RESUMEN
Crude oil is a hazardous pollutant that poses significant and lasting harm to human health and ecosystems. In this study, Moesziomyces aphidis XM01, a biosurfactant mannosylerythritol lipids (MELs)-producing yeast, was utilized for crude oil degradation. Unlike most microorganisms relying on cytochrome P450, XM01 employed two extracellular unspecific peroxygenases, MaUPO.1 and MaUPO.2, with preference for polycyclic aromatic hydrocarbons (PAHs) and n-alkanes respectively, thus facilitating efficient crude oil degradation. The MELs produced by XM01 exhibited a significant emulsification activity of 65.9% for crude oil and were consequently supplemented in an "exogenous MELs addition" strategy to boost crude oil degradation, resulting in an optimal degradation ratio of 72.3%. Furthermore, a new and simple "pre-MELs production" strategy was implemented, achieving a maximum degradation ratio of 95.9%. During this process, the synergistic up-regulation of MaUPO.1, MaUPO.1 and the key MELs synthesis genes contributed to the efficient degradation of crude oil. Additionally, the phylogenetic and geographic distribution analysis of MaUPO.1 and MaUPO.1 revealed their wide occurrence among fungi in Basidiomycota and Ascomycota, with high transcription levels across global ocean, highlighting their important role in biodegradation of crude oil. In conclusion, M. aphidis XM01 emerges as a novel yeast for efficient and eco-friendly crude oil degradation.
Asunto(s)
Biodegradación Ambiental , Glucolípidos , Oxigenasas de Función Mixta , Petróleo , Tensoactivos , Petróleo/metabolismo , Tensoactivos/metabolismo , Tensoactivos/química , Glucolípidos/metabolismo , Oxigenasas de Función Mixta/metabolismo , Oxigenasas de Función Mixta/genética , Hidrocarburos Policíclicos Aromáticos/metabolismo , Hidrocarburos Policíclicos Aromáticos/química , Alcanos/metabolismoRESUMEN
Ndt80-like transcription factor Ron1 is best known for its essential role in the regulation of N-acetylglucosamine (GlcNAc) catabolism. Ron1 was again found to be essential for sensing GlcNAc in Beauveria bassiana. Importantly, our study revealed that Ron1 is involved in the metabolic processes of chitin and asexual development. To further investigate the novel functions of Ron1 in B. bassiana, extracellular chitinase activity in the ΔRon1 mutant was found to decrease by 84.73% compared with wild type. The deletion of Ron1 made it difficult for the fungus to accumulate intracellular GlcNAc. Furthermore, transcriptomic analysis revealed that Ron1 exerted a significant effect on global transcription and positively regulated genes encoding chitin metabolism in respond to chitin nutrition. Yeast one-hybrid assay confirmed that Ron1 could bind to specific cis-acting elements in the promoters of chitinase and hexokinase. In addition, ΔRon1 displayed an impaired chitin component of the cell wall, with a chitin synthetase (ChsVII) predicted to function downstream of Ron1. Finally, the virulence of ΔRon1 mutant was significantly reduced in the Galleria mellonella insect model through cuticle infection or cuticle bypassing infection. These data functionally characterize Ron1 in B. bassiana and expand our understanding of how the transcription factor Ron1 works in pathogens.
Asunto(s)
Beauveria , Quitinasas , Quitina/metabolismo , Quitinasas/genética , Quitinasas/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Factores de Transcripción/metabolismo , Virulencia/genéticaRESUMEN
The mannosyltransferase Alg9 plays a vital role in N-linked protein glycosylation in Saccharomyces cerevisiae, but its function in most filamentous fungi is not clear. The present study characterized BbAlg9 (an ortholog of S. cerevisiae Alg9) in Beauveria bassiana to determine the roles of N-mannosyltransferase in biological control potential of the filamentous entomopathogenic fungus. The disruption of BbAlg9 led to slower fungal growth in media with various nutrition compositions. The conidiation of ΔBbAlg9 was less than that of the wild type from the third to the fifth day but showed no significant difference on the sixth day, suggesting that BbAlg9 affects the development of conidia rather than conidial yield of late stage. ΔBbAlg9 showed defects in conidial germination, multiple stress tolerances and the yield of blastospores, with altered size and density, and virulence in hosts infected via the immersion and injection methods. The deletion of BbAlg9 resulted in defects in cell wall integrity, including increased mannoprotein and glucan content and decreased chitin content, which were accompanied by transcriptional activation or suppression of genes related to cell wall component biosynthesis. Notably, deletion of the N-mannosyltransferase BbAlg9 altered the transcription levels of O-mannosyltransferase genes (Pmt and Ktr family). These data show that BbAlg9 is involved in the fungal development, conidial stress tolerance, cell wall integrity and virulence of B. bassiana.
