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Spontaneous fermentation is still used in numerous wineries, but in the last twenty years, yeast- manufacturing companies have developed selected starter cultures. These strains are now widely used to reduce the risk of wine spoilage, prevent stuck fermentation and improve wine quality. At present, four main strategies can be used to obtain such optimized strains. Firstly, pure strain clones may be isolated from spontaneous fermenting must. Manual Finder; Needle Guide; Troubleshooter; Timing Test; E-Book; Contact Us; Timing Test. Example of the timing test performed on a machine with a top-loading bobbin. Find great deals on eBay for Minnesota Sewing Machine in Antique Sewing Machines. Shop with confidence. ![]() ![]() Previous article in issue: Expression of stress response genes in wine strains with different fermentative behavior Previous article in issue: Expression of stress. This is the main strategy used at present . 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Although very efficient, the low level of acceptance of this technology by consumers has limited its impact . Thirdly, mutagenesis may be used to widen the natural genetic polymorphism. This strategy is an essential approach to optimize bacteria and fungi, and has proved to be very efficient for industrial purposes. As wine yeast strains are not haploid and since most mutations are recessive, it is not likely to be efficient in the short term, but efforts in this direction have been made. Fourthly, as yeasts are eukaryotes with standard Mendelian genetic behavior, the strategies which have been successfully developed for plants and animals could be applied to yeasts. Specifically, crosses and progeny analysis could theoretically be used to improve genotypes, thereby accumulating general and specific properties in a strain. Unfortunately many wine yeasts are homothallic. Homothallic haploid cells (HO) are able to switch their mating type and to conjugate with cells of the same single spore colony. Consequently, hybridization techniques for developing new strains have proved elusive and have required the use of a micromanipulator to achieve direct spore- to- spore mating . Several attempts have already been made to this end . To rationalize the latter strategy, the first requirement is to try to establish the importance of the genetic determinism of the enological parameters of yeast. The availability of relevant and reliable phenotypic tests to screen a large population of yeast strains in laboratory conditions is the prerequisite condition to appreciate the contribution of genetics in different characters. Simple phenotypes like hydrogen sulphide production, flocculation or killer activity . However, more complex traits such as ethanol production . The continuous quantitative variation of these traits within wine yeast populations can in part be tentatively explained by a polygenic determinism. However, the environmental conditions of fermentation (i. For this purpose, reliable and relevant laboratory fermentations measuring volatile- acidity production and ethanol tolerance in enological conditions were established. We also showed how the meiotic segregation of industrial yeast strains can provide new and interesting genetic variants without screening natural isolates. These strains presenting unique karyotypes could be readily commercialized or used in breeding programs. Materials and methods. Strains, media and growth conditions. All yeast strains are referenced as Saccharomyces cerevisiae (Table 1). Laboratory strains X2. A (MAT a) and X- 2. B (MAT . The wine yeasts were strains isolated from native microflora of spontaneous fermentations. Yeast was grown at 3. Ascospores were isolated by a micromanipulator Singer MSM Manual on YPD- agar. Ascus wall was digested using cytohelicase (Sigma) adjusted to 2 mg ml. Germination efficiency was expressed as the percentage of isolated spores forming a colony after three days at 3. Mating types were defined by microscopic observation of zygote formation with either tester strain X2. A or X2. 18. 0 1. B. For this purpose, cells of analyzed strains were mixed with each tester and then incubated on YPD- agar for 6–1. Pulse- field gel electrophoresis. Chromosomal DNA was prepared from overnight cultures of yeasts in agarose plugs as described by Bellis et al. Chromosomes were separated with a CHEF DRII apparatus (Bio- Rad, Richmond, California) on a 1% agarose gel (Qbiogene, Carlsbad, CA, USA). Electrophoresis was carried out at 2. V and 1. 4 . Chromosomal DNA of X2. A was used as S. Five levels of color were used: 1 – white, 2 – light brown, 3 – brown, 4 – dark brown, 5 – black. 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