酿酒酵母1450产孢培养基的筛选及单倍体的分离
摘要
诱导酵母产孢进而获得单倍体是进行酵母遗传学和育种工作极为关键的一步,它既是性状遗传分析所必需,又可为杂交作好亲本的准备。孢子的获得必需要提供适当的产孢条件,酿酒酵母营养体在营养缺乏等饥饿条件下,经过减数分裂由二倍体变为单倍体,形成子囊。在常用的酵母产孢培养基中没有一种产孢培养基能够适用于所有的酵母产孢。为此,本文在几种不同生孢培养基的条件下进行酿酒酵母1450的生孢实验,选择一种最适培养基进行酿酒酵母1450单倍体的分离,并采用发酵法筛选出一株性能良好的单倍体,供随后的育种实验使用。
主要研究结果如下:
1.通过对四种常用于酵母产孢的培养基的优选,确认Mcclary培养基为适宜的酿酒酵母1450的生孢培养基。
2.用Mcclary培养基对1450进行产孢培养的最适温度为25℃-28℃,培养时间为5-7d。温度对酵母生孢能力的影响与相关酶的活性有关。
3.在测定菌株的发酵力试验中,发现单倍体菌株的酒精发酵力差异较大,推测原因是酒酵母的发酵能力由多个基因控制。并且通过对酒精发酵力和酒精发酵性能的综合分析,得到一株性能良好的单倍体S-3,可以作为以后杂交实验的亲本。
4.对单倍体S-3进行了生长曲线的测定,结果表明在进行育种工作时选取9h左右的菌体较好。
The process of inducing yeast to produce spores and then obtaining its haploid is crucial in Yeast Genetics and Breeding. This is indispensable for analyzing the inheritance of acquired characters and preparing the parent strains for hybridization. The spores can be obtained only in fitting sporulation condition. On the barren condition of nutrition, the trophosome of S.Cerevisiae transits into haploid from the former diploid through the process of meiosis, forming sporangium. None of the present common yeast spores-breeding culture media was suitable for all the yeasts to produce their spores. Therefore, the spores-breeding experiment of S.Cerevisiae 1450 was undertaken under different spores-breeding conditions in this essay. Then, the optimum culture medium was selected for the separation of the haploid of S.Cerevisiae 1450. Next, a haploid strain with good biological function was screened out through zymotechnics method so as to be applied in the next breeding experiment.
The main research results are as follows.
1. After the selective preference of four common yeast spores-breeding culture media, Mcclary culture medium was confirmed as the optimum one for S.Cerevisiae 1450.
2. The optimum temperature and time for S.Cerevisiae 1450 in the Mcclary culture medium were 25℃-28℃and 5-7d, separately. The influence of temperature over the spores-breeding capability of S.Cerevisiae 1450 was connected with the activity of concerning enzymes.
3. In the experiment of identifying the ferment capability of strains, the ferment capability varies largely among different haploid strains. The supposed reason was speculated that the ferment capability of S.Cerevisiae was controlled by multiple genes. And through the comprehensive analysis of the fermentation capability and performance, a haploid strain S-3 with good biological function was obtained and could be used as the parent strains in the future hybridization experiment.
4. The growth curve of the haploid S-3 was determined, and the results showed that it’s better to choose the strains around 9h in the breeding job.
主要研究结果如下:
1.通过对四种常用于酵母产孢的培养基的优选,确认Mcclary培养基为适宜的酿酒酵母1450的生孢培养基。
2.用Mcclary培养基对1450进行产孢培养的最适温度为25℃-28℃,培养时间为5-7d。温度对酵母生孢能力的影响与相关酶的活性有关。
3.在测定菌株的发酵力试验中,发现单倍体菌株的酒精发酵力差异较大,推测原因是酒酵母的发酵能力由多个基因控制。并且通过对酒精发酵力和酒精发酵性能的综合分析,得到一株性能良好的单倍体S-3,可以作为以后杂交实验的亲本。
4.对单倍体S-3进行了生长曲线的测定,结果表明在进行育种工作时选取9h左右的菌体较好。
The process of inducing yeast to produce spores and then obtaining its haploid is crucial in Yeast Genetics and Breeding. This is indispensable for analyzing the inheritance of acquired characters and preparing the parent strains for hybridization. The spores can be obtained only in fitting sporulation condition. On the barren condition of nutrition, the trophosome of S.Cerevisiae transits into haploid from the former diploid through the process of meiosis, forming sporangium. None of the present common yeast spores-breeding culture media was suitable for all the yeasts to produce their spores. Therefore, the spores-breeding experiment of S.Cerevisiae 1450 was undertaken under different spores-breeding conditions in this essay. Then, the optimum culture medium was selected for the separation of the haploid of S.Cerevisiae 1450. Next, a haploid strain with good biological function was screened out through zymotechnics method so as to be applied in the next breeding experiment.
The main research results are as follows.
1. After the selective preference of four common yeast spores-breeding culture media, Mcclary culture medium was confirmed as the optimum one for S.Cerevisiae 1450.
2. The optimum temperature and time for S.Cerevisiae 1450 in the Mcclary culture medium were 25℃-28℃and 5-7d, separately. The influence of temperature over the spores-breeding capability of S.Cerevisiae 1450 was connected with the activity of concerning enzymes.
3. In the experiment of identifying the ferment capability of strains, the ferment capability varies largely among different haploid strains. The supposed reason was speculated that the ferment capability of S.Cerevisiae was controlled by multiple genes. And through the comprehensive analysis of the fermentation capability and performance, a haploid strain S-3 with good biological function was obtained and could be used as the parent strains in the future hybridization experiment.
4. The growth curve of the haploid S-3 was determined, and the results showed that it’s better to choose the strains around 9h in the breeding job.
引文
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[5] Erten H. Relations between elevated temperatures and fermentation behaviour of Kloeckera apiculata and Saccharomyces cerevisiae associated with winemaking in mixed cultures [J]. World J. Microbiol. Biotechnol. 2002. 18, 373-378.
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