Asunto(s)
Beauveria , Beauveria/genética , Pared Celular , Proteínas Fúngicas/genética , Eliminación de Gen , Manosiltransferasas/genética , Saccharomyces cerevisiae , Esporas Fúngicas , Estrés Fisiológico , VirulenciaRESUMEN
Beauveria bassiana is an insect pathogenic fungus that serves as a model system for exploring the mechanisms of fungal development and host-pathogen interactions. Clinical and experimental studies have indicated that SND1 is closely correlated with the progression and invasiveness of common cancers as a potential oncogene, but this gene has rarely been studied in fungi. Here, we characterized the contributions of an SND1 ortholog (Tdp1) by constructing a BbTdp1 deletion strain and a complemented strain of B. bassiana. Compared with the wild-type (WT) strain, the ΔBbTdp1 mutant lost conidiation capacity (â¼87.7%) and blastospore (â¼96.3%) yields, increased sensitivity to chemical stress (4.4 to 54.3%) and heat shock (â¼44.2%), and decreased virulence following topical application (â¼24.7%) and hemocoel injection (â¼40.0%). Flow cytometry readings showed smaller sizes of both conidia and blastospores for ΔBbTdp1 mutants. Transcriptomic data revealed 4,094 differentially expressed genes (|log2 ratio| > 2 and a q value of <0.05) between ΔBbTdp1 mutants and the WT strain, which accounted for 41.6% of the total genes, indicating that extreme fluctuation in the global gene expression pattern had occurred. Moreover, deletion of BbTdp1 led to an abnormal cell cycle with a longer S phase and shorter G2/M and G0/G1 phases of blastospores, and enzyme-linked immunosorbent assay confirmed that the level of phosphorylated cyclin-dependent kinase 1 (Cdk1) in the ΔBbTdp1 strain was â¼31.5% lower than in the WT strain. In summary, our study is the first to report that BbTdp1 plays a vital role in regulating conidia and blastospore yields, fungal morphological changes, and pathogenicity in entomopathogenic fungi. IMPORTANCE In this study, we used Beauveria bassiana as a biological model to report the role of BbTdp1 in entomopathogenic fungi. Our findings indicated that BbTdp1 contributed significantly to cell development, the cell cycle, and virulence in B. bassiana. In addition, deletion of BbTdp1 led to drastic fluctuations in the transcriptional profile. BbTdp1 can be developed as a novel target for B. bassiana development and pathogenicity, which also provides a framework for the study of Tdp1 in other fungi.
Asunto(s)
Beauveria/crecimiento & desarrollo , Beauveria/metabolismo , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Insectos/microbiología , Animales , Beauveria/genética , Beauveria/patogenicidad , Ciclo Celular , Proteínas Fúngicas/genética , Regulación Fúngica de la Expresión Génica , Transcriptoma , Dominio Tudor , VirulenciaRESUMEN
Morphological transition is an important adaptive mechanism in the host invasion process. Wor1 is a conserved fungal regulatory protein that controls the phenotypic switching and pathogenicity of Candida albicans. By modulating growth conditions, we simulated three models of Beauveria bassiana morphological transitions, including CTH (conidia to hyphae), HTC (hyphae to conidia), and BTB (blastospore to blastospore). Disruption of BbWor1 (an ortholog of Wor1) resulted in a distinct reduction in the time required for conidial germination (CTH), a significant increase in hyphal growth, and a decrease in the yield of conidia (HTC), indicating that BbWor1 positively controls conidium production and negatively regulates hyphal growth in conidium-hypha switching. Moreover, ΔBbWor1 prominently decreased blastospore yield, shortened the G0/G1 phase, and prolonged the G2/M phase under the BTB model. Importantly, BbWor1 contributed to conidium-hypha switching and blastospore propagation via different genetic pathways, and yeast one-hybrid testing demonstrated the necessity of BbWor1 to control the transcription of an allergen-like protein gene (BBA_02580) and a conidial wall protein gene (BBA_09998). Moreover, the dramatically weakened virulence of ΔBbWor1 was examined by immersion and injection methods. Our findings indicate that BbWor1 is a vital participant in morphological transition and pathogenicity in entomopathogenic fungi. IMPORTANCE As a well-known entomopathogenic fungus, Beauveria bassiana has a complex life cycle and involves transformations among single-cell conidia, blastospores, and filamentous hyphae. This study provides new insight into the regulation of the fungal cell morphological transitions by simulating three models. Our research identified BbWor1 as a core transcription factor of morphological differentiation that positively regulates the production of conidia and blastospores but negatively regulates hyphal growth. More importantly, BbWor1 affects fungal pathogenicity and the global transcription profiles within three models of growth stage transformation. The present study lays a foundation for the exploration of the transition mechanism of entomopathogenic fungi and provides material for the morphological study of fungi.
