Male sterility is an essential trait in hybrid seed production,especially for monoclinous and autogamous food crops.Soybean male-sterile ms1 mutant has been known for more than 50 years and could be instrumental in making hybrid seeds.However,the gene responsible for the male-sterile phenotype has remained unknown.Here,we report the map-based cloning and char-acterization of the MS1 gene in soybean.MS1 encodes a kinesin protein and localizes to the nucleus,where it is required for the male meiotic cytokinesis after telophase Ⅱ.We further substantiated that MS1 colocalizes with microtubules and is essential for cell plate formation in soybean male gametogenesis through immunostaining.Both ms1 and CRISPR/Cas9 knockout mutants show complete male sterility but are otherwise phenotypically normal,making them perfect tools for producing hybrid seeds.The identification of MS1 has the practical potential for assembling the sterility system and speeding up hybrid soybean breeding.