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1.
Commun Biol ; 7(1): 1184, 2024 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-39300313

RESUMO

Major symbiotic organisms have evolved to establish beneficial relationships with hosts. However, understanding the interactions between symbionts and insect hosts, particularly for their roles in defense against pathogens, is still limited. In a previous study, we proposed that the fungus Metarhizium anisopliae can infect the brown planthopper Nilaparvata lugens, a harmful pest for rice crops. To expand on this, we investigated changes in N. lugens' intestinal commensal community after M. anisopliae infection and identified key gut microbiotas involved. Our results showed significant alterations in gut microbiota abundance and composition at different time points following infection with M. anisopliae. Notably, certain symbionts, like Acinetobacter baumannii, exhibited significant variations in response to the fungal infection. The decrease in these symbionts had a considerable impact on the insect host's survival. Interestingly, reintroducing A. baumannii enhanced the host's resistance to M. anisopliae, emphasizing its role in pathogen defense. Additionally, A. baumannii stimulated host immune responses, as evidenced by increased expression of immune genes after reintroduction. Overall, our findings highlight the significance of preserving a stable gut microbial community for the survival of insects. In specific conditions, the symbiotic microorganism A. baumannii can enhance the host's ability to resist entomopathogenic pathogens through immune regulation.


Assuntos
Acinetobacter baumannii , Microbioma Gastrointestinal , Hemípteros , Metarhizium , Simbiose , Animais , Metarhizium/fisiologia , Metarhizium/patogenicidade , Acinetobacter baumannii/fisiologia , Hemípteros/microbiologia , Hemípteros/imunologia , Interações Hospedeiro-Patógeno , Resistência à Doença
2.
J Invertebr Pathol ; 207: 108204, 2024 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-39313093

RESUMO

Microcycle conidiation has displayed the greater potential than normal conidiation in large-scale production of mycopesticides. Fungi require partial hydrolysis of the cell wall to achieve the necessary plasticity during their morphological changes. Therefore, various cell wall-associated hydrolases are crucial for fungal morphogenesis. Eng1, as an endo-ß-1,3-glucanase, is involved in the cell separation of fungi, but its role in morphological changes of entomopathogenic fungi is not yet clear. Here, the endo-ß-1,3-glucanase gene MaEng1 was characterized in the model entomopathogenic fungi M. acridum. MaEng1 possesses a typical carbohydrate hydrolase domain and belongs to the GH81 family. The functions of MaEng1 in fungal growth, stress tolerance, pathogenicity, and conidiation capacity were analyzed using targeted gene disruption. The results displayed that the absence of MaEng1 does not affect the fungal growth, stress tolerances, and pathogenicity in M. acridum. However, the knockout of MaEng1 led to the normal conidiation of M. acridum on the SYA medium, which can induce the microcycle conidiation. Moreover, the content of ß-1,3-glucan in the cell wall of the MaEng1-disruption strain were significantly reduced and the exposures of ß-1,3-glucan on the surface of the mature conidia and mycelia in ΔMaEng1 were declined, indicating that MaEng1 contributes to the conversion of conidiation mode in M. acridum by affecting the cell wall structure.

3.
J Fungi (Basel) ; 10(8)2024 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-39194870

RESUMO

Insect pathogenic fungi have shown great potential in agricultural pest control. Conidiation is crucial for the survival of filamentous fungi, and dispersal occurs through two methods: normal conidiation, where conidia differentiate from mycelium, and microcycle conidiation, which involves conidial budding. The conidiation process is related to cell separation. The forkhead box gene Sep1 in Schizosaccharomyces pombe plays a crucial role in cell separation. Nevertheless, the function of Sep1 has not been clarified in filamentous fungi. Here, MaSep1, the homolog of Sep1 in Metarhizium acridum, was identified and subjected to functional analysis. The findings revealed that conidial germination of the MaSep1-deletion strain (ΔMaSep1) was accelerated and the time for 50% germination rate of conidial was shortened by 1 h, while the conidial production of ΔMaSep1 was considerably reduced. The resistances to heat shock and UV-B irradiation of ΔMaSep1 were enhanced, and the expression of some genes involved in DNA damage repair and heat shock response was significantly increased in ΔMaSep1. The disruption of MaSep1 had no effect on the virulence of M. acridum. Interestingly, ΔMaSep1 conducted the normal conidiation on the microcycle conidiation medium, SYA. Furthermore, 127 DEGs were identified by RNA-Seq between the wild-type and ΔMaSep1 strains during microcycle conidiation, proving that MaSep1 mediated the conidiation pattern shift by governing some genes associated with conidiation, cell division, and cell wall formation.

