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Antonie Van Leeuwenhoek Oct 2019This study set out to isolate and identify epiphytic yeasts producing pulcherrimin, and to evaluate their potential as biological control agents (BCAs). We isolated...
This study set out to isolate and identify epiphytic yeasts producing pulcherrimin, and to evaluate their potential as biological control agents (BCAs). We isolated Metschnikowia sp. strains from flowers and fruits collected in Poland. The plant material had been collected between April to September 2017 from two small orchards where traditional organic management is employed. We identified the essential phenotypic features of the yeast, including assimilation and enzymatic profiles, stress resistance, adhesion properties, and antimicrobial activity against various fungi involved in crop and/or food spoilage. Yeast screening was performed using YPD agar supplemented with chloramphenicol and Fe(III) ions. Taxonomic classification was determined by sequence analysis of the D1/D2 domains of the large subunit rRNA gene. The isolates were identified as Metschnikowia andauensis and Metschnikowia sinensis. The yeast isolates were further characterized based on their enzymatic and assimilation profiles, as well as their growth under various stress conditions. In addition, the hydrophobicity and adhesive abilities of the Metschnikowia isolates were determined using a MATH test and luminometry. Their antagonistic action against molds representing typical crop spoiling microflora was also evaluated. The assimilation profiles of the wild isolates were similar to those displayed by collection strains of M. pulcherrima. However, some of the isolates displayed more beneficial phenotypic properties, especially good growth under stress conditions. Several of the epiphytes grew well over a wider range of temperatures (8-30 °C) and pH levels (3-9), and additionally showed elevated tolerance to ethanol (8%), glucose (30%), and peroxides (50 mM). The hydrophobicity and adhesion of the yeast cells were strain- and surface-dependent. The tested yeasts showed potential for use as BCAs, with some exhibiting strong antagonism against molds belonging to the genera Alternaria, Botrytis, Fusarium, Rhizopus, and Verticillium, as well as against yeasts isolated as food spoilage microbiota.
Topics: Amino Acids, Sulfur; Antibiosis; Cluster Analysis; DNA, Fungal; DNA, Ribosomal; Flowers; Fruit; Fungi; Hydrogen-Ion Concentration; Metschnikowia; Mycological Typing Techniques; Pest Control, Biological; Phylogeny; Piperidines; Poland; RNA, Ribosomal; Sequence Analysis, DNA; Temperature
PubMed: 31111331
DOI: 10.1007/s10482-019-01272-w -
Frontiers in Microbiology 2023Eukaryotic life depends on the functional elements encoded by both the nuclear genome and organellar genomes, such as those contained within the mitochondria. The...
INTRODUCTION
Eukaryotic life depends on the functional elements encoded by both the nuclear genome and organellar genomes, such as those contained within the mitochondria. The content, size, and structure of the mitochondrial genome varies across organisms with potentially large implications for phenotypic variance and resulting evolutionary trajectories. Among yeasts in the subphylum Saccharomycotina, extensive differences have been observed in various species relative to the model yeast , but mitochondrial genome sampling across many groups has been scarce, even as hundreds of nuclear genomes have become available.
METHODS
By extracting mitochondrial assemblies from existing short-read genome sequence datasets, we have greatly expanded both the number of available genomes and the coverage across sparsely sampled clades.
RESULTS
Comparison of 353 yeast mitochondrial genomes revealed that, while size and GC content were fairly consistent across species, those in the genera and trended larger, while several species in the order Saccharomycetales, which includes , exhibited lower GC content. Extreme examples for both size and GC content were scattered throughout the subphylum. All mitochondrial genomes shared a core set of protein-coding genes for Complexes III, IV, and V, but they varied in the presence or absence of mitochondrially-encoded canonical Complex I genes. We traced the loss of Complex I genes to a major event in the ancestor of the orders Saccharomycetales and Saccharomycodales, but we also observed several independent losses in the orders Phaffomycetales, Pichiales, and Dipodascales. In contrast to prior hypotheses based on smaller-scale datasets, comparison of evolutionary rates in protein-coding genes showed no bias towards elevated rates among aerobically fermenting (Crabtree/Warburg-positive) yeasts. Mitochondrial introns were widely distributed, but they were highly enriched in some groups. The majority of mitochondrial introns were poorly conserved within groups, but several were shared within groups, between groups, and even across taxonomic orders, which is consistent with horizontal gene transfer, likely involving homing endonucleases acting as selfish elements.
