The purpose of this study is to develop eco-printing method using natural dyes combined with chitosan treatment to impart various pattern effect on cotton fabric. It was examined whether tone-on-tone pattern effects could be produced by screen printing with chitosan paste and subsequently dip-dyed in natural dye bath. For polychromic colorants, fabrics were pre-mordanted and then screen printed with chitosan, while only chitosan printing treatment was applied for monochromic colorants. Color, dye uptake, washing, light and rubbing fastnesses of printed fabrics were measured in relation to the presence of chitosan printing. Pre-mordanting and chitosan printing treatment were applied to such polychromic colorants as madder, sappanwood, onion and cochineal. Also, printing pattern formation on fabrics was evaluated by differences in dye uptake and color. The results suggested that madder, onion and cochineal were effective on pattern formation. Regarding sappanwood, since dye uptake was increased only by pre-mordanting without chitosan printing treatment, there was little color difference and thus patterns were not clearly distinguished. Chitosan printing treatment tended to increase washing, light and rubbing fastnesses of dyed fabrics with polychromic colorants. Chitosan printing treatment was applied to monochromic colorants such as chlorophyll, gardenia and indigo and the effect of printing formation was examined according to the difference in dye uptake. Among monochromic dyes tried, chlorophyll was effective and chitosan printing improved washing and light fastnesses.