Kiwifruit (Actinidia chinensis), a recently commercialized horticultural crop, is rich in various nutrient compounds. However, the regulatory networks controlling the dynamic changes in key metabolites among different tissues remain largely unknown. Here, high-resolution spatiotemporal datasets obtained by ultraperformance liquid chromatography-tandem mass spectrometry methodology and RNA-seq were employed to investigate the dynamic changes in the metabolic and transcriptional landscape of major kiwifruit tissues across different developmental stages, including from fruit skin, outer pericarp, inner pericarp, and fruit core. Kiwifruit spatiotemporal regulatory networks (KSRN) were constructed by integrating the 1,243 identified metabolites and co-expressed genes into 10 different clusters and 11 modules based on their biological functions. These networks allowed the generation of a global map for the major metabolic and transcriptional changes occurring throughout the life cycle of different kiwifruit tissues and discovery of the underlying regulatory networks. KSRN predictions confirmed previously established regulatory networks, including the spatiotemporal accumulation of anthocyanin and ascorbic acid (