DISCUSSION
As the number of available fungal nuclear genomes continues to expand, the methods described here to retrieve mitochondrial genome sequences from these datasets will prove invaluable to ensuring that studies of fungal mitochondrial genomes keep pace with their nuclear counterparts.
PubMed: 38075892
DOI: 10.3389/fmicb.2023.1268944 -
Environmental Microbiology Feb 2015Microbial habitats that contain an excess of carbohydrate in the form of sugar are widespread in the microbial biosphere. Depending on the type of sugar, prevailing... (Review)
Review
Microbial habitats that contain an excess of carbohydrate in the form of sugar are widespread in the microbial biosphere. Depending on the type of sugar, prevailing water activity and other substances present, sugar-rich environments can be highly dynamic or relatively stable, osmotically stressful, and/or destabilizing for macromolecular systems, and can thereby strongly impact the microbial ecology. Here, we review the microbiology of different high-sugar habitats, including their microbial diversity and physicochemical parameters, which act to impact microbial community assembly and constrain the ecosystem. Saturated sugar beet juice and floral nectar are used as case studies to explore the differences between the microbial ecologies of low and higher water-activity habitats respectively. Nectar is a paradigm of an open, dynamic and biodiverse habitat populated by many microbial taxa, often yeasts and bacteria such as, amongst many others, Metschnikowia spp. and Acinetobacter spp., respectively. By contrast, thick juice is a relatively stable, species-poor habitat and is typically dominated by a single, xerotolerant bacterium (Tetragenococcus halophilus). A number of high-sugar habitats contain chaotropic solutes (e.g. ethyl acetate, phenols, ethanol, fructose and glycerol) and hydrophobic stressors (e.g. ethyl octanoate, hexane, octanol and isoamyl acetate), all of which can induce chaotropicity-mediated stresses that inhibit or prevent multiplication of microbes. Additionally, temperature, pH, nutrition, microbial dispersion and habitat history can determine or constrain the microbiology of high-sugar milieux. Findings are discussed in relation to a number of unanswered scientific questions.
Topics: Acinetobacter; Carbohydrate Metabolism; Carbohydrates; Ecology; Enterococcaceae; Environment; Flowers; Fruit; Metschnikowia
PubMed: 25041632
DOI: 10.1111/1462-2920.12570 -
Frontiers in Bioscience (Elite Edition) Jan 2023The vineyard is a great reservoir of autochthonous yeast strains whose composition is defined by different regional (edaphology, orography or climatology) and...
BACKGROUND
The vineyard is a great reservoir of autochthonous yeast strains whose composition is defined by different regional (edaphology, orography or climatology) and anthropological factors (cultivation systems or cultural practices). Most of this yeast diversity corresponds to non- strains, some of which have potential use in winemaking.
METHODS
The oenological potential of 29 different native non- strains belonging to 4 species (, , and spp.) was evaluated, using the autochthonous XG3 strain as a control. Microfermentations with pure culture of each strain were performed in duplicate and the basic parameters and major volatiles of wines were analysed following official methodology. The best strain within each species was selected using a quantification matrix including the relevant oenological characteristics.
RESULTS
The fermentative ability of non- was lower than in all cases, but with differences among species. and showed higher fermentation rates than , whereas spp. presented a low fermentative power. At chemical level all non- strains reduced the alcoholic content, the higher alcohols and the volatile acidity of wines and increased the content of glycerol, with differences among strains within a given species. and increased the total acidity of wines. The latter and spp. strains produced lactic acid, which decreased the wine pH in the case of . According to their oenological traits the best rated strains of each species were Lt93, Td315, Mf278 and Sb474. In addition, the data obtained in pure fermentations were correlated to those chemical and aromatic compounds obtained with these non- strains in sequential fermentations.
CONCLUSIONS
Autochthonous strains of non- yeast species contribute distinctive chemical characteristics to the wines. The correlations observed between wines fermented with the different non- indigenous strains in pure and sequential fermentations suggest that their contribution to wine properties remains stable regardless of must composition or winemaking techniques.
Topics: Saccharomyces cerevisiae; Vitis; Wine; Glycerol
PubMed: 36959102
DOI: 10.31083/j.fbe1501001 -
Antonie Van Leeuwenhoek Nov 2018From comparisons of ITS1-5.8S-ITS2 and gene sequences for nuclear D1/D2 LSU rRNA, nuclear SSU (18S) rRNA, translation elongation factor 1-α (EF1-α) and RNA polymerase...
