Yarrowia lipolytica is a nonconventional, heterothallic, oleaginous yeast with wide range of industrial applications. Increasing ploidy can improve advantageous traits for industrial applications including genetic stability, stress resistance, and productivity, but the construction of knock-out mutant strains from polyploid cells requires significant effort due to the increased copy numbers of target genes. The goal of this study was to evaluate the effectiveness of a mating-type switching strategy by single step transformation without a genetic manipulation vestige, and to optimize the conventional method for increasing ploidy (mating) in Y. lipolytica. In this study, mating-type genes in haploid Y. lipolytica cells were scarlessly converted into the opposite type genes by site-specific homologous recombination, and the resulting MATB-type cells were mated at low temperature (22°C) with addition of sodium citrate with each MATA-type haploid cell to yield a MATA/MATB-type diploid strain with genetic information from both parental strains. The results of this study can be used to increase ploidy and for whole genome engineering of a yeast strain with unparalleled versatility for industrial application.

中文翻译：

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杂藻酵母解脂耶氏酵母中增强的交配型转换和有性杂交。

解脂耶氏酵母是非常规的，杂藻的，油质酵母，具有广泛的工业应用。倍数性的提高可以改善工业应用的有利性状，包括遗传稳定性，抗逆性和生产力，但是从多倍体细胞构建敲除突变株需要大量的努力，因为靶基因的拷贝数增加。这项研究的目的是通过无遗传操作痕迹的单步转化来评估交配型转换策略的有效性，并优化在解脂耶氏酵母中增加倍性（交配）的常规方法。在这项研究中，单倍体解脂耶氏酵母细胞中的交配型基因通过位点特异性同源重组毫不留情地转化为相反型基因，然后将所得的MATB型细胞在低温（22°C）下与柠檬酸钠与每个MATA型单倍体细胞进行交配，以产生具有两个亲本菌株的遗传信息的MATA / MATB型二倍体菌株。这项研究的结果可用于增加倍性，并用于酵母菌株的全基因组工程，具有无与伦比的多功能性，可用于工业应用。