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The adaptation to the different biotic and abiotic factors of wine fermentation has led to the accumulation of numerous genomic hallmarks in Saccharomyces cerevisiae wine strains. IRC7, a gene encoding a cysteine-S-β-lyase enzyme related volatile thiols production in wines, has two alleles: a full-length allele (IRC7F ) and a mutated one (IRC7S ), harbouring a 38 bp-deletion. Interestingly, IRC7S -encoding a less active enzyme - appears widespread amongst wine populations. Studying the global distribution of the IRC7S allele in different yeast lineages, we confirmed its high prevalence in the Wine clade and demonstrated a minority presence in other domesticated clades (Wine-PDM, Beer and Bread) while it is completely missing in wild clades. Here, we show that IRC7S -homozygous (HS) strains exhibited both fitness and competitive advantages compared with IRC7F -homozygous (HF) strains. There are some pieces of evidence of the direct contribution of the IRC7S allele to the outstanding behaviour of HS strains (i.e., improved response to oxidative stress conditions and higher tolerance to high copper levels); however, we also identified a set of sequence variants with significant co-occurrence patterns with the IRC7S allele, which can be co-contributing to the fitness and competitive advantages of HS strains in wine fermentations.

A cryophilic basidiomycetous yeast unable to grow at 18 °C or higher temperatures was isolated from a subsurface permafrost layer collected in the Eastern Swiss Alps and from melted sea ice collected in the Artic at Frobisher Bay, Nunavut, Canada. Phylogenetic analyses employing combined sequences of the D1/D2 domain and ITS region indicated that the two new isolates belong to the family Camptobasidiaceae of the class Microbotryomycetes but are distantly related to any of the currently recognized species and genera. Consequently, the novel genus Cryolevonia, and the novel species Cryolevonia schafbergensis (type strain PYCC 8347T=CBS 16055T) are proposed to accommodate this cryophilic yeast. Although sparse hyphae and teliospore-like stuctures were observed upon prolonged incubation, a sexual cycle was not observed and therefore C. schafbergensis is documented solely from its asexual stage.

The Arxan-Chaihe volcanic field of the Da Hinggan mountains in north-East PR China hosts various typical crater lakes. In this study we performed a yeast diversity survey using water sampled from five crater lakes and a total of 122 yeast strains belonging to 33 species of 25 genera were isolated. Three strains, TFL1-L, TFL2B and ATC4C, were identified as three novel species belonging to the Tremellomycetes based on a multiple gene phylogeny and on the comparison of physiological data. A phylogenetic study employing the sequences of seven genes indicated that the new species were more related to three separated phylogenetic lineages of the Tremellomycetes and their closest relatives were Heterocephalacria arrabidensis, Phaeotremella skinneri and Solicoccozyma keelungensis. The divergence values of the D1/D2 domain of LSU sequences of strains TFL1-L, TFL2B and ATC4C from H. arrabidensis CBS 8678T, P. skinneri CBS 5029T and S. keelungensisSN-82T were 4.8，3.4，2.1 %， respectively. The divergence values of the sequences of ITS regions between strains TFL1-L, TFL2B and ATC4C and their close relatives (H. arrabidensis, P. skinneri and S. keelungensis) were 16.1, 5.9 and 8.1 %, respectively. Moreover, the three strains differed from their phylogenetic neighbours by the ability to grow on distinct carbon and nitrogen sources. On the basis of these findings, it is suggested that these strains represent three novel species for which the names Heterocephalacria sinensis sp. nov. (holotype CGMCC 2.5595), Phaeotremella lacus sp. nov. (holotype CGMCC 2.5580) and Solicoccozyma aquatica sp. nov. (holotype CGMCC 2.5574) are proposed.

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.

Phaffia is an orange-colored basidiomycetous yeast genus of the order Cystofilobasidiales that contains a single species, P. rhodozyma. This species is the only fungus known to produce the economically relevant carotenoid astaxanthin. Although Phaffia was originally found in the Northern hemisphere, its diversity in the southern part of the globe has been shown to be much greater. Here we analyze the genomes of two Australasian lineages that are markedly distinct from P. rhodozyma. The two divergent lineages were investigated within a comprehensive phylogenomic study of representatives of the Cystofilobasidiales that supported the recognition of two novel Phaffia species, for which we propose the names of P. australis sp. nov. and P. tasmanica sp. nov. Comparative genomics and other analyses confirmed that the two new species have the typical Phaffia hallmark&mdash, the six genes necessary for the biosynthesis of astaxanthin could be retrieved from the draft genome sequences, and this carotenoid was detected in culture extracts. In addition, the organization of the mating-type (MAT) loci is similar to that of P. rhodozyma, with synteny throughout most regions. Moreover, cases of trans-specific polymorphism involving pheromone receptor genes and pheromone precursor proteins in the three Phaffia species, together with their shared homothallism, provide additional support for their classification in a single genus.

In recent years, ‘multi-omic’ sciences have affected all aspects of fundamental and applied biological research. Yeast taxonomists, though somewhat timidly, have begun to incorporate complete genomic sequences into the description of novel taxa, taking advantage of these powerful data to calculate more reliable genetic distances, construct more robust phylogenies, correlate genotype with phenotype and even reveal cryptic sexual behaviors. However, the use of genomic data in formal yeast species descriptions is far from widespread. The present review examines published examples of genome-based species descriptions of yeasts, highlights relevant bioinformatic approaches, provides recommendations for new users and discusses some of the challenges facing the genome-based systematics of yeasts.

An inventory of culturable yeasts from the soil and water of natural CO2 springs (mofettes) in northeast Slovenia is presented. In mofettes, CO2 of geological origin reaches the soil surface causing temporarily and spatially stable hypoxic environments in soil and water. In total, 142 yeast strains were isolated and identified from high CO2 and control meadow soil, meadow ground-water, forest pond and stream water. All water locations showed below-ground CO2 release. They were assigned to six basidiomycetous yeast genera (six species) and 11 ascomycetous genera (18 species). All ascomycetous yeasts, with the exception of Debaryomyces hansenii, were able to grow under elevated CO2 and fermented glucose. Candida sophiae-reginae, Pichia fermentans and Candida vartiovaarae were the dominating species in meadow and forest high CO2 exposed water. Meyerozyma guilliermondii and Wickerhamomyces anomalus predominated in high CO2 exposed soils. Using high dilution plating of a mofette soil sample, four strains of an unknown basidiomycetous species were isolated and are here newly described as Occultifur mephitis based on molecular phylogenetic and phenotypic criteria. The type strain of Occultifur mephitis is EXF-6436T[CBS 14611=PYCC 7049, LT594852 (D1/D2), KX929055 (ITS)]. An additional three isolated strains are EXF-6437 (LT594853, KX929056), EXF-6473 (LT594863, KX929057) and EXF-6482 (LT594867, KX929054), as well as a strain reported from previous studies isolated from a leaf of Cistus albidus in Portugal (CBS 10223=PYCC 6067), EU002842 (D1/D2), KY308183 (ITS).

