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Fungal Biology Jun 2024Volatile organic compounds (VOCs) produced by yeasts can positively affect crops, acting as antifungals or biostimulants. In this study, Aureobasidium pullulans and...
Volatile organic compounds (VOCs) produced by yeasts can positively affect crops, acting as antifungals or biostimulants. In this study, Aureobasidium pullulans and Metschnikowia pulcherrima were evaluated as potential antagonists of Trichoderma spp., common fungal pathogen in mushroom cultivation. To assess the biocontrol ability and biostimulant properties of the selected yeast species, in vitro co-culture and VOCs exposure assays were conducted. In both assays, VOCs produced by Aureobasidium spp. showed the stronger antifungal activity with a growth inhibition up to 30 %. This result was further confirmed by the higher volatilome alcohol content revealed by solid phase microextraction-gas chromatography mass spectrometry (SPME/GC-MS). Overall, Aureobasidium strains can be potentially used as biocontrol agent in Pleorotus ostreatus and Cyclocybe cylindracea mycelial growth, without affecting their development as demonstrated by VOCs exposure assay and Fourier-transform infrared spectroscopy (FT-IR). Conversely, M. pulcherrima was characterized by a lower or absent antifungal properties and by a volatilome composition rich in isobutyl acetate, an ester often recognized as plant growth promoter. As confirmed by FT-IR, Lentinula mycelia exposed to M. pulcherrima VOCs showed a higher content of proteins and lipids, suggesting an improvement of some biochemical properties. Our study emphasizes that VOCs produced by specific yeast strains are potentially powerful alternative to synthetic fungicide in the vegetative growth of mushroom-forming fungi and also able to modify their biochemical composition.
Topics: Volatile Organic Compounds; Mycelium; Agaricales; Gas Chromatography-Mass Spectrometry; Antifungal Agents; Biological Control Agents; Metschnikowia; Antibiosis; Aureobasidium; Trichoderma; Solid Phase Microextraction
PubMed: 38876538
DOI: 10.1016/j.funbio.2024.05.007 -
Foods (Basel, Switzerland) May 2024This study explores the potential probiotic properties of yeasts isolated from various Chilean honeys, focusing on Ulmo, Quillay, and Mountain honeys. Six yeast strains...
This study explores the potential probiotic properties of yeasts isolated from various Chilean honeys, focusing on Ulmo, Quillay, and Mountain honeys. Six yeast strains were identified, including , sp., , , , and . Phenotypic characterization involved assessing their fermentative performance, ethanol and hops resistance, and cross-resistance. Ethanol concentration emerged as a limiting factor in their fermentative performance. The probiotic potential of these yeasts was evaluated based on resistance to high temperatures, low pH, auto-aggregation capacity, survival in simulated digestion (INFOGEST method), and antimicrobial activity against pathogens like , , and . Three yeasts, , , and , exhibited potential probiotic characteristics by maintaining cell concentrations exceeding 10 CFU/mL after digestion. They demonstrated fermentative abilities and resistance to ethanol and hops, suggesting their potential as starter cultures in beer production. Despite revealing promising probiotic and technological aspects, further research is necessary to ascertain their viability in producing fermented foods. This study underscores the innovative potential of honey as a source for new probiotic microorganisms and highlights the need for comprehensive investigations into their practical applications in the food industry.
PubMed: 38790882
DOI: 10.3390/foods13101582 -
Current Microbiology May 2024Postharvest fungal diseases cause serious fruit losses and food safety issues worldwide. The trend in preventing food loss and waste has shifted to environmentally...
