scholarly journals Extracellular Vesicles from Fusarium graminearum Contain Protein Effectors Expressed during Infection of Corn

2021 ◽  
Vol 7 (11) ◽  
pp. 977
Author(s):  
Donovan Garcia-Ceron ◽  
Rohan G. T. Lowe ◽  
James A. McKenna ◽  
Linda M. Brain ◽  
Charlotte S. Dawson ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Extracellular Vesicles ◽  
Plant Pathogens ◽  
Control Mechanisms ◽  
Pathogenic Yeast ◽  
Fungal Virulence ◽  
Fungal Plant Pathogens ◽  
Low Efficiency ◽  
Plant Pathogenesis

Fusarium graminearum (Fgr) is a devastating filamentous fungal pathogen that causes diseases in cereals, while producing mycotoxins that are toxic for humans and animals, and render grains unusable. Low efficiency in managing Fgr poses a constant need for identifying novel control mechanisms. Evidence that fungal extracellular vesicles (EVs) from pathogenic yeast have a role in human disease led us to question whether this is also true for fungal plant pathogens. We separated EVs from Fgr and performed a proteomic analysis to determine if EVs carry proteins with potential roles in pathogenesis. We revealed that protein effectors, which are crucial for fungal virulence, were detected in EV preparations and some of them did not contain predicted secretion signals. Furthermore, a transcriptomic analysis of corn (Zea mays) plants infected by Fgr revealed that the genes of some of the effectors were highly expressed in vivo, suggesting that the Fgr EVs are a mechanism for the unconventional secretion of effectors and virulence factors. Our results expand the knowledge on fungal EVs in plant pathogenesis and cross-kingdom communication, and may contribute to the discovery of new antifungals.

Localization and Visualization of Specific Gene Products in Fungal Plant Pathogens

2000 ◽  
Vol 6 (S2) ◽  
pp. 680-681 ◽  
Author(s):  
T. M. Bourett ◽  
K. J. Czymmek ◽  
T. M. Dezwaan ◽  
J. A. Sweigard ◽  
R. J. Howard
Keyword(s):  
Fungal Pathogens ◽  
Plant Pathogens ◽  
Protein Localization ◽  
Infected Plant ◽  
Cell Interaction ◽  
Specific Gene ◽  
Gene Products ◽  
Tem Analysis ◽  
Fungal Plant Pathogens ◽  
Plant Pathogenesis

Specific gene products of both pathogens and hosts have been implicated as decisive elements during plant pathogenesis. While expression of some of these genes is constitutive, that of others is likely ephemeral and activated only during a particular stage of the interaction. Because the relative timing of expression may be critical, transcription and translation have often been addressed by extracting mRNA and proteins from infected plant tissue. This approach, however, cannot readily detect proteins of low abundance in bulk samples nor offer much useful information on cell-cell interaction. Only a cytological analysis that employs microscopy can resolve the temporal and spatial details of gene expression. Typically, such protein localization studies have required specific antibodies, but these large probe molecules do not diffuse into living or conventionally fixed cells of either fungal pathogens or plant hosts. For TEM analysis, these permeability-imposed limitations have been reduced by thin sectioning to render accessible antibody binding sites.

Regulation and Dynamics of Gene Expression During the Life Cycle of Fusarium graminearum

2020 ◽  
Vol 110 (8) ◽  
pp. 1368-1374
Author(s):  
Elizabeth K. Brauer ◽  
Rajagopal Subramaniam ◽  
Linda J. Harris
Keyword(s):  
Gene Expression ◽  
Fusarium Graminearum ◽  
Fungal Pathogens ◽  
Plant Pathogens ◽  
Transcriptional Profiling ◽  
Fungal Plant Pathogens ◽  
Essential Components ◽  
Host Infection

