Improved redox homeostasis owing to the up-regulation of one-carbon metabolism and related pathways is crucial for yeast heterosis at high temperature.

Heterosis or hybrid vigor is a common phenomenon in plants and animals; however, the molecular mechanisms underlying heterosis remain elusive, despite extensive studies on the phenomenon for more than a century. Here we constructed a large collection of F1 hybrids of Saccharomyces cerevisiae by spore-to-spore mating between homozygous wild strains of the species with different genetic distances and compared growth performance of the F1 hybrids with their parents. We found that heterosis was prevalent in the F1 hybrids at 40°C. A hump-shaped relationship between heterosis and parental genetic distance was observed. We then analyzed transcriptomes of selected heterotic and depressed F1 hybrids and their parents growing at 40°C and found that genes associated with one-carbon metabolism and related pathways were generally up-regulated in the heterotic F1 hybrids, leading to improved cellular redox homeostasis at high temperature. Consistently, genes related with DNA repair, stress responses, and ion homeostasis were generally down-regulated in the heterotic F1 hybrids. Furthermore, genes associated with protein quality control systems were also generally down-regulated in the heterotic F1 hybrids, suggesting a lower level of protein turnover and thus higher energy use efficiency in these strains. In contrast, the depressed F1 hybrids, which were limited in number and mostly shared a common aneuploid parental strain, showed a largely opposite gene expression pattern to the heterotic F1 hybrids. We provide new insights into molecular mechanisms underlying heterosis and thermotolerance of yeast and new clues for a better understanding of the molecular basis of heterosis in plants and animals.

Highly diverged lineages of Saccharomyces paradoxus in temperate to subtropical climate zones in China.

The wild yeast Saccharomyces paradoxus has become a new model in ecology and evolutionary biology. Different lineages of S. paradoxus have been recognized across the world, but the distribution and genetic diversity of the species remain unknown in China, where the origin of its sibling species S. cerevisiae lies. In this study, we investigated the ecological and geographic distribution of S. paradoxus through an extensive field survey in China and performed population genomic analysis on a set of S. paradoxus strains, including 27 strains, representing different geographic and ecological origins within China, and 59 strains representing all the known lineages of the species recognized in the other regions of the world so far. We found two distinct lineages of S. paradoxus in China. The majority of the Chinese strains studied belong to the Far East lineage, and six strains belong to a novel highly diverged lineage. The distribution of these two lineages overlaps ecologically and geographically in temperate to subtropical climate zones in China. With the addition of the new China lineage, the Eurasian population of S. paradoxus exhibits higher genetic diversity than the American population. We observed more possible lineage-specific introgression events from the Eurasian lineages than from the American lineages. Our results expand the knowledge on ecology, genetic diversity, biogeography, and evolution of S. paradoxus.

Diversity and distribution of yeasts in indigenous fermented foods and beverages of Ethiopia.

BACKGROUND: As one of the origin centers of domesticated plants in the world, Ethiopia is rich in diversified fermented foods and beverages, in which yeasts are usually among the essential functional microorganisms. This study aims to investigate yeast species diversity and distribution in indigenous fermented products in Ethiopia using conventional isolation and molecular identification methods. RESULT: Yeast cell counts in 221 samples of various Ethiopian traditional fermented products, including fermented staple foods, alcoholic beverages, dairy products, and condiments, were compared using the typical dilution plating method. A total of 475 yeast isolates were recovered from these samples and 41 yeast species belonging to 25 genera were identified from yeast isolates using the sequence analysis of the D1/D2 domain of the large subunit of rRNA gene. Candida humilis, Wickerhamomyces anomalus, Meyerozyma guilliermondii, Pichia kudriavzevii, and Saccharomyces cerevisiae were the most dominant species that were widely distributed among the majority of the fermented products analyzed in the current study. CONCLUSION: Significant variations were encountered both in yeast cell counts, diversity, and distribution of yeast species among different types of fermented products and even among different samples of the same types of fermented products. It calls for a more extensive and systematic microbiological study of Ethiopian indigenous fermented foods, beverages and other related products that can be helpful for standardization and large-scale production of these foods in Ethiopia. © 2020 Society of Chemical Industry.

P68 RNA helicase promotes invasion of glioma cells through negatively regulating DUSP5.

