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The inebriation of wild African ele phants fr om eating the ripened and rotting fruit of the marula tree is a persistent myth in Southern Africa. However, the y easts r esponsib le for alcoholic fermentation to intoxicate the elephants remain poorly documented. In this study, we consider ed Botsw ana, a countr y with the world's largest population of wild elephants, and where the marula tree is indigenous, a bundant and pr otected, to assess the occurr ence and biodiversity of yeasts with a potential to ferment and subsequently inebriate the wild elephants. We collected marula fruits from over a stretch of 800 km in Botswana and isolated 106 yeast strains representing 24 yeast species. Over 93% of these isolates, typically known to ferment simple sugars and produce ethanol comprising of high ethanol producers belonging to Sacchar om yces, Br ettanom yces, and Pichia, and intermediate ethanol producers Wickerhamom yces, Zygotorulaspora, Candida, Hanseniaspora, and Kluyver om yces. Fermentation of marula juice r ev ealed convincing fermentative and ar omatic bouquet credentials to suggest the potential to influence foraging behaviour and inebriate elephants in nature. There is insufficient evidence to refute the aforementioned myth. This work serves as the first work to war ds understanding the biodiversity marula associated yeasts to debunk the myth or appr ov e the facts. [ABSTRACT FROM AUTHOR]

This confusion prevailed in the literature until recently and resulted in the inaccurate use of the name I S. bayanus i or I S. bayanus i variety I uvarum i to refer to I S. uvarum i (e.g. Caudy et al. [8], Cliften et al. [9], Kellis et al. [16]). That study also showed that I S. uvarum i was the sister taxon of I S. eubayanus i and, importantly, was not a synonym of I S. bayanus i , thus confirming earlier reports (e.g. Nguyen and Gaillardin [25]). Therefore, the finding of I S. eubayanus i in South America solved one mystery but created another - the Atlantic Ocean and 12 000 km separated the region where I S. eubayanus i was found from the region where, plausibly, the formation of the hybrid I S. pastorianus i took place. The genomes of I S. eubayanus i and the non-cerevisiae subgenome of I S. pastorianus i had 99.56% identity, which demonstrated that I S. eubayanus i was the elusive non-cerevisiae progenitor of lager yeasts. [Extracted from the article]

The study of microbe domestication has witnessed major advances that contribute to a better understanding of the emergence of artificially selected phenotypes and set the foundations of their rational improvement for biotechnology. Several features make Saccharomyces cerevisiae an ideal model for such a study, notably the availability of a catalogue of signatures of artificial selection and the extensive knowledge available on its biological processes. Here, we investigate with population and comparative genomics a set of strains used for cachaça fermentation, a Brazilian beverage based on the fermentation of sugar cane juice. We ask if the selective pressures posed by this fermentation have given rise to a domesticated lineage distinct from the ones already known, like wine, beer, bread, and sake yeasts. Our results show that cachaça yeasts derive from wine yeasts that have undergone an additional round of domestication, which we define as secondary domestication. As a consequence, cachaça strains combine features of wine yeasts, such as the presence of genes relevant for wine fermentation and advantageous gene inactivations, with features of beer yeasts like resistance to the effects of inhibitory compounds present in molasses. For other markers like those related to sulfite resistance and biotin metabolism our analyses revealed distributions more complex than previously reported that support the secondary domestication hypothesis. We propose a multilayered microbe domestication model encompassing not only transitions from wild to primarily domesticated populations, as in the case of wine yeasts, but also secondary domestications like those of cachaça yeasts. [ABSTRACT FROM AUTHOR]

The budding yeast Saccharomyces cerevisiae can be found in the wild and is also frequently associated with human activities. Despite recent insights into the phylogeny of this species, much is still unknown about how evolutionary processes related to anthropogenic niches have shaped the genomes and phenotypes of S. cerevisiae. To address this question, we performed population-level sequencing of 82 S. cerevisiae strains from wine, flor, rum, dairy products, bakeries, and the natural environment (oak trees). These genomic data enabled us to delineate specific genetic groups corresponding to the different ecological niches and revealed high genome content variation across the groups. Most of these strains, compared with the reference genome, possessed additional genetic elements acquired by introgression or horizontal transfer, several of which were population-specific. In addition, several genomic regions in each population showed evidence of nonneutral evolution, as shown by high differentiation, or of selective sweeps including genes with key functions in these environments (e.g. amino acid transport for wine yeast). Linking genetics to lifestyle differences and metabolite traits has enabled us to elucidate the genetic basis of several niche-specific population traits, such as growth on galactose for cheese strains. These data indicate that yeast has been subjected to various divergent selective pressures depending on its niche, requiring the development of customized genomes for better survival in these environments. These striking genome dynamics associated with local adaptation and domestication reveal the remarkable plasticity of the S. cerevisiae genome, revealing this species to be an amazing complex of specialized populations. [ABSTRACT FROM AUTHOR]

