This study examines the effects of Evening primrose on colors, color fastness, and the antimicrobial activity of dyed fabrics. The results are as follows. The dyeing conditions of Evening primrose on cotton and mercerized cotton were optimized at 50℃, 60 minutes, and 200% (o.w.f.). In addition, Evening primrose dyeing on silk was determined at 90℃, 60 minutes and 200% (o.w.f.). The pre-mordant concentration of chemicals of cotton, mercerized cotton and silk was optimized at 1% (o.w.f.). The post-mordant concentration on mercerized cotton, silk and cotton was determined at 1% (o.w.f.) and 3% (o.w.f.), respectively. The mordant methods (such as pre-mordant and post-mordant) were slightly affected on the hue of dye-fabrics. Wet cleaning fastness of cotton was improved by post-mordant; otherwise, the wet cleaning fastness of mercerized cotton and silk was improved by a pre-mordant. The dry cleaning fastness of cotton and silk was excellent regardless of mordant methods. The dry cleaning fastness of mercerized cotton was improved by a post-mordant compared to a pre-mordant. The antimicrobial activity of Evening primrose-dyed fabrics was shown at 99.9%. The excellent antimicrobial activity of dyed fabrics remained after the mordant as well as wet and dry cleaning.