Gliomas are the most common central nervous system tumors. They show malignant characteristics indicating rapid proliferation and a high invasive capacity and are associated with a poor prognosis. In our previous study, p68 was overexpressed in glioma cells and correlated with both the degree of glioma differentiation and poor overall survival. Downregulating p68 significantly suppressed proliferation in glioma cells. Moreover, we found that the p68 gene promoted glioma cell growth by activating the nuclear factor-κB signaling pathway by a downstream molecular mechanism that remains incompletely understood. In this study, we found that dual specificity phosphatase 5 (DUSP5) is a downstream target of p68, using microarray analysis, and that p68 negatively regulates DUSP5. Upregulating DUSP5 in stably expressing cell lines (U87 and LN-229) suppressed proliferation, invasion, and migration in glioma cells in vitro, consistent with the downregulation of p68. Furthermore, upregulating DUSP5 inhibited ERK phosphorylation, whereas downregulating DUSP5 rescued the level of ERK phosphorylation, indicating that DUSP5 might negatively regulate ERK signaling. Additionally, we show that DUSP5 levels were lower in high-grade glioma than in low-grade glioma. These results suggest that the p68-induced negative regulation of DUSP5 promoted invasion by glioma cells and mediated the activation of the ERK signaling pathway.

Increased expression of ABCB6 enhances protoporphyrin IX accumulation and photodynamic effect in human glioma.

BACKGROUND: Glioma recurrence usually occurs close to the tumor resection margins as a result of residual infiltrating glioma cells. 5-aminolevulinic acid (ALA) fluorescence-guided resection of gliomas has been demonstrated to enhance discrimination of tumor tissue and to improve survival. ALA-based photodynamic therapy is an effective albeit still experimental adjuvant treatment option for gliomas. However, insufficient protoporphyrin IX (PpIX) accumulation may limit the benefits of fluorescence-guided resection and photodynamic therapy. METHODS: We investigated the expression of the ATP-binding cassette transporter ABCB6, which regulates porphyrin synthesis, in surgical specimens from human gliomas and manipulated ABCB6 in human glioma cell lines. RESULTS: Our findings demonstrated that expression levels of ABCB6 were greatly elevated in human gliomas compared with normal brain tissues and correlated with World Health Organization histologic grade. A previously undescribed finding was that ABCB6 mRNA expression in solidly fluorescing tumor tissues was higher than that in vaguely fluorescing tumors, suggesting that ABCB6 may be at least in part responsible for PpIX accumulation in glioma cells. Accordingly, ABCB6 overexpression in glioma cell lines caused a marked increase in intracellular levels of PpIX, and was more sensitive to ALA-induced photodynamic therapy-events that could be prevented by silencing ABCB6 via siRNA treatment. CONCLUSIONS: Our findings indicate a crucial role of ABCB6 in ALA metabolism and accumulation of PpIX in glioma. ABCB6 overexpression is a potential approach to enhance accumulation of PpIX for optimizing the subjective discrimination of vague fluorescence and improving the efficacy of ALA-based photodynamic therapy.

Fungal-Algal Association Drives Lichens' Mutualistic Symbiosis: A Case Study with Trebouxia-Related Lichens.

Biotic and abiotic factors influence the formation of fungal-algal pairings in lichen symbiosis. However, the specific determinants of these associations, particularly when distantly related fungi are involved, remain poorly understood. In this study, we investigated the impact of different drivers on the association patterns between taxonomically diverse lichenized fungi and their trebouxioid symbiotic partners. We collected 200 samples from four biomes and identified 41 species of lichenized fungi, associating them with 16 species of trebouxioid green algae, of which 62% were previously unreported. The species identity of both the fungal and algal partners had the most significant effect on the outcome of the symbiosis, compared to abiotic factors like climatic variables and geographic distance. Some obviously specific associations were observed in the temperate zone; however, the nestedness value was lower in arid regions than in cold, polar, and temperate regions according to interaction network analysis. Cophylogenetic analyses revealed congruent phylogenies between trebouxioid algae and associated fungi, indicating a tendency to reject random associations. The main evolutionary mechanisms contributing to the observed phylogenetic patterns were "loss" and "failure to diverge" of the algal partners. This study broadens our knowledge of fungal-algal symbiotic patterns in view of Trebouxia-associated fungi.

Microbial communities and their correlation with flavor compound formation during the mechanized production of light-flavor Baijiu.

