Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang Province, China.
Appl Microbiol Biotechnol. 2014 Apr;98(7):3059-70. doi: 10.1007/s00253-013-5423-7. Epub 2013 Dec 18.
Whole-genome shuffling (WGS) is a powerful technology of improving the complex traits of many microorganisms. However, the molecular mechanisms underlying the altered phenotypes in isolates were less clarified. Isolates with significantly enhanced stress tolerance and ethanol titer under very-high-gravity conditions were obtained after WGS of the bioethanol Saccharomyces cerevisiae strain ZTW1. Karyotype analysis and RT-qPCR showed that chromosomal rearrangement occurred frequently in genome shuffling. Thus, the phenotypic effects of genomic structural variations were determined in this study. RNA-Seq and physiological analyses revealed the diverse transcription pattern and physiological status of the isolate S3-110 and ZTW1. Our observations suggest that the improved stress tolerance of S3-110 can be largely attributed to the copy number variations in large DNA regions, which would adjust the ploidy of yeast cells and expression levels of certain genes involved in stress response. Overall, this work not only constructed shuffled S. cerevisiae strains that have potential industrial applications but also provided novel insights into the molecular mechanisms of WGS and enhanced our knowledge on this useful breeding strategy.
全基因组重排(WGS)是一种强大的技术,可以改善许多微生物的复杂特性。然而,对于在分离物中改变表型的分子机制还不太清楚。在对生物乙醇酿酒酵母菌株 ZTW1 进行 WGS 后,获得了在超高糖条件下具有显著增强的应激耐受性和乙醇产量的分离物。核型分析和 RT-qPCR 表明,染色体重排在基因组重排中经常发生。因此,本研究确定了基因组结构变异的表型效应。RNA-Seq 和生理分析揭示了分离物 S3-110 和 ZTW1 的转录模式和生理状态的多样性。我们的观察表明,S3-110 应激耐受性的提高在很大程度上归因于大 DNA 区域的拷贝数变异,这将调整酵母细胞的倍性和参与应激反应的某些基因的表达水平。总的来说,这项工作不仅构建了具有潜在工业应用价值的重排酿酒酵母菌株,而且为 WGS 的分子机制提供了新的见解,并加深了我们对这一有用的育种策略的认识。