The presence of yeasts at different steps of Vietnamese soy paste production was studied. Yeast growth occurred during primary soybean fermentation, with the cell density reaching 4.106 c.f.u. ml−1, and terminated during brine fermentation. The dominant species were Pichia kudriavzevii and Millerozyma farinosa. Over the span of 14 years, nine strains of Moniliella were isolated. The strains had identical PCR fingerprints generated with primer (GAC)5 and identical D1/D2 and internal transcribed spacer (ITS) sequences. A D1/D2-based phylogeny indicated that the strains were closest to a group of four previously assigned as Moniliella suaveolens strains. Together they form a new lineage that is well separated from all known species, including M. suaveolens (over 12.7 % divergence). ITS sequences indicated the presence of four species differing from each other by 9–57 nt. The name Moniliella sojae sp. nov. is proposed to accommodate the strains isolated from Vietnamese soy paste, Moniliella pyrgileucina sp. nov. is proposed for PYCC 6800 and Moniliella casei sp. nov. is proposed for CBS 157.58. An emended combination Moniliella macrospora is proposed for CBS 221.32 and CBS 223.32. The type strains and MycoBank numbers are: M. sojae sp. nov., SS 4.2T=CBS 126448T=NRRL Y-48680T and MB 822871; M. pyrgileucina sp. nov., PYCC 6800T=CBS 15203T and MB 823030; M. casei sp. nov., CBS 157.58T=IFM 60348T and MB 822872; M. macrospora emend. comb. nov., CBS 221.32T (=MUCL 11527T) and MB 822874.

Background: Lignocellulosic biomass is a common resource across the globe, and its fermentation offers a promising option for generating renewable liquid transportation fuels. The deconstruction of lignocellulosic biomass releases sugars that can be fermented by microbes, but these processes also produce fermentation inhibitors, such as aromatic acids and aldehydes. Several research projects have investigated lignocellulosic biomass fermentation by the baker's yeast Saccharomyces cerevisiae. Most projects have taken synthetic biological approaches or have explored naturally occurring diversity in S. cerevisiae to enhance stress tolerance, xylose consumption, or ethanol production. Despite these efforts, improved strains with new properties are needed. In other industrial processes, such as wine and beer fermentation, interspecies hybrids have combined important traits from multiple species, suggesting that interspecies hybridization may also offer potential for biofuel research. Results: To investigate the efficacy of this approach for traits relevant to lignocellulosic biofuel production, we generated synthetic hybrids by crossing engineered xylose-fermenting strains of S. cerevisiae with wild strains from various Saccharomyces species. These interspecies hybrids retained important parental traits, such as xylose consumption and stress tolerance, while displaying intermediate kinetic parameters and, in some cases, heterosis (hybrid vigor). Next, we exposed them to adaptive evolution in ammonia fiber expansion-pretreated corn stover hydrolysate and recovered strains with improved fermentative traits. Genome sequencing showed that the genomes of these evolved synthetic hybrids underwent rearrangements, duplications, and deletions. To determine whether the genus Saccharomyces contains additional untapped potential, we screened a genetically diverse collection of more than 500 wild, non-engineered Saccharomyces isolates and uncovered a wide range of capabilities for traits relevant to cellulosic biofuel production. Notably, Saccharomyces mikatae strains have high innate tolerance to hydrolysate toxins, while some Saccharomyces species have a robust native capacity to consume xylose. Conclusions: This research demonstrates that hybridization is a viable method to combine industrially relevant traits from diverse yeast species and that members of the genus Saccharomyces beyond S. cerevisiae may offer advantageous genes and traits of interest to the lignocellulosic biofuel industry. Fil: Peris, David. University of Wisconsin; Estados Unidos Fil: Moriarty, Ryan V.. University of Wisconsin; Estados Unidos Fil: Alexander, William G.. University of Wisconsin; Estados Unidos Fil: Baker, EmilyClare. University of Wisconsin; Estados Unidos Fil: Sylvester, Kayla. University of Wisconsin; Estados Unidos Fil: Sardi, Maria. University of Wisconsin; Estados Unidos Fil: Langdon, Quinn K.. University of Wisconsin; Estados Unidos Fil: Libkind Frati, Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales. Universidad Nacional del Comahue. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales.; Argentina Fil: Wang, Qi-Ming. University of Wisconsin; Estados Unidos. Institute Of Microbiology Chinese Academy Of Sciences; China Fil: Bai, Feng-Yan. Institute Of Microbiology Chinese Academy Of Sciences; China Fil: Leducq, Jean Baptiste. University of Montreal; Canadá. Laval University; Canadá Fil: Charron, Guillaume. Laval University; Canadá Fil: Landry, Christian R.. Laval University; Canadá Fil: Sampaio, José Paulo. New University Of Lisbon; Portugal Fil: Gonçalves, Paula. New University Of Lisbon; Portugal Fil: Hyma, Katie E.. Washington University in St. Louis; Estados Unidos Fil: Fay, Justin C.. Washington University in St. Louis; Estados Unidos Fil: Sato, Trey K.. University of Wisconsin; Estados Unidos Fil: Hittinger, Chris Todd. University of Wisconsin; Estados Unidos

Saccharomyces eubayanus was described less than 10 years ago and its discovery settled the long-lasting debate on the origins of the cold-tolerant yeast responsible for lager beer fermentation. The largest share of the genetic diversity of S. eubayanus is located in South America, and strains of this species have not yet been found in Europe. One or more hybridization events between S. eubayanus and S. cerevisiae ale beer strains gave rise to S. pastorianus, the allopolyploid yeasts responsible for lager beer production worldwide. The identification of the missing progenitor of lager yeast opened new avenues for brewing yeast research. It allowed not only the selective breeding of new lager strains, but revealed also a wild yeast with interesting brewing abilities so that a beer solely fermented by S. eubayanus is currently on the market.

During a survey of yeast populations associated with grape and apple musts used for wine and cider fermentation, respectively, six pink-coloured ballistoconidia-forming yeasts belonging to the order Sporidiobolales (Basidiomycota) were isolated. Phylogenetic analysis inferred using sequences of the internal transcribed spacer (ITS), the D1/D2 domain of the large subunit rRNA gene, the small subunit (SSU) rRNA gene and DNA-directed RNA polymerase II subunit (RPB2) indicated that the six isolates were separated in two novel species. One of the new species, Sporobolomyces agrorum sp. nov., isolated from grape must, had Sporobolomyces roseus and Sporobolomyces metaroseus as its closest relatives, but showed four/two and 16 nucleotide substitutions in the D1/D2 and ITS regions, respectively, to these two species. The other novel species, Sporobolomyces sucorum sp. nov., was found in apple must and was closely related to Sporobolomyces pararoseus and Sporobolomyces patagonicus, but showed two/three and five substitutions in those two regions for its closest relatives. We detected additional representatives of this species, most of them isolated from grapes whose sequences were already available on public databases. A sexual stage could not be observed for the novel species. publishersversion published

Fundacao para a Ciencia e a Tecnologia (Portugal) grants PTDC/BIA-MIC/30785/2017. SFRH/BD/136462/2018. Slovenian Research Agency Grants P4-0116 and MRIC-ULZIM, IP-0510 (NC). The adaptation of the yeast Saccharomyces cerevisiae to man-made environments for the fermentation of foodstuffs and beverages illustrates the scientific, social, and economic relevance of microbe domestication. Here we address a yet unexplored aspect of S. cerevisiae domestication, that of the emergence of lineages harboring some domestication signatures but that do not fit completely in the archetype of a domesticated yeast, by studying S. cerevisiae strains associated with processed olives, namely table olives, olive brine, olive oil, and alpechin. We confirmed earlier observations that reported that the Olives population results from a hybridization between S. cerevisiae and S. paradoxus. We concluded that the olive hybrids form a monophyletic lineage and that the S. cerevisiae progenitor belonged to the wine population of this species. We propose that homoploid hybridization gave rise to a diploid hybrid genome, which subsequently underwent the loss of most of the S. paradoxus sub-genome. Such a massive loss of heterozygosity was probably driven by adaptation to the new niche. We observed that olive strains are more fit to grow and survive in olive brine than control S. cerevisiae wine strains and that they appear to be adapted to cope with the presence of NaCl in olive brine through expansion of copy number of ENA genes. We also investigated whether the S. paradoxus HXT alleles retained by the Olives population were likely to contribute to the observed superior ability of these strains to consume sugars in brine. Our experiments indicate that sugar consumption profiles in the presence of NaCl are different between members of the Olives and Wine populations and only when cells are cultivated in nutritional conditions that support adaptation of their proteome to the high salt environment, which suggests that the observed differences are due to a better overall fitness of olives strains in the presence of high NaCl concentrations. Although relic olive hybrids exhibit several characteristics of a domesticated lineage, tangible benefits to humans cannot be associated with their phenotypes. These strains can be seen as a case of adaptation without positive or negative consequences to humans, that we define as a quasi-domestication. publishersversion published

