Wild Yeast Isolates Research Articles

Molecular Mechanisms of Genotype-Dependent Lifespan Variation Mediated by Caloric Restriction: Insight from Wild Yeast Isolates.

Caloric restriction (CR) is known to extend lifespan across different species and holds great promise for preventing human age-onset pathologies. However, two major challenges exist. First, despite extensive research, the mechanisms of lifespan extension in response to CR remain elusive. Second, genetic differences causing variations in response to CR and genetic factors contributing to variability of CR response on lifespan are largely unknown. Here, we took advantage of natural genetic variation across 46 diploid wild yeast isolates of Saccharomyces species and the lifespan variation under CR conditions to uncover the molecular factors associated with CR response types. We identified genes and metabolic pathways differentially regulated in CR-responsive versus non-responsive strains. Our analysis revealed that altered mitochondrial function and activation of GCN4-mediated environmental stress response are inevitably linked to lifespan variation in response to CR and a unique mitochondrial metabolite might be utilized as a predictive marker for CR response rate. In sum, our data suggests that the effects of CR on longevity may not be universal, even among the closely related species or strains of a single species. Since mitochondrial-mediated signaling pathways are evolutionarily conserved, the dissection of related genetic pathways will be relevant to understanding the mechanism by which CR elicits its longevity effect.

Isolation of wild yeasts and the application of <i>Lachancea thermotolerans</i> for alcohol fermentation

Isolation of wild yeasts and the application of <i>Lachancea thermotolerans</i> for alcohol fermentation

Molecular mechanisms of genotype-dependent lifespan variation mediated by caloric restriction: insight from wild yeast isolates.

Caloric restriction (CR) is known to extend lifespan across different species and holds great promise for preventing human age-onset pathologies. However, two major challenges exist. First, despite extensive research, the mechanisms of lifespan extension in response to CR remain elusive. Second, genetic differences causing variations in response to CR and genetic factors contributing to variability of CR response on lifespan are largely unknown. Here, we took advantage of natural genetic variation across 46 diploid wild yeast isolates of Saccharomyces species and the lifespan variation under CR conditions to uncover the molecular factors associated with CR response types. We identified genes and metabolic pathways differentially regulated in CR-responsive versus non-responsive strains. Our analysis revealed that altered mitochondrial function and activation of GCN4-mediated environmental stress response are inevitably linked to lifespan variation in response to CR and a unique mitochondrial metabolite might be utilized as a predictive marker for CR response rate. In sum, our data suggests that the effects of CR on longevity may not be universal, even among the closely related species or strains of a single species. Since mitochondrial-mediated signaling pathways are evolutionarily conserved, the dissection of related genetic pathways will be relevant to understanding the mechanism by which CR elicits its longevity effect.

Isolation of wild yeasts from Olympic National Park and Moniliella megachiliensis ONP131 physiological characterization for beer fermentation

Thousands of yeasts have the potential for industrial application, though many were initially considered contaminants in the beer industry. However, these organisms are currently considered important components in beers because they contribute new flavors. Non-Saccharomyces wild yeasts can be important tools in the development of new products, and the objective of this work was to obtain and characterize novel yeast isolates for their ability to produce beer. Wild yeasts were isolated from environmental samples from Olympic National Park and analyzed for their ability to ferment malt extract medium and beer wort. Six different strains were isolated, of which Moniliella megachiliensis ONP131 displayed the highest levels of attenuation during fermentations. We found that M. megachiliensis could be propagated in common yeast media, tolerated incubation temperatures of 37 °C and a pH of 2.5, and was able to grow in media containing maltose as the sole carbon source. Yeast cultivation was considerably impacted (p < 0.05) by lactic acid, ethanol, and high concentrations of maltose, but ONP131 was tolerant to high salinity and hop acid concentrations. This is one of the first physiological characterizations of M. megachiliensis, which has potential for the production of beer and other fermented beverages.