4.
J Fungi (Basel) ; 10(8)2024 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-39194890

RESUMO

The entomopathogenic fungus (EPF) Metarhizium acridum is a typical filamentous fungus and has been used to control migratory locusts (Locusta migratoria manilensis). This study examines the impact of the Zn(II)2Cys6 transcription factor, MaAzaR, in the virulence of M. acridum. Disruption of MaAzaR (ΔMaAzaR) diminished the fungus's ability to penetrate the insect cuticle, thereby decreasing its virulence. The median lethal time (LT50) for the ΔMaAzaR strain increased by approximately 1.5 d compared to the wild-type (WT) strain when topically inoculated, simulating natural infection conditions. ΔMaAzaR compromises the formation, turgor pressure, and secretion of extracellular hydrolytic enzymes in appressoria. However, the growth ability of ΔMaAzaR within the hemolymph is not impaired; in fact, it grows better than the WT strain. Moreover, RNA-sequencing (RNA-Seq) analysis of ΔMaAzaR and WT strains grown for 20 h on locust hindwings revealed 87 upregulated and 37 downregulated differentially expressed genes (DEGs) in the mutant strain. Pathogen-host interaction database (PHI) analysis showed that about 40% of the total DEGs were associated with virulence, suggesting that MaAzaR is a crucial transcription factor that directly regulates the expression of downstream genes. This study identifies a new transcription factor involved in EPF cuticle penetration, providing theoretical support and genetic resources for the developing highly virulent strains.

5.
PLoS Pathog ; 20(7): e1012431, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-39078845

RESUMO

Reactive carbonyl and oxygen species (RCS/ROS), often generated as metabolic byproducts, particularly under conditions of pathology, can cause direct damage to proteins, lipids, and nucleic acids. Glyoxal oxidases (Gloxs) oxidize aldehydes to carboxylic acids, generating hydrogen peroxide (H2O2). Although best characterized for their roles in lignin degradation, Glox in plant fungal pathogens are known to contribute to virulence, however, the mechanism underlying such effects are unclear. Here, we show that Glox in the insect pathogenic fungus, Metarhizium acridum, is highly expressed in mycelia and during formation of infection structures (appressoria), with the enzyme localizing to the cell membrane. MaGlox targeted gene disruption mutants showed RCS and ROS accumulation, resulting in cell toxicity, induction of apoptosis and increased autophagy, inhibiting normal fungal growth and development. The ability of the MaGlox mutant to scavenge RCS was significantly reduced, and the mutant exhibited increased susceptibility to aldehydes, oxidative and cell wall perturbing agents but not toward osmotic stress, with altered cell wall contents. The ΔMaGlox mutant was impaired in its ability to penetrate the host cuticle and evade host immune defense resulting in attenuated pathogenicity. Overexpression of MaGlox promoted fungal growth and conidial germination, increased tolerance to H2O2, but had little to other phenotypic effects. Transcriptomic analyses revealed downregulation of genes related to cell wall synthesis, conidiation, stress tolerance, and host cuticle penetration in the ΔMaGlox mutant. These findings demonstrate that MaGlox-mediated scavenging of RCS is required for virulence, and contributes to normal fungal growth and development, stress resistance.


