Dynamic genome-scale modeling of Saccharomyces cerevisiae unravels mechanisms for ester formation during alcoholic fermentation

Fermentation employing Saccharomyces cerevisiae has produced alcoholic beverages and bread for millennia. More recently, S. cerevisiae has been used to manufacture specific metabolites for the food, pharmaceutical, and cosmetic industries. Among the most important of these metabolites are compounds associated with desirable aromas and flavors, including higher alcohols and esters. Although the physiology of yeast has been well-studied, its metabolic modulation leading to aroma production in relevant industrial scenarios such as winemaking is still unclear. Here we ask what are the underlying metabolic mechanisms that explain the conserved and varying behavior of different yeasts regarding aroma formation under enological conditions? We employed dynamic flux balance analysis (dFBA) to answer this key question using the latest genome-scale metabolic model (GEM) of S. cerevisiae. The model revealed several conserved mechanisms among wine yeasts, for example, acetate ester formation is dependent on intracellular metabolic acetyl-CoA/CoA levels, and the formation of ethyl esters facilitates the removal of toxic fatty acids from cells using CoA. Species-specific mechanisms were also found, such as a preference for the shikimate pathway leading to more 2-phenylethanol production in the Opale strain as well as strain behavior varying notably during the carbohydrate accumulation phase and carbohydrate accumulation inducing redox restrictions during a later cell growth phase for strain Uvaferm. In conclusion, our new metabolic model of yeast under enological conditions revealed key metabolic mechanisms in wine yeasts, which will aid future research strategies to optimize their behavior in industrial settings.     

The prevalence of root surface caries amongst Irish adults

In a national survey of adult dental health conducted in the Republic of Ireland in 1989/90 a total of 1,527 subjects aged 25 and older were examined for root surface caries. It was found that the prevalence of root surface caries was highest in older age groups and also amongst males, residents of non-fluoridated communities and those earning low incomes. Tooth loss masked the potential prevalence of root surface caries. With more persons retaining their natural teeth into middle and old age the prevalence of root surface caries is likely to increase in the future.     

Future directions for root caries research

Interest among researchers in the diagnosis, aetiology, prevention, and treatment of root caries has increased substantially over the past two decades. However, there are some fundamental problems impeding the advancement of the field which remain to be addressed and resolved. A universally acceptable definition of root caries is not yet available. The relationship of root caries to coronal caries has not been established. The underlying disease process is still not clearly understood. The optimal utilisation of preventive/therapeutic agents for the treatment or prevention of root caries has not been determined. New treatment materials and preventive agents have not yet been tested in controlled clinical trials. These are a few of the issues and problems which we address in this paper.     

Anti-CRISPR Discovery: Using Magnets to Find Needles in Haystacks

CRISPR-Cas immune systems in bacteria and archaea protect against viral infection, which has spurred viruses to develop dedicated inhibitors of these systems called anti-CRISPRs (Acrs). Like most host-virus arms races, many diverse examples of these immune and counter-immune proteins are encoded by the genomes of bacteria, archaea, and their viruses. For the case of Acrs, it is almost certain that just a small minority of nature’s true diversity has been described. In this review, I discuss the various approaches used to identify these Acrs and speculate on the future for Acr discovery. Because Acrs can determine infection outcomes in nature and regulate CRISPR-Cas activities in applied settings, they have a dual importance to both host-virus conflicts and emerging biotechnologies. Thus, revealing the largely hidden world of Acrs should provide important lessons in microbiology that have the potential to ripple far beyond the field.     

