We have used bulked segregant analysis to screen the strain-specific DNA marker associated psychrophilic strain of Pleurotus eryngii. Bulked genomic DNAs of Pleurotus eryngii were amplified by randomly amplified polymorphic DNA (RAPD) using OP-A, OP-B, OP-L, OP-P, OP-R and OP-S primers to screen the strain-specific DNA marker. A unique DNA fragment of 490 bp was amplified with OP-L18 primer from the psychrophilic strain and sequenced. A sequence characterized amplified region (SCAR) marker was designed on the basis of the determined sequence and named as OP-L18-1. The PCR analysis with the OP-L18-1 primer showed that this SCAR marker clearly distinguish the psychrophilic strains from the control strains.