Assuntos
Oxirredutases do Álcool , Proteínas Fúngicas , Metarhizium , Virulência , Oxirredutases do Álcool/metabolismo , Oxirredutases do Álcool/genética , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/genética , Metarhizium/patogenicidade , Metarhizium/genética , Metarhizium/metabolismo , Animais , Espécies Reativas de Oxigênio/metabolismo , Estresse Fisiológico , Estresse Oxidativo
6.
J Fungi (Basel) ; 10(7)2024 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-39057353

RESUMO

Entomopathogenic fungi are valuable sources of biological pesticides, with conidial yield and quality being pivotal factors determining their broad applications. AzaR, a fungus-specific zinc-cluster transcription factor, is known to regulate the biosynthesis of polyketone secondary metabolites in Aspergillus niger; however, its role in pathogenic fungi remains unclear. This study investigated the role of MaAzaR in the growth, development, and environmental tolerance of Metarhizium acridum. MaAzaR deletion slowed down conidial germination rate, caused reduction in conidial yield, lowered fungal tolerance to UV radiation, did not affect fungal heat-shock tolerance, and increased fungal sensitivity to the cell-wall-destructive agent calcofluor white. Furthermore, MaAzaR deletion transformed microcycle conidiation to normal conidiation on the microcycle conidiation medium. Transcription profile analysis demonstrated that MaAzaR could regulate transformation of the conidiation pattern by controlling the expression of genes related to cell division, mycelium growth and development, and cell wall integrity. Thus, this study identified a new gene related to fungal conidiation and environmental tolerance, enriching our understanding of the molecular mechanism of microcycle conidiation and providing theoretical support and genetic resources for the development of high-yielding strains.

7.
Pest Manag Sci ; 80(2): 820-836, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37794279

RESUMO

BACKGROUND: The fungal genera Metarhizium contain many important multiple species that are used as biocontrol agents and as model organisms for exploring insect-fungal interactions. Metarhizium spp. exhibit different traits of pathogenicity, suggesting that the pathogenesis can be quite distinctive. However, the underlying differences in their pathogenesis remain poorly understood. RESULTS: Pathogenicity analysis showed that Metarhizium anisopliae (strain CQMa421) displayed higher virulence against oriental migratory locusts, Locusta migratoria manilensis (Meyen), than the acridid-specific specie Metarhizium acridum (strain CQMa102). Relative to M. acridum, M. anisopliae possessed a higher conidial hydrophobicity, increased ability to penetrate the host, accelerated growth under hypoxia and enhanced ability for the utilization of different carbon sources. Different distributions of carbohydrate epitopes at cell wall surface of M. anisopliae might also contribute to successful evasion of host immune defenses. Comparative genomics showed that M. anisopliae has 98 more virulence-related secreted proteins (133) than M. acridum (35), which can be functionally classified as hydrolases, virulence effectors, cell wall degradation and stress tolerance-related proteins, and helpful to the cuticle penetration and host internal environment adaption. In addition, differences in genomic clusters specifically related to secondary metabolites, including the clusters of Indole-NRPS hybrid, T1PKS-NRPS like hybrid, Betalactone, Fungal-Ripp and NRPS-Terpene hybrid, may lead to differences in core virulence-related secondary metabolite genes in M. acridum (18) and M. anisopliae (36). CONCLUSION: The comparative study provided new insights into the different infection strategies between M. anisopliae and M. acridum, and further facilitate the identification of virulence-related genes for the improvement of mycoinsecticides. © 2023 Society of Chemical Industry.


Assuntos
Metarhizium , Virulência , Metarhizium/fisiologia , Genômica
8.
Int J Biol Macromol ; 253(Pt 6): 127389, 2023 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-37827395