Seasonal dynamics of the association between sweet potato and vesicular-arbuscular mycorrhizal fungi

To better understand the behavior of selected vesicular-arbuscular mycorrhizal (VAM) isolates in the field, we documented the growth of roots, root hairs, and VAM colonization of inoculated and noninoculated sweet potato plants (Ipomea batatas (L.) Lam. cv White Star) over a growing season. We also determined the seasonal dynamics of P and Zn uptake, and shoot and storage-root growth. Shoot cuttings were inoculated with an isolate of either Glomus etunicatum Becker and Gerdemann or Acaulospora rugosa Mortan, or were not inoculated, and were harvested 2, 4, 8, 13, 20, and 27 weeks after planting (WAP). At each harvest, roots were sampled at 0 to 30, 30 to 60, and 60 to 90 cm depths and at 0, 23, 83, and 116 cm from the base of the shoot. At the end of the study, the roots of three non-inoculated plants were sampled by soil horizon. Inoculation had no affect on shoot growth or total shoot uptake of P and Zn; shoot dry mass and P and Z content increased rapidly up to 20 WAP, while shoot length continued to increase through 27 WAP. Shoot-P concentration of plants inoculated with A. rugosa at 2 and 8 WAP were higher than the noninoculated plants, while shoot-Zn concentration was not affected by inoculation. Storage-root yields of inoculated plants were higher than yields for noninoculated plants. Root length density, and percentage of root length with root hairs and VAM colonization were highest and most dynamic near the base of the plant. Percentage of root length colonization by VAM fungi was highest in the E2 horizon, intermediate in the Bh horizon, and lowest in the Ap horizon. Percentage of root length with root hairs had the opposite pattern. Intensive measurements of root characteristics close to the base of the plant, and shoot P-content and concentration during the period of rapid yield production, provided the most useful data for evaluating the activity of effective isolates.Published as Florida Agricultural Experimental Station Journal Series No. R-02576     

乌云伞的解剖学及花粉形态学研究

八角莲属(Dysosma)是小檗科(Berberidaceae)一个重要属,全国共有7个种,大约分布于北纬23—32°,东经94—122°之间。湖北八角莲属有4个种,其中乌云伞(Dysosma lichuanensis Z. Cheng, sp. nev., ined.)为内定种,是多年生草本,它的形态特征与八角莲(Dysosma     

Nitrogen and energy losses and methane emissions from beef cattle fed diets with gradual replacement of maize silage and concentrate with grass silage and corn-cob mix

Diets based on large proportions of grassland-based feed are uncommon in forage-based intensive beef production, thus contradicting governmental or commercial strategies to promote the use of grassland-based feed in ruminant production systems. Compared with typical maize silage/concentrate diets, grassland-based diets are associated with impaired nitrogen (N) and energy utilisation because of the comparably lower energy and higher CP content of these feeds. However, quantitative studies concerning the effects of increased dietary proportions of grassland-derived feeds on N and energy losses and utilisation and on methane emissions are missing and the compensation potential of using a limited proportion of an energy-rich forage is unknown. Therefore, we tested five diets with varying types and proportions of forage and concentrate. Three diets consisted of grass silage, maize silage, and concentrate in ratios of, g/kg DM, 100:600:300 (G100; control), 300:500:200 (G300), and 500:300:200 (G500), respectively. Two diets were composed of grass silage, corn-cob mix (CCM), and concentrate in ratios of, g/kg DM, 500:300:200 (G500_CCM), and 750:150:100 (G750_CCM), respectively. A high-protein concentrate (270 g CP/kg DM) was fed to G100, whereas a low-protein concentrate (140 g CP/kg DM) was used in the remaining diets. Diets were fed throughout the entire fattening period to groups of six Limousin-crossbred bulls each. When weighing 246 ± 18 kg, each animal underwent a 7-day total daily faeces and urine collection, which was followed by measuring methane emissions in respiration chambers for 48 h. Total DM intake was similar across all diets, whereas the N intake varied (P < 0.05). Urinary N loss (g/day) was the highest for G750_CCM (28.2) and G100 (26.6) and lowest for G500_CCM (15.2) and G300 (16.9) (P < 0.001). Energy utilisation was comparable among all groups. Metabolisable energy intake decreased numerically only with increasing proportions of grass silage in the diet. Substituting maize silage with CCM counteracted the loss in metabolisable energy intake. Absolute methane emissions were not different across the groups, but methane emission intensity (mg/g body protein retention) varied (P < 0.05), being numerically lower for G100 (349) and G500_CCM (401) compared with the other groups (488 on average). In conclusion, the results show that the grass silage proportion in beef cattle diets can be substantially increased when strategically combined with energy-dense forages, such as CCM. This also limits the need for concentrate and additional protein sources; in addition, the associated urinary N emissions, which are potentially noxious to the environment, are avoided.     