Fungal pathogens survive harsh environments and overcome physical, temporal, and chemical barriers to colonize their hosts and reproduce. Fusarium graminearum was one of the first fungal plant pathogens for which transcriptomic tools were developed, making analysis of gene expression a cornerstone approach in studying its biology. The analysis of gene expression in diverse in vitro conditions and during infection of different cereal crops has revealed subsets of both unique and shared transcriptionally regulated genes. Together with genetic studies, these approaches have enhanced our understanding of the development and infection cycle of this economically important pathogen. Here, we will outline recent advances in transcriptional profiling during sporogenesis, spore germination, vegetative growth, and host infection. Several transcriptional regulators have been identified as essential components in these responses and the role of select transcription factors will be highlighted. Finally, we describe some of the gaps in our understanding of F. graminearum biology and how expression analysis could help to address these gaps.

Adaptation and response to mycotoxin presence in pathogen-pathogen interactions within the Fusarium genus

2016 ◽  
Vol 9 (4) ◽  
pp. 565-575 ◽  
Author(s):  
A. Dawidziuk ◽  
G. Koczyk ◽  
D. Popiel
Keyword(s):  
Fusarium Graminearum ◽  
Plant Pathogens ◽  
Fusarium Verticillioides ◽  
Warning Signal ◽  
Resistance Mechanisms ◽  
Growth Patterns ◽  
Fungal Plant Pathogens ◽  
Low Concentrations ◽  
Dose Dependent

The ability of fungal plant pathogens to exude bioactive compounds is an important element of competition in a changing environment. The filamentous fungi usually retain a number of adaptations related not only to the production of toxic compounds by themselves but also to the mitigation of exogenous influences by toxins present in the environment. We examined a distinct effect of toxins on morphology, growth patterns and gene expression after stimulation in mycotoxin-producing and nonproducing isolates representing four evolutionarily divergent species (and chemotypes) within the Fusarium genus (Fusarium graminearum, Fusarium oxysporum, Fusarium proliferatum and Fusarium verticillioides). The aim of our work was to investigate the influence of mycotoxins present in the environment on fungal isolates belonging to evolutionarily divergent complexes within Fusarium genus. The results point to retention of resistance mechanisms in non-producer isolates (F. oxysporum) and specific dose-dependent differences in response to other mycotoxins. In particular, the growth of Fusarium graminearum (confirmed zearalenone and trichothecene producer) was shown to be significantly inhibited by fumonisin B1 and deoxynivalenol. Conversely, spread of Fusarium verticillioides was accelerated by low concentrations (0.5 mg/l) of nivalenol and zearalenone and deoxynivalenol addition resulted in upregulation of the fumonisin poliketyde synthase (FUM1). The basics of competition between divergent fusaria can be described by ‘rock-paper-scissors’ theory, but some of the effects can be explained by other interactions, e.g. autotoxicity of deoxynivalenol and the potential role of low doses of trichothecenes and zearalenone acting as a ‘warning signal’ for competing species.

scholarly journals Barrage Zone Formation Between Vegetatively Incompatible Fusarium graminearum (Gibberella zeae) Isolates

2004 ◽  
Vol 94 (5) ◽  
pp. 432-437 ◽  
Author(s):  
B. D. McCallum ◽  
A. Tekauz ◽  
J. Gilbert
Keyword(s):  
Fusarium Graminearum ◽  
Plant Pathogens ◽  
Vegetative Compatibility ◽  
Gibberella Zeae ◽  
Constant Light ◽  
Constant Darkness ◽  
Zone Formation ◽  
Vegetative Compatibility Groups ◽  
Auxotrophic Mutants ◽  
Fungal Plant Pathogens