RESUMO

Locusts (Locusta migratoria) are one of the most destructive insect pests worldwide. Entomopathogenic fungi can infect and kill locusts, with Metarhizium acridum having evolved as a specialized acridid pathogen. However, locusts have evolved countermeasures to limit or avoid microbial pathogens, although the underlying molecular mechanisms behind these defenses remain obscure. Here, we demonstrate that L. migratoria exhibit avoidance behaviors towards M. acridum contaminated food via recognition of fungal volatiles, with locust perception of the volatile mediated by the LmigCSP60 chemosensory protein. RNAi-knockdown of LmigCSP60 lowered locust M. acridum avoidance behavior and increased infection and mortality. The fungal volatile, 2-phenylethanol (PEA), was identified to participate in locust behavioral avoidance. RNAi-knockdown of LmigCSP60 reduced antennal electrophysiological responses to PEA and impaired locust avoidance to the compound. Purified LmigCSP60 was able to bind a set of fungal volatiles including PEA. Furthermore, reduction of PEA emission by M. acridum via construction of a targeted gene knockout mutant of the alcohol dehydrogenase gene (ΔMaAdh strain) that contributes to PEA production reduced locust avoidance behavior towards the pathogen. These findings identify an olfactory circuit used by locusts to detect and avoid potential microbial pathogens before they are capable of initiating infection and highlight behavioral and olfactory adaptations affecting the co-evolution of host-pathogen interactions.


Assuntos
Gafanhotos , Locusta migratoria , Animais , Gafanhotos/genética , Proteínas de Insetos/genética , Locusta migratoria/genética , Olfato , Alimentos
9.
FEMS Microbiol Ecol ; 99(11)2023 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-37742208

RESUMO

The highly destructive southern rice black-streaked dwarf virus (SRBSDV) causes significant losses in rice production. To understand its impact on rice root, we studied fibrous root development and root microbiota variation (rhizosphere and endosphere) after SRBSDV infection. SRBSDV infection reduced the number and length of fibrous roots in rice. Interestingly, the rhizosphere had higher bacterial diversity and abundance at the initial (0 days) and 30-day postinfection stages, while 30-day-old roots showed increased diversity and abundance. However, there were no significant differences in microbiota diversity between infected and noninfected rice plants. The major rhizosphere microbiota included Proteobacteria, Bacteroidota, Acidobacteriota, and Planctomycetota, comprising about 80% of the community. The endosphere was dominated by Proteobacteria and Cyanobacteria, constituting over 90%, with Bacteroidota as the next most prominent group. Further, we identified differentially expressed genes related to plant-pathogen interactions, plant hormone signal, and ABC transporters, potentially affecting root morphology. Notably, specific bacteria (e.g. Inquilinus and Actinoplanes) showed correlations with these pathways. In conclusion, SRBSDV primarily influences root growth through host metabolism, rather than exerting direct effects on the root microbiota. These insights into the interactions among the pathogen, rice plant, and associated microbiota could have implications for managing SRBSDV's detrimental effects on rice production.

10.
J Fungi (Basel) ; 9(7)2023 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-37504734

RESUMO

Fungal diseases are widespread among insects and play a crucial role in naturally regulating insect populations. Mosquitoes, known as vectors for numerous infectious diseases, pose a significant threat to human health. Entomopathogenic fungi (EPF) have emerged as highly promising alternative agents to chemical mosquitocides for controlling mosquitoes at all stages of their life cycle due to their unique infection pathway through direct contact with the insect's cuticle. In recent years, significant advancements have been made in understanding the infection pathways and pathogenic mechanisms of EPF against mosquitoes. Various strategies involving the use of EPF alone or combinations with other approaches have been employed to target mosquitoes at various developmental stages. Moreover, the application of genetic technologies in fungi has opened up new avenues for enhancing the mosquitocidal efficacy of EPF. This review presents a comprehensive summary of recent advancements in our understanding the pathogenic mechanisms of EPF, their applications in mosquito management, and the combination of EPF with other approaches and employment of transgenic technologies. The biosafety concerns associated with their use and the corresponding approaches are also discussed. The recent progress suggests that EPF have the potential to serve as a future biorational tool for controlling mosquito vectors.

