The flanking sequence of a rice T-DNA inserted mutant G48 with a blight bacteria resistance was analyzed via PCR-walking.Results showed the T-DNA was inserted into the clone OSJNBa0076E06 (chromosome 3,AC132215.1) in Oryza sativa L.japonica cv.nipponbare.The left and right border of T-DNA inserted was 93750th and 93824th nucleotide of OSJNBa0076E06,respectively.PCR-walking provided a simple and effective method for analyzing flanking sequence of T-DNA insertion.