Asunto(s)
Beauveria/crecimiento & desarrollo , Proteínas Fúngicas/metabolismo , Hifa/metabolismo , Esporas Fúngicas/metabolismo , Factores de Transcripción/metabolismo , Animales , Beauveria/genética , Beauveria/metabolismo , Beauveria/patogenicidad , Proteínas Fúngicas/genética , Regulación del Desarrollo de la Expresión Génica , Regulación Fúngica de la Expresión Génica , Hifa/genética , Hifa/crecimiento & desarrollo , Mariposas Nocturnas/microbiología , Esporas Fúngicas/genética , Esporas Fúngicas/crecimiento & desarrollo , Factores de Transcripción/genética , VirulenciaRESUMEN
The p53-like transcription factor (TF) NDT80 plays a vital role in the regulation of pathogenic mechanisms and meiosis in certain fungi. However, the effects of NDT80 on entomopathogenic fungi are still unknown. In this paper, the NDT80 orthologue BbTFO1 was examined in Beauveria bassiana, a filamentous entomopathogenic fungus, to explore the role of an NDT80-like protein for fungal pest control potential. Disruption of BbTFO1 resulted in impaired resistance to oxidative stress (OS) in a growth assay under OS and a 50% minimum inhibitory concentration experiment. Intriguingly, the oxidation resistance changes were accompanied by transcriptional repression of the two key antioxidant enzyme genes cat2 and cat5. ΔBbTFO1 also displayed defective conidial germination, virulence and heat resistance. The specific supplementation of BbTFO1 reversed these phenotypic changes. As revealed by this work, BbTFO1 can affect the transcription of catalase genes and play vital roles in the maintenance of phenotypes associated with the biological control ability of B. bassiana.
Asunto(s)
Beauveria/metabolismo , Proteínas Fúngicas/metabolismo , Insectos/microbiología , Factores de Transcripción/metabolismo , Virulencia/genética , Animales , Beauveria/aislamiento & purificación , Beauveria/patogenicidad , Catalasa/genética , Catalasa/metabolismo , Proteínas Fúngicas/clasificación , Proteínas Fúngicas/genética , Regulación Fúngica de la Expresión Génica , Mutación , Estrés Oxidativo/genética , Fenotipo , Filogenia , Estrés Fisiológico , Temperatura , Factores de Transcripción/clasificación , Factores de Transcripción/genéticaRESUMEN
Chitin is one of the major components of the fungal cell wall and contributes to the mechanical strength and shape of the fungal cell. Zn(II)2Cys6 transcription factors are unique to the fungal kingdom and have a variety of functions in some fungi. However, the mechanisms by which Zn(II)2Cys6 proteins affect entomopathogenic fungi are largely unknown. Here, we characterized the Zn(II)2Cys6 transcription factor BbTpc1 in the insect pathogenic fungus Beauveria bassiana. Disruption of BbTpc1 resulted in a distinct changes in vegetative growth and septation patterns, and a significant decrease in conidia and blastospore yield. The ΔBbTpc1 mutant displayed impaired resistance to chemical stresses and heat shock and attenuated virulence in topical and intrahemocoel injection assays. Importantly, the ΔBbTpc1 mutant had an abnormal cell wall with altered wall thickness and chitin synthesis, which were accompanied by transcriptional repression of the chitin synthetase family genes. In addition, comparative transcriptomics revealed that deletion of BbTpc1 altered fungal asexual reproduction via different genetic pathways. These data revealed that BbTpc1 regulates fungal development, chitin synthesis and biological control potential in B. bassiana.