Evolution of Natural Lifespan Variation and Molecular Strategies of Extended Lifespan

To understand the genetic basis and the selective forces acting on longevity, it is useful to employ ecologically diverse individuals of the same species, widely different in lifespan. This way, we may capture the experiment of Nature that modifies the genotype arriving at different lifespans. Here, we analyzed 76 ecologically diverse wild yeast isolates and discovered wide diversity of lifespan. We sequenced the genomes of these organisms and analyzed how their replicative lifespan is shaped by nutrients and transcriptional and metabolite patterns. By identifying genes, proteins and metabolites that correlate with longevity across these isolates, we found that long-lived strains elevate intermediary metabolites, differentially regulate genes involved in NAD metabolism and adjust control of epigenetic landscape through conserved, rare histone modifier. Our data further offer insights into the evolution and mechanisms by which caloric restriction regulates lifespan by modulating the availability of nutrients without decreasing fitness.

Evolution of natural lifespan variation and molecular strategies of extended lifespan in yeast.

To understand the genetic basis and selective forces acting on longevity, it is useful to examine lifespan variation among closely related species, or ecologically diverse isolates of the same species, within a controlled environment. In particular, this approach may lead to understanding mechanisms underlying natural variation in lifespan. Here, we analyzed 76 ecologically diverse wild yeast isolates and discovered a wide diversity of replicative lifespan (RLS). Phylogenetic analyses pointed to genes and environmental factors that strongly interact to modulate the observed aging patterns. We then identified genetic networks causally associated with natural variation in RLS across wild yeast isolates, as well as genes, metabolites, and pathways, many of which have never been associated with yeast lifespan in laboratory settings. In addition, a combined analysis of lifespan-associated metabolic and transcriptomic changes revealed unique adaptations to interconnected amino acid biosynthesis, glutamate metabolism, and mitochondrial function in long-lived strains. Overall, our multiomic and lifespan analyses across diverse isolates of the same species shows how gene-environment interactions shape cellular processes involved in phenotypic variation such as lifespan.

Isolation of Wild Yeasts from Freshwaters and Soils in Nonsan Stream and Sapgyoho in Chungcheongnam-do, Korea, and Microbiological Characteristics of the Unrecorded Wild Yeasts

Isolation of Wild Yeasts from Freshwaters and Soils in Nonsan Stream and Sapgyoho in Chungcheongnam-do, Korea, and Microbiological Characteristics of the Unrecorded Wild Yeasts

Isolation of Wild Yeasts from Riversides in Geumgang Upstream, Taean and Suncheonman Seashores and Microbiological Characteristics of the Unrecorded Wild Yeasts

Isolation of Wild Yeasts from Riversides in Geumgang Upstream, Taean and Suncheonman Seashores and Microbiological Characteristics of the Unrecorded Wild Yeasts

Isolation of Wild Yeasts and Characterization of Physiological Functionalities of Unrecorded Wild Yeasts Obtained from Flowers and Soils of the Wolpyung Park, Daejeon City and Gykpo Beach, Buan, Jeollabuk-do in Korea

Isolation of Wild Yeasts and Characterization of Physiological Functionalities of Unrecorded Wild Yeasts Obtained from Flowers and Soils of the Wolpyung Park, Daejeon City and Gykpo Beach, Buan, Jeollabuk-do in Korea

Molecular identification and performance evaluation of wild yeasts from different Ethiopian fermented products.

Different types of yeasts and lactic acid bacteria dominate in spontaneously fermented products (food, beverages, and condiments) that are commonly consumed in Ethiopia. The aim of this study was to identify efficient fermentative yeasts from fermented foods, fermented beverages, honey and molasses using genotypic methods. Out of the 70 samples tested, 180 distinct wild yeast isolates were recovered. A total of 23 isolates were selected for genomic analysis based on their basis of biomass yield, fermentation capacity, and leavening performance. The nucleotide sequence analysis of the internal transcribed spacer ITS-5.8S rDNA region revealed that the indigenous yeast isolates had close relatedness to Saccharomyces cerevisiae, Candida humilis, Kazachstania bulderi, Pichia fermentans and Pichia kudriavzevii with greater than 97% nucleotide similarity. The study shows a high diversity of indigenous wild yeasts in fermented products and that potent strains had higher biomass yield, good gas production and remarkable leavening capacity that indicates their inherent potential for use in the baking industry.