The most common fermented beverage, lager beer, is produced by interspecies hybrids of the brewing yeast Saccharomyces cerevisiae and its wild relative S. eubayanus. Lager-brewing yeasts are not the only example of hybrid vigour or heterosis in yeasts, but the full breadth of interspecies hybrids associated with human fermentations has received less attention. Here we present a comprehensive genomic analysis of 122 Saccharomyces hybrids and introgressed strains. These strains arose from hybridization events between two to four species. Hybrids with S. cerevisiae contributions originated from three lineages of domesticated S. cerevisiae, including the major wine-making lineage and two distinct brewing lineages. In contrast, the undomesticated parents of these interspecies hybrids were all from wild Holarctic or European lineages. Most hybrids have inherited a mitochondrial genome from a parent other than S. cerevisiae, which recent functional studies suggest could confer adaptation to colder temperatures. A subset of hybrids associated with crisp flavour profiles, including both lineages of lager-brewing yeasts, have inherited inactivated S. cerevisiae alleles of critical phenolic off-flavour genes and/or lost functional copies from the wild parent through multiple genetic mechanisms. These complex hybrids shed light on the convergent and divergent evolutionary trajectories of interspecies hybrids and their impact on innovation in lager brewing and other diverse fermentation industries., Work supported by the National Science Foundation (grant nos. DEB-1253634 to C.T.H. and DGE-1256259 (Graduate Research Fellowship to Q.K.L.), the USDA National Institute of Food and Agriculture Hatch Project (nos. 1003258 and 1020204 to C.T.H.) and in part by the DOE Great Lakes Bioenergy Research Center (DOE BER Office of Science nos. DE-SC0018409 and DE-FC02-07ER64494). Q.K.L. was also supported by the Predoctoral Training Program in Genetics, funded by the National Institutes of Health (grant no. 5T32GM007133). D.P. is a Marie Sklodowska-Curie fellow of the European Union’s Horizon 2020 research and innovation programme (grant no. 747775).

Two yeast strains that are members of the same species were isolated from different marine habitats, i.e. one from Mid-Atlantic Ridge ocean water samples located in the direct vicinity of black smokers near the Rainbow deep-sea hydrothermal vent and one from Brazilian marine water samples off the Ipanema beach. Strains CLIB 1964(T) and CLIB 1965 are anamorphic ascomycetous yeasts affiliated to the Yamadazyma clade of Saccharomycetales. Interestingly, these strains were phylogenetically and distinctly positioned into a group of species comprising all species of the genus Yamadazyma isolated from marine habitats including deep-sea hydrothermal vents, i.e. Candida atmosphaerica, C. spencermartinsiae, C. atlantica, C. oceani and C. taylorii. These strains differed significantly in their D1/D2 domain sequences of the LSU rRNA gene from the closely related species mentioned above, by 2.6, 3.0, 3.4, 3.8 and 6.0 %, respectively. Internal transcribed spacer region sequence divergence was also significant and corresponded to 4.6, 4.7, 4.7, 12.0 and 24.7% with C. atlantica, C. atmosphaerica, C. spencermartinsiae, C. oceani and C. taylorii, respectively. Phenotypically, strains CLIB 1964(T) and CLIB 1965 could be distinguished from closely related species by their inability to assimilate L-sorbose. CLIB 1964(T) (= CBS 14301(T) = UBOCC-A-214001(T)) is the designated type strain for Yamadazyma barbieri sp. nov. The MycoBank number is MB 815884.

This work was supported by Fundacao para a Ciencia e a Tecnologia, Portugal, grants PTDC/BIA-EVF/118618/2010 (J.P.S., P.A., P.G.), PTDC/AGR-ALI/118590/2010 (J.P.S., P.A., P.G., R.B.), UID/Multi/04378/2013 (J.P.S., P.G.), and SFRH/BD/77390/2011 (P.A.), by Conselho Nacional de Desenvolvimento Cientifico e Tecnologico-CNPq (CAR, process numbers 560715/2010-2 and 457499/2014-1, PBM process number 457443/2012-0) and Fundacao de Amparo a Pesquisa de Minas Gerais FAPEMIG and VALE S.A (CAR, process number RCP-00094-10). Work of C.R.L. on this project was supported by a NSERC Discovery grant. C.R.L. holds the Canada Research Chair in Evolutionary Cell and Systems Biology. The authors thank Dr. Siu Mui Tsai, Universidade de Sao Paulo, Brazil, for making available strain UFMG-CM-Y640. 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, atropical 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. publishersversion published

Yeasts are unicellular fungi that do not form fruiting bodies.Although the yeast lifestyle has evolved multiple times, mostknown species belong to the subphylum Saccharomycotina(syn. Hemiascomycota, hereafter yeasts). This diverse groupincludes the premier eukaryotic model system,Saccharomyces cerevisiae; the common human commensaland opportunistic pathogen, Candida albicans; and over1000 other known species (with more continuing to bediscovered). Yeasts are found in every biome and continent andare more genetically diverse than angiosperms or chordates.Ease of culture, simple life cycles, and small genomes (10?20 Mbp) have made yeasts exceptional models for moleculargenetics, biotechnology, and evolutionary genomics. Herewe discuss recent developments in understanding thegenomic underpinnings of the making of yeast biodiversity,comparing and contrasting natural and human-associatedevolutionary processes. Only a tiny fraction of yeastbiodiversity and metabolic capabilities has been tapped byindustry and science. Expanding the taxonomic breadth ofdeep genomic investigations will further illuminate how genomefunction evolves to encode their diverse metabolisms andecologies. Fil: Hittinger, Chris Todd. University Of Wisconsin; Estados Unidos. University Of Wisconsin; Estados Unidos Fil: Rokas, Antonis. Vanderbilt University; Estados Unidos Fil: Bai, Feng Yan. Chinese Academy Of Sciences; República de China Fil: Boekhout, Teun. Chinese Academy Of Sciences; República de China. CBS-KNAW Fungal Biodiversity Centre; Países Bajos. Second Military Medical University; China Fil: Gonçalves, Paula. Universidade Nova de Lisboa; Portugal Fil: Jeffries, Thomas W.. University Of Wisconsin; Estados Unidos Fil: Kominek, Jacek. University Of Wisconsin; Estados Unidos Fil: Lachance, Marc Andre. University of Western Ontario; Canadá Fil: Libkind Frati, Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Patagonia Norte. Instituto de Investigación en Biodiversidad y Medioambiente; Argentina. Universidad Nacional del Comahue; Argentina Fil: Rosa, Carlos A.. Universidade Federal do Minas Gerais; Brasil Fil: Sampaio, José Paulo. Universidade Nova de Lisboa; Portugal Fil: Kurtzman, Cletus P.. United States Department of Agriculture; Argentina