Postharvest fungal diseases cause serious fruit losses and food safety issues worldwide. The trend in preventing food loss and waste has shifted to environmentally friendly and sustainable methods, such as biological control. Penicillium expansum is a common postharvest contaminant fungus that causes blue mould disease and patulin formation on apples. This study aimed to provide biocontrol using Metschnikowia pulcherrima isolates against P. expansum, and to understand their antagonistic action mechanisms. In vitro, 38.77-51.69% of mycelial growth inhibition of P. expansum was achieved by M. pulcherrima isolates with the dual culture assay, while this rate was 69.45-84.89% in the disc diffusion assay. The disease symptoms of P. expansum on wounds were reduced by M. pulcherrima, on Amasya apples. The lesion diameter, 41.84 mm after 12 d of incubation in control, was measured as 24.14 mm when treated with the most effective M. pulcherrima DN-MP in vivo. Although the antagonistic mechanisms of M. pulcherrima isolates were similar, there was a difference between their activities. In general, DN-HS and DN-MP isolates were found to be more effective. In light of all these results, it can be said that M. pulcherrima isolates used in the study have an antagonistic effect against the growth of P. expansum both in vitro and in vivo in Amasya apples, therefore, when the appropriate formulation is provided, they can be used as an alternative biocontrol agent to chemical fungicides in the prevention of postharvest diseases.
Topics: Penicillium; Malus; Plant Diseases; Antibiosis; Metschnikowia; Fruit; Biological Control Agents
PubMed: 38761223
DOI: 10.1007/s00284-024-03700-1 -
Scientific Reports May 2024In molecular systematics, the delimitation of yeast species is based on the notion that the barcode differences are smaller within species than between them. The most...
In molecular systematics, the delimitation of yeast species is based on the notion that the barcode differences are smaller within species than between them. The most widely used barcodes are segments of the chromosomal repeats coding for ribosomal RNAs that are homogenised in yeasts. The analysis of these segments of the type strains of ten species recently merged in Metschnikowia pulcherrima and 37 new isolates demonstrated that this is not the case in this species. The intragenomic diversity significantly exceeded the threshold gaps used to differentiate related yeast species. Large segments of the D1/D2 domains were not diverse within the genomes and could therefore be used to determine the taxonomic affiliation of the isolates. The genome structures of the isolates were compared by RAPD and the RFLP of the mitochondrial DNA. Both patterns were highly heterogeneous. The sequence analysis of the PUL4 gene (a member of the PUL gene cluster involved in pulcherrimin production) revealed very high intragenomic differences, suggesting that the genomes may be chimerised. Three phenotypic traits related to the antimicrobial antagonism characteristic of the species were also highly diverse and prone to reversible segregation resembling epigenetic processes (silencing and reactivation of regulators) rather than mutations and back-mutations. These features make M. pulcherrima unique among yeasts and indicate that it evolves in a non-standard way.
Topics: Metschnikowia; Genome, Fungal; Evolution, Molecular; Phylogeny; Genetic Variation; Phenotype; DNA, Mitochondrial
PubMed: 38714828
DOI: 10.1038/s41598-024-61335-5 -
Journal of Chemical Ecology May 2024Chemical control of Drosophila suzukii (Diptera: Drosophilidae) based on the use of insecticides is particularly challenging as the insect attacks ripening fruits...
Chemical control of Drosophila suzukii (Diptera: Drosophilidae) based on the use of insecticides is particularly challenging as the insect attacks ripening fruits shortly before harvest. An alternative strategy may rely on the use of yeasts as phagostimulants and baits, applied on canopy as attract-and-kill formulations. The aim of this research was to identify the most attractive among six yeast species for D. suzukii: Saccharomyces cerevisiae, Hanseniaspora uvarum, Clavispora santaluciae, Saccharomycopsis vini, Issatchenkia terricola, and Metschnikowia pulcherrima. The volatile profile of C. santaluciae was described for the first time. Behavioural experiments identified H. uvarum and S. vini as the most attractive yeasts. The characterization of yeast headspace volatiles using direct headspace (DHS) and solid-phase microextraction (SPME) revealed several strain-specific compounds. With DHS injection, 19 volatiles were characterised, while SPME revealed 71 compounds constituting the yeast headspace. Both analyses revealed terpenoids including β-ocimene, citronellol, (Z)-geraniol (nerol), and geranial as distinct constituents of S. vini. H. uvarum and S. vini were further investigated using closed-loop stripping analysis (CSLA) and electroantennography. Out of 14 compounds quantified by CSLA, ethyl acetate, isoamyl acetate, β-myrcene, benzaldehyde and linalool were detected by D. suzukii antennae and might generate the strong attractiveness of S. vini and H. uvarum. Our results highlight a strong attraction of D. suzukii to various yeasts associated with both the flies and their habitat and demonstrate how different sampling methods can impact the results of volatile compound characterization. It remains to be demonstrated whether the distinct attraction is based on special adaptations to certain yeasts and to what extent the metabolites causing attraction are interchangeable.
