Fungal biology最新文献

The coconut rhinoceros beetle (CRB; Oryctes rhinoceros) is one of the most destructive insect pests of coconut and oil palms in tropical Asia and the Pacific islands. Members of a new variant, known as CRB-G (clade I), have recently spread into the Pacific islands, causing significant damage. Biopesticides containing Metarhizium spp. are the strongest candidates for inundative biological control against the emerging CRB threat. Selection of the most virulent and robust isolate may determine the impact of this control option on the pest. In this work, CRB specimens with natural fungal infection were collected in Papua New Guinea (PNG) and Solomon Islands (SI). Putative entomopathogenic fungi were isolated and identified. These new isolates and some previously obtained from other Pacific countries were molecularly identified, characterized, and tested for virulence against CRB larval populations in PNG and SI in laboratory bioassays. Of the new isolates collected, four obtained from SI were identified as Metarhizium majus (conidia length ⁓11–15 μm), and four from PNG were identified as Metarhizium pingshaense (conidia length ⁓4–6 μm). The most virulent isolate was M. majus AgR-F717, which caused 100 % mortality in 20–23 days against a CRB variant from the CRB-S grouping (clade II) in laboratory bioassays carried out in PNG. Isolates of M. pingshaense did not show pathogenicity against CRB larvae. M. majus AgR-F717 was also the most virulent in laboratory bioassays using the mixed SI population (from both CRB-S and CRB-G groupings) and was selected for further evaluation using artificial breeding sites. Under field conditions, this isolate demonstrated its ability to infect CRB, dispersal up to 100 m from treated artificial breeding sites, and persistence in soil for at least four months. The new isolate AgR-F717 of M. majus has demonstrated potential as an augmentative biological control agent for CRB in PNG and SI.

An intense black pigmented halotolerant yeast GUBPC1, was obtained from the solar salterns of Nerul, Goa-India. The isolate could tolerate 0 to 20 % NaCl. FE-SEM analysis revealed its polymorphic nature, exhibiting oval cells at higher salt concentrations and filamentous spindle like shapes at lower concentrations. Initially, the cells appear oval, yeast like in shape but gradually after 15 days of incubation, it becomes elongated and undergoes budding, exhibiting various budding patterns, from single polar bud to bipolar buds with annellidic ring, to lateral buds and eventually forming filamentous hyphae. The intracellular black pigment was identified as melanin based on ultraviolet–visible spectroscopy analysis. The molecular identification of the culture showed closest similarity with Hortaea werneckii. Plant polymer-degrading enzymatic activities such as cellulase, laccase, chitinase, xylanase, pectinase, amylase and protease were exhibited by the isolate GUBPC1. To further understand and explore its biotechnological potential, we performed whole-genome sequencing and analysis. The obtained genome size was 26.93 Mb with 686 contigs and a GC content of 53.24 %. We identified 9383 protein-coding genes, and their functional annotation revealed the presence of 435 CAZyme genes and 16 functional genes involved in secondary metabolite synthesis, thus providing a basis for its potential value in various biotechnological applications.

Arbuscular mycorrhizal (AM) fungi can sequester different potentially toxic elements, such as trace elements (TEs), within their structures to alleviate the toxicity for its host plant and themselves. To elucidate the role of AM fungi in TEs immobilization in the rhizosphere of host plants, it is important to know the TEs distribution in AM fungal structures. In the present study, we investigated the distribution and concentration of TEs within extraradical spores and mycelium of the AM fungus Rhizophagus intraradices, collected from the rhizosphere of Senecio bonariensis plants grown in a soil polluted with multiple TEs, by using Particle-Induced X-ray Emission with a micro-focused beam (micro PIXE). This technique enabled the simultaneous micrometric mapping of elements in a sample. The calculated values were compared with those in the polluted substrate, measured by the Wavelength Dispersive X-ray Fluorescence technique. The highest concentrations of Fe, P, Ti, Mn, Cr, Cu and Zn were found in AM fungal spores, where they were accumulated, while extraradical mycelium was enriched in Cu. Finally, we demonstrated that AM fungi can simultaneously accumulate high amounts of different TEs in their structures, thus reducing the toxicity of these elements to its host plant.

