The stolon is the main asexual reproductive organ of the medicinal plant Tulipa edulis and has special morphology. In the greenhouse experiment presented herein, the dynamic changes in carbohydrates and related enzymes, proteins, and endogenous hormones of stolons during T. edulis stolon formation were investigated. The results showed that the content of total soluble sugar, sucrose, reducing sugar, fructose, and starch were all significantly enhanced in the middle period when stolon emerged and maintained at relatively high levels until the later period of stolon formation, while protein content decreased during stolon formation. The activities of amylase (AMY), sucrose phosphate synthase (SPS), and sucrose synthase (SS) peaked in the initial period and were negatively correlated with soluble sugars. However, adenosine diphosphoglucose pyrophosphorylase (AGPase) activity increased as stolon formation progressed, and the changes in soluble starch synthase (SSS), granule-bound starch synthase (GBSS) activities presented a single peak, reaching their maximums in the middle period. AGPase, SSS, and GBSS activities were all positively related to starch content. Moreover, quantitative real-time polymerase chain reaction (qRT-PCR) verified the changes in SS and SSS activities via the expression levels of the SS, SSSI, and SSSII genes. The gibberellin (GA) and zeatin riboside (ZR) content attained their maximum in the initial period of stolon formation. Indole-3-acetic acid (IAA) and abscisic acid (ABA) remained at high levels during the initial and middle period and decreased significantly during the later period of stolon formation, inversely to the ratio of ABA:IAA. Analysis of the physiological changes in T. edulis stolon indicated that the accumulation of soluble sugars and starch via various enzymes, a high level of IAA and a low ABA to IAA ratio mainly contributed to stolon development of T. edulis. This paper explored carbohydrate levels and endogenous hormones profiles during stolon formation, which provided the theory basis for further regulating stolon growth of T. edulis.