The invention relates to a saccharomyces cerevisiae Y3401 strain for high-yielding ethanol and a symbiotic fermentation culture method of the saccharomyces cerevisiae Y3401 strain and a Wickerhamomyces anomalus Y3604 strain for high-yielding ethyl acetate as well as application of the saccharomyces cerevisiae Y3401 strain. The saccharomyces cerevisiae Y3401 was preserved in China General Microbiological Culture Collection Center on October 20th, 2017, with a preservation number of CGMCC No.14828; the similarity of 26S rDNA D1/D2 sequence of the saccharomyces cerevisiae Y3401 strain to 26S rDNA D1/D2 sequence of other saccharomyces cerevisiae strains; symbiotic fermentation of two yeast strains in a sorghum enzymatic hydrolysate culture medium by a standing or shaking mode has the advantages that the content of the ethyl acetate is favorably improved and the contents of flavor substances such as total ester, beta-phenylethanol and isoamyl alcohol also can be improved. In a solid brewing system, the symbiotic fermentation of the two strains can enhance the characteristics of ester-improving and flavor-enhancing of yeast. The symbiotic fermentation method of the two strains, disclosed by the invention, can be applied to the brewing industries, having demands on the ethyl acetate, such as Baijiu, yellow rice wine and soy sauce.