GmMs1 encodes a kinesin-like protein essential for male fertility in soybean (Glycine max L.).

The application of heterosis is a promising approach for greatly increasing yield in soybean (Glycine max L.). Nuclear male sterility is essential for hybrid seed production and the utilization of heterosis. Here we report the cloning of the gene underlying the soybean male-sterile mutant ms-1, which has been widely used for recurrent selection in soybean breeding programs. We initially delimited the ms1 locus to a 16.15 kb region on chromosome 13, based on SLAF_BSA sequencing followed by genotyping of an F2 population segregating for the locus. Compared to the same region in fertile plants, the mutant region lack s a sequence of ~38.7 kb containing five protein-coding genes, including an ortholog of the kinesin-like protein gene NACK2, named GmMs1. The GmMs1 knockout plants generated via CRISPR/Cas-mediated gene editing displayed a complete male-sterile phenotype. Metabolic profiling showed that fertile anthers accumulated starch and sucrose normally, whereas sterile anthers had higher anthocyanin levels and lower flavonoid levels and lower antioxidant enzyme activities. These results provide insights into the molecular mechanisms governing male sterility and demonstrate that GmMs-1 could be used to create male-sterile lines through targeted mutagenesis. These findings pave the way for designing seed production technology (SPT) and an intelligent male-sterile line system to utilize heterosis in soybean. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.