectrometry (LC-MS) and electron spin resonance (EPR). The superoxide radical (·O2-) is the main active species and the MO molecule could be decomposition completely.

12.
Langmuir ; 39(29): 10098-10111, 2023 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-37432980

RESUMO

Adsorption materials have demonstrated huge potential in treating sewage; however, it is a great challenge to fabricate an adsorbent effectively adsorbing multiple dyestuffs and heavy metal ions simultaneously. Here, a magnetic core@shell Fe3O4@polypyrrole@sodium dodecyl sulfate (Fe3O4@PPy@SDS) composite is prepared through the combination of a hydrothermal method, an in situ polymerization method, and modification, exhibiting enhanced selective removal of five dyestuffs (methylene blue (MB), malachite green (MG), rhodamine B (RhB), Congo red (CR), acid red 1 (AR1)), and heavy metal ions (Mn(VII)). The effects of adsorbent type, time, initial concentration of the adsorbate, and temperature on adsorption performances are investigated in detail. Kinetics and isotherm studies indicate that all adsorption processes are more in line with the pseudo-second-order kinetic model and the Langmuir model, the diffusion behavior is controlled by intraparticle diffusion and liquid film diffusion, and research of thermodynamics reveals a spontaneous endothermic behavior. The removal efficiency after five desorption-adsorption cycles can still reach more than 90%. The prepared Fe3O4@PPy@SDS composite is an efficient and promising renewable adsorbent for the treatment of dyestuffs and Mn(VII), exhibiting a wide range of applications in the field of adsorption.

13.
Microbiol Spectr ; 11(4): e0440322, 2023 08 17.
Artigo em Inglês | MEDLINE | ID: mdl-37367437

RESUMO

Signal peptidase (SPase) is responsible for cleavage of N-terminal signal peptides in most secretory precursor proteins and many membrane proteins during maturation. In this study, we identified four components of the SPase complex (FoSec11, FoSpc1, FoSpc2, and FoSpc3) in the banana wilt fungal pathogen Fusarium odoratissimum. We proved that interactions exist among the four SPase subunits by bimolecular fluorescence complementation (BiFC) and affinity purification and mass spectrometry (AP-MS) assays. Among the four SPase genes, FoSPC2 was successfully deleted. FoSPC2 deletion caused defects in vegetative growth, conidiation, and virulence. Loss of FoSPC2 also affected the secretion of some pathogenicity-related extracellular enzymes, suggesting that SPase without FoSpc2 may have a lower efficiency in managing the maturation of the extracellular enzymes in F. odoratissimum. In addition, we found that the ΔFoSPC2 mutant had increased sensitivity to light, and the colonies of the mutant grew faster under all-dark conditions than under all-light conditions. We further observed that deletion of FoSPC2 affected expression of the blue light photoreceptor gene FoWC2, leading to cytoplasmic accumulation of FoWc2 under all-light conditions. Since FoWc2 has signal peptides, FoSpc2 may regulate the expression and subcellular localization of FoWc2 indirectly. Contrary to its response to light, the ΔFoSPC2 mutant displayed a significant decreased sensitivity to osmotic stress, and culturing the mutant under osmotic stress conditions restored both the localization of FoWc2 and light sensitivity of the ΔFoSPC2, suggesting that a cross talk between osmotic stress and light response pathways in F. odoratissimum and FoSpc2 takes part in these processes. IMPORTANCE In this study, we identified four components of SPase in the banana wilt pathogen Fusarium odoratissimum and characterized the SPase FoSpc2. Loss of FoSPC2 affected the secretion of extracellular enzymes, suggesting that SPase without FoSpc2 may have a lower efficiency in managing the maturation of the extracellular enzymes in F. odoratissimum. In addition, this is the first time that we have found a relationship between the SPase and fungal light response. Deletion of FoSPC2 resulted in decreased sensitivity to the osmotic stresses but with increased sensitivity to light. Continuous light inhibited the growth rate of the ΔFoSPC2 mutant and affected the cellular localization of the blue light photoreceptor FoWc2 in this mutant, but culturing the mutant under osmotic stress both restored the localization of FoWc2 and eliminated the light sensitivity of the ΔFoSPC2 mutant, suggesting that loss of FoSPC2 may affect a cross talk between the osmotic stress and light response pathways in F. odoratissimum.