From comparisons of ITS1-5.8S-ITS2 and gene sequences for nuclear D1/D2 LSU rRNA, nuclear SSU (18S) rRNA, translation elongation factor 1-α (EF1-α) and RNA polymerase II subunit 2 (RPB2), the following four new ascosporogenous yeast species were resolved and are described as Metschnikowia anglica (NRRL Y-7298 [type strain], CBS 15342, MycoBank MB 823167), Metschnikowia leonuri (NRRL Y-6546, CBS 15341, MB 823166), Metschnikowia peoriensis (NRRL Y-5942, CBS 15345, MB 823164) and Metschnikowia rubicola (NRRL Y-6064, CBS 15344, MB 823165). The following six species of Candida are members of the Metschnikowia clade and are proposed for transfer to Metschnikowia as new combinations: Candida chrysomelidarum (NRRL Y-27749, CBS 9904, MB 823223), Candida gelsemii (NRRL Y-48212, CBS 10509, MB 823192), Candida kofuensis (NRRL Y-27226, CBS 8058, MB 823195), Candida picachoensis (NRRL Y-27607, CBS 9804, MB 823197), Candida pimensis (NRRL Y-27619, CBS 9805, MB 823205) and Candida rancensis (NRRL Y-48702, CBS 8174, MB 823224). Candida fructus (NRRL Y-17072, CBS 6380, MB 823206) is transferred to Clavispora as a new combination, and Candida musae is shown to be a synonym of C. fructus. Apparent multiple alleles for ITS, D1/D2, EF1-α and RPB2 were detected in strains of some species.
Topics: Candida; DNA, Fungal; DNA, Ribosomal Spacer; Metschnikowia; Phylogeny; Saccharomycetales
PubMed: 29754318
DOI: 10.1007/s10482-018-1095-8 -
Frontiers in Microbiology 2023Finding alternatives to the use of chemical inputs to preserve the sanitary and organoleptic quality of food and beverages is essential to meet public health... (Review)
Review
Finding alternatives to the use of chemical inputs to preserve the sanitary and organoleptic quality of food and beverages is essential to meet public health requirements and consumer preferences. In oenology, numerous manufacturers already offer a diverse range of bio-protection yeasts to protect must against microbiological alterations and therefore limit or eliminate sulphites during winemaking. Bio-protection involves selecting non- yeasts belonging to different genera and species to induce negative interactions with indigenous microorganisms, thereby limiting their development and their impact on the matrix. Although the effectiveness of bio-protection in the winemaking industry has been reported in numerous journals, the underlying mechanisms are not yet well understood. The aim of this review is to examine the current state of the art of field trials and laboratory studies that demonstrate the effects of using yeasts for bio-protection, as well as the interaction mechanisms that may be responsible for these effects. It focuses on the yeast , particularly recommended for the bio-protection of grape musts.
PubMed: 37664122
DOI: 10.3389/fmicb.2023.1252973 -
FEMS Microbiology Ecology Jul 2023Bee-fungus associations are common, and while most studies focus on entomopathogens, emerging evidence suggests that bees associate with a variety of symbiotic fungi... (Review)
Review
Bee-fungus associations are common, and while most studies focus on entomopathogens, emerging evidence suggests that bees associate with a variety of symbiotic fungi that can influence bee behavior and health. Here, we review nonpathogenic fungal taxa associated with different bee species and bee-related habitats. We synthesize results of studies examining fungal effects on bee behavior, development, survival, and fitness. We find that fungal communities differ across habitats, with some groups restricted mostly to flowers (Metschnikowia), while others are present almost exclusively in stored provisions (Zygosaccharomyces). Starmerella yeasts are found in multiple habitats in association with many bee species. Bee species differ widely in the abundance and identity of fungi hosted. Functional studies suggest that yeasts affect bee foraging, development, and pathogen interactions, though few bee and fungal taxa have been examined in this context. Rarely, fungi are obligately beneficial symbionts of bees, whereas most are facultative bee associates with unknown or ecologically contextual effects. Fungicides can reduce fungal abundance and alter fungal communities associated with bees, potentially disrupting bee-fungi associations. We recommend that future study focus on fungi associated with non-honeybee species and examine multiple bee life stages to document fungal composition, abundance, and mechanistic effects on bees.
Topics: Bees; Animals; Fungicides, Industrial; Ecosystem; Mycobiome; Porifera; Metschnikowia; Fungi
PubMed: 37422442
DOI: 10.1093/femsec/fiad077 -
Frontiers in Microbiology 2018The internal transcribed spacer (ITS) region (ITS1, 5.8S rDNA, and ITS2) separates the genes coding for the SSU 18S and the LSU 26S genes in the rDNA units which are...