Horizontal gene transfer (HGT) and introgression are increasingly recognized as a prominent source of adaptation in eukaryotes. Over the past decade, several cases of HGT from prokaryotes to the yeast Saccharomyces strains have been demonstrated. In addition, we have recently identified eukaryote-to-eukaryote gene transfers and some of them have been shown to be involved in the adaptation of the yeast S. cerevisiae to environmental conditions (1;2;3). Three large genomic regions, (A, B and C), were acquired by wine yeasts strains from more distant yeast species (1;3). The yeasts Zygosaccharomyces bailii and Torulaspora microellipsoides, species found in wine fermentations, were identified as the donors of regions B and C, respectively (1;3). We obtained evidence for the amplification of the region B (17 kb) in the genome of S. cerevisiae wine strains (4). The organization of this region differ considerably between strains and the identification of an autonomously replicating sequence functional in S. cerevisiae strongly suggest an expansion mechanism in yeast genomes involving an extrachromosomal circular DNA molecule. We also showed that region C have undergone several rearrangements and carries genes playing a key role in the adaptation of wine yeasts to the nitrogen-limited wine fermentation environment (3). The analysis of 82 S. cerevisiae strains from various ecological origins allowed identification of 33 HGT or introgression for which we have proposed potential donors using phylogenetic topology test. Among these events, we observed, the replacement of a GAL cluster in cheese strains which enables a faster switch from glucose to galactose and improves growth speed in a media containing a mix of the two hexoses such as fermented milk products. These data support the vision of customized yeast genomes associated to specific ecological niches and highlight the key role of HGT in the adaptation of fungal species to their environments.1. Novo et al. 2009, PNAS, 106: 16333-16338.2. Damon et al 2011, ISME Journal 9: 67.3. Marsit et al 2015, Mol. Biol. Evol., 32:1695–1707 4. Galeote et al 2011, PLoS One 6: e17872.

The gut of insects harbors a yeast community that is still poorly understood. Here, a novel species of the ascomycetous genus Blastobotrys is proposed based on a yeast strain isolated from the larval gut of the silkworm Bombyx mori (Order Lepidoptera). The novel species is closely related to Blastobotrys aristata and Blastobotrys elegans on the basis of the results of molecular phylogenetic analyses. A preliminary screening revealed that it produces 1.5 g l−1 ethanol by fermenting 5 % d-xylose. The novel species, that represents the first report, to our knowledge, of yeast isolation from silkworms, is described as Blastobotrys bombycis sp. nov. (type strain RAAB001T=CBS 15274T=PYCC 8105T=MCC 1427T; MycoBank accession number MB 825095).

International audience; During a survey of the yeast community associated with the phylloplane of corn in Thailand, a basidiomycetous yeast strain belonging to the genus Papiliotrema was isolated. Analyses of the D1/D2 domains of the 26S (LSU) rRNA gene and complete ITS region supported the recognition of a novel species, for which the name Papiliotrema plantarum sp. nov. is proposed (type strain DMKU-CP801(T) =CBS 15220(T) =PYCC 7257(T)). Another strain of P. plantarum sp. nov., isolated in French Guiana, was found to be sexually compatible with the Thai isolate and mycelium with clamp connections, basidia and basidiospores were observed in culture. The basidial morphology of P. plantarum combined features previously observed for Papiliotrema bandonii and Papiliotrema fuscus, which represent the only sexual species hitherto known in the genus, i.e. transversely septate basidia, with sexual structures of the Tremella type.

The diversity of yeast species collected from the bromeliad tanks of Vriesea minarum, an endangered bromeliad species, and their ability to produce extracellular enzymes were studied. Water samples were collected from 30 tanks of bromeliads living in a rupestrian field site located at Serrada Piedade, Minas Gerais state, Brazil, during both the dry and rainy seasons. Thirty-six species were isolated, representing 22 basidiomycetous and 14 ascomycetous species. Occultifur sp., Cryptococcus podzolicus and Cryptococcus sp. 1 were the prevalent basidiomycetous species. The yeast-like fungus from the order Myriangiales, Candida silvae and Aureobasidium pullulans were the most frequent ascomycetous species. The diversity of the yeast communities obtained between seasons was not significantly different, but the yeast composition per bromeliad was different between seasons. These results suggest that there is significant spatial heterogeneity in the composition of populations of the yeast communities within bromeliad tanks, independent of the season. Among the 352 yeast isolates tested, 282 showed at least one enzymatic activity. Protease activity was the most widely expressed extracellular enzymatic activity, followed by xylanase, amylase, pectinase and cellulase activities. These enzymes may increase the carbon and nitrogen availability for the microbial food web in the bromeliad tank of V. minarum. Sequence analyses revealed the existence of 10 new species, indicating that bromeliad tanks are important sources of new yeasts. The novel species Occultifur brasiliensis, f.a., sp. nov., is proposed to accommodate the most frequently isolated yeast associated with V. minarum. The type strain of O. brasiliensis, f.a., sp. nov. is UFMG-CM-Y375(T) (= CBS 12687(T)). The Mycobank number is MB 809816.

The brewing industry is experiencing a period of change and experimentation largely driven by customer demand for product diversity. This has coincided with a greater appreciation of the role of yeast in determining the character of beer and the widespread availability of powerful tools for yeast research. Genome analysis in particular has helped clarify the processes leading to domestication of brewing yeast and has identified domestication signatures that may be exploited for further yeast development. The functional properties of non-conventional yeast (both Saccharomyces and non- Saccharomyces) are being assessed with a view to creating beers with new flavours as well as producing flavoursome non-alcoholic beers. The discovery of the psychrotolerant S. eubayanus has stimulated research on de novo S. cerevisiae × S. eubayanus hybrids for low-temperature lager brewing and has led to renewed interest in the functional importance of hybrid organisms and the mechanisms that determine hybrid genome function and stability. The greater diversity of yeast that can be applied in brewing, along with an improved understanding of yeasts' evolutionary history and biology, is expected to have a significant and direct impact on the brewing industry, with potential for improved brewing efficiency, product diversity and, above all, customer satisfaction. Fil: Gibson, B.. Vtt Technical Research Centre Of Finland; Finlandia Fil: Geertman, J. M. A.. Heineken Supply Chain; Países Bajos Fil: Hittinger, C. T.. University of Wisconsin; Estados Unidos Fil: Krogerus, K.. Vtt Technical Research Centre Of Finland; Finlandia. Aalto University; Finlandia Fil: Libkind Frati, Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales. Universidad Nacional del Comahue. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales.; Argentina Fil: Louis, E. J.. University of Leicester; Reino Unido Fil: Magalhães, F.. Vtt Technical Research Centre Of Finland; Finlandia. Aalto University; Finlandia Fil: Sampaio, J.P.. Universidade Nova de Lisboa; Portugal

Horizontal Gene Transfer is a key mechanism permitting adaptation of [i]S. cerevisiae[/i][i][/i] to its ecological niches. Reunión anual conjunta genética y evolución