The natural biology of Saccharomyces cerevisiae, the best known unicellular model eukaryote, remains poorly documented and understood although recent progress has started to change this situation. Studies carried out recently in the Northern Hemisphere revealed the existence of wild populations associated with oak trees in North America, Asia, and in the Mediterranean region. However, in spite of these advances, the global distribution of natural populations of S. cerevisiae, especially in regions were oaks and other members of the Fagaceae are absent, is not well understood. Here we investigate the occurrence of S. cerevisiae in Brazil, a tropical region where oaks and other Fagaceae are absent. We report a candidate natural habitat of S. cerevisiae in South America and, using whole-genome data, we uncover new lineages that appear to have as closest relatives the wild populations found in North America and Japan. A population structure analysis revealed the penetration of the wine genotype into the wild Brazilian population, a first observation of the impact of domesticated microbe lineages on the genetic structure of wild populations. Unexpectedly, the Brazilian population shows conspicuous evidence of hybridization with an American population of Saccharomyces paradoxus. Introgressions from S. paradoxus were significantly enriched in genes encoding secondary active transmembrane transporters. We hypothesize that hybridization in tropical wild lineages may have facilitated the habitat transition accompanying the colonization of the tropical ecosystem. [ABSTRACT FROM AUTHOR]

Soil yeasts represent a poorly known fraction of the soil microbiome due to limited ecological surveys. Here, we provide the first comprehensive inventory of cultivable soil yeasts in a Mediterranean ecosystem, which is the leading biodiversity hotspot for vascular plants and vertebrates in Europe. We isolated and identified soil yeasts from forested sites of Serra da Arrábida Natural Park (Portugal), representing the Mediterranean forests, woodlands and scrub biome. Both cultivation experiments and the subsequent species richness estimations suggest the highest species richness values reported to date, resulting in a total of 57 and 80 yeast taxa, respectively. These values far exceed those reported for other forest soils in Europe. Furthermore, we assessed the response of yeast diversity to microclimatic environmental factors in biotopes composed of the same plant species but showing a gradual change from humid broadleaf forests to dry maquis. We observed that forest properties constrained by precipitation level had strong impact on yeast diversity and on community structure and lower precipitation resulted in an increased number of rare species and decreased evenness values. In conclusion, the structure of soil yeast communities mirrors the environmental factors that affect aboveground phytocenoses, aboveground biomass and plant projective cover. [ABSTRACT FROM AUTHOR]

Species of Leucosporidiales are a group of psychrotolerant yeasts with biotechnological potential. In the present work, we studied the phenotypic, genetic and sexual characteristics of three species of this genus (Leucosporidium scottii, Leucosporidiella creatinivora and Le. yakutica) to clarify the evolutionary relationship among these closely related taxa. From the results obtained, it becomes clear that these yeasts can interbreed. Although genetic delimitation is possible for the three species, the extent of nucleotide substitutions and phenotypic differences observed between them are lower than that expected for species that have ended the speciation process. Our taxonomic conclusion is to maintain the three taxa until further genomic data are gathered. However, the concept of L. scottii species complex is proposed for this group of species. Finally, we transfer all Leucosporidiella and Mastigobasidium species to Leucosporidium (Leucosporidiales), and, in order to end the polyphyly condition of these taxa, we propose the new genus Pseudoleucosporidium gen. nov. and the new combination Peudoleucosporidium fasciculatum comb. nov. [ABSTRACT FROM AUTHOR]

Mountain lakes located at a high elevation are typically exposed to high UV radiation (UVR). Little is known about the ecology and diversity of yeasts inhabiting these extreme environments. We studied yeast occurrence (with special emphasis on those producing carotenoid pigments) at five high-altitude (>1400 m a.s.l.) water bodies located in the Nahuel Huapi National Park (Bariloche, Argentina). Isolates were identified using a polyphasic approach. Production of photoprotective compounds (carotenoids and mycosporines) by yeast isolates, and UVB resistance of selected species were studied. All water samples contained viable yeast cells in variable numbers, generally ranging from 49 to 209 cells L−1. A total of 24 yeast species was found; at least four represented novel species. Carotenogenic yeasts prevailed in lakes with low water conductivity and higher transparency and chlorophyll a levels. Apparently, the ability to produce photoprotective compounds in yeasts was related to the transparency of mountain lake waters, and strains from more transparent waters developed increased UVB resistance. Our results indicate that UVR is an important environmental factor affecting the yeast community structure in aquatic habitats. [ABSTRACT FROM AUTHOR]