Fusarium verticillioides is both an endophyte and pathogen of maize. During growth on maize, the fungus often synthesizes the mycotoxins fumonisins, which have been linked to a variety of diseases, including cancer in some animals. How F. verticillioides responds to other fungi, such as Fusarium proliferatum, Aspergillus flavus, Aspergillus niger, and Penicillium oxalicum, that coinfect maize, has potential to impact mycotoxin synthesis and disease. We hypothesize that low molecular weight acids produced by these fungi play a role in communication between the fungi in planta/nature. To address this hypothesis, we exposed 48-h maize kernel cultures of F. verticillioides to oxalic acid, citric acid, fusaric acid, or kojic acid and then compared transcriptomes after 30 min and 6 h. Transcription of some genes were affected by multiple chemicals and others were affected by only one chemical. The most significant positive response was observed after exposure to fusaric acid which resulted in >2-fold upregulation of 225 genes, including genes involved in fusaric acid synthesis. Exposure of cultures to the other three chemicals increased expression of only 3–15 genes. The predicted function and frequent co-localization of three sets of genes support a role in protecting the fungus from the chemical or a role in catabolism. These unique transcriptional responses support our hypothesis that these chemicals can act as signaling molecules. Studies with gene deletion mutants will further indicate if the initial transcriptional response to the chemicals benefit F. verticillioides.

Bacillus spp. produce numerous antimicrobial metabolites. Among these metabolites, cyclic lipopeptides (CLP) including fengycins, iturins, and surfactins are known to have varying antifungal activity against phytopathogenic fungi. The differential activities of CLP have been attributed to diverse mechanisms of action on fungal membranes. However, the precise biochemical determinants driving their antifungal modes of action have not been conclusively identified. In this study, three plant pathogenic fungi of varying lipopeptide sensitivities, Alternaria solani, Cladosporium cucumerinum, and Fusarium sambucinum, were studied to determine how their cell membrane lipid compositions may confer sensitivity and/or tolerance to fengycin, iturin, and surfactin. Results indicated that sensitivity to all three lipopeptides correlated with lower ergosterol content and elevated phospholipid fatty acid unsaturation. Fungal sensitivity to surfactin was also notably different than fengycin and iturin, as surfactin was influenced more by lower phosphatidylethanolamine amounts, higher levels of phosphatidylinositol, and less by phospholipid fatty acyl chain length. Results from this study provide insight into the fungal membrane composition of A. solani, F. sambucinum, and C. cucumerinum and the specific membrane characteristics influencing the antifungal effectiveness of fengycin, iturin, and surfactin. Understanding of these determinants should enable more accurate prediction of sensitivity-tolerance outcomes for other fungal species exposed to these important CLP.

Cordyceps chanhua, an important cordycipitoid medical mushroom with wide use in Asia, has gained attention for its bioactive component beauvericin (BEA), which is of medicinal value as a drug lead, but also of food safety risk. Recent observations by our group revealed a significant decrease of BEA content in C. chanhua when exposed to light, but the underlying regulatory mechanisms remain elusive. In this study, a comprehensive approach combining metabolomics and transcriptomics was employed to investigate the effects of white light on the secondary metabolism of C. chanhua for elucidation of the influence of light on BEA biosynthesis in this fungus. The result showed that the genes and metabolites involved in the synthesis of D-hydroxyisovaleric acid, a precursor of BEA synthesis, were down-regulated under light exposure, while those associated with the synthesis of phenylalanine, another precursor of BEA synthesis, were up-regulated leading to elevated phenylalanine levels. It suggested that the suppressive effect of light on BEA synthesis in C. chanhua occurred primarily through the inhibition of D-hydroxyisovaleric acid synthesis, while the enhanced phenylalanine biosynthesis likely directed towards other metabolic pathway such as pigment synthesis. These results contributed to a better understanding on how light modulates the secondary metabolism of C. chanhua and provided valuable guidance for optimizing BEA production in cultivation practices.

The Oomycetes fungus Phytophthora spp. which causes Abnormal leaf fall (ALF) disease poses a significant threat as one of the most devastating diseases affecting rubber trees in India. A total of 30 Phytophthora isolates were obtained from ALF-affected samples collected during the Southwest monsoon season of Kerala. The colony morphology of Phytophthora isolates revealed eight different types of growth patterns, with stellate, stellate striated, and petaloid patterns growing rapidly, whereas chrysanthemum pattern grew slowly. Sporangia were papillate to non-papillate in various shapes, and sporangiophores exhibited simple, simple sympodial, or irregularly branching patterns. Highly virulent isolates exhibited petaloid morphology and rapid growth rates. Regardless of their virulence, all isolates showed susceptibility to the fungicide metalaxyl. Under in vitro conditions, the highly virulent isolate (R17) from rubber caused severe infections in chili, brinjal, and tomato with brown water-soaked lesions. Sequence analysis and multi-locus phylogeny of Internal transcribed spacer (ITS), cCytochrome c oxidase 1 (COX 1), Heat shock protein 90 (HSP 90), and Ribosomal protein L10 (RPL 10) confirmed the pathogen as Phytophthora meadii. A comprehensive understanding of both morphological and molecular traits of P. meadii is crucial for precise identification and future genetic variability studies.

