Quinoa is a plant with special nutritional and biological properties and tolerant in strict environmental conditions, which can be considered to a strategic product for the future food supply of mankind, as a pseudocereal. Quinoa root is rich in bioactive compounds with antimicrobial, antioxidant activity and diverse medicinal and nutritional properties that can have a potential for commercialization. The use of cultivation and propagation techniques in controlled laboratory conditions, such as the induction of differentiated tissues of hairy roots, allows the production of plants and the rapid and massive access to secondary metabolites. In this research, the induction of hairy roots was evaluated using four Rhizobium rhizogenes strains, including A4, A7, 11325, and ATCC 15834, in quinoa and the effect of different concentrations of acetosyringone and sucrose and removal of macro elements in the culture medium in increasing the rooting performance of R. rhizogenes was investigated. The results showed the different effects of the mutual effect of different concentrations of sucrose and acetosyringone, as well as the commonly used elements of the half strength MS culture medium on the biomass of hairy roots. The most optimal conditions for the induction of hairy roots culture were obtained in half strength MS culture medium, containing 30 g/l sucrose and 1 mM acetosyringone and without NH4NO3 salt. This efficient protocol provides a basis for future studies to be able to use the induced hairy root as a source of valuable secondary metabolites of quinoa root and create a new approach for exploiting the nutritional, medicinal and health compounds of the quinoa plant.