Spine grape (Vitis davidii Foex) is an important wild plant species in South China. To provide economical and environmentally safe ways to promote the precocious maturation of spine grape berries, the effects of riboflavin were investigated. Riboflavin affected the reactive oxygen species metabolism in spine grape berries by increasing superoxide radical production and the hydrogen peroxide content, and it impaired the activities of the antioxidant enzymes superoxide dismutase and catalase. Riboflavin also induced the upregulated expression of maturation-related genes in advance, and the earlier accumulation of anthocyanin and total soluble solids. Phenological observations revealed that the treated grape berries underwent a color-turning stage 9 days earlier than the control, and the maturation stage occurred 7 days earlier than the control. Thus, riboflavin may significantly promote the precocious maturation of spine grape berries.