Metabolomic analysis was carried out to investigate the metabolic differences of diploid (α/a) and homogenous haploid (α,a) yeasts, and further assess their response to ethanol stress. The dynamic metabolic variations of diploid and haploid caused by 3 and 7% (v/v) ethanol stress were evaluated by gas chromatography coupled to time-of-flight mass spectrometry combined with statistical analysis. Metabolite profiles originating from three strains in presence/absence of ethanol stress were distinctive and could be distinguished by principal components analysis. Results showed that the divergence among the strains with ethanol stress was smaller than without it. Furthermore, the levels of most glycolytic intermediates and amino acids in haploid were lower than these in diploid with/without ethanol stress, which was considered as species-specific behaviors. The increases of protective metabolites including polyols, amino acids, precursors of phospholipids, and unsaturated fatty acids under ethanol stress in three strains revealed the ethanol stress-specific responses. Higher fold change in most of these protectants in haploid indicated that haploid was more susceptible to ethanol stress than diploid. These findings provided underlying basis for better understanding diploid and haploid yeasts, and further breeding tolerant strains for efficient ethanol fermentation.
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