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High-quality and disease-resistant male sterile lines have great potential for applications in hybrid rice breeding.We introduced specific mutations into the TMS5,Pi21,and Xa13 genes in Pinzhan intermediate breeding material using the CRISPR/Cas9 multiplex genome editing system.We found that the transgenefree homozygous triple tms5/pi21/xa13 mutants obtained in the T1 generation displayed characteristics of thermosensitive genic male sterility (TGMS) with enhanced resistance to rice blast and bacterial blight.Our study provides a convenient and effective way of converting breeding intermediate material into TGMS lines through multiplex gene editing,which could significantly accelerate the breeding of sterile lines.