酿酒酵母萌发突变株的筛选

Isolation of Germination Mutants inSaccharomyces cerevisiae

DOI：10.3969/j.issn.1673-1689.2019.12.013

中文关键词: 酿酒酵母 萌发 孢子 筛选

英文关键词: Saccharomyces cerevisiae,germination,spore,isolate

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中文摘要:

酵母细胞孢子作为微胶囊首次运用于酶的固定化，但是由于其孢子遇到营养物质即萌发的性质，导致固定化酶的重复利用率受到限制，因此筛选有萌发缺陷的酵母细胞孢子突变株对微胶囊的应用至关重要。利用EMS对酵母细胞进行诱变再经过乙醚的筛选获得4株萌发突变株AN120-m5、AN120-m8、AN120-m10和AN120-m17。验证后发现他们均产孢正常，除了AN120-m5生长有些弱之外，其余3株生长都正常。后对这些突变株进行了液体培养基以及固体培养基内的验证，实验发现所有突变株都表现出明显的萌发缺陷，其中AN120-m10孢子在液体培养基中萌发率只有1%，而AN120-m5单个孢子在固体培养基上则完全不萌发。由于AN120-m5单个孢子不能萌发因此得不到杂交后代，而其余3株突变株在与野生型杂交三代后，发现野生型孢子与突变型孢子萌发状况相比始终呈现2∶2的萌发优劣比，从而可以推测突变株是单基因突变菌株，为后续研究确定突变基因从而筛选得到完全不萌发菌株打下基础。

英文摘要:

The spores ofSaccharomyces cerevisiaefirstly were applied for enzyme immobilization as microcapsule. The utilization of the enzyme has been limited due to germination of spores when treated with nutrients，thus finding germination mutant yeast spore has a significant effect on applying of microcapsule. We used EMS as a mutagen for yeast cells and screened by ether and isolated 4 mutants，AN120-m5，AN120-m8，AN120-m10 and AN120-m17. They all can sporulate and grew well except that AN120-m5 was slightly deficient in cell growth. When verifying the germination efficiency of 4 mutants in the liquid medium and on solid medium，4 mutants all exhibited obvious germination defect，however only 1% AN120-m10 spores germinated in the liquid medium，and single spore of AN120-m5 completely cannot germinate on solid medium. Hybrid cells of AN120-m5 cannot be collected because of its defect on solid medium，and other 3 mutant haploid cells crossed with wide type haploid cells after 3 generations，and spores of wild type exhibited a germination eudismic ratio of 2∶2 compared with mutant spores. This suggested that a single gene was mutated in mutant strains will facilitate to find mutated gene in later screening in order to isolate a strain that completely doesn’t germinate.

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