Here we report on the results of a survey of the yeast populations occurring on submerged leaves (alder, eucalyptus and oak) in a natural mountain stream, during different phases of their decomposition and through two consecutive years. Leaf litter mass loss, total yeast counts, Shannon–Weiner index ( H′), yeast community structure and physiologic abilities were analyzed to evaluate the dynamics of yeast communities during decay. Seventy-two yeast taxa were recorded, and in all litter types, species of basidiomycetous affinity predominated over ascomycetous ones. Discriminant analysis of presence/absence data (yeast species) showed significant differences both among substrate types ( P<0.0026) and with decomposition time ( P<0.0001). Carbon and nitrogen source utilization by yeast strains also varied with the substrate ( P<0.0001) and decomposition time ( P<0.0001). Further conclusions were that: (1) all litter types have in common ubiquitous yeast species, such as Cryptococcus albidus, Debaryomyces hansenii and Rhodotorula glutinis, among the common 20 yeast species; (2) only a few species were dominant, and most species were rare, being recorded once or twice throughout decomposition; and (3) the order of yeast appearance, and their substrate assimilation patterns, strongly suggest a succession phenomenon. Finally, explanations for the distribution patterns and variations in yeast communities are discussed. [ABSTRACT FROM AUTHOR]

The Iberian Pyrite Belt extends from Portugal to Spain and is one of the most important pyrite regions in the world. Its aquatic reservoirs display extreme conditions characterized by low pH and high concentrations of heavy metals. In this study, the diversity of microeukaryotes was analysed at the abandoned mines of São Domingos (Portugal) and at Rio Tinto (Spain). DNA was extracted from water samples and a set of eukaryotic universal primers directed to the small subunit rRNA genes (rDNA) was used. The amplicons were analysed by molecular cloning and temperature gradient gel electrophoresis (TGGE). In addition, a fungi-specific primer set was also used in TGGE experiments. The fungi-specific primers contributed to a substantial increase in the number of fungal taxa found due, probably, to the relative low density of fungal structures. Several microorganisms, belonging (or closely related) to the ascomycetous yeast Pichia acaciae, the basidiomycetous yeasts Cryptococcus humicola and Cystofilobasidium bisporidii, the green algae Chlamydomonas noctigama and Chlorella protothecoides var. acidicola and some uncultured microeukaryotes were present at both localities, which suggests that specific microorganisms are adapted to the peculiar conditions of the Iberian Pyrite Belt extreme environments. However, in spite of the similarities, a higher algal richness was observed at S. Domingos, whereas for R. Tinto the richness of fungi was more prominent. [ABSTRACT FROM AUTHOR]

Abstract: Temperature gradient gel electrophoresis (TGGE) was employed for the assessment of yeast diversity in the estuary of the Tagus river (Portugal). The molecular detection of yeasts was carried out directly from water samples and, in parallel, a cultivation approach by means of an enrichment step was employed. A nested PCR was employed to obtain a fungal amplicon containing the D2 domain of the 26S rRNA gene. For identification the TGGE bands were extracted, re-amplified, and sequenced. Fourteen fungal taxa were detected and all except one were yeasts. Most yeast sequences corresponded to members of the Ascomycota and only three belonged to the Basidiomycota. Five yeasts (four ascomycetes and one basidiomycete) could not be identified to the species level due to the uniqueness of their sequences. The number of species detected after enrichment was higher than the number of taxa found using the direct detection method. This suggests that some yeast populations are present in densities that are below the detection threshold of the method. With respect to the analysis of the yeast community structure, our results indicate that the dominant populations belong to Debaryomyces hansenii, Rhodotorula mucilaginosa, Cryptococcus longus, and to an uncultured basidiomycetous yeast phylogenetically close to Cr. longus. The combined analysis of direct detection and cultivation approaches indicates a similar community structure at the two sampled sites since nine species were present at both localities. [Copyright &y& Elsevier]

The phenotypic and genetic heterogeneity of the basidiomycetous yeast species Rhodotorula glutinis was investigated in a group of 109 isolates. A polyphasic taxonomic approach was followed which included PCR fingerprinting, determination of sexual compatibility, 26S and ITS rDNA sequence analysis, DNA–DNA reassociation experiments and reassessment of micromorphological and physiological attributes. The relationships with species of the teleomorphic genus Rhodosporidium were studied and isolates previously identified as Rh. glutinis were found to belong to Rhodosporidium babjevae, Rhodosporidium diobovatum and Rhodosporidium sphaerocarpum. Other isolates included in the study were found to belong to Rh. glutinis var. dairenensis, which is elevated to the species level, or to undescribed species. The concept of Rh. glutinis sensu stricto is proposed due to the close phenetic and phylogenetic proximity detected for Rh. glutinis, Rhodotorula graminis and R. babjevae. [ABSTRACT FROM AUTHOR]

