GR24, a synthetic strigolactone analogue, plays a crucial role in a wide range of life processes. Studying the effects of the pre-harvest application of GR24 on strawberry ripening and fruit quality provides a scientific basis for the application of GR24 in horticultural crop production. GR24 solutions at concentrations of 0, 0.5, 1 and 2 µmol/L were applied to strawberries at the de-greening stage. The results showed that pre-harvest treatment with 1 µmol/L GR24 increased fruit weight and size; promoted fruit softening; increased the content of sugars, total flavonoids, total phenolics, and anthocyanins, and increased antioxidant activity. These results proved that GR24 effectively accelerated fruit ripening and improved fruit quality. In addition, to gain more insight into the biological mechanism of GR24 in fruit ripening, we conducted a combined transcriptomic and metabolomic analysis. It was found that the differences were related to sugar and flavonoid biosynthesis, particularly in the later stage of fruit ripening. qRT-PCR analysis revealed that GR24 increased gene expression activity and metabolite accumulation in the anthocyanin and sugar biosynthetic pathway, thereby promoting strawberry color and flavor.