Light-flavor Baijiu fermentation is a typical spontaneous solid-state fermentation process fueled by a variety of microorganisms. Mechanized processes have been increasingly employed in Baijiu production to replace traditional manual operation processes, however, the microbiological and physicochemical dynamics in mechanized processes remain largely unknown. Here, we investigated the microbial community succession and flavor compound formation during a whole mechanized fermentation process of light-flavor Baijiu using the conventional dilution plating method, PacBio single-molecule real-time (SMRT) sequencing and headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry. The results showed that largely different fungal and bacterial communities were involved in the soaking and fermentation processes. A clear succession from Pantoea agglomerans to Bacillus (B.) smithii and B. coagulans in dominant bacterial species and from Cladosporium exasperatum to Saccharomyces cerevisiae and Lichtheimia ramosa in dominant fungal species occurred in the soaking processes. In the fermentation process, the most dominant bacterial species was shifted from Pantoea agglomerans to Lactobacillus (La.) acetotolerans and the most dominant fungal species were shifted from Lichtheimia ramose and Rhizopus arrhizus to Saccharomyces cerevisiae. The bacterial and fungal species positively associated with acidity and the formation of ethanol and different flavor compounds were specified. The microbial species exhibited strong co-occurrence or co-exclusion relationships were also identified. The results are helpful for the improvement of mechanized fermentation process of light-flavor Baijiu production.

Microbial Community Affects Daqu Quality and the Production of Ethanol and Flavor Compounds in Baijiu Fermentation.

Daqu is a traditional starter for Baijiu fermentation and is produced by spontaneous fermentation of ground and moistened barley or wheat. The quality of Daqu is traditionally evaluated based on physicochemical and subjective sensory parameters without microbiological analysis. Here, we compared the physicochemical characteristics of qualified (QD) and inferior (ID) Daqu, their microbial communities based on plate counting and PacBio SMRT sequencing of rRNA gene libraries, and their impacts on Baijiu fermentation. The results showed that the glucoamylase and α-amylase activities of QD were significantly higher than those of ID. The counts of yeasts and relative abundances of functional microbes, especially the amylolytic bacterium Bacillus licheniformis and fungi Saccharomycopsis fibuligera and Lichtheimia ramosa, were significantly higher in QD than in ID. The laboratory-scale Baijiu fermentation tests showed that the relative abundances of the amylolytic microbes were higher in the QD than the ID fermentation set, resulting in more efficient fermentation, as indicated by more weight loss and higher moisture content in the former. Consequently, more glycerol, acetic acid, ethanol, and other volatile compounds were produced in the QD than in the ID fermentation set. The results suggest that Daqu quality is determined by, and can be evaluated based on, its microbial community.

The Ecology and Evolution of the Baker's Yeast Saccharomyces cerevisiae.

The baker's yeast Saccharomyces cerevisiae has become a powerful model in ecology and evolutionary biology. A global effort on field survey and population genetics and genomics of S. cerevisiae in past decades has shown that the yeast distributes ubiquitously in nature with clearly structured populations. The global genetic diversity of S. cerevisiae is mainly contributed by strains from Far East Asia, and the ancient basal lineages of the species have been found only in China, supporting an 'out-of-China' origin hypothesis. The wild and domesticated populations are clearly separated in phylogeny and exhibit hallmark differences in sexuality, heterozygosity, gene copy number variation (CNV), horizontal gene transfer (HGT) and introgression events, and maltose utilization ability. The domesticated strains from different niches generally form distinct lineages and harbor lineage-specific CNVs, HGTs and introgressions, which contribute to their adaptations to specific fermentation environments. However, whether the domesticated lineages originated from a single, or multiple domestication events is still hotly debated and the mechanism causing the diversification of the wild lineages remains to be illuminated. Further worldwide investigations on both wild and domesticated S. cerevisiae, especially in Africa and West Asia, will be helpful for a better understanding of the natural and domestication histories and evolution of S. cerevisiae.

Genetic diversity and population structure of the amylolytic yeast Saccharomycopsis fibuligera associated with Baijiu fermentation in China.

The amylolytic yeast Saccharomycopsis fibuligera is a predominant species in starters and the early fermentation stage of Chinese liquor (Baijiu). However, the genetic diversity of the species remains largely unknown. Here we sequenced the genomes of 97 S. fibuligera strains from different Chinese Baijiu companies. The genetic diversity and population structure of the strains were analyzed based on 1,133 orthologous genes and the whole genome single nucleotide polymorphisms (SNPs). Four main lineages were recognized. One lineage contains 60 Chinese strains which are exclusively homozygous with relatively small genome sizes (18.55-18.72 Mb) and low sequence diversity. The strains clustered in the other three lineages are heterozygous with larger genomes (21.85-23.72 Mb) and higher sequence diversity. The genomes of the homozygous strains showed nearly 100% coverage with the genome of the reference strain KPH12 and the sub-genome A of the hybrid strain KJJ81 at the above 98% sequence identity level. The genomes of the heterozygous strains showed nearly 80% coverage with both the sub-genome A and the whole genome of KJJ81, suggesting that the Chinese heterozygous strains are also hybrids with nearly 20% genomes from an unidentified source. Eighty-three genes were found to show significant copy number variation between different lineages. However, remarkable lineage specific variations in glucoamylase and α-amylase activities and growth profiles in different carbon sources and under different environmental conditions were not observed, though strains exhibiting relatively high glucoamylase activity were mainly found from the homozygous lineage.