Background: The class Tremellomycete (Agaricomycotina) encompasses more than 380 fungi. Although there are a few edible Tremella spp., the only species with current biotechnological use is the astaxanthin-producing yeast Phaffia rhodozyma (Cystofilobasidiales). Besides astaxanthin, a carotenoid pigment with potent antioxidant activity and great value for aquaculture and pharmaceutical industries, P. rhodozyma possesses multiple exceptional traits of fundamental and applied interest. The aim of this study was to obtain, and analyze two new genome sequences of representative strains from the northern (CBS 7918T, the type strain) and southern hemispheres (CRUB 1149) and compre them to a previously published genome sequence (strain CBS 6938). Photoprotection and antioxidant related genes, as well as genes involved in sexual reproduction were analyzed. Results: Both genomes had ca. 19Mb and 6000 protein coding genes, similar to CBS 6938. Compared to other fungal genomes P. rhodozyma strains and other Cystofilobasidiales have the highest number of intron-containing genes and highest number of introns per gene. The Patagonian strain showed 4.4% of nucleotide sequence divergence compared to the European strains which differed from each other by only 0.073%. All known genes related to the synthesis of astaxanthin were annotated. A hitherto unknown gene cluster potentially responsible for photoprotection (mycosporines) was found in the newly sequenced P. rhodozyma strains but was absent in the non-mycosporinogenic strain CBS 6938. A broad battery of enzymes that act as scavengers of free radical oxygen species were detected, including catalases and superoxide dismutases (SODs). Additionally, genes involved in sexual reproduction were found and annotated. Conclusions: A draft genome sequence of the type strain of P. rhodozyma is now available, and comparison with that of the Patagonian population suggests the latter deserves to be assigned to a distinct variety. An unexpected genetic trait regarding high occurrence of introns in P. rhodozyma and other Cystofilobasidiales was revealed. New genomic insights into fungal homothallism were also provided. The genetic basis of several additional photoprotective and antioxidant strategies were described, indicating that P. rhodozyma is one of the fungi most well-equipped to cope with environmental oxidative stress, a factor that has probably contributed to shaping its genome. Fil: Bellora, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales. Universidad Nacional del Comahue. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales.; Argentina Fil: Moline, Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales. Universidad Nacional del Comahue. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales.; Argentina Fil: David Palma, Márcia. Universidade Nova de Lisboa; Portugal Fil: Coelho, Marco A.. Universidade Nova de Lisboa; Portugal Fil: Hittinger, Chris Todd. University of Wisconsin; Estados Unidos Fil: Sampaio, José P.. Universidade Nova de Lisboa; Portugal Fil: Gonçalves, Paula. Universidade Nova de Lisboa; Portugal Fil: Libkind Frati, Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales. Universidad Nacional del Comahue. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales.; Argentina

Beer is one of the oldest alcoholic beverages and is produced by the fermentation of sugars derived from starches present in cereal grains. Contrary to lager beers, made by bottom-fermenting strains of Saccharomyces pastorianus, a hybrid yeast, ale beers are closer to the ancient beer type and are fermented by S. cerevisiae, a top-fermenting yeast. Here, we use population genomics to investigate (1) the closest relatives of top-fermenting beer yeasts; (2) whether top-fermenting yeasts represent an independent domestication event separate from those already described; (3) whether single or multiple beer yeast domestication events can be inferred; and (4) whether top-fermenting yeasts represent non-recombinant or recombinant lineages. Our results revealed that top-fermenting beer yeasts are polyphyletic, with a main clade composed of at least three subgroups, dominantly represented by the German, British, and wheat beer strains. Other beer strains were phylogenetically close to sake, wine, or bread yeasts. We detected genetic signatures of beer yeast domestication by investigating genes previously linked to brewing and using genome-wide scans. We propose that the emergence of the main clade of beer yeasts is related with a domestication event distinct from the previously known cases of wine and sake yeast domestication. The nucleotide diversity of the main beer clade more than doubled that of wine yeasts, which might be a consequence of fundamental differences in the modes of beer and wine yeast domestication. The higher diversity of beer strains could be due to the more intense and different selection regimes associated to brewing. Fil: Gonçalves, Margarida. Universidade Nova de Lisboa; Portugal Fil: Pontes, Ana. Universidade Nova de Lisboa; Portugal Fil: Almeida, Pedro. Universidade Nova de Lisboa; Portugal Fil: Barbosa, Raquel. Universidade Nova de Lisboa; Portugal Fil: Serra, Marta. Universidade Nova de Lisboa; Portugal Fil: Libkind Frati, Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales. Universidad Nacional del Comahue. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales.; Argentina Fil: Hutzler, Mathias. Research Center Weihenstephan for Brewing and Food Quality; Alemania Fil: Gonçalves, Paula. Universidade Nova de Lisboa; Portugal Fil: Sampaio, José Paulo. Universidade Nova de Lisboa; Portugal

A novel species in the genus Candida was obtained from deep-sea hydrothermal fields on the Mid-Atlantic Ridge. Strains Mo39, MARY089 and CBS 5307, respectively, isolated from an unidentified deep-sea coral collected near Rainbow hydrothermal vent, from water samples near Menez Gwen hydrothermal field and from the stomach of a marine fish are considered as a novel taxon. Sequence similarities in the D1/D2 region of the 26S rRNA gene indicated that strains Mo39, MARY089 and CBS 5307 have for closest neighbors Candida spencermartinsiae, Candida taylorii, Candida atmosphaerica and Candida atlantica. The strains, respectively, differ from C. spencermartinsiae, C. taylorii, C. atmosphaerica andCandida atlantica by 4, 4.3, 4.3 and 4.7% in the D1/D2 domain. Strains Mo39, MARY089 and CBS 5307 were differentiated from others by differences in the ability to assimilate d-Gluconate and in the ability to grow at relatively high temperature. Only strain Mo39 displays an optimal growth at 3% sea salts, indicating that this strain is clearly adapted to live in marine conditions. Sequence similarities between strains Mo39, MARY089 and CBS 5307 and related species and differences in the ability to utilize specific carbon compounds revealed that these strains represent a hitherto unknown species. Sexual reproduction was not observed in strains Mo39, MARY089 and CBS 5307. An anamorphic name Candida oceani sp. nov. is proposed for the type strain Mo39T (= CBS 11857T = DSM 23777T) and the two other strains MARY089 and CBS 5307. To our knowledge, this is the first description of a micro-eukaryotic organism including a strain isolated from a deep-sea coral near a hydrothermal ecosystem.

A basidiomycetous yeast, strain CRUB 1032T, which formed salmon-pink colonies, was isolated from glacial meltwater in Patagonia, Argentina. Morphological, physiological and biochemical characterization indicated that this strain belonged to the genus Rhodotorula. Molecular taxonomic analysis based on the 26S rDNA D1/D2 domain and internal transcribed spacer region sequences showed that strain CRUB 1032T represents an undescribed yeast species, for which the name Rhodotorula meli sp. nov. is proposed (type strain is CRUB 1032T=CBS 10797T=JCM 15319T). Phylogenetic analysis showed that Rhodotorula lamellibrachii was the closest known species, which, together with R. meli, formed a separate cluster related to the Sakaguchia clade within the Cystobasidiomycetes. Additional Patagonian yeast isolates of the class Cystobasidiomycetes are also investigated in the present work.

