Kengyilia hirsuta is an important pioneer plant distributed on the desertified grassland of the Qinghai-Tibet Plateau. It has strong adaptability to alpine desert habitats, so it can be used as a sand-fixing plant on sandy alpine land. To study the response mechanisms of root morphological and physiological characteristics of K. hirsuta to sandy soil moisture, 10%, 25% and 40% moisture levels were set up through potted weighing water control method. The biomass, root-shoot ratio, root architecture parameters, and biochemical parameters malondialdehyde, free proline, soluble protein, indole-3-acetic acid, abscisic acid, cytokinin, gibberellin, relative conductivity and antioxidant enzyme activities were measured in the trefoil stage, and the response mechanisms of roots at different moisture levels were analyzed. The results showed that with the increase of soil moisture, root morphological indexes such as root biomass, total root length, total root volume and total root surface increased, while the root topological index decreased continuously. The malondialdehyde content, relative conductivity, superoxide dismutase activity, peroxidase activity, catalase activity, free proline content, soluble protein content, abscisic acid content and cytokinin content at the 25% and 40% moisture levels were significantly decreased compared with the 10% level (P< 0.05). Thus, the root growth of K. hirsuta was restricted by the 10% moisture level, but supported by the 25% and 40% moisture levels. An artificial neural network revealed that total root length, total root surface area, root link average length, relative conductivity, soluble protein, free proline and moisture level were the key factors affecting root development. These research results could contribute to future agricultural sustainability.