To breed a new Korean Pleurotus eryngii cultivar with high quality and yield, single crosses between 24×46 and KNR2539 were performed, and a new cultivar, 6×13, was selected based on the days to harvest (14.6 days), quality (7.2), and yield (85.9 g/850 cc bottle). The strain was named Aeryni 5 and cultivated on a large scale at the mushroom farms to compare with Keuneutari 2ho. The yield of Aeryni 5 (82.2 g) was 122.7% of Keuneutari 2ho, and the quality of the new cultivar was 7.7 while reference cultivar was 6.3. The yield and quality of the two cultivars were statistically different. The lightness of the pileus of Aeryni 5 (61.7) was greater than that of Keuneutari 2ho by 3.4 points; thus, the pileus of Aeryni 5 looked brighter. PCR with URP2 was used to discriminate between Aeryni 5 and Kenneutari 2ho.