酿酒酵母单倍体1450原生质体制备的研究

Study on the Preparation of Protoplast on Haploid of Saccharomyces cerevisiae 1450

通过研究菌龄、菌体预处理、渗透压稳定剂及含有不同渗稳剂的再生平板等不同因素对酿酒酵母1450原生质体制备的影响,确定了其原生质体制备的条件:菌龄为6h,预处理剂为PB+0.1%EDTANa2+3%β-巯基乙醇,采用1mol/L山梨醇为渗透压稳定剂,再生培养基添加0.5mol/L蔗糖为稳定剂,在此条件下,其原生质体形成率达99.85%,再生率达14.31%。

The effects of different factors including culture time, preculture and osmotic stabilizers on the preparation and the regeneration of haploid ofsaccharomyces cerevisiae 1450 were studied. The optimum technical parameters were summed up as follows： 6 h culture of 1450 ceils in logarithmic growth phase, PB＋0.1% EDTANa2＋3 % β-mercaploethanol used in the prestreatment, 0.5 mol/L sucrose used as osmotic stabilizer during the protoplastformation and 1 mol/L D-sorbitol sorbitol used in the regeneration. Under the above conditions, the formation rate of the protoplast was 99.85 % and regeneration rate reached up to 14.31%.