Krogerus Kristoffer, Arvas Mikko, De Chiara Matteo, Magalhães Frederico, Mattinen Laura, Oja Merja, Vidgren Virve, Yue Jia-Xing, Liti Gianni, Gibson Brian
VTT Technical Research Centre of Finland, Tietotie 2, P.O. Box 1000, FI-02044, Espoo, Finland.
Department of Biotechnology and Chemical Technology, Aalto University, School of Chemical Technology, Kemistintie 1, Aalto, P.O. Box 16100, FI-00076, Espoo, Finland.
Appl Microbiol Biotechnol. 2016 Aug;100(16):7203-22. doi: 10.1007/s00253-016-7588-3. Epub 2016 May 17.
The genomes of hybrid organisms, such as lager yeast (Saccharomyces cerevisiae × Saccharomyces eubayanus), contain orthologous genes, the functionality and effect of which may differ depending on their origin and copy number. How the parental subgenomes in lager yeast contribute to important phenotypic traits such as fermentation performance, aroma production, and stress tolerance remains poorly understood. Here, three de novo lager yeast hybrids with different ploidy levels (allodiploid, allotriploid, and allotetraploid) were generated through hybridization techniques without genetic modification. The hybrids were characterized in fermentations of both high gravity wort (15 °P) and very high gravity wort (25 °P), which were monitored for aroma compound and sugar concentrations. The hybrid strains with higher DNA content performed better during fermentation and produced higher concentrations of flavor-active esters in both worts. The hybrid strains also outperformed both the parent strains. Genome sequencing revealed that several genes related to the formation of flavor-active esters (ATF1, ATF2¸ EHT1, EEB1, and BAT1) were present in higher copy numbers in the higher ploidy hybrid strains. A direct relationship between gene copy number and transcript level was also observed. The measured ester concentrations and transcript levels also suggest that the functionality of the S. cerevisiae- and S. eubayanus-derived gene products differs. The results contribute to our understanding of the complex molecular mechanisms that determine phenotypes in lager yeast hybrids and are expected to facilitate targeted strain development through interspecific hybridization.
诸如拉格酵母(酿酒酵母×真贝氏酵母)等杂交生物的基因组包含直系同源基因,其功能和效应可能因其起源和拷贝数的不同而有所差异。拉格酵母中的亲本亚基因组如何对发酵性能、香气产生和胁迫耐受性等重要表型性状产生影响,目前仍知之甚少。在此,通过杂交技术而非基因改造产生了三种不同倍性水平(异源二倍体、异源三倍体和异源四倍体)的新生拉格酵母杂交体。这些杂交体在高浓度麦芽汁(15°P)和超高浓度麦芽汁(25°P)发酵过程中进行了表征,并监测了香气化合物和糖分浓度。DNA含量较高的杂交菌株在发酵过程中表现更佳,且在两种麦芽汁中均产生了更高浓度的风味活性酯。杂交菌株的表现也优于亲本菌株。基因组测序显示,与风味活性酯形成相关的几个基因(ATF1、ATF2、EHT1、EEB1和BAT1)在较高倍性的杂交菌株中以更高的拷贝数存在。还观察到了基因拷贝数与转录水平之间的直接关系。所测得的酯浓度和转录水平还表明,酿酒酵母和真贝氏酵母来源的基因产物的功能存在差异。这些结果有助于我们理解决定拉格酵母杂交体表型的复杂分子机制,并有望通过种间杂交促进定向菌株的开发。