The internal transcribed spacer (ITS) region (ITS1, 5.8S rDNA, and ITS2) separates the genes coding for the SSU 18S and the LSU 26S genes in the rDNA units which are organized into long tandem arrays in the overwhelming majority of fungi. As members of a multigenic family, these units are subject of concerted evolution, which homogenizes their sequences. Exceptions have been observed in certain groups of plants and in a few fungal species. In our previous study we described exceptionally high degree of sequence diversity in the D1/D2 domains of two pulcherrimin-producing (Saccharomycotina) species which appeared to evolve by reticulation. The major goals of this study were the examination of the diversity of the ITS segments and their evolution. We show that the ITS sequences of these species are not homogenized either, differ from each other by up to 38 substitutions and indels which have dramatic effects on the predicted secondary structures of the transcripts. The high intragenomic diversity makes the D1/D2 domains and the ITS spacers unsuitable for barcoding of these species and therefore the taxonomic position of strains previously assigned to them needs revision. By analyzing the genome sequence of the type strain, we also show that the rDNA of this species is fragmented, contains pseudogenes and thus evolves by the birth-and-death mechanism rather than by homogenisation, which is unusual in yeasts. The results of the network analysis of the sequences further indicate that the ITS regions are also involved in reticulation. and can form interspecies hybrids and their hybrids segregate, providing thus possibilities for reticulation of the rDNA repeats.
PubMed: 29946303
DOI: 10.3389/fmicb.2018.01193 -
International Journal of Molecular... Jan 2021The surfaces of grapes are covered by different yeast species that are important in the first stages of the fermentation process. In recent years, non- yeasts such as ,... (Review)
Review
The surfaces of grapes are covered by different yeast species that are important in the first stages of the fermentation process. In recent years, non- yeasts such as , , , and have become popular with regard to winemaking and improved wine quality. For that reason, several manufacturers started to offer commercially available strains of these non- species. stands out, mainly due to its contribution to wine aroma, glycerol, ethanol yield, and killer factor. The metabolism of the yeast allows it to increase volatile molecules such as esters and varietal thiols (aroma-active compounds), which increase the quality of specific varietal wines or neutral ones. It is considered a low- or non-fermentative yeast, so subsequent inoculation of a more fermentative yeast such as is indispensable to achieve a proper fermented alcohol. The impact of is not limited to the grape wine industry; it has also been successfully employed in beer, cider, durian, and tequila fermentation, among others, acting as a promising tool in those fermentation processes. Although no species other than is available in the regular market, several recent scientific studies show interesting improvements in some wine quality parameters such as aroma, polysaccharides, acid management, and color stability. This could motivate yeast manufacturers to develop products based on those species in the near future.
Topics: Bioengineering; Ecology; Fermentation; Food Quality; Industrial Microbiology; Pichia; Vitis; Wine
PubMed: 33530422
DOI: 10.3390/ijms22031196 -
Archives of Microbiology Aug 2023Recently, there has been growing interest in the characterization of native yeasts for their use in production of wines with regional characteristics. This study aimed...
Recently, there has been growing interest in the characterization of native yeasts for their use in production of wines with regional characteristics. This study aimed to investigate Saccharomyces and non-Saccharomyces yeasts present in the spontaneous fermentation of Tannat and Marselan grape musts collected from Concordia (Entre Ríos, Argentina) over 2019, 2020, and 2021 vintages. The evolution of these fermentative processes was carried out by measuring total soluble solids, total acidity, volatile acidity, pH, ethanol concentration, and total carbon content. Isolated Saccharomyces and non-Saccharomyces yeasts were identified based on colony morphology in WL medium, 5.8S-ITS-RFLP analysis, and 26S rDNA D1/D2 gene sequencing. Two hundred and ten yeast colonies were isolated and identified as Pichia kudriavzevii, Saccharomyces cerevisiae, Hanseniaspora uvarum, Metschnikowia pulcherrima, Candida albicans, Candida parapsilosis, Pichia occidentalis, Pichia bruneiensis, Hanseniaspora opuntiae, Issatchenkia terricola, and Hanseniaspora vineae. P. kudriavzevii isolated from all vintages was associated with the spontaneous fermentation of grape musts from the Concordia region.
Topics: Vitis; Fermentation; Yeasts; Wine; Saccharomyces cerevisiae; DNA, Ribosomal
PubMed: 37550458
DOI: 10.1007/s00203-023-03646-1