Adaptive Gene Content and Allele Distribution Variations in the Wild and Domesticated Populations of Saccharomyces cerevisiae.

Recent studies on population genomics of Saccharomyces cerevisiae have substantially improved our understanding of the genetic diversity and domestication history of the yeast. However, the origin of the domesticated population of S. cerevisiae and the genomic changes responsible for ecological adaption of different populations and lineages remain to be fully revealed. Here we sequenced 64 African strains from various indigenous fermented foods and forests in different African countries and performed a population genomic analysis on them combined with a set of previously sequenced worldwide S. cerevisiae strains representing the maximum genetic diversity of the species documented so far. The result supports the previous observations that the wild and domesticated populations of S. cerevisiae are clearly separated and that the domesticated population diverges into two distinct groups associated with solid- and liquid-state fermentations from a single ancestor. African strains are mostly located in basal lineages of the two domesticated groups, implying a long domestication history of yeast in Africa. We identified genes that mainly or exclusively occur in specific groups or lineages and genes that exhibit evident group or lineage specific allele distribution patterns. Notably, we show that the homing endonuclease VDE is generally absent in the wild but commonly present in the domesticated lineages of S. cerevisiae. The genes with group specific allele distribution patterns are mostly enriched in functionally similar or related fundamental metabolism processes, including the evolutionary conserved TOR signaling pathway.

Occurrence and Molecular Identification of Wild Yeasts from Jimma Zone, South West Ethiopia.

Yeasts are common inhabitants of most fruit trees' rhizospheres and phyllospheres. Wild yeasts are the major driving force behind several modern industrial biotechnologies. This study focused on determining the occurrence and frequency of wild yeasts associated with domestic and wild edible tree barks, fruits, and rhizosphere soil samples collected over two seasons (i.e., spring and summer) in South West Ethiopia. A total of 182 yeast strains were isolated from 120 samples. These strains belonged to 16 genera and 27 species as identified based on the sequence analysis of the D1/D2 domain of the large subunit (26S) ribosomal RNA gene. Candida blattae, Pichia kudriavzevii, Candida glabrata, Saccharomyces cerevisiae, and Candida humilis were the most dominant yeast species isolated from the bark samples. Only Pichia kudriavzevii was regularly detected from the bark, rhizosphere, fruit, and sugarcane samples. The retrieval of yeasts from bark samples was more frequent and diverse than that of soil, fruits, and sugarcane. The frequency of detection of yeasts during the spring was significantly higher than in the summer season. However, there was no significant seasonal variation in the frequency of detection of yeast species between the rhizosphere and phyllosphere samples.

[Study on fitting growth density and best harvest time of Tribulus terrestris].

OBJECTIVE: To study the suitable growth density and the optimal harvest time of Tribulus terrestris. METHOD: Four growth densities were set with 60 cm breadth ridge and individual distance of 10, 20, 30 and 50 cm. The yield per individual and per unit area under the different growth densities were determined. Using yam saponin as a standard substance, the total saponin of T. terrestris was determined by UV spectrophotometry. RESULT: The individual yield decreased with the density increase, but the difference between 30 cm and 50 cm individual distance was not substantial. The yield per unit area increased with density increase, and the difference between all densities was significant. The yield peak was in the last ten-day of August. The best leaves area index was 1.4 at the growth peak time. The total saponins content reached peaks respectively in the last ten-day of June and August, but the peak in last ten-day of August was consistent with the one of yield per unit area, and the total ashes content was the lowest at the same time. CONCLUSION: The suitable growth density is 10 cm individual distance on the 60 cm breadth ridge. The optimal harvest time is in the last ten-day of August. The yield per unit area is 1 400 - 2 000 kg x hm(-2).

[Degradation characteristics of swamps in Zoige Plateau induced by drainage based on quantitative classification of vegetation].

