摘要
【目的】产甘油假丝酵母作为一株优良高产甘油菌株,已成功应用于工业生产15年。近年来由于产甘油假丝酵母染色体倍性尚不明确,限制了对其进行遗传改造的研究进展,因而我们对产甘油假丝酵母染色体倍性进行研究,分析确定其染色体倍性。【方法】选用酿酒酵母细胞进行生孢,制备酿酒酵母单倍体细胞作对照,并选用热带假丝酵母作为二倍体酵母细胞对照,利用血球计数板得到热带假丝酵母、产甘油假丝酵母、单倍体及二倍体酿酒酵母细胞数,提取染色体,通过二苯胺检测法测定DNA含量。由于在相同紫外照射条件下单倍体细胞比二倍体细胞更容易死亡,因而进行紫外照射试验进行进一步验证。【结果】通过筛选得到所需的酿酒酵母单倍体细胞,并测定得到以上4种酵母单位细胞染色体DNA的含量,通过分析得出单细胞产甘油假丝酵母的染色体DNA含量为单倍体酿酒酵母细胞的1.82倍,为二倍体酿酒酵母的0.848倍,为热带假丝酵母的0.97倍,因而初步确定产甘油假丝酵母为二倍体;通过紫外照射进一步得到验证其为二倍体。【结论】本文通过测定细胞染色体DNA含量并经紫外照射验证分析得出Candida glycerinogenes细胞为二倍体,为对其进行进一步研究和改造奠定了一定的遗传学基础。
[Objective]To determine the chromosome DNA ploidy of Candida glycerinogenes.[Methods]We screened the haploid cell of Saccharomyces cerevisiae and the diploid of Candida tropicalis as the reference cell to identify the ploidy of Candida glycerinogenes.The concentrations of chromosome DNA extracted from both haploid and diploid cells were determined by the diphenylamine reaction method.Meanwhile,the cell number of Candida tropicalis,Candida glycerinogenes,Saccharomyces cerevisiae and its haploid were accounted by hemocytometer measurement method,respectively.Because in the same UV irradiation conditions,the haploid cells was more easy died than diploid cells,so we used the UV irradiation test to further verification.[Results]The results of the content of chromosome DNA in a single cell showed that Candida glycerinogenes was a diploid cell.Moreover,it was also further confirmed because the haploid cell of Saccharomyces cerevisiae was more sensitive than Candida glycerinogenes under UV treatment.[Conclusion]By this study,we confirmed that Candida glycerinogenes was a diploid cell.
出处
《微生物学报》
CAS
CSCD
北大核心
2011年第3期326-331,共6页
Acta Microbiologica Sinica
基金
国家“863计划”(2006AA020103,2009AA02Z210)~~