PubMed: 38691267
DOI: 10.1007/s10886-024-01494-x -
FEMS Microbiology Ecology Apr 2024The Melipona gut microbes differ from other social bees, with the absence of crucial corbiculate core gut symbionts and the high occurrence of environmental strains. We...
The Melipona gut microbes differ from other social bees, with the absence of crucial corbiculate core gut symbionts and the high occurrence of environmental strains. We studied the microbial diversity and composition of three Melipona species and their honey to understand which strains are obtained by horizontal transmission (HT) from the pollination environment; or represent symbionts co-evolved with Melipona by HT from the hive/food stores or vertical transmission (VT) via social interactions. Bees harbored higher microbial alpha diversity and a different and more species-specific bacterial composition than honey. Otherwise, the fungal communities of bee and honey samples are less dissimilar. As expected, the core symbionts Snodgrassella and Gilliamella were absent in bees that had a prevalence of Lactobacillus Firm-5, environmental Lactobacillaceae, Bifidobacteriaceae and Acetobacteraceae. Also, Pectinatus and Floricoccus have habitat preferences for bees, putatively representing novel symbionts from the environment that co-evolved via VT among generations. Fructilactobacillus found in bees possibly had HT to bees from honey stores. Metschnikowia yeasts, consistent in all bees and honey samples, might have HT to bees from food stores. Similarly, Saccharomycetales might have HT from honey or plants/flowers to bees. This work contributes to the understanding of Melipona symbionts and their modes of transmission.
PubMed: 38650068
DOI: 10.1093/femsec/fiae063 -
Folia Microbiologica Apr 2024Patients with burn injury and inhalation injury are highly susceptible to infectious complications, including opportunistic pathogens, due to the loss of skin cover and...
Patients with burn injury and inhalation injury are highly susceptible to infectious complications, including opportunistic pathogens, due to the loss of skin cover and mucosal damage of respiratory tract as well as the disruption of homeostasis. This case report, a 34-year-old man suffered critical burns, provides the first literature description of triple-impact immunoparalysis (critical burns, inhalation injury, and SARS-CoV-2 infection), leading to a lethal multifocal infection caused by several fungi including very rare environmental representatives Metschnikowia pulcherrima and Wickerhamomyces anomalus. The co-infection by these common environmental yeasts in a human is unique and has not yet been described in the literature. Importantly, our patient developed refractory septic shock and died despite targeted antifungal therapy including the most potent current antifungal agent-isavuconazole. It can be assumed that besides immunoparalysis, effectiveness of therapy by isavuconazole was impaired by the large distribution volume in this case. As this is a common situation in intensive care patients, routine monitoring of plasmatic concentration of isavuconazole can be helpful in personalization of the treatment and dose optimization. Whatmore, many fungal species often remain underdiagnosed during infectious complications, which could be prevented by implementation of new methods, such as next-generation sequencing, into clinical practice.
PubMed: 38647991
DOI: 10.1007/s12223-024-01165-0 -
Ecology and Evolution Apr 2024Secondary metabolites produced by primary producers have a wide range of functions as well as indirect effects outside the scope of their direct target. Research...
