The textile industry is one of the major environmental polluters. It has been increased the researcher interest to explore eco-friendly ways and replace hazardous chemical materials with natural products. In the present work, for the first time, Z. Spina-christ root and Sichka gall plant by-products were employed for sustainable dyeing of wool yarns. Sichka is an asexual gall that results from the sting of the wasp Andricus curtisii (Cynips mediterranea Trott) on the buds of the oak (Quercus infectoria). Bio-compounds of Z. Spina-christ and gall were extracted by ultrasonic method and the extracts were used as a natural dye and bio-mordant for the dyeing process, respectively. Response surface method and central composite design were used to evaluate the effect of the main process parameters and dyeing optimization. Results showed that the selected materials contain phenolic and tannin bio-constitute and have a sufficient ability to use in the eco-dyeing process of textiles. The analysis of the variance of the quadratic model showed that the dye concentration, bio-mordant concentration, and pH were significant model terms and had the highest effects on the dyeing performance, respectively. Results indicate that for the conditions used in our experiments, the optimum dyeing conditions are dye conc. =96%, bio-mordant conc. =12%, and pH=7.5. Dye fiber bonding stability of the dyed samples was evaluated in terms of colorfastness to wash and light. Using Sichka as a bio- mordant in the dyeing process with Z. Spina-christ root extract can improve the color strength of wool yarns and obtain acceptable dye fastness properties. As a result, Z. Spina-christ root and Sichka bio-wastes, may, thus, be considered good replacements for the toxic dyes and mordants, especially because they improve the natural dyeing efficiency of wool yarns without adverse environmental effects and produce biodegradable byproducts with value-added properties.