Over 80% of the Earth’s environments are permanently or periodically exposed to temperatures below 5 °C. Cold habitats harbour a wide diversity of psychrophilic and psychrotolerant yeasts. During ecological studies of yeast communities carried out in cold ecosystem in the Italian Alps, Svalbard (Norway, Arctic region), and Portugal, 23 yeast strains that could not be assigned to any known fungal taxa were isolated. In particular, two of them were first identified as Rhodotorula sp., showing the highest degree of D1/D2 sequence identity with Cystobasidum laryngis accounted to only 97% with the type strain (C. laryngis CBS 2221). The other 21 strains, exhibiting identical D1/D2 sequences, had low identity (97%) with Rhodosporidiobolus lusitaniae and Rhodosporidiobolus colostri. Similarly, ITS sequences of the type strains of the most closely related species (93–94%). In a 2-genes multilocus D1/D2 and ITS ML phylogenetic tree, the studied strains pooled in two well separated and supported groups. In order to classify the new 23 isolates based on phylogenetic evidences, we propose the description of two novel species Cystobasidium alpinum sp. nov. and Rhodosporidiobolus oreadorum sp. nov.

Yeasts are common inhabitants of different types of aquatic habitats, including marine and estuarine waters and rivers. Although numerous studies have surveyed yeast occurrence in these habitats, the identification of autochthonous populations has been problematic because several yeast species seem to be very versatile and therefore mere presence is not sufficient to establish an ecological association. In the present study we investigated the dynamics of the yeast community in the Tagus river estuary (Portugal) by combining a microbiological study involving isolation, quantification, and molecular identification of dominant yeast populations with the analysis of hydrological and hydrographical data. We set out to test the hypothesis of the multiple origins of estuarine yeast populations in a transect of the Tagus estuary and we postulate four possible sources: open sea, terrestrial, gastrointestinal and the estuary itself in the case of populations that have become resident. Candida parapsilosis and Pichia guilliermondii were correlated with Escherichia coli, which indicated an intestinal origin. Other cream-colored yeasts like Debaryomyces hansenii and Candida zeylanoides had similar dynamics, but no association with E. coli and quite distinct ecological preferences. They might represent a group of resident estuarine populations whose primary origin is diverse and can include marine, terrestrial, and gastrointestinal habitats. Another major yeast population was represented by Rhodotorula mucilaginosa. The cosmopolitan nature of that species and its moderate association with E. coli point to terrestrial sources as primary habitats.

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.

The Rio Agrio and Lake Caviahue system (RAC), in Northwestern Patagonia, is a natural acidic environment. The aims of this study were to characterize the yeast community and to provide the first ecological assessment of yeast diversity of this extreme aquatic environment. Yeast occurrence and diversity were studied at seven sites where the water pH varied between 1.8 and 6.7. Yeast CFU counts in the river ranged from 30 to 1200 CFU L(-1), but in the Lake the values were lower (30-60 CFU L(-1)). A total of 25 different yeast species were found, 11 of which belonged to undescribed taxa. Among these was an unusual strongly acidophilic Cryptococcus species. The RAC yeast community resembles that of acidic aquatic environments resulting from anthropic activities such as the Sao Domingos mines in Portugal and the Rio Tinto in Spain, respectively. The isolated yeast species were organized into different grades of adaptation to the RAC aquatic system. Based on the proposed grades, Rhodotorula mucilaginosa, Rhodosporidium toruloides and two novel Cryptococcus species were the most adapted species. These Cryptococcus species are apparently specialists of acidic aquatic environments, and might bear physiological features that possibly account for their ability to thrive in such extreme environments.

The relationship between Sporidiobolus johnsonii and S. salmonicolor was investigated using rDNA sequence data. Two statistically well-supported clades were obtained. One clade included the type strain of S. johnsonii and the other included the type strain of S. salmonicolor. However, some mating strains of S. salmonicolor were found in the S. johnsonii group. These strains belonged to mating type A2 and were sexually compatible with mating type A1 strains from the S. salmonicolor group. DNA–DNA reassociation values were high within each clade and moderate between the two clades. In the re-investigation of teliospore germination, we observed that the basidia of S. salmonicolor were two-celled. In S. johnsonii, basidia were not formed and teliospore germination resulted in direct formation of yeast cells. We hypothesize that the S. johnsonii clade is becoming genetically isolated from the S. salmonicolor group and that a speciation process is presently going on. We suspect that the observed sexual compatibility between strains of the S. johnsonii and S. salmonicolor groups and the possible genetic flow between the two species has little biological relevance because distinct phenotypes have been fixed in the two taxa and intermediate (hybrid) sequences for LSU and ITS rDNAs have not been detected.

The yeast Saccharomyces cerevisiae is one of the most important microorganisms for food and drink production and it is as well a model for biology. Surprisingly, the bases of yeast population structure have been unraveled only recently [1, 2] and the first genomic population approaches have failed to point adaptation to ecological niches. However, as many groups present highly contrasted lifestyle (e.g. anaerobic growth on grape must for wine yeast, aerobic growth as a biofilm on ethanol and glycerol for flor strains, growth on low amount of sugars for oak strains) different adaptations are expected. In this project we have obtained high quality genome sequences of 82 yeast strains: 35 from wine environment (27 wine strains, 8 flor strains) as well as 47 other strains from rum, fermented dairy product, bakery including 8 from oaks trees. Our genomic data enable us to delineate specific genetic groups corresponding to the different ecological niches, and indicate different life cycles. We first detected the amplification of several genes with critical function in their environment (e.g. CUP1 for wine yeast, IMA2 and SUC2 for bakery yeast…). We then established a catalog of genes potentially impacted per population. Several tests revealed a non-neutral evolution at several loci, and especially the presence of selective sweep for wine yeast, suggesting positive selection. The comparison of flor and wine yeast revealed allelic variation associated to growth under velum, as well as the fructophily of flor strains, or a specific metal homeostasis. Last among the three large chromosomal regions originated from horizontal gene transfer (HGT) from distant yeasts first discovered in EC1118 [3] were encountered mainly in wine or flor yeasts but the complete region C was only found among flor strains. These genomic data represent a unique resource for understanding the adaptation of yeast to wine making niches and thus for elucidating the bases of technological properties.

Exploring the adaptation of yeast to wine making environment using comparative genomics . SMBE Satellite meeting SMBEBA 2015 "Investigating biological adaptation with NGS: data and models"

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. Fromthe results obtained, it becomes clear that these yeasts can interbreed. Although genetic delimitation is possible for thethree species, the extent of nucleotide substitutions and phenotypic differences observed between them are lower thanthat expected for species that have ended the speciation process. Our taxonomic conclusion is to maintain the three taxauntil further genomic data are gathered. However, the concept of L. scottii species complex is proposed for this group ofspecies. Finally, we transfer all Leucosporidiella and Mastigobasidium species to Leucosporidium (Leucosporidiales), and, inorder to end the polyphyly condition of these taxa, we propose the new genus Pseudoleucosporidium gen. nov. and the newcombination Peudoleucosporidium fasciculatum comb. nov. Fil: de Garcia, Virginia. Universidad Nacional del Comahue. Centro Regional Universitario Bariloche; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Patagonia Norte. Instituto de Investigación en Biodiversidad y Medioambiente; Argentina Fil: Coelho, Marco A.. Universidade Nova de Lisboa; Portugal Fil: Maia, Teresa M.. Universidade Nova de Lisboa; Portugal Fil: Rosa, Luiz H.. Universidade Federal do Minas Gerais; Brasil Fil: Vaz, Aline Martin. Universidade Federal do Minas Gerais; Brasil Fil: Rosa, Carlos A.. Universidade Federal do Minas Gerais; Brasil Fil: Sampaio, José Paulo. Universidade Nova de Lisboa; Portugal Fil: Goncalves, Paula. Universidade Nova de Lisboa; Portugal Fil: Van Broock, María. Universidad Nacional del Comahue. Centro Regional Universitario Bariloche; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Patagonia Norte. Instituto de Investigación en Biodiversidad y Medioambiente; Argentina Fil: Libkind Frati, Diego. Universidad Nacional del Comahue. Centro Regional Universitario Bariloche; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Patagonia Norte. Instituto de Investigación en Biodiversidad y Medioambiente; Argentina

