The internal stipe necrosis of cultivated mushrooms (Agaricus bisporus) is caused by the bacterium Ewingella americana, a species of the Enterobacteriaceae. Recently, Ewingella americana was isolated from cultivated white button mushrooms in Korea evidencing symptoms of internal stipe browning. Its symptoms are visible only at harvest, and appear as a variable browning reaction in the center of the stipes. From these lesions, we isolated one bacterial strain (designated CH4). Inoculation of the bacterial isolate into mushroom sporocarps yielded the characteristic browning symptoms that were distinguishable from those of the bacterial soft rot that is well known to mushroom growers. The results of Gram stain, flagellal staining, and biochemical tests identified these isolates as E. americana. This was verified by pathogenicity, physiological and biochemical characteristics, and the results of an analysis of the 16S rRNA gene sequences and the fatty acids profile. This is the first report of the isolation of E. americana from cultivated white button mushrooms in Korea.