11.
Front Cell Infect Microbiol ; 13: 1140765, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36936763

RESUMO

Dendritic cells (DCs) are crucial for the initiation and regulation of adaptive immune responses. When encountering immune stimulus such as bacterial and viral infection, parasite invasion and dead cell debris, DCs capture antigens, mature, acquire immunostimulatory activity and transmit the immune information to naïve T cells. Then activated cytotoxic CD8+ T cells directly kill the infected cells, while CD4+ T helper cells release cytokines to aid the activity of other immune cells, and help B cells produce antibodies. Thus, detailed insights into the DC maturation process are necessary for us to understand the working principle of immune system, and develop new medical treatments for infection, cancer and autoimmune disease. This review summarizes the DC maturation process, including environment sensing and antigen sampling by resting DCs, antigen processing and presentation on the cell surface, DC migration, DC-T cell interaction and T cell activation. Application of advanced imaging modalities allows visualization of subcellular and molecular processes in a super-high resolution. The spatiotemporal tracking of DCs position and migration reveals dynamics of DC behavior during infection, shedding novel lights on DC biology.


Assuntos
Linfócitos T CD8-Positivos , Células Dendríticas , Citocinas/metabolismo , Apresentação de Antígeno , Diferenciação Celular
12.
Appl Microbiol Biotechnol ; 107(9): 2969-2982, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36941435

RESUMO

Carbon sources and their utilization are vital for fungal growth and development. C4-dicarboxylic acids are important carbon and energy sources that function as intermediate products of the tricarboxylic acid cycle. Transport and regulation of C4-dicarboxylic acid uptake are mainly dependent on tetracarboxylic acid transporters (Dcts) in many microbes, although the roles of Dct genes in fungi have only been partially characterized. Here, we report on the functions of two Dct genes (Dct1 and Dct2) in the entomopathogenic fungus Metarhizium acridum. Our data showed that loss of the MaDct1 gene affected utilization of tetracarboxylic acids and other carbon sources. ΔMaDct1 mutants showed larger colony sizes with extensive mycelial growth but were delayed in conidiation with decreased conidia yield as compared to the wild-type parental strain. On the nutrient-deficient medium, SYA, the wild-type strain produced microcycle conidia, whereas the ΔMaDct1 mutant produced (normal) aerial conidia. In addition, ΔMaDct1 had decreased tolerance to cell wall perturbing agents, but increased tolerances to UV-B radiation and osmotic stress. Insect bioassays indicated that loss of MaDct1 did not affect pathogenicity. In contrast, no distinct phenotypic change was observed for the MaDct2 mutant in terms of growth and biocontrol characteristics. Transcriptomic profiling between wild type and ΔMaDct1 showed that differentially expressed genes were enriched in carbohydrate and amino acid metabolism, transport and catabolism, and signal transduction. These results demonstrate that MaDct1 regulates the conidiation pattern shift and mycelial growth by affecting utilization of carbon sources. These findings are helpful for better understanding the effect of intermediates of carbon metabolism on fungal growth and conidiation. KEY POINTS: • MaDct1 influences fungal growth and conidiation by affecting carbon source utilization. • MaDct1 regulates conidiation pattern shift under nutrient deficiency condition. • MaDct1 is involved in stress tolerance and has no effect on virulence. • MaDct2 has no effect on growth and biocontrol characteristic.


Assuntos
Regulação Fúngica da Expressão Gênica , Metarhizium , Perfilação da Expressão Gênica , Virulência , Proteínas de Membrana Transportadoras/metabolismo , Esporos Fúngicos , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo
13.
Appl Microbiol Biotechnol ; 107(4): 1257-1268, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36640205