Assuntos
Proteínas Fúngicas , Fotofobia , Humanos , Virulência/genética , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Serina Endopeptidases/metabolismo , Proteínas de Membrana/metabolismo , Sinais Direcionadores de Proteínas
14.
New Phytol ; 239(4): 1384-1403, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-37291895

RESUMO

Secretion is a fundamental process that plant pathogens utilize to deliver effectors into the host to downregulate immunity and promote infection. Here, we uncover a fascinating membrane trafficking and delivery route that originates from vacuolar membranes in Magnaporthe oryzae and conduits to the host interface and plasma membrane. To perform such secretory/trafficking function, MoRab7 first recruits the retromer complex to the vacuolar membrane, enabling recognition of a family of SNARE proteins, including MoSnc1. Live-cell imaging confirmed a highly dynamic vesicular trafficking of the retromer complex component(s) and MoSnc1 toward and across the host interface or plasma membrane, and subsequent fusion with target membranes. Interestingly, disruption of the MoRab7/Retromer/MoSnc1-based endolysosomal cascade affects effector secretion and fungal pathogenicity. Taken together, we discovered an unconventional protein and membrane trafficking route starting from the fungal endolysosomes to the M. oryzae-rice interaction interface and dissect the role of MoRab7/Retromer/MoSnc1 sorting machinery in effector secretion during biotrophy and invasive growth in rice blast fungus.


Assuntos
Magnaporthe , Oryza , Endossomos/metabolismo , Transporte Proteico , Vacúolos/metabolismo , Transporte Biológico , Membrana Celular/metabolismo , Oryza/metabolismo , Proteínas Fúngicas/metabolismo , Doenças das Plantas/microbiologia
15.
Environ Sci Pollut Res Int ; 30(29): 73560-73569, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37191748

RESUMO

In shal low eutrophic lakes, submersed macrophytes are essential for maintaining a clear water state and they are significantly affected by benthic fish disturbance, light availability, and sediment types. We conducted a mesocosm experiment with benthic fish (Misgurnus anguillicaudatus), two light regimes, and submerged macrophyte (Vallisneria natans) growing in two sediment types to investigate the ecological effects of benthic fish and light regimes on water quality and the growth of submersed macrophyte. Our findings indicated that the benthic fish increased the concentrations of total nitrogen, total phosphorus, and total dissolved phosphorus in the overlying water. The effects of benthic fish on ammonia-nitrogen (NH4+-N) and chlorophyll a (Chl-a) contents were related to light regimes. Fish disturbance indirectly promoted the growth of macrophytes growing in sand by increasing NH4+-N content in overlying water. However, the increasing Chl-a content stimulated by fish disturbance and high light regime reduced the growth of submersed macrophytes growing in clay due to shading. Macrophytes with different sediments had different strategies coping with light. Plants growing in sand responded to low light mainly by adjusting the leaf and root biomass allocation, whereas plants growing in clay responded to low light by physiologically adjusting the soluble carbohydrate content. The findings of this study might help restore lake vegetation to some degree, and using nutrient-poor sediment might be an appropriate method to avoid the detrimental effects of fish-mediated disturbances on the growth of submerged macrophytes.


Assuntos
Hydrocharitaceae , Qualidade da Água , Animais , Clorofila A , Areia , Argila , Lagos , Peixes , Hydrocharitaceae/fisiologia , Fósforo/análise , Nitrogênio/análise
16.
Microbiol Spectr ; 11(3): e0505822, 2023 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-37158744

RESUMO

The RNA exosome complex is a conserved, multisubunit RNase complex that contributes to the processing and degradation of RNAs in mammalian cells. However, the roles of the RNA exosome in phytopathogenic fungi and how it relates to fungal development and pathogenicity remain unclear. Herein, we identified 12 components of the RNA exosome in the wheat fungal pathogen Fusarium graminearum. Live-cell imaging showed that all the components of the RNA exosome complex are localized in the nucleus. FgEXOSC1 and FgEXOSCA were successfully knocked out; they are both involved in the vegetative growth, sexual reproduction, and pathogenicity of F. graminearum. Moreover, deletion of FgEXOSC1 resulted in abnormal toxisomes, decreased deoxynivalenol (DON) production, and downregulation of the expression levels of DON biosynthesis genes. The RNA-binding domain and N-terminal region of FgExosc1 are required for its normal localization and functions. Transcriptome sequencing (RNA-seq) showed that the disruption of FgEXOSC1 resulted in differential expression of 3,439 genes. Genes involved in processing of noncoding RNA (ncRNA), rRNA and ncRNA metabolism, ribosome biogenesis, and ribonucleoprotein complex biogenesis were significantly upregulated. Furthermore, subcellular localization, green fluorescent protein (GFP) pulldown, and coimmunoprecipitation (co-IP) assays demonstrated that FgExosc1 associates with the other components of the RNA exosome to form the RNA exosome complex in F. graminearum. Deletion of FgEXOSC1 and FgEXOSCA reduced the relative expression of some of the other subunits of the RNA exosome. Deletion of FgEXOSC1 affected the localization of FgExosc4, FgExosc6, and FgExosc7. In summary, our study reveals that the RNA exosome is involved in vegetative growth, sexual reproduction, DON production, and pathogenicity of F. graminearum. IMPORTANCE The RNA exosome complex is the most versatile RNA degradation machinery in eukaryotes. However, little is known about how this complex regulates the development and pathogenicity of plant-pathogenic fungi. In this study, we systematically identified 12 components of the RNA exosome complex in Fusarium head blight fungus Fusarium graminearum and first unveiled their subcellular localizations and established their biological functions in relation to the fungal development and pathogenesis. All the RNA exosome components are localized in the nucleus. FgExosc1 and FgExoscA are both required for the vegetative growth, sexual reproduction, DON production and pathogenicity in F. graminearum. FgExosc1 is involved in ncRNA processing, rRNA and ncRNA metabolism process, ribosome biogenesis and ribonucleoprotein complex biogenesis. FgExosc1 associates with the other components of RNA exosome complex and form the exosome complex in F. graminearum. Our study provides new insights into the role of the RNA exosome in regulating RNA metabolism, which is associated with fungal development and pathogenicity.