The domestication of the wine yeast Saccharomyces cerevisiae is thought to be contemporary with the development and expansion of viticulture along the Mediterranean basin. Until now, the unavailability of wild lineages prevented the identification of the closest wild relatives of wine yeasts. Here, we enlarge the collection of natural lineages and employ whole-genome data of oak-associated wild isolates to study a balanced number of anthropic and natural S. cerevisiae strains. We identified industrial variants and new geographically delimited populations, including a novel Mediterranean oak population. This population is the closest relative of the wine lineage as shown by a weak population structure and further supported by genomewide population analyses. A coalescent model considering partial isolation with asymmetrical migration, mostly from the wild group into the Wine group, and population growth, was found to be best supported by the data. Importantly, divergence time estimates between the two populations agree with historical evidence for winemaking. We show that three horizontally transmitted regions, previously described to contain genes relevant to wine fermentation, are present in the Wine group but not in the Mediterranean oak group. This represents a major discontinuity between the two populations and is likely to denote a domestication fingerprint in wine yeasts. Taken together, these results indicate that Mediterranean oaks harbour the wild genetic stock of domesticated wine yeasts.

In the Iberian Pyrite Belt (IPB), acid rock drainage gives rise to aquatic habitats with low pH and high concentrations of heavy metals, a situation that causes important environmental problems. We investigated the occurrence and diversity of yeasts in two localities of the IPB: São Domingos (Portugal) and Rio Tinto (Spain). Yeast isolation was performed on conventional culture media (MYP), acidified (pH 3) media (MYP3), and on media prepared with water from the study sites (MYPw). The main goal of the study was to determine the structure of the yeast community; a combination of molecular methods was used for accurate species identifications. Our results showed that the largest fraction of the yeast community was recovered on MYPw rather than on MYP and MYP3. Twenty-seven yeast species were detected, 48% of which might represent undescribed taxa. Among these, an undescribed species of the genus Cryptococcus required low pH for growth, a property that has not been observed before in yeasts. The communities of S. Domingos and R. Tinto showed a considerable resemblance, and eight yeast species were simultaneously found in both localities. Taking into consideration the physicochemical parameters studied, we propose a hierarchic organization of the yeast community in terms of high-, intermediate-, or low-stress conditions of the environment. According to this ranking, the acidophile yeast Cryptococcus sp. 5 is considered the most tolerant species, followed by Cryptococcus sp. 3 and Lecytophora sp. Species occurring in situations of intermediate environmental stress were Candida fluviatilis, Rhodosporidium toruloides, Williopsis californica, and three unidentified yeasts belonging to Rhodotorula and Cryptococcus.

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.

The occurrence and diversity of yeasts in seawater was investigated in a study site located 20 Km off Faro, Portugal, above the Alvares Cabral Trench. A total of 43 water samples from different layers (above the permanent thermocline, under the thermocline and near the bottom) and directly from the surface, originated 234 isolates. All the isolates were identified using a molecular approach that included, in a first stage, MSP-PCR fingerprinting. A total of 31 MSP-PCR classes were formed, 8 for the pigmented yeasts and 23 for the non-pigmented yeasts. The pink coloured isolates were identified by direct comparison of the new fingerprints with those obtained for representative strains of the various species. For identification of the non-pigmented yeasts, a representative isolate of each MSP-PCR class was selected for sequence analysis and compared with reference sequences. The five most abundant yeast species were Sakaguchia dacryoidea, Pseudozyma aphidis, Rhodosporidium babjevae, R. diobovatum and Debaryomyces hansenii. The distribution of isolates and species in the major taxonomic groups indicated that the number of basidiomycetous yeasts and their diversity are prevalent in relation to their ascomycetous counterpart. Diversity indices were determined and superficial water and water near the bottom had the highest diversity. The sampling effort effectiveness was estimated, and found to correspond to approximately 60% of the species present. MSP-PCR identification proved suitable for pigmented basidiomycetous yeasts and, when used in conjunction with sequence analysis, was effective for the characterization of non-pigmented populations. Our results indicate that the MSP-PCR fingerprinting method is appropriate for the characterization of large groups of isolates due to its simplicity and good reproducibility.

Fifteen aquatic environments (lakes, lagoons and rivers) of glacial origin in the northern Andean Patagonia (Argentina) were surveyed for the occurrence of red yeasts. Subsurface water samples were filtered and used for colony counting and yeast isolation. A preliminary quantitative analysis indicated that total yeast counts ranged between 0 and 250 cells l−1. A polyphasic approach including physiological and molecular methods was used for the identification of 64 carotenogenic yeast strains. The molecular characterisation of the isolates was based on the mini/microsatellite-primed PCR technique (MSP-PCR) employing the (GTG)5 and the M13 primers. Comparison of representative fingerprints of each group with those of the type strains of pigmented yeasts allowed the expeditious identification of 87.5% isolates. The sequence analysis of the D1/D2 domains of the 26S rDNA was employed to confirm identifications and in the characterization of the unidentified MSP-PCR groups. Teleomorphic yeast species were detected by performing sexual compatibility assays. The isolates corresponded to 6 genera and 15 yeast species, including four new yeast species of the genera Cryptococcus (1), Rhodotorula (1) and Sporobolomyces (2). Rhodotorula mucilaginosa was found in the majority of the samples and represented ca. 50% of the total number of isolates. However, this yeast was not detected in aquatic environments with very low anthropic influence. Other frequent yeast isolates were teleomorphic yeast species of Rhodosporidium babjevae, R. kratochvilovae and Sporidiobolus salmonicolor. This study represents the first report on red yeast occurrence and biodiversity in northwestern Patagonia.

A culture was obtained from a spore print of a basidiocarp of Mycogloea nipponica collected in Taiwan. A yeast stage and basidia identical to those of M. nipponica developed in laboratory media. The Taiwanese specimen of M. nipponica and its yeast anamorph were characterised in the present study. Comparative morphological, molecular, and ultrastructural studies indicated that the yeast stage can be assigned to the genus Kurtzmanomyces. The revealed connection between the sexual species Mycogloea nipponica and the asexual genus Kurtzmanomyces demonstrates the importance of anamorphic characteristics in the modern systematics of heterobasidiomycetous fungi.

A new anamorphic heterobasidiomycetous yeast species, Kurtzmanomyces insolitus, is described using a polyphasic taxonomic approach. The new species has the salient characteristics of the genus Kurtzmanomyces and, additionally, the ability to produce ballistoconidia. Data derived from comparative micromorphological studies, physiological characterisation, ultrastructure and nucleic acid analyses led to assigning the new species to Kurtzmanomyces rather than to the currently accepted genera of ballistoconidia-forming fungi. An emendation of the genus Kurtzmanomyces is proposed to allow the inclusion of the new species.