RESUMO

Microcycle conidiation commonly exists in filamentous fungi and has great potential for mass production of mycoinsecticides. L-Arginine metabolism is essential for conidiation and conditional growth and virulence, but its role in microcycle conidiation has not been explored. Here, a unique putative arginase (MaAGA) was characterized in the entomopathogenic fungus Metarhizium acridum. Conidial germination and thermotolerance were facilitated by the disruption of MaAGA. Despite little impact on fungal growth and virulence, the disruption resulted in normal conidiation after a 60-h incubation on microcycle conidiation medium (SYA) under normal culture conditions. In the MaAGA-disruption mutant (ΔMaAGA), intracellular arginine accumulation was sharply increased. Replenishment of the direct metabolites of arginase, namely ornithine and/or urea, was unable to restore the disruption mutant's microcycle conidiation on SYA. Interestingly, nitric oxide synthase (NOS) activity and nitric oxide (NO) levels of the ΔMaAGA strain were markedly decreased in the 60-h-old SYA cultures. Finally, adding Nω-nitro-L-arginine, an inhibitor of NOS, into the SYA converted the microcycle conidiation of the wild-type strain to normal conidiation. In contrast, adding sodium nitroprusside, an NO donor, into the SYA recovered the mutant's microcycle conidiation. The results indicate that arginine metabolism controls microcycle conidiation by changing the content of NO. KEY POINTS: • The MaAGA-disruption led to normal conidiation on microcycle conidiation medium SYA. • Nitric oxide (NO) level of the ΔMaAGA strain was markedly decreased. • Adding an NO donor into the SYA recovered the microcycle conidiation of ΔMaAGA.


Assuntos
Metarhizium , Óxido Nítrico , Óxido Nítrico/metabolismo , Arginase/genética , Arginase/metabolismo , Arginina/metabolismo , Esporos Fúngicos
14.
Dev Comp Immunol ; 138: 104530, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36084754

RESUMO

The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is one of the most highly polyphagous invasive pests causing serious damage to maize crops in China. However, little is known about the gut immune responses to the environment, particularly along the migration routes in Jianghuai, China, throughout the autumn and winter. In this study, high-throughput sequencing and real-time quantitative PCR (RT-qPCR) were employed to examine the variations in immune genes and gut microbiome communities between captive and wild fall armyworm populations. Results showed that the diversity and community of the gut's microbes were higher in wild populations, and the average weighted UniFrac distance between bacterial taxa varied. A wide variety of immune genes were more abundant in the wild populations than in others. Results indicated that diets and different survival conditions impacted the gut microbiota and immune system of S. frugiperda, which was crucial for environmental adaptation. These differences in gut microbiota and immune responses between wild and captive Fall armyworms are critical for comprehending the symbiotic relationship between microbes, immune genes, and hosts. They also highlight the need for increased focus on developing more effective and environmentally friendly pest control methods.


Assuntos
Microbioma Gastrointestinal , Animais , China , Sistema Imunitário , Larva , Spodoptera/genética , Zea mays/genética
15.
J Adv Res ; 48: 1-16, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-36064181

RESUMO

INTRODUCTION: Odorant-binding proteins (OBPs) are a class of small molecular weight soluble proteins that exist as expanded gene families in all insects, acting as ligand carriers mediating olfaction and other physiological processes. During fungal infection, a subset of insect OBPs were shown to be differentially expressed. OBJECTIVES: We tested whether the altered expression of insect OBPs during pathogenic infection plays a role in behavioral or immune interactions between insect hosts and their pathogens. METHODS: A wide range of techniques including RNAi-directed knockdown, heterologous protein expression, electrophysiological/behavioral analyses, transcriptomics, gut microbiome analyses, coupled with tandem mass spectrometry ion monitoring, were used to characterize the function of a locust OBP in host behavioral and immune responses. RESULTS: The entomopathogenic fungus Metarhizium anisopliae produces the volatile compound phenylethyl alcohol (PEA) that causes behavioral avoidance in locusts. This is mediated by the locust odorant binding protein 11 (LmOBP11). Expression of LmOBP11 is induced by M. anisopliae infection and PEA treatment. LmOBP11 participates in insect detection of the fungal-produced PEA and avoidance of PEA-contaminated food, but the upregulation of LmOBP11 upon M. anisopliae infection negatively affects the insect immune responses to ultimately benefit successful mycosis by the pathogen. RNAi knockdown of LmOBP11 increases the production of antimicrobial peptides and enhances locust resistance to M. anisopliae infection, while reducing host antennal electrophysiological responses to PEA and locust avoidance of PEA treated food. Also, transcriptomic and gut microbiome analyses reveal microbiome dysbiosis and changes in host genes involved in behavior and immunity. These results are consistent with the elevated expression of LmOBP11 leading to enhanced volatile detection and suppression of immune responses. CONCLUSION: These findings suggest a crosstalk between olfaction and immunity, indicating manipulation of host OBPs as a novel target exploited by fungal pathogens to alter immune activation and thus promote the successful infection of the host.