Assuntos
Fusarium , Tricotecenos , Fusarium/genética , Virulência/genética , Complexo Multienzimático de Ribonucleases do Exossomo/genética , Complexo Multienzimático de Ribonucleases do Exossomo/metabolismo , Tricotecenos/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Ribonucleoproteínas/metabolismo
17.
J Fungi (Basel) ; 9(4)2023 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-37108875

RESUMO

Soil-borne beneficial microbes establish symbioses with plant hosts and play key roles during growth and development therein. In this study, two fungal strains, FLP7 and B9, were isolated from the rhizosphere microbiome associated with Choy Sum (Brassica rapa var. parachinensis) and barley (Hordeum vulgare), respectively. Sequence analyses of the internal transcribed spacer and 18S ribosomal RNA genes combined with colony and conidial morphology identified FLP7 and B9 to be Penicillium citrinum strains/isolates. Plant-fungus interaction assays revealed that isolate B9 showed significant growth promotion effects in Choy Sum plants cultivated in normal soil, as well as under phosphate-limiting conditions. In comparison to the mock control, B9-inoculated plants showed a 34% increase in growth in aerial parts and an 85% upsurge in the fresh weight of roots when cultivated in sterilized soil. The dry biomass of such fungus-inoculated Choy Sum increased by 39% and 74% for the shoots and roots, respectively. Root colonization assays showed that P. citrinum associates directly with the root surface but does not enter or invade the root cortex of the inoculated Choy Sum plants. Preliminary results also indicated that P. citrinum can promote growth in Choy Sum via volatile metabolites too. Interestingly, we detected relatively higher amounts of gibberellins and cytokinins in axenic P. citrinum culture filtrates through liquid chromatography-mass spectrometry analyses. This could plausibly explain the overall growth induction in P. citrinum-inoculated Choy Sum plants. Furthermore, the phenotypic growth defects associated with the Arabidopsis ga1 mutant could be chemically complemented by the exogenous application of P. citrinum culture filtrate, which also showed accumulation of fungus-derived active gibberellins. Our study underscores the importance of transkingdom beneficial effects of such mycobiome-assisted nutrient assimilation and beneficial fungus-derived phytohormone-like metabolites in the induction of robust growth in urban farmed crops.

18.
Transl Psychiatry ; 13(1): 135, 2023 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-37100779

RESUMO

Carboxypeptidase E (CPE) is a multifunctional protein with many nonenzymatic functions in various systems. Previous studies using CPE knock-out mice have shown that CPE has neuroprotective effects against stress and is involved in learning and memory. However, the functions of CPE in neurons are still largely unknown. Here we used a Camk2a-Cre system to conditionally knockout CPE in neurons. The wild-type, CPEflox/-, and CPEflox/flox mice were weaned, ear-tagged, and tail clipped for genotyping at 3 weeks old, and they underwent open field, object recognition, Y-maze, and fear conditioning tests at 8 weeks old. The CPEflox/flox mice had normal body weight and glucose metabolism. The behavioral tests showed that CPEflox/flox mice had impaired learning and memory compared with wild-type and CPEflox/- mice. Surprisingly, the subiculum (Sub) region of CPEflox/flox mice was completely degenerated, unlike the CPE full knockout mice, which exhibit CA3 region neurodegeneration. In addition, doublecortin immunostaining suggested that neurogenesis in the dentate gyrus of the hippocampus was significantly reduced in CPEflox/flox mice. Interestingly, TrkB phosphorylation in the hippocampus was downregulated in CPEflox/flox mice, but brain-derived neurotrophic factor levels were not. In both the hippocampus and dorsal medial prefrontal cortex, we observed reduced MAP2 and GFAP expression in CPEflox/flox mice. Taken together, the results of this study demonstrate that specific neuronal CPE knockout leads to central nervous system dysfunction in mice, including learning and memory deficits, hippocampal Sub degeneration and impaired neurogenesis.


