This chapter reviews the various biotic and abiotic elicitors applied to hairy root cultures and their stimulating effects on the accumulation of secondary metabolites. Elicitors generally refer to the agents that stimulate the defense responses of plants. As a major response of plants to biotic and abiotic stress, the accumulation of secondary metabolites in plant tissue cultures can be stimulated by the elicitors. Among the many elicitors applied to hairy root cultures as well as plant cell suspension cultures, the most common and effective elicitors are fungal cell extracts, polysaccharides from fungal and plant cells, and heavy metal salts. With the crude fungal cell extracts, it is essential to observe the preparation conditions carefully for achieving reproducible effects. In addition to the chemical agents, UV-radiation, hyperosmotic stress and temperature shift have been shown effective for some plant species/metabolites. Elicitor type, dose, and treatment schedule are major factors determining the effects on the secondary metabolite production. In addition to the accumulation of products in roots, elicitor treatments often stimulate the release of intracellular products. Although elicitation is mainly effective to increase specific product yield on per unit mass of roots, the incorporation of nutrient feeding strategies can be applied to enhance the volumetric product yield. The integration of in situ product recovery from the roots/liquid medium is another synergistic strategy with the elicitor treatment to improve the process.