Assuntos
Gafanhotos , Metarhizium , Micoses , Animais , Odorantes , Insetos/microbiologia , Gafanhotos/microbiologia , Metarhizium/fisiologia , Imunidade Inata
16.
Biology (Basel) ; 11(12)2022 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-36552240

RESUMO

Entomopathogenic fungi are promising biocontrol agents of insect-mediated crop damage. Microcycle conidiation has shown great potential in enhancing the conidial yield and quality of entomopathogenic fungi. Homologs of Cts1, an endochitinase of Saccharomyces cerevisiae, participate in cell separation in several fungal spp. and may contribute to the morphological differences that occur during the shift to microcycle conidiation. However, the precise functions of Cts1 in entomopathogenic fungi remain unclear. Herein, the endochitinase gene, MaCts1, was characterized in the model entomopathogen, Metarhizium acridum. A loss of function line for MaCts1 led to a delay of 1 h in the median germination time, a 28% reduction in conidial yield and significant defects in fungal resistances to UV-irradiation (18%) and heat-shock (15%), while fungal tolerances to cell wall stressors, oxidative and hyperosmotic stresses and virulence remained unchanged. The MaCts1-disruption strain displayed typical conidiation on the microcycle conidiation induction medium, SYA. In contrast, deletion of key genes in the morphogenesis-related NDR kinase network (MOR pathway)/regulation of Ace2 and morphogenesis (RAM pathway) did not affect the SYA-induction of microcycle conidiation. This indicates that MaCts1 makes contributions to the microcycle conidiation, which may not be dependent on the MOR/RAM pathway in M. acridum.

17.
J Fungi (Basel) ; 8(9)2022 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-36135667

RESUMO

Zinc finger proteins are an important class of multifunctional regulators. Here, the roles of a C2H2 zinc finger protein MaNCP1 (Metarhizium acridum nitrate-related conidiation pattern shift regulatory factor 1) in nitrogen utilization and conidiation were explored in the entomopathogenic fungus M. acridum. The results showed that MaNCP1-disruption mutant (ΔMaNCP1) impaired the ability to utilize nitrate, ammonium and glutamine and reduced the expression of nitrate assimilation-related genes, suggesting that MaNCP1 was involved in governing nitrogen utilization. In addition, the conidial yield of the ΔMaNCP1 strain, cultured on the microcycle conidiation medium (SYA), was significantly decreased, which could be restored or even enhanced than that of the WT strain through increasing the nitrate content in SYA medium. Further study showed that MaAreA, a core regulator in the nitrogen catabolism repression (NCR) pathway, was a downstream target gene of MaNCP1. Screening the differential expression genes between WT and ΔMaNCP1 strains revealed that the conidial yield of M. acridum regulated by nitrate might be related to NCR pathway on SYA medium. It could be concluded that MaNCP1 contributes to the nitrate assimilation and conidiation, which will provide further insights into the relationship between the nitrogen utilization and conidiation in fungi.