This report presents the description of a new Sporobolomyces species, Sp. odoratus sp. nov. The new species was characterized by DNA fingerprinting using the micro/minisatellite-primed PCR approach (MSP-PCR). A phylogenetic analysis using the D1/D2 region of the 26S rDNA revealed that Sp. odoratus belongs to a clade that includes Rhodosporidium fluviale and Sporidiobolus ruineniae. An integrated comparison with related species is presented. A number of presumptive strains of S. ruineniae were also investigated using the MSP-PCR method and 26S rDNA sequence analysis. Mating experiments revealed, for the first time, that S. ruineniae includes heterothallic strains, besides those already known to be self-fertile. [ABSTRACT FROM AUTHOR]

Although sugarcane produces a large amount of energy per hectare, its ensiled storage leads to losses, which makes it one of the most expensive options for animal feeding. These losses occur due to fermentation of sugars and organic acids by yeasts, producing alcohol and gas. Inoculants containing Lactobacillus buchneri increase acetic acid production, which has effect against yeast. In this study, we produced egg yolk antibodies (IgY) against yeasts that ferment lactate (Candida glabrata, Pichia kudriavzevii, and Pichia manshurica) and sugar (Saccharomyces cerevisiae, Torulospora delbrueckii, Schizosaccharomyces pombe and Debaryomyces etchellsii) isolated in sugarcane silage. Shortly before ensiling, doses of 0, 175, 350, and 700 g/t IgY, and Lactobacillus buchneri were applied to chopped forage. Gas losses were calculated using experimental silos in triplicate, according to the equation: G = (PCf - PCa)/(MFf) × 100, in which: G = gas losses (kg/t); PCf = weight of full silo (equipped with a Bunsen valve to allow gas outflow) at sealing (kg); PCa = weight of full bucket at opening (kg); MFf = forage mass at sealing (kg). The data were analyzed as a completely randomized design and submitted to analysis of variance and regression using the SAS (SAS, 1998), which was chosen because of the significance of the regression parameters, tested by Tukey's test (P < 0.05) and the values of the coefficients of determination. Thirty days after sealing the L. buchneri inoculant and doses of 350 and 700 g/t IgY promoted less gas losses than control that had no any product (12.74; 15.59 and 15.28 versus 27.74, respectively, P < 0.05) and the L. buchneri did not differ from IgY doses. The gas losses decreased quadratically with the anti-yeast addition, estimating maximum of 530 g/t fresh forage. We conclude that anti-yeast use reduces gas production during fermentation. With continuity of this research, we may obtain a roughage with higher nutritional value, as its mode of action in yeast control does not involve consumption of sugars or organic acids. [ABSTRACT FROM AUTHOR]

In most fungi, sexual reproduction is bipolar; that is, two alternate sets of genes at a single mating-type (MAT) locus determine two mating types. However, in the Basidiomycota, a unique (tetrapolar) reproductive system emerged in which sexual identity is governed by two unlinked MAT loci, each of which controls independent mechanisms of self/nonself recognition. Tetrapolar-to-bipolar transitions have occurred on multiple occasions in the Basidiomycota, resulting, for example, from linkage of the two MAT loci into a single inheritable unit. Nevertheless, owing to the scarcity of molecular data regarding tetrapolar systems in the earliest-branching lineage of the Basidiomycota (subphylum Pucciniomycotina), it is presently unclear if the last common ancestor was tetrapolar or bipolar. Here, we address this question, by investigating the mating system of the Pucciniomycotina yeast Leucosporidium scottii. Using whole-genome sequencing and chromoblot analysis, we discovered that sexual reproduction is governed by two physically unlinked gene clusters: a multiallelic homeodomain (HD) locus and a pheromone/receptor (P/R) locus that is biallelic, thereby dismissing the existence of a third P/R allele as proposed earlier. Allele distribution of both MAT genes in natural populations showed that the two loci were in strong linkage disequilibrium, but independent assortment of MAT alleles was observed in the meiotic progeny of a test cross. The sexual cycle produces fertile progeny with similar proportions of the four mating types, but approximately 2/3 of the progeny was found to be nonhaploid. Our study adds to others in reinforcing tetrapolarity as the ancestral state of all basidiomycetes. [ABSTRACT FROM AUTHOR]