Astragalus hoantchy, a widely cultivated medicinal plant species in traditional Chinese and Mongolian medicine, has been often hampered by replant failure during cultivation, like many other herbs of the genus Astragalus. Root aqueous extracts of Astragalus herbs were reported to exhibit allelopathic activity against other plants and autotoxic activity on their own seedlings, but the allelochemicals released by Astragalus plants have not been specified so far. Ten compounds were isolated from the rhizosphere soil extract of cultivated A. hoantchy and elucidated by spectroscopic analysis. Compounds 1–6 observably showed allelopathic activity against Lactuca sativa seedlings and autotoxic activity against A. hoantchy seedlings. The isolated compounds were further confirmed and quantified by high-performance liquid chromatography (HPLC) in the rhizosphere soil, with a total concentration of 9.78 μg/g (dry weight). These results specify and verify the allelochemicals released by cultivated A. hoantchy into the soil environment, which may provide new insights into the allelopathic mechanisms of this medicinal plant and probably assist in clarifying the replant problems of Astragalus plants.