Ethanol resistance in Drosophila melanogaster has increased in parallel cold-adapted populations and shows a variable genetic architecture within and between populations

Understanding the genetic properties of adaptive trait evolution is a fundamental crux of biological inquiry that links molecular processes to biological diversity. Important uncertainties persist regarding the genetic predictability of adaptive trait change, the role of standing variation, and whether adaptation tends to result in the fixation of favored variants. Here, we use the recurrent evolution of enhanced ethanol resistance in Drosophila melanogaster during this species’ worldwide expansion as a promising system to add to our understanding of the genetics of adaptation. We find that elevated ethanol resistance has evolved at least three times in different cooler regions of the species’ modern range - not only at high latitude but also in two African high altitude regions - and that ethanol and cold resistance may have a partially shared genetic basis. Applying a bulk segregant mapping framework, we find that the genetic architecture of ethanol resistance evolution differs substantially not only between our three resistant populations, but also between two crosses involving the same European population. We then apply population genetic scans for local adaptation within our quantitative trait locus regions, and we find potential contributions of genes with annotated roles in spindle localization, membrane composition, sterol and alcohol metabolism, and other processes. We also apply simulation-based analyses that confirm the variable genetic basis of ethanol resistance and hint at a moderately polygenic architecture. However, these simulations indicate that larger-scale studies will be needed to more clearly quantify the genetic architecture of adaptive evolution, and to firmly connect trait evolution to specific causative loci.

Effect of Polymerizable Emulsifiers on the Properties of Polyacrylate Latexes and the Relative Waterborne Inks

Polyacrylate latexes made from non-polymerizable emulsifiers and their inks typically suffer from poor ethanol resistance stability and low adhesion on biaxially oriented polypropylene (BOPP) and polyethylene (PE) films. In this contribution, a composite emulsifier system containing a polymerizable anionic and a polymerizable non-ionic emulsifier was used to synthesize core-shell polyacrylate latexes. Additionally, a control latex was also prepared using a traditional emulsifier TX-30 to replace the polymerizable non-ionice mulsifier in the above composite emulsifiers. The effect of the polymerizable emulsifier on ethanol resistance, Ca 2+ resistance stability, and adhesion on PE and BOPP films of the latexes and the inks, and water resistance of the latex films and ink films were studied. The results show that, when compared with the control latex, the one made from double polymerizable compound emulsifier system and its ink demonstrates better ethanol resistance, higher stability of calcium ions and higher adhesion on BOPP and PE. When the ratio of anionic emulsifier to non-ionic emulsifier is 1.5/1 and the total dosage is 2.5 wt%, the latex showed the best comprehensive performance. The calcium ion resistance stability of the latex increased from 5% of the control latex to 25%. Accordingly, the adhesion of yellow ink on BOPP film increased from 92% of the ink based on the control latex to 99% and increased from 99% to 100% on PE film. The adhesion of blue ink on BOPP film increased from 92% to 99%, and from 99% to 100% on PE film. These results indicate that the fully polymerizable emulsifiers can effectively improve the properties of latex.

Adaptive Laboratory Evolution of Native Torulaspora delbrueckii YCPUC10 With Enhanced Ethanol Resistance and Evaluation in Co-inoculated Fermentation

Torulaspora delbrueckii is a yeast species typically present in the early stages of the fermentation process. T. delbrueckii positively modifies the aromatic properties of wines. However, its contribution to the final quality of the wine is restricted by its low tolerance to ethanol. T. delbrueckii is capable of fermenting and tolerating an ethanol concentration ranging from 7.4% (v/v) to slightly higher than 9% (v/v). For this reason, it cannot complete fermentation, when alcohol reach levels higher than 12% (v/v), limiting their use in the industry. The objective of this work was to obtain new variants of T. delbrueckii with improved resistance to ethanol through adaptive laboratory evolution. Variants capable of tolerating ethanol levels of 11.5% (v/v) were obtained. These presented improved kinetic parameters, and additionally showed an increase in resistance to SO2 in ethanol compared to the original strain. Co-inoculated fermentations were performed with the original strain (FTd/Sc) and with the evolved strain (FTdF/Sc), in addition to a control fermentation using only Saccharomyces cerevisiae EC1118 (FSc). The results obtained show that FTdF/Sc present higher levels of 2-Ethylhexanol, compared to FTd/Sc and FSc. Furthermore, FTdF/Sc presents higher levels of total alcohols, total aldehydes, total phenolic derivatives, and total sulfur compounds with significant differences with FSc. These results provide a T. delbrueckii YCPUC10-F yeast with higher resistance to ethanol, which can be present throughout the fermentation process and be used in co-inoculated fermentations. This would positively impact the performance of T. delbrueckii by allowing it to be present not only in the early stages of fermentation but to remain until the end of fermentation.

Preliminary Characterization of Yeasts from Bombino Bianco, a Grape Variety of Apulian Region, and Selection of an Isolate as a Potential Starter

Eighty-seven yeasts were isolated from Bombino bianco, a white grape variety from Apulian Region (Southern Italy). The isolates were characterized for the splitting of arbutin, the hydrolysis of pectins, sulphite production, the resistance to acetic acid, SO2, and ethanol. An enhanced arbutin splitting (β-glucosidase) and a moderate pectolytic activity were found. Concerning ethanol resistance, the most of yeast population showed a low-to-moderate resistance, but some isolates, identified as Saccharomyces cerevisiae, were able to grow in presence of 15% v/v of ethanol. Four isolates were selected (coded as 43D, 44D, 45D, and 46D), studied for their ability to decarboxylate amino acids and used in small-scale fermentation trial; for this last experiment a reference strain was used (S. cerevisiae EC1118). This experiment suggested the existence of an isolate (S. cerevisiae 46D) with interesting traits and performances, which could be potentially proposed as a starter for Bombino bianco.

Heterologous expression of ctsR from Oenococcus oeni enhances the acid-ethanol resistance of Lactobacillus plantarum

Oenococcus oeni is a lactic acid bacterium that is widely used in wine-making to conduct malolactic fermentation (MLF). During MLF, O. oeni undergoes acid and ethanol stress that impairs its growth. In order to investigate the role that the ctsR gene plays in acid-ethanol stress, the ctsR gene from O. oeni was expressed heterologously in L. plantarum. The transcription level of the ctsR gene and 10 additional stress response genes in L. plantarum were analyzed by RT-qPCR. Physiological assays to assess ROS accumulation, cell membrane integrity, intracellular ATP and GSH levels, Ca2+/Mg2+-ATPase and Na+/K+-ATPase activities were also performed. Results showed that the recombinant strain WCFS1-CtsR exhibited stronger growth performance than the control strain WCFS1-Vector, and the expression of ctsR, clp, and hsp genes were significantly increased under acid-ethanol stress. Furthermore, WCFS1-CtsR displayed 1.08-, and 1.39-fold higher ATP and GSH concentrations, respectively, compared to the corresponding values for WCFS1-Vector under acid-ethanol stress. ROS accumulation and PI value of WCFS1-CtsR were decreased by 46.52% and 42.80%, respectively, compared to the control strain. In addition, the two ATPase activities in WCFS1-CtsR increased significantly compared with WCFS1-Vector. This is the first report demonstrating that ctsR gene enhances the acid-ethanol tolerance of L. plantarum.