Based on the field survey of swamp ecological characteristics and environmental quality and the ecological investigation of drained swamp transects in Zoige Plateau of Tibet in 2009, twenty typical swamp plots in the Plateau were classified into three types by TWINSPAN, i. e., primary swamp, long-term drained degraded swamp, and short-term drained degraded swamp, and each type of the degraded swamps was divided into three degradation grades, i. e., light degradation, moderate degradation, and severe degradation, with the degradation characteristics of vegetation and soil along the swamp degraded gradient studied. The swamp degradation in the Plateau was mainly driven by drainage pattern, drainage intensity, and soil moisture gradient, and the vegetation degradation was more obvious than the soil degradation. In the vegetation degradation, the structural change of hydro-type functional assemblage was most obvious, e. g., the importance value of helophytes under the stress of long-term drainage and short-term drainage decreased from 0.920 to 0.183 and 0.053, while that of mesophytes increased from 0.029 to 0.613 and 0.686, respectively. The soil response to the swamp degradation was in hysteresis, i. e., the soil physical and chemical properties presented definite variations but the differences were not significant among the swamps with different grades of degradation. The results of CCA indicated that soil moisture and nitrogen and potassium contents were the most important factors affecting the plant species distribution in drained degraded swamps in Zoige Plateau.

[Spatial patterns of plant species diversity in a degraded successional series of fragmented Leymus chinensis meadow in Songnen Plain of Northeast China].

Species diversity pattern is a hot topic in the forward field of international biodiversity research. Taking a degraded successional series (six communities distributed in 144 isolated patches) of fragmented Leymus chinensis meadow in Songnen Plain as test objects, this paper studied the spatial patterns of alpha, beta, and gamma diversities and related mechanisms. Eighty seven plant species were recorded, but none of them were distributed across all patches. L. chinensis + Kalimeris integrifolia community or L. chinensis community had higher alpha, beta, and gamma diversities, with more rare species and endemic species, while Kochia sieversiana community had lower alpha, beta, and gamma diversities, with few rare species and no endemic species. The gamma diversity showed significant positive correlation with a diversity, but no correlation with beta diversity. There was a significant power function correlation between the a diversity and the area of single patch of the six communities, but no correlation between the beta diversity (Bray-Curtis similarity index, Sjk ) and the area of single patch of the communities except for L. chinensis + K. integrifolia. Both the alpha angamma diversity showed significant correlations with the mean patch area and total patch area, but the p3beta diversity did not. The higher the species richness in the communities, the more the rare species and endemic species, the larger the possibility of species depopulation in local patches was. The importance of beta diversity for shaping species diversity pattern was related to the degree of habitat fragmentation.

Differential Islet-1 expression among lumbosacral spinal motor neurons in prenatal mouse.

Onuf's nucleus in the lumbosacral spinal cord, comprising somatic motoneurons that innervate the pelvic floor muscles via the pudendal nerve, shares some characteristics with the autonomic preganglionic neurons and functions in coordination with the autonomic nervous system. In mouse, neurons projecting to the urethral sphincter and ischiocavernosus muscles form the dorsolateral (DL) nucleus at the caudal lumbar levels, whereas neurons projecting to the limb and hip joint muscles comprise the retrodorsolateral and ventral nucleus, as well as the DL nucleus at the rostral lumbar levels. The results of the present study in mouse revealed that the expression pattern of a LIM homeodomain protein Islet-1, an embryonic marker for motoneurons in the spinal cord, was different among motoneuronal groups at the prenatal stage (embryonic days 13.5-15.5); the highest expression was observed in the DL at the caudal lumbar cord, whereas there was little expression in the lateral part of the rostral DL. Islet-1 expression was also observed in the parasympathetic preganglionic neurons at the sacral spinal cord. These findings provide evidence that the DL neurons at the caudal lumbar cord, corresponding to Onuf's nucleus, are chemically distinct among the motoneuronal groups at the prenatal stages. This differential Islet-1 expression among the motoneuronal groups suggests that Islet-1 not only leads to a motoneuronal lineage, but also to the differentiation of motoneuronal subsets in the lumbosacral spinal cord.

[Structural characteristics of Hemiptelea davidii community on Kerqin sandy land].

An investigation was made on the species composition and eco-type of Hemiptelea davidii community on Kerqin sandy land. The results showed that in study area, no shrub layer existed in H. davidii forest, while arbor layer could be divided into two sub-layers, with a height of 4.05-7.86 m for the upper layer, and of 2.05-3.20 m for the lower layer. In the community, there were 32 herbal species belonging to 27 genera of 13 families and dominant by Poaceae, Fabaceae and Compositae, and 11 areal types, among which, the species number in Mongolian-Northeastern China-Dahuricia-North China areal type was the highest (34.38%), followed by that in Northeastern China-North China areal type (12.5%). Among three water ecological types, mesophytes occupied 59.37% of the total, and mesoxerophytes and xerophytes occupied 25% and 15.63%, respectively. Hemicryptophytes had a larger amount in six life forms, accounting for 31.25% of the total. H. davidii community had the typical life form characteristics of temperate steppe.