热带假丝酵母产生子囊孢子的条件及单倍体分离的研究  

Study on Sporulation and Haploid Separation Conditions of Candida tropicalis

作　　者：严小琼[1,2] 黄思思[1,2] 覃香香[1] 周玉恒[1] 蔡爱华[1] 陈海珊[1] 张厚瑞[1] 

机构地区：[1]中国科学院广西植物研究所,广西植物功能物质研究与利用重点实验室,桂林541006 [2]广西师范大学,生命科学学院,桂林541004

出　　处：《基因组学与应用生物学》2013年第2期201-206,共6页Genomics and Applied Biology

基　　金：广西青年基金(桂科青0991074);广西自然科学基金项目(2010GXNSFA013060;2010GXNSFB013043)共同资助

摘　　要：本文研究了诱导热带假丝酵母子囊孢子产生和子囊破壁的条件,以及单倍体分离和鉴别方法。结果表明:NaAc、KCl这两种成分即可满足热带假丝酵母的产孢营养要求,在含有NaAc8.2g/L、KCl1.8g/L的琼脂培养基中,28℃培养7d,热带假丝酵母细胞的产孢率可达到47.5%;单纯使用蜗牛酶对破除热带假丝酵母子囊壁的效率甚低,酶解液加入适量的石英砂同时振荡处理4h左右,可以显著提高对热带假丝酵母子囊破壁速率。用这种方法处理,绝大多数子囊壁均可破除。子囊破壁处理后再以52℃处理10min杀死残余的营养体细胞,分离物的单倍体孢子比例可由灭活处理前的48%,提高到76%以上。The conditions of sporulation and enzymatic hydrolysis of Candida tropicalis as well as the separation and identification methods of haploid were researched in this research. The results showed that two ingredients, which are NaAc and KC1, can meet the nutritional requirements of the sporulation, when Candida tropicalis was cultured in the medium containing the NaAc 8.2 g/L, KC11.8 g/L, 7 days at 28℃, sporulation rate could reach 47.5%; It is ineffectiveness when only uses helicase to break the ascus wall of Candida tropicalis, while if right amount of quartz sand is added, and shaking culture for 4 hours, the sporulation rate was improved significant. With this method, the majority ofascus wall ofCandida tropicalis could be broken. Heat treatment 10 min at 52℃ after enzy- matic hydrolysis and the remanent vegetative cells were killed, the proportion of isolates haploid spore could reach from 48% to 76%.

关 键 词：热带假丝酵母 产孢条件 子囊孢子 

分 类 号：Q93-33[生物学—微生物学]