Abstract: The present invention uses the CRISPR/Cas9 genome editing technology to rapidly create a male sterile line of tomato and application thereof, The present invention uses the CRISPR/Cas9 genome editing technology to edit the Solyc03g053130 gene of tomato, and then obtains a male sterile mutant which is homozygous and does not contain the CAS9 transgene by self-crossing. The present invention also discloses a method for assisting in identification of a male sterile plant, which is to detect the genotype of SNP1606 in the Solyc03g053130 gene in the genome of tomato, and if the genotype of SNP1606 of the genome of the tomato to be tested is homozygous T/T, the tomato to be tested is a male sterile plant or is a candidate male sterile plant. The male sterile line of tomato and the method for detecting male sterile plants created by the present invention can be applied to other tomato strains, and have great application prospect and economic value in breeding.