Vegetative compatibility has been used to assess the population biology of many fungal plant pathogens. However, for many species, including Fusarium graminearum, this has meant making auxotrophic mutants to force heterokaryon formation. A method was developed to observe barrage zones of thick, raised mycelium at the junctions of vegetatively incompatible F. graminearum isolates. The appearance of the barrage zones was influenced by the growth medium and the light. Barrage zones on V8 agar were thicker and better defined than those on potato dextrose agar, Spezieller Nahrstoffarmer agar, and water agar. The addition of ground wheat kernels to V8 agar enhanced barrage zone formation. Incubating the cultures under constant light at 2,150 lx produced more distinct barrage zones than constant light at 3,400 lx, constant darkness, or ambient room light. Forty-three F. graminearum isolates from 34 vegetative compatibility groups, determined previously using nit auxotrophic mutants, were paired in all combinations using these optimized conditions. Isolates in different vegetative compatibility groups typically formed distinct, thick barrage zones at their junctions. Pairs of isolates in the same vegetative compatibility group had a very slight or no visible reaction, or rarely, a distinct “line gap” of sparse mycelium. Subcultures from the same isolate typically had no visible reaction at their colony junctions; however, subcultures from some isolates had thin, slight barrage zones. This method was used to identify the proportion of each of four F. graminearum isolates from infected barley spikes in the field, inoculated previously with a mixture of conidia from these four isolates. Barrage zone formation represents a rapid method to screen vegetative compatibility groups in F. graminearum and may be useful for other Fusarium species.

scholarly journals Near-Complete Genomes of Two Trichoderma Species: A Resource for Biological Control of Plant Pathogens

2020 ◽  
Vol 33 (8) ◽  
pp. 1036-1039 ◽  
Author(s):  
Yi Zhou ◽  
Yilian Wang ◽  
Kai Chen ◽  
Yuanzheng Wu ◽  
Jindong Hu ◽  
...  
Keyword(s):  
Biological Control ◽  
Enzyme Production ◽  
Plant Pathogens ◽  
Plant Diseases ◽  
Effective Control ◽  
Control Mechanisms ◽  
Trichoderma Spp ◽  
Trichoderma Species ◽  
Genome Data ◽  
Fungal Plant Pathogens

Trichoderma species are widely used to control fungal and nematode diseases of crops. To date, only one complete Trichoderma genome has been sequenced, T. reesei QM6a, a model fungus for industrial enzyme production, while the species or strains used for biological control of plant diseases are only available as draft genomes. Previously, we demonstrated that two Trichoderma strains (T. afroharzianum and T. cyanodichotomus) provide effective control of nematode and fungal plant pathogens. Based on deep sequencing using Illumina and Pacbio platforms, we have assembled high-quality genomes of the above two strains, with contig N50 reaching 4.2 and 1.7 Mbp, respectively, which is greater than those of published draft genomes. The genome data will provide a resource to assist research on the biological control mechanisms of Trichoderma spp.

scholarly journals Unravelling the role of alcohol copper radical oxidases in fungal plant pathogens

2021 ◽  
Author(s):  
Bastien Bissaro ◽  
Sayo Kodama ◽  
Hayat Hage ◽  
David Ribeaucourt ◽  
Mireille Haon ◽  
...  
Keyword(s):  
Plant Pathogens ◽  
Crop Protection ◽  
Redox Partner ◽  
Fungal Plant Pathogens ◽  
Plant Cuticles ◽  
Penetration Ability ◽  
Molecular Signals

Abstract Copper radical oxidases (CRO) form a class of enzymes with a longstanding history encompassing diverse substrate specificities. While the biological function of most CROs remains unknown, we observed that CROs active on aliphatic alcohols are found only in fungal plant pathogens. Here, we unveil the role of these CROs and the identity of their natural redox partner, a haem-iron peroxidase. Combining multiscale approaches, we report that Colletotrichum and Magnaporthe appressoria (specialized cells that puncture the plant cuticles) co-secrete this pair of metalloenzymes early during penetration. We show in vivo that mutant appressoria lacking either or both enzymes have impaired penetration ability and pathogenicity. We reveal in vitro a finely-tuned enzyme interplay is responsible for the oxidation of plant cuticular long-chain alcohols into aldehyde products, suggested to act as key molecular signals in the fungal infection machinery. Our results open new avenues to design oxidase-specific inhibitors as anti-penetrants for crop protection.