Widespread Prion-Based Control of Growth and Differentiation Strategies in Saccharomyces cerevisiae.

Theory and experiments suggest that organisms would benefit from pre-adaptation to future stressors based on reproducible environmental fluctuations experienced by their ancestors, but the mechanisms driving pre-adaptation remain enigmatic. We report that the [SMAUG+] prion allows yeast to anticipate nutrient repletion after periods of starvation, providing a strong selective advantage. By transforming the landscape of post-transcriptional gene expression, [SMAUG+] regulates the decision between two broad growth and survival strategies: mitotic proliferation or meiotic differentiation into a stress-resistant state. [SMAUG+] is common in laboratory yeast strains, where standard propagation practice produces regular cycles of nutrient scarcity followed by repletion. Distinct [SMAUG+] variants are also widespread in wild yeast isolates from multiple niches, establishing that prion polymorphs can be utilized in natural populations. Our data provide a striking example of how protein-based epigenetic switches, hidden in plain sight, can establish a transgenerational memory that integrates adaptive prediction into developmental decisions.

Pervasive function and evidence for selection across standing genetic variation in S. cerevisiae

Quantitative genetics aims to map genotype to phenotype, often with the goal of understanding how organisms evolved. However, it remains unclear whether the genetic variants identified are exemplary of evolution. Here we analyzed progeny of two wild Saccharomyces cerevisiae isolates to identify 195 loci underlying complex metabolic traits, resolving 107 to single polymorphisms with diverse molecular mechanisms. More than 20% of causal variants exhibited patterns of emergence inconsistent with neutrality. Moreover, contrary to drift-centric expectation, variation in diverse wild yeast isolates broadly exhibited this property: over 30% of shared natural variants exhibited phylogenetic signatures suggesting that they are not neutral. This pattern is likely attributable to both homoplasy and balancing selection on ancestral polymorphism. Variants that emerged repeatedly were more likely to have done so in isolates from the same ecological niche. Our results underscore the power of super-resolution mapping of ecologically relevant traits in understanding adaptation and evolution.

Widespread Prion-Based Control of Growth and Differentiation Strategies in &lt;i&gt;Saccharomyces Cerevisiae&lt;/i&gt;

Theory and experiment suggest that organisms can pre-adapt to future stressors based on reproducible environmental fluctuations experienced by their ancestors. Yet the mechanisms driving such pre-adaptation remain enigmatic. We report that the [SMAUG+] prion allows yeast to anticipate nutrient repletion after periods of starvation, providing a strong selective advantage. By transforming the landscape of post-transcriptional gene expression, [SMAUG+] regulates the decision between two broad growth and survival strategies: mitotic proliferation or meiotic differentiation into a stress-resistant state. [SMAUG+] is common in laboratory yeast strains, where standard propagation practice produces regular cycles of nutrient scarcity followed by repletion. Distinct [SMAUG+] variants are also widespread in wild yeast isolates from multiple niches, establishing that prion polymorphs can be utilized in natural populations. Our data provide a striking example of how protein-based epigenetic switches can be hidden in plain sight and can establish a transgenerational memory that integrates adaptive prediction into developmental decisions.

Performance of non-conventional yeasts in co-culture with brewers' yeast for steering ethanol and aroma production.