In the present work, we investigated the phylogenetic position and phenotypic characteristics of eight yeast isolates collected from migratory birds on the island of Ustica, Italy. A phylogenetic analysis based on the D1/D2 region of the large-subunit rRNA gene showed that all isolates clustered as a single separate lineage within the Wickerhamomyces clade. They exhibited distinct morphological and physiological characteristics and were clearly separated from their closest relatives, Wickerhamomyces lynferdii, Wickerhamomyces anomalus and Wickerhamomyces subpelliculosus, in blastn searches. On the basis of the isolation source, physiological features and molecular strain typing carried out with randomly amplified polymorphic DNA (RAPD)-PCR and minisatellite-primed (MSP)-PCR analysis, the isolates were identified as strains of the same species. The name Wickerhamomyces sylviae f.a., sp. nov. is proposed to accommodate these novel strains; the type strain is U88A2T ( = PYCC 6345T = CBS 12888T). The MycoBank number is MB 804762.

Microbes establish very diverse but still poorly understood associations with other microscopic or macroscopic organisms that do not follow the more conventional modes of competition or mutualism. Phaffia rhodozyma, an orange-coloured yeast that produces the biotechnologically relevant carotenoid astaxanthin and grows in sugar-rich tree exudates, exhibits an Holarctic association with birch trees in temperate forests that contrasts with the more recent finding of a South American population associated with Nothofagus (southern beech) and with stromata of its biotrophic fungal parasite Cyttaria spp. Here we first investigated if the association of Phaffia with Nothofagus-Cyttaria could be expanded to Australasia, the other region of the world where Nothofagus are endemic, and then studied the genetic structure of populations representing the known worldwide distribution of Phaffia and analyzed the evolution of the association with tree hosts. The phylogenetic analysis of concatenated partial sequences of seven genes from forty strains revealed that Phaffia diversity in Australasia is much higher than in other regions of the globe and that two endemic and markedly divergent lineages seem to represent new species. Moreover, the observed genetic diversity correlates with host tree genera rather than with geography, which suggests that adaptation to the different niches is driving population structure in this yeast. The high genetic diversity and endemism in Australasia indicates that the genus Phaffia evolved in this region and that the association with Nothofagus is the ancestral tree-association. Our estimates of the divergence times of the various Phaffia lineages point to splits that are much more recent than the breakup of Gondwana. This suggests that long distance dispersal rather than vicariance is responsible for the presence of P. rhodozyma in Australasia, South America and in the Holarctic region. Fil: David Palma, Márcia. Universidade Nova de Lisboa; Portugal Fil: Libkind Frati, Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Patagonia Norte. Instituto de Investigación en Biodiversidad y Medioambiente; Argentina. Universidad Nacional del Comahue; Argentina Fil: Sampaio, José Paulo. Universidade Nova de Lisboa; Portugal

The synthesis of the UV Absorbing compounds named mycosporines (MYCs) has been detected in a few basidiomycetous yeast groups. Conspicuous accumulation of mycosporine-glutaminol-glucoside (MGG) in yeasts requires photo-induction and its photoprotective function has been postulated. The distribution of the ability to produce MYCs appeared to be related to the yeast taxonomic affiliation. In view of the potential significance of MYCs in yeast taxonomy, we here studied the distribution of this trait among dimorphic basidiomycetes of the Pucciniomycotina. Of the 94 fungal species (377 strains and 33 genera) tested, almost half were MYC-positive and MGG was the main compound produced. MGG synthesis was observed for representatives of five of seven Pucciniomycotina classes, indicating that this trait is widely distributed in this group. MGG detection proved useful for the differentiation of species of the polyphyletic genera, such as Rhodotorula and Sporobolomyces ,t hat are phylogenetically separated. MGG quantification and UV tolerance studies in Cystobasidiomycetes supported the idea that the habitat of origin of each strains is important in the level of MGG synthesis and that MYCs have a photoprotective function in yeasts. The taxonomic value of this trait in fungal systematics is discussed. Copyright  2011 John Wiley & Sons, Ltd.

Publisher Summary This chapter discusses the genus Cystofilobasidium. It provides a systematic discussion of the species and concludes with comments on the genus. The diagnosis of the genus is done on the basis of asexual, sexual, physiology and biochemistry, and phylogenetic placement. In asexual reproduction yeast cells are ovoid or ellipsoid. Asexual reproduction is predominantly by polar budding. Poorly developed pseudohyphae may be present in some species. Ballistoconidia are absent. The cultures have an orange to pinkish color that darkens upon aging, but nonpigmented variants are known in some species. Pigmentation is due to the presence of carotenoid pigments. In sexual reproduction species may be self-sterile (heterothallic) or self-fertile (homothallic). The self-sterile compatibility systems are tetrapolar or bipolar. Depending on the species, clamp connections are present or absent. The septal pore is a dolipore without parenthesomes. Teliospores are formed, which are spherical and dark pigmented, and upon germination form slender, tubular or short, nonseptate basidia with terminal and sessile basidiospores that germinate by budding.

Publisher Summary This chapter studies the genus Papiliotrema. In asexual reproduction yeast cells are ovoid and budding is polar. In sexual reproduction it is seen that the sexual structures are formed in the filamentous state and have only been observed in nature. This chapter further examines physiology, biochemistry, and phylogenetic placement. The type species and accepted species mentioned is Papiliotema bandonii. The systematic discussion of the species includes growth on 5% malt extract agar, Dalmau plate culture on corn meal agar, sexual state, fermentation, gene sequence accession numbers, cell carbohydrates, origin of the strains studied, type strain, systematics, ecology, biotechnology, and agriculture and food.

The genus Saccharomyces currently includes eight species in addition to the model yeast Saccharomyces cerevisiae, most of which can be consistently isolated from tree bark and soil. We recently found sympatric pairs of Saccharomyces species, composed of one cryotolerant and one thermotolerant species in oak bark samples of various geographic origins. In order to contribute to explain the occurrence in sympatry of Saccharomyces species, we screened Saccharomyces genomic data for protein divergence that might be correlated to distinct growth temperature preferences of the species, using the dN/dS ratio as a measure of protein evolution rates and pair-wise species comparisons. In addition to proteins previously implicated in growth at suboptimal temperatures, we found that glycolytic enzymes were among the proteins exhibiting higher than expected divergence when one cryotolerant and one thermotolerant species are compared. By measuring glycolytic fluxes and glycolytic enzymatic activities in different species and at different temperatures, we subsequently show that the unusual divergence of glycolytic genes may be related to divergent evolution of the glycolytic pathway aligning its performance to the growth temperature profiles of the different species. In general, our results support the view that growth temperature preference is a trait that may have undergone divergent selection in the course of ecological speciation in Saccharomyces.

Publisher Summary This chapter studies the genus Sporidiobolus. In the determination of asexual reproduction it is found that reproduction is by budding and by formation of ballistoconidia. Yeast cells are ovoid, ellipsoidal, or elongate. Visible carotenoid pigments are formed and the cultures are pink to orange in color. In sexual reproduction it is seen that hyphae have clamp connections. Spherical teliospores are formed and germinate to produce transversely septate basidia with ovoid to bacilliform basidiospores, which are passively released and germinate by budding. In physiology and biochemistry it is seen that fermentation is absent. This chapter further examines phylogenetic placement. The type species mentioned is Sporidiobolus johnsonii. The species accepted are Sporidiobolus johnsonii, Sporidiobolus longiusculus, Sporidiobolus metaroseus, Sporidiobolus microsporus, Sporidiobolus pararoseus, Sporidiobolus ruineniae, and Sporidiobolus salmonicolor. The systematic discussion of the species includes synonyms, growth on 5% malt extract agar, Dalmau plate culture on corn meal agar, sexual state, CoQ, fermentation, Mol% G + C, gene sequence accession number, type strain, cell carbohydrates, origin of the strains studied, systematics, ecology, biotechnology, and agriculture and food.