scholarly journals Small RNA Functions Are Required for Growth and Development of Magnaporthe oryzae

2017 ◽  
Vol 30 (7) ◽  
pp. 517-530 ◽  
Author(s):  
Vidhyavathi Raman ◽  
Stacey A. Simon ◽  
Feray Demirci ◽  
Mayumi Nakano ◽  
Blake C. Meyers ◽  
...  
Keyword(s):  
Magnaporthe Oryzae ◽  
Small Rna ◽  
Plant Pathogens ◽  
Single Gene ◽  
Fungal Growth ◽  
Fungal Species ◽  
Fungal Virulence ◽  
Fungal Plant Pathogens ◽  
Intergenic Regions ◽  
Barley Leaves

RNA interference (RNAi) is conserved in eukaryotic organisms, and it has been well studied in many animal and plant species and some fungal species, yet it is not well studied in fungal plant pathogens. In the rice blast fungus Magnaporthe oryzae, we examined small RNA (sRNA) and their biogenesis in the context of growth and pathogenicity. Through genetic and genomic analyses, we demonstrate that loss of a single gene encoding Dicer, RNA-dependent RNA polymerase, or Argonaute reduces sRNA levels. These three proteins are required for the biogenesis of sRNA-matching genome-wide regions (coding regions, repeats, and intergenic regions). The loss of one Argonaute reduced both sRNA and fungal virulence on barley leaves. Transcriptome analysis of multiple mutants revealed that sRNA play an important role in transcriptional regulation of repeats and intergenic regions in M. oryzae. Together, these data support that M. oryzae sRNA regulate developmental processes including, fungal growth and virulence.

scholarly journals Selective packaging of mitochondrial proteins into extracellular vesicles prevents the release of mitochondrial DAMPs

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Kiran Todkar ◽  
Lilia Chikhi ◽  
Véronique Desjardins ◽  
Firas El-Mortada ◽  
Geneviève Pépin ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Mitochondrial Proteins ◽  
Control Mechanisms ◽  
Mitochondrial Content ◽  
Sorting Nexin ◽  
Inflammatory Conditions ◽  
Selective Targeting ◽  
Molecular Patterns ◽  
Damage Associated Molecular Patterns ◽  
Insight Into

AbstractMost cells constitutively secrete mitochondrial DNA and proteins in extracellular vesicles (EVs). While EVs are small vesicles that transfer material between cells, Mitochondria-Derived Vesicles (MDVs) carry material specifically between mitochondria and other organelles. Mitochondrial content can enhance inflammation under pro-inflammatory conditions, though its role in the absence of inflammation remains elusive. Here, we demonstrate that cells actively prevent the packaging of pro-inflammatory, oxidized mitochondrial proteins that would act as damage-associated molecular patterns (DAMPs) into EVs. Importantly, we find that the distinction between material to be included into EVs and damaged mitochondrial content to be excluded is dependent on selective targeting to one of two distinct MDV pathways. We show that Optic Atrophy 1 (OPA1) and sorting nexin 9 (Snx9)-dependent MDVs are required to target mitochondrial proteins to EVs, while the Parkinson’s disease-related protein Parkin blocks this process by directing damaged mitochondrial content to lysosomes. Our results provide insight into the interplay between mitochondrial quality control mechanisms and mitochondria-driven immune responses.