Asunto(s)
Beauveria/crecimiento & desarrollo , Beauveria/patogenicidad , Quitina/biosíntesis , Proteínas Fúngicas/metabolismo , Insectos/microbiología , Factores de Transcripción/metabolismo , Secuencia de Aminoácidos , Animales , Autofagia , Beauveria/genética , Pared Celular/metabolismo , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Eliminación de Gen , Hifa/crecimiento & desarrollo , Mutación/genética , Filogenia , Reproducción Asexuada , Esporas Fúngicas/crecimiento & desarrollo , Estrés Fisiológico , Transcriptoma/genética , VirulenciaRESUMEN
To evaluate the traits and rheological properties of thermosensitive in situ gel of Yihuang Decoction and its common gel for vaginal use, and predict the release behavior of Yihuang Decoction in situ gel in vitro. Poloxamer was used as thermosensitive material to prepare Yihuang Decoction vaginal in situ gel, and Yihuang Decoction common gel was prepared with carbopol. Then the differences of the two gels before and after diluting with vaginal fluid were compared. The rheological parameters of Yihuang Decoction in situ gel and its common gel were determined with Anton Paar MCR102 rheometer. In addition, berberine hydrochloride was selected as an index component to evaluate the in vitro release properties of Yihuang Decoction vaginal thermosensitive in situ gel. Yihuang Decoction vaginal thermosensitive in situ gel was Newtonian fluid under low-temperature conditions, which was yellow and transparent. After reaching the gelling temperature of 24.5 â, it became semi-solid, pseudoplastic fluid. The gelling temperature was predicted to be 37 â, and the phase transition time was 30 s after diluting with simulated vaginal fluid. However, the rheological properties of Yihuang Decoction common gel had no significant changes with temperature. Compared with in situ gel, the color of common gel was darker and more translucent. Besides, its mobility was stronger after diluting with simulated vaginal fluid. The in vitro release study showed that the kinetic behavior of berberine hydrochloride in Yihuang Decoction vaginal thermosensitive in situ gel was matched with the Higuchi equation. Through simulation of vaginal administration, physical properties and dynamic rheological parameters were used to intuitively and scientifically evaluate the two gels. Compared with the common gel, the thermosensitive in situ gel could quickly attached to the vaginal mucosa and release drug, and thus was more suitable for developing vaginal administration of Yihuang Decoction, which also provides references for studying new vaginal preparation of Yihuang Decoction.
Asunto(s)
Administración Intravaginal , Medicamentos Herbarios Chinos/química , Geles/química , Femenino , Humanos , Poloxámero , Reología , Temperatura , ViscosidadRESUMEN
The aim of the study was to examine the effect of complex prescription on the pharmacokinetics of baicalin, rhynochophylline and gastrodin in Tiangou antihypertensive capsule. After administration, rat plasma was collected at different time-points. High-performance liquid chromatography was used to determine the content of baicalin, rhynochophylline and gastrodin in plasma. Two peaks occurred in the baicalin concentration-time curve. No significant difference was found for the peak concentration time (tmax) and area under concentration-time curve (AUC) of baicalin between the complex prescription and baicalin groups. The peak concentration (Cmax) of baicalin in the complex prescription group was significantly decreased, while no significant difference was found for the absorption factor (Ka) and AUC of baicalin between the complex prescription and gastrodin groups. The elimination factor (Ke) of gastrodin in the complex prescription group was significantly decreased, while the apparent volume of distribution (Vd) of gastrodin was significantly increased. No significant difference was found for AUC of baicalin between the complex prescription and rhynochophylline groups. Levels of Ka and Ke of rhynochophylline were lower in the complex prescription group while Vd was higher. Thus, complex prescription made plasma concentration-time curve more smooth. By contrast, the Tiangou antihypertensive capsule improved the distribution of baicalin, rhynochophylline and gastrodin in vivo.