SummaryIncreasing interest in new beer types has stimulated the search for approaches to extend the metabolic variation of brewers’ yeast. Therefore, we tested two approaches using non‐conventional yeast to create a beer with lower ethanol content and a complex aroma bouquet. First, the mono‐culture performance was monitored of 49 wild yeast isolates of Saccharomyces cerevisiae (16 strains), Cyberlindnera fabianii (9 strains) and Pichia kudriavzevii (24 strains). Interestingly, both C. fabianii and P. kudriavzevii isolates produced relatively more esters compared with S. cerevisiae isolates, despite their limited fermentation capacity. Next, one representative strain of each species (Sc131, Cf65 and Pk129) was applied as co‐culture with brewers’ yeast (ratio 1:1). Co‐cultures with Cf65 and Pk129 resulted in a beer with lower alcohol content (3.5, 3.8 compared with 4.2% v/v) and relatively more esters. At higher inoculum ratios of Cf65 over brewers’ yeast, growth inhibition of brewers’ yeast was observed, most likely caused by competition for oxygen between brewers’ yeast and Cf65 resulting in a reduced level of ethanol and altered aroma profiles. With this study, we demonstrate the feasibility of using non‐conventional yeast species in co‐cultivation with traditional brewers’ yeast to tailor aroma profiles as well as the final ethanol content of beer.

&lt;b&gt;Use of wild yeasts as a biocontrol agent against toxigenic fungi and OTA production

This study evaluated the antagonistic potential of 32 wild yeast isolates from coffee and cocoa bean fermentation. These yeasts were inoculated in co-cultivation with Aspergillus carbonarius (CCDCA 10608 and CCDCA 10408) and Aspergillus ochraceus (CCDCA 10612) isolated from grapes and coffee beans. The mycelial growth and ochratoxin A (OTA) production were evaluated, and the spores were counted after cultivation at 28°C for seven days. The yeasts presented higher inhibitory effects (53% in relation to the control) over the mycelial growth of the isolated A. ochraceus (CCDCA10612) . Pichia anomala CCMA0148 and Saccharomyces cerevisiae CCMA0159 provided the greatest inhibition of the growth of all fungal strains. All Pichia species presented the highest inhibitory effects on the production of spores, and S. cerevisiae CCMA 0159 at concentrations of both 10 4 and 10 7 mL -1 cells inhibited the production of spores by 100%. Rhodotorula mucilaginosa was effective at inhibiting OTA production by the three isolates of Aspergillus . S. cerevisiae CCMA0159 and Pichia anomala CCMA0148 showed high potential as biocontrol agents in the conditions tested.

Isolation of Wild Yeasts from Soils of Reed Fields in Seocheon-gun County, Chungcheongnam-do, South Korea, and Characterization of Unrecorded Yeasts

충남 서천군 한산면 신성리 금강하구 갈대밭 토양의 야생효모 분포특성을 알아보기 위하여 갈대밭 토양 20점을 채취하여 야생효모들을 분리하여 총 11종 20균주의 야생효모를 동정하였다. Candida속의 효모가 10균주로 가장 많이 분리되었고, 그 중 Candida subhashii 균이 6균주로 가장 많았다. 분리된 균주 중에서 Candida tropicalis (Y5-1,Y5-2, Y6-1), Candida subhashii (J7-1, J7-3, J8-1, Y9-1, Y9-2, Y9-3), Lachancea thermotolerans (MJS 007, MJS 019) 균주 등 12균주들이 알코올 발효능을 보였다. Bullera japonica YJ10-1, C. subhashii J7-1, Kluyveromyces yarrowii YJ11-1, Ustilago shanxiensis Y10-1등 4균주들이 국내 미기록 효모 균주들로 최종 선별되었다. 이들 국내 미기록 균주들은 구형으로 Candida subhashii J7-1만이 자낭포자와 의균사를 형성하였다. Kluyveromyces yarrowii YJ11-1은 비타민을 함유하지 않은 배지에서도 생육하였고 Candida subhashii J7-1는 YM과 YPD 배지 및 5% NaCl을 함유한 YPD 배지에서 생장하지 않았다. The goal of this study was to isolate wild yeasts from reed fields in the Seocheon-gun county in Chungcheongnam-do, South Korea. Molecular analysis of the D1/D2 domain of the 26S rDNA of the yeasts isolated from soil samples was performed using Basic Local Alignment Search Tool (BLAST). In total, 20 yeast strains from 11 species were isolated from 20 different soil samples. Candida species were observed to be the most common yeasts. Of these 20 strains, Candida subhashii (6 strains), Candida tropicalis (3 strains), Candida boleticola MS008, and Lachancea thermotolerans (2 strains) were identified as alcohol-fermenting yeasts. Further, Bullera japonica YJ10-1, Candida subhashii J7-1, Kluyveromyces yarrowii YJ11-1, and Ustilago shanxiensis Y10-1 were newly recorded yeast strains in Korea, and therefore, their microbiological characteristics were investigated further. All of these unrecorded yeast strains had oval cells; only Candida subhashii J7-1 formed ascospores and pseudomycelia. Kluyveromyces yarrowii YJ11-1 grew in vitamin-free medium, and all of these strains, except Candida subhashii J7-1, grew in 5% NaCl-containing YPD broth.