18.
Int J Biol Macromol ; 216: 426-436, 2022 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-35809667

RESUMO

C2H2 zinc finger proteins (ZFPs) are a class of important transcriptional regulators in eukaryotes involved in multiple biological regulation processes. Here, MaNCP1, a C2H2 ZFP, was functionally characterized in the model entomopathogenic fungus Metarhizium acridum. Deletion of MaNCP1 delayed conidial germination and hyphal growth, decreased the conidial yield and reduced the tolerances to UV-B irradiation and heat-shock. The N-terminal zinc fingers (ZFs) of MaNCP1 made the main contributions to these traits. In addition, disruption of MaNCP1 altered the conidial surface structure and decreased the conidial hydrophobicity. Bioassays showed that the virulence of the MaNCP1-disruption strain (ΔMaNCP1) was reduced in topical inoculation compared to the WT or the mutant complemented strain (CP), and the N-terminal C2H2 ZFs made a major contribution to virulence. Furthermore, the ΔMaNCP1 and C2H2 ZFs deletion mutants (MaNCP1∆N and MaNCP1∆N+C) impaired cuticular penetration. RNA-seq showed that several cuticle-degrading genes were down-regulated in the ΔMaNCP1 background, suggesting that MaNCP1 plays vital roles in regulating insect cuticle penetration. In summary, MaNCP1 affected the growth, stress tolerances and virulence of M. acridum, and the N-terminal C2H2 ZFs played indispensable roles in these important biocontrol traits. These results provide further insights into the functions of C2H2 ZFPs in entomopathogenic fungi.


Assuntos
Metarhizium , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Metarhizium/metabolismo , Metarhizium/patogenicidade , Esporos Fúngicos , Virulência , Dedos de Zinco
19.
J Fungi (Basel) ; 8(6)2022 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-35736070

RESUMO

Opy2 is an important membrane-anchored protein upstream of the HOG-MAPK signaling pathway and plays important roles in both the HOG-MAPK and Fus3/Kss1 MAPK. In this study, the roles of MaOpy2 in Metarhizium acridum were systematically elucidated. The results showed that the MaOpy2 disruption significantly reduced fungal tolerances to UV, heat shock and cell-wall-disrupting agents. Bioassays showed that the decreased fungal pathogenicity by topical inoculation mainly resulted from the impaired penetration ability. However, the growth ability of ∆MaOpy2 was enhanced in insect hemolymph. Importantly, MaOpy2 deletion could significantly increase the conidial yield of M. acridum by shifting the conidiation pattern from normal conidiation to microcycle conidiation on the 1/4SDAY medium. Sixty-two differentially expressed genes (DEGs) during the conidiation pattern shift, including 37 up-regulated genes and 25 down-regulated genes in ∆MaOpy2, were identified by RNA-seq. Further analysis revealed that some DEGs were related to conidiation and hyphal development. This study will provide not only the theoretical basis for elucidating the regulation mechanism for improving the conidial yield and quality in M. acridum but also theoretical guidance for the molecular improvement of entomopathogenic fungi.

20.
J Fungi (Basel) ; 8(6)2022 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-35736077

RESUMO

Conidium is the main infection unit and reproductive unit of pathogenic fungi. Exploring the mechanism of conidiation and its regulation contributes to understanding the pathogenicity of pathogenic fungi. Vib-1, a transcription factor, was reported to participate in the conidiation process. However, the regulation mechanism of Vib-1 in conidiation is still unclear. In this study, we analyzed the function of Vib-1 and its regulation mechanism in conidiation through knocking out and overexpression of Vib-1 in entomopathogenic fungus Metarhizium acridum. Results showed that the colonial growth of Mavib-1 disruption mutant (ΔMavib-1) was significantly decreased, and conidiation was earlier compared to wild type (WT), while overexpression of Mavib-1 led to a delayed conidiation especially when carbon or nitrogen sources were insufficient. Overexpression of Mavib-1 resulted in a conidiation pattern shift from microcycle conidiation to normal conidiation on nutrient-limited medium. These results indicated that Mavib-1 acted as a positive regulator in vegetative growth and a negative regulator in conidiation by affecting utilization of carbon and nitrogen sources in M. acridum. Transcription profile analysis demonstrated that many genes related to carbon and nitrogen source metabolisms were differentially expressed in ΔMavib-1 and OE strains compared to WT. Moreover, Mavib-1 affects the conidial germination, tolerance to UV-B and heat stresses, cell wall integrity, conidial surface morphology and conidial hydrophobicity in M. acridum. These findings unravel the regulatory mechanism of Mavib-1 in fungal growth and conidiation, and enrich the knowledge to conidiation pattern shift of filamentous fungi.

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