Assuntos
Hipocampo , Aprendizagem , Camundongos , Animais , Camundongos Knockout , Carboxipeptidase H/genética , Carboxipeptidase H/metabolismo , Hipocampo/metabolismo , Transtornos da Memória/genética , Transtornos da Memória/metabolismo , Aprendizagem em Labirinto/fisiologia
19.
PLoS Pathog ; 19(3): e1011255, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-36928713

RESUMO

The mitotic exit is a key step in cell cycle, but the mechanism of mitotic exit network in the wheat head blight fungus Fusarium graminearum remains unclear. F. graminearum infects wheat spikelets and colonizes the entire head by growing through the rachis node at the bottom of each spikelet. In this study, we found that a small GTPase FgTem1 plays an important role in F. graminearum pathogenicity and functions in regulating the formation of infection structures and invasive hyphal growth on wheat spikelets and wheat coleoptiles, but plays only little roles in vegetative growth and conidiation of the phytopathogen. FgTem1 localizes to both the inner nuclear periphery and the spindle pole bodies, and negatively regulates mitotic exit in F. graminearum. Furthermore, the regulatory mechanisms of FgTem1 have been further investigated by high-throughput co-immunoprecipitation and genetic strategies. The septins FgCdc10 and FgCdc11 were demonstrated to interact with the dominant negative form of FgTem1, and FgCdc11 was found to regulate the localization of FgTem1. The cell cycle arrest protein FgBub2-FgBfa1 complex was shown to act as the GTPase-activating protein (GAP) for FgTem1. We further demonstrated that a direct interaction exists between FgBub2 and FgBfa1 which crucially promotes conidiation, pathogenicity and DON production, and negatively regulates septum formation and nuclear division in F. graminearum. Deletion of FgBUB2 and FgBFA1 genes caused fewer perithecia and immature asci formations, and dramatically down-regulated trichothecene biosynthesis (TRI) gene expressions. Double deletion of FgBUB2/FgBFA1 genes showed that FgBUB2 and FgBFA1 have little functional redundancy in F. graminearum. In summary, we systemically demonstrated that FgTem1 and its GAP FgBub2-FgBfa1 complex are required for fungal development and pathogenicity in F. graminearum.


Assuntos
Fusarium , Proteínas Monoméricas de Ligação ao GTP , Virulência , Proteínas Monoméricas de Ligação ao GTP/genética , Proteínas Monoméricas de Ligação ao GTP/metabolismo , Divisão do Núcleo Celular , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Doenças das Plantas/microbiologia , Esporos Fúngicos
20.
Nat Commun ; 14(1): 1289, 2023 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-36894574

RESUMO

Arboviruses and symbiotic viruses can be paternally transmitted by male insects to their offspring for long-term viral persistence in nature, but the mechanism remains largely unknown. Here, we identify the sperm-specific serpin protein HongrES1 of leafhopper Recilia dorsalis as a mediator of paternal transmission of the reovirus Rice gall dwarf virus (RGDV) and a previously undescribed symbiotic virus of the Virgaviridae family, Recilia dorsalis filamentous virus (RdFV). We show that HongrES1 mediates the direct binding of virions to leafhopper sperm surfaces and subsequent paternal transmission via interaction with both viral capsid proteins. Direct interaction of viral capsid proteins mediates simultaneously invasion of two viruses into male reproductive organs. Moreover, arbovirus activates HongrES1 expression to suppress the conversion of prophenoloxidase to active phenoloxidase, potentially producing a mild antiviral melanization defense. Paternal virus transmission scarcely affects offspring fitness. These findings provide insights into how different viruses cooperatively hijack insect sperm-specific proteins for paternal transmission without disturbing sperm functions.


Assuntos
Arbovírus , Hemípteros , Reoviridae , Animais , Masculino , Proteínas do Espermatozoide , Proteínas do Capsídeo , Sêmen , Insetos , Reoviridae/fisiologia
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