GENETIC EVIDENCE FOR MULTIPLE ORIGINS OF DIOECY IN THE HAWAIIAN SHRUB WIKSTROEMIA (THYMELAEACEAE)

Dioecy is unusually common in the Hawaiian Islands, yet little is known about the evolutionary biology of this breeding system. A native shrub, Wikstroemia, has an unusually diverse array of breeding systems: two forms of dioecy, cryptic and morphological dioecy, as well as hermaphroditism (perfect flowers). The existence of two forms of dioecy is significant for three reasons: 1) the presence of cryptic unisexuals that are functionally unisexual, but retain the appearance of hermaphroditism in both sexes, is strong evidence for the ancestral status of hermaphroditism; 2) the production of nonfunctional pollen, by female cryptic unisexuals, is a new instance of a phenomenon which has previously been reported for a few other species; 3) the two forms of dioecy are morphological markers which are useful in hybridization studies for tracing the genetic basis of their inheritance. Crosses were made between cryptically unisexual individuals (C), between morphologically unisexual individuals (M), and between the two types of unisexuality. The offspring of crosses between individuals with the same sex type usually resulted in offspring with that sex type, but most of the progeny of between-sex type crosses were, unexpectedly, perfect-flowered hermaphrodites. These results show that genetic control of sex determination is not homologous in all populations, suggesting that dioecy has evolved at least twice in Hawaiian Wikstroemia. The genetic data further suggest that males are the heterozygous sex.     

Impact of cell disruption and polymer recycling upon aqueous two-phase processes for protein recovery

A practical study is presented of the influence of cell debris and polymer recycling upon the operation of two-stage acqueous two-phase systems (ATPS) for the recovery of yeast bulk protein, pyruvate kinase and fumarase. Brewers' yeast was disrupted using one of two types of high-pressure homogenisers or a bead mill. The different cell debris suspensions were partitioned in a single PEG-phosphate ATPS extraction and the efficiency of solid-liquid separation was examined. A continuously operated two-stage ATPS process, using spray columns, is presented and practical problems of polymer recycling are discussed. Conclusions are drawn concerning the generic implementation and operational stability of ATPS in practical protein recoveries.     

Transpiration and drought resistance of Douglas-fir seedlings exposed to excess ammonium

 In a pot trial growth and transpiration of 3-year-old Douglas-fir seedlings on an acid, sandy soil was examined at a deficient (30 kg N ha^{–  1} year^{–  1}) and an excessive level (120 kg N ha^{–  1} year^{–  1}) of NH₄ application. Dissolved ammonium sulphate was applied to the pots weekly for two growing seasons. In half of the pots a complete set of other nutrients was applied in optimal proportions to the applied nitrogen. Water supply was optimal and transpiration was recorded. At the end of the second treatment season irrigation was stopped for 2 weeks during dry and sunny weather. Both high application of NH₄ and additional nutrients increased shoot growth and transpiration demand in the first treatment year. The root system was smaller at higher N level and this reduced water uptake accordingly. In the second year the combination of high NH₄ ⁺ and additional nutrients affected root functioning predominantly due to salinity effects and this seriously decreased water uptake capacity and shoot water potentials, finally resulting in tree death. Without addition of other nutrients the high NH₄ ⁺ application resulted in a high degree of soil acidification, which damaged the roots, that showed a decrease in water uptake capacity. At the low NH₄ supply level soil acidification was lower, and root functioning was not affected, and the trees recovered quickly from the imposed drought. Higher needle K and P status depressed transpiration rates at the low NH₄ application rate. Received: 9 January 1995 / Accepted: 18 September 1995