Asynchronous spore germination in isogenic natural isolates of Saccharomyces paradoxus.

Spores from wild yeast isolates often show great variation in the size of colonies they produce, for largely unknown reasons. Here we measure the colonies produced from single spores from six different wild Saccharomyces paradoxus strains. We found remarkable variation in spore colony sizes, even among spores that were genetically identical. Different strains had different amounts of variation in spore colony sizes, and variation was not affected by the number of preceding meioses, or by spore maturation time. We used time-lapse photography to show that wild strains also have high variation in spore germination timing, providing a likely mechanism for the variation in spore colony sizes. When some spores from a laboratory strain make small colonies, or no colonies, it usually indicates a genetic or meiotic fault. Here, we demonstrate that in wild strains spore colony size variation is normal. We discuss and assess potential adaptive and non-adaptive explanations for this variation.

Isolation of wild yeasts having alcohol fermentation ability from plants in "Dewasanzan (Mt. Yudono, Mt. Haguro, Mt. Gassan)"

Isolation of wild yeasts having alcohol fermentation ability from plants in "Dewasanzan (Mt. Yudono, Mt. Haguro, Mt. Gassan)"

Dosage compensation can buffer copy-number variation in wild yeast.

Aneuploidy is linked to myriad diseases but also facilitates organismal evolution. It remains unclear how cells overcome the deleterious effects of aneuploidy until new phenotypes evolve. Although laboratory strains are extremely sensitive to aneuploidy, we show here that aneuploidy is common in wild yeast isolates, which show lower-than-expected expression at many amplified genes. We generated diploid strain panels in which cells carried two, three, or four copies of the affected chromosomes, to show that gene-dosage compensation functions at >30% of amplified genes. Genes subject to dosage compensation are under higher expression constraint in wild populations-but they show elevated rates of gene amplification, suggesting that copy-number variation is buffered at these genes. We find that aneuploidy provides a clear ecological advantage to oak strain YPS1009, by amplifying a causal gene that escapes dosage compensation. Our work presents a model in which dosage compensation buffers gene amplification through aneuploidy to provide a natural, but likely transient, route to rapid phenotypic evolution.

Phenotypic heterogeneity is a selected trait in natural yeast populations subject to environmental stress

Populations of genetically uniform microorganisms exhibit phenotypic heterogeneity, where individual cells have varying phenotypes. Such phenotypes include fitness-determining traits. Phenotypic heterogeneity has been linked to increased population-level fitness in laboratory studies, but its adaptive significance for wild microorganisms in the natural environment is unknown. Here, we addressed this by testing heterogeneity in yeast isolates from diverse environmental sites, each polluted with a different principal contaminant, as well as from corresponding control locations. We found that cell-to-cell heterogeneity (in resistance to the appropriate principal pollutant) was prevalent in the wild yeast isolates. Moreover, isolates with the highest heterogeneity were consistently observed in the polluted environments, indicating that heterogeneity is positively related to survival in adverse conditions in the wild. This relationship with survival was stronger than for the property of mean resistance (IC50) of an isolate. Therefore, heterogeneity could be the major determinant of microbial survival in adverse conditions. Indeed, growth assays indicated that isolates with high heterogeneities had a significant competitive advantage during stress. Analysis of yeasts after cultivation for ≥ 500 generations additionally showed that high heterogeneity evolved as a heritable trait during stress. The results showed that environmental stress selects for wild microorganisms with high levels of phenotypic heterogeneity.