A paddy rice EPSPS mutant, and an encoding gene and use thereof, relating to the technical field of genetic engineering. The paddy rice EPSPS mutant has an amino acid sequence shown as SEQ ID NO.1. The paddy rice EPSPS mutant has glyphosate resistance and can resist glyphosate having a concentration of 100mM. The paddy rice EPSPS mutant has very wide use prospect in the field of cultivating glyphosate-resistant plants.