A Systematic Review on Extracellular Vesicles-Enriched Fat Grafting: A Shifting Paradigm

2020 ◽  
Author(s):  
Mohammad Ghiasloo ◽  
Laura De Wilde ◽  
Kashika Singh ◽  
Patrick Tonnard ◽  
Alexis Verpaele ◽  
...  
Keyword(s):  
Systematic Review ◽  
Graft Survival ◽  
Extracellular Vesicles ◽  
Retention Rate ◽  
Fat Grafting ◽  
Retention Rates ◽  
Fat Graft ◽  
Cochrane Central Register

Abstract Background Recent evidence confirms that mesenchymal stem cells (MSCs) facilitate angiogenesis mainly through paracrine function. Extracellular vesicles (EVs) are regarded as key components of the cell secretome, possessing functional properties of their source cells. Subsequently, MSC-EVs have emerged as a novel cell-free approach to improve fat graft retention rate. Objectives To provide a systematic review of all studies reporting the use of MSC-EVs to improve graft retention rate. Methods A systematic search was undertaken using the Embase, PubMed and the Cochrane Central Register of Controlled Trials databases. Outcome measures included donor/receptor organism of the fat graft, study model, intervention groups, evaluation intervals, EV research data, in vitro and in vivo results. Results Of the total 1717 articles, 62 full-texts were screened. Seven studies reporting on 294mice were included. Overall, EV treated groups showed higher graft retention rates compared to untreated groups. Notably, retention rate was similar following EV- and MSC-treatment. In addition to reduced inflammation, graft enrichment with EVs resulted in early revascularization and better graft integrity. Interestingly, hypoxic preconditioning of MSCs improved their beneficial paracrine effects and led to a more proangiogenic EV population, as observed by both in vitro and in vivo results. Conclusions MSC-EVs appear to offer an interesting cell-free alternative to improve fat graft survival. While their clinical relevance remains to be determined, it is clear that not the cells, but their secretome is essential for graft survival. Thus, a paradigm shift from cell-assisted lipotransfer towards ‘secretome-assisted lipotransfer’ is well on its way.

scholarly journals microRNA-186 in extracellular vesicles from bone marrow mesenchymal stem cells alleviates idiopathic pulmonary fibrosis via interaction with SOX4 and DKK1

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jing Zhou ◽  
Yang Lin ◽  
Xiuhua Kang ◽  
Zhicheng Liu ◽  
Wei Zhang ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Idiopathic Pulmonary Fibrosis ◽  
Pulmonary Fibrosis ◽  
Extracellular Vesicles ◽  
Luciferase Assay ◽  
Therapeutic Effects ◽  
Loss Of Function ◽  
Fibroblast Activation ◽  
Promising Therapeutic Approach

Abstract Background Previous reports have identified that human bone marrow mesenchymal stem cell-derived extracellular vesicles (BMSC-EVs) with their cargo microRNAs (miRNAs) are a promising therapeutic approach for the treatment of idiopathic pulmonary fibrosis (IPF). Therefore, we explored whether delivery of microRNA-186 (miR-186), a downregulated miRNA in IPF, by BMSC EVs could interfere with the progression of IPF in a murine model. Methods In a co-culture system, we assessed whether BMSC-EVs modulated the activation of fibroblasts. We established a mouse model of PF to evaluate the in vivo therapeutic effects of BMSC-EVs and determined miR-186 expression in BMSC-EVs by polymerase chain reaction. Using a loss-of-function approach, we examined how miR-186 delivered by BMSC-EVs affected fibroblasts. The putative relationship between miR-186 and SRY-related HMG box transcription factor 4 (SOX4) was tested using luciferase assay. Next, we investigated whether EV-miR-186 affected fibroblast activation and PF by targeting SOX4 and its downstream gene, Dickkopf-1 (DKK1). Results BMSC-EVs suppressed lung fibroblast activation and delayed IPF progression in mice. miR-186 was downregulated in IPF but enriched in the BMSC-EVs. miR-186 delivered by BMSC-EVs could suppress fibroblast activation. Furthermore, miR-186 reduced the expression of SOX4, a target gene of miR-186, and hence suppressed the expression of DKK1. Finally, EV-delivered miR-186 impaired fibroblast activation and alleviated PF via downregulation of SOX4 and DKK1. Conclusion In conclusion, miR-186 delivered by BMSC-EVs suppressed SOX4 and DKK1 expression, thereby blocking fibroblast activation and ameliorating IPF, thus presenting a novel therapeutic target for IPF.

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