In this study, we selected the annual grapevine cv. Muscat Hamburg as the research material to explore the influence of root restriction on the growth and development of grape root system. By comparing with the traditional cultivation, we can understand the changes in grape root cell structure, IAA content and its metabolism and the expression of genes related to root growth and development after root restriction, thus explore the internal mechanism of root restriction cultivation affecting the changes of grape root system architecture. The results showed that: (1) after root restriction treatment, the architecture of grape root system changed significantly, which was mainly manifested in the appearance of a large number of clustered roots at the root tip, continuous adventitious roots, and the appearance of spiraling roots. In addition, the arrangement of root cells became looser, the root cap area of the young root tip was thicker, and the cells were larger and longer after root restriction. So, endothelial cells of the capillary roots were thickened, and the pericytes and cortical cells were larger. (2) Root restriction can significantly inhibit the content of IAA in grape roots. At 40, 100 and 205 d after planting, the changes in IAA content were consistent with the expression of key synthetic genes VvYUC6 and VvYUC8. Root restriction treatment induced the expression levels of IAA synthesisrelated genes VvTAR2, VvYUC2 and VvYUC4 to significantly upregulated during most sampling periods. At 70, 85, 130 and 205 d after planting, root restriction resulted in a significant upregulation of the expression levels of 8 genes related to IAA transport, such as VvAUX1 and VvLAX1, which coincided with the period when a high incidence of root system under root restriction. (3) Under root restriction, the expression of VvcycA1, VvcycB1, VvcycD3 and Vvcdc2 related to cell cycle regulation were upregulated from midAugust to early November. Our studies have shown that root restriction leads to changes in the configuration and cell structure of the grapevine roots, and the IAA content also changes significantly. In addition, the induction of auxin synthesis and polar transportrelated gene expression abundance changes, thereby affecting the synthesis and transport of auxin in the root system, thus adapting grapes to the root restriction cultivation environment.