Secondary metabolites produced by primary producers have a wide range of functions as well as indirect effects outside the scope of their direct target. Research suggests that protease inhibitors produced by cyanobacteria influence grazing by herbivores and may also protect against parasites of cyanobacteria. In this study, we asked whether those same protease inhibitors produced by cyanobacteria could also influence the interactions of herbivores with their parasites. We used the - zooplankton host-fungal parasite system to address this question because it is well documented that cyanobacteria protease inhibitors suppress trypsin and chymotrypsin in the gut of , and because it is known that infects via the gut. We tested the hypothesis that gut proteases are necessary for spores to be released from their asci. We then also tested whether diets that decrease trypsin and chymotrypsin activity in the guts of lead to lower levels of infection. Our results show that chymotrypsin promotes the release of the fungal spores from their asci. Moreover, a diet that strongly inhibited chymotrypsin activity in decreased infection levels, particularly in the most susceptible clones. Our results support the growing literature that cyanobacterial diets can be beneficial to zooplankton hosts when challenged by parasites and uncover a mechanism that contributes to the protective effect of cyanobacterial diets. Specifically, we demonstrate that host chymotrypsin enzymes promote the dehiscence of spores; when cyanobacteria inhibit the activity of chymotrypsin in hosts, this most likely traps the spore inside the ascus, preventing the parasite from puncturing the gut and beginning the infection process. This study illustrates how secondary metabolites of phytoplankton can protect herbivores against their own enemies.
PubMed: 38646007
DOI: 10.1002/ece3.11340 -
Molecular Biology Reports Apr 2024Yeast biosynthesizes fusel alcohols in fermentation through amino acid catabolism via the Ehrlich pathway. ARO8 and ARO9 genes are involved in the first step of the...
BACKGROUND
Yeast biosynthesizes fusel alcohols in fermentation through amino acid catabolism via the Ehrlich pathway. ARO8 and ARO9 genes are involved in the first step of the Ehrlich pathway, while ADH2 and ADH5 genes are involved in the last step. In this study, we describe RT-qPCR methods to determine the gene expression level of genes (ARO8, ARO9, ADH2, ADH5) found in Saccharomyces cerevisiae (Sc) and Metschnikowia pulcherrima (Mp) strains growth pasteurized white grape juice.
METHODS AND RESULTS
We used RNA extraction and cDNA synthesis protocols. The RT-qPCR efficiency of primer pairs was evaluated by generating a standard curve through serial dilution of yeast-derived cDNA. Method performance criteria were determined for each RT-qPCR assay. Then, we evaluated the gene expression levels of the four genes in all samples. RNA extraction and cDNA synthesis from yeast samples demonstrated the method's capability to generate high-yield, high-purity nucleic acids, supporting further RT-qPCR analysis. The highest normalized gene expression levels of ARO8 and ARO9 were observed in SC1, SC4, and SC5 samples. No significant difference in ADH2 gene expression among Mp strains was observed during the examination of ADH2 and ADH5 genes (p < 0.05). We observed no expression of the ADH5 gene in Mp strains except MP6 strain. The expression of ADH2 and ADH5 genes was higher in Sc strains compared to Mp strains.
CONCLUSIONS
The results suggest that the proposed RT-qPCR methods can measure gene expression of ARO8, ARO9, ADH2, and ADH5 in Sc and Mp strains growing in pasteurized white grape juice.
Topics: Saccharomyces cerevisiae; Vitis; DNA, Complementary; Transaminases; Fermentation; RNA; Metschnikowia
PubMed: 38642187
DOI: 10.1007/s11033-024-09444-2 -
FEMS Yeast Research Jan 2024Although filamentous Ascomycetes may produce structures that are interpreted as male and female gametangia, ascomycetous yeasts are generally not considered to possess...
Although filamentous Ascomycetes may produce structures that are interpreted as male and female gametangia, ascomycetous yeasts are generally not considered to possess male and female sexes. In haplontic yeasts of the genus Metschnikowia, the sexual cycle begins with the fusion of two morphologically identical cells of complementary mating types. Soon after conjugation, a protuberance emerges from one of the conjugants, eventually maturing into an ascus. The originating cell can be regarded as an ascus mother cell, hence as female. We tested the hypothesis that the sexes, female or male, are determined by the mating types. There were good reasons to hypothesize further that mating type α cells are male. In a conceptually simple experiment, we observed the early stages of the mating reaction of mating types differentially labeled with fluorescent concanavalin A conjugates. Three large-spored Metschnikowia species, M. amazonensis, M. continentalis, and M. matae, were examined. In all three, the sexes were found to be independent of mating type, cautioning that the two terms should not be used interchangeably.
Topics: Genes, Mating Type, Fungal; Metschnikowia
PubMed: 38632043
DOI: 10.1093/femsyr/foae014