The objective of this research was to determine effects of growth medium composition on the propagation of cultured milk vetch (Astragalus adsurgens Pall) roots. An orthogonal design was used to test multiple components in the growth medium. A second objective was to measure allelopathic activities of cultured milk vetch roots by conducting a bioassay using filtered extracts from the growth medium after culturing the roots. The results indicated that among all the nutritional components, Fe2+ had the greatest effect on the propagation of cultured milk vetch roots; the effects of sucrose,H2PO+4, Mg2+, Mn2+, Cu2+, Zn2+, BO3-3, Co2+, I- and C8H12ClNO3 + C12H18Cl2N4OS + C6H5O2N + C6H12O6 were intermediate; and the effects of nitrogen, Ca2+, MoO2-4 and NAA were lowest. A recommendation was made regarding the optimum nutrient content of growth medium for the propagation of cultured milk vetch roots. A bioassay using filtered extracts from the growth medium indicated that milk vetch roots might have produced allelopathic chemicals during their culture. Differences in the degree of allelopathic effects suggest that nutrient stress may influence the production of allelopathic chemicals. This study provides a basis for the improvement of milk root regeneration and propagation and suggests that nutrition may influence the production of allelopathic chemicals by this plant.