Ectomycorrhizal (ECM) fungi play a major role in forest ecosystems and managed tree plantations. Particularly, they facilitate mineral weathering and nutrient transfer towards colonized roots. Among nutrients provided by these fungi, potassium (K) has been understudied compared to phosphorus (P) or nitrogen (N). The ECM fungus Paxillus ammoniavirescens is a generalist species that interacts with the root of many trees and can directly transfer K to them, including loblolly pine. However, the forms of K that ECM fungi can store is still unknown. Here, we used synchrotron potassium X-ray fluorescence (XRF) and K-edge X-ray Absorption Near Edge Structure (XANES) spectroscopy on P. ammoniavirescens growing in axenic conditions to investigate the K chemistries accumulating in the center and the edge of the mycelium. We observed that various K forms accumulated in different part of the mycelium, including K-nitrate (KNO₃), K-C-O compounds (such as K-tartrate K₂(C₄H₄O₆) and K-oxalate (K₂C₂O₄)), K-S and K-P compounds. Saprotrophic fungi have been shown to excrete carboxylic acids, which in turn play a role in soil mineral weathering. Our finding of several K counter-ions to carboxylic acids may suggest that, besides their direct transfer to colonized roots, K ions can also be involved in the production of compounds necessary for sourcing nutrients from their surrounding environment by ECM fungi. Additionally, this work reveals that XANES spectroscopy can be used to identify the various forms of K accumulating in biological systems.

Mutations have underpinned research into gene characterization across all domains of life. This includes the discovery of the genes involved in the development of asexual spores in filamentous fungi. Mutants in the ascomycete Paecilomyces variotii were isolated with impaired biosynthesis of the characteristic yellow pigment produced by this filamentous fungus. The affected genes were identified as pvpP, encoding the polyketide synthase that is required for synthesis of the pigment YWA1, and abaA and wetA that are two genes that encode components of the AbaA-BrlA-WetA module required for the development of asexual spores in species in the Eurotiales order. WetA was further characterized. A strain expressing a functional WetA-GFP fusion was created and used to find that WetA is expressed primarily in spores and concentrated in their nuclei, providing evidence that this conserved protein likely functions as a regulator of transcription in conidia. Analysis of the phenotypes of the P. variotii wetA mutant suggests that how this three-protein module impacts fungal biology will vary from species-to-species, despite being conserved amongst filamentous Ascomycete species.

Eucalyptus spp. in plantations are negatively affected by canker and wilt diseases caused by several species of Ceratocystis, particularly those in the Latin American Clade (LAC). Ceratocystis eucalypticola and Ceratocystis manginecans are of particular concern where disease epidemics are reported globally, with recent outbreaks emerging in South African and Indonesian Eucalyptus plantations. Consequently, a rapid screening protocol is required for these pathogens. In this study, a high-resolution melting curve analysis (HRMA) was developed to detect C. eucalypticola and C. manginecans that bypasses time-consuming isolation and post-PCR procedures. Primers targeting a 172 bp region of the cerato-platanin (CP) gene were designed. Using these primers, the accuracy of HRMA to detect and distinguish between these two LAC species was assessed using pure fungal DNA, and DNA extracted directly from Eucalyptus samples naturally infected with C. eucalypticola. The assay accurately detected the presence of C. eucalypticola and C. manginecans and quantifies their DNA, both from cultures, and directly from wood samples. HRMA further differentiated these two species from all other tested LAC individuals. This assay was also able to detect the presence of all the tested LAC species and distinguish seven of these, including C. fimbriata, to species level. Ceratocystis polyconidia was the only non-LAC off-target species detected. Based on these results, the developed assay can be used to rapidly identify C. eucalypticola and C. manginecans directly from infected plant material or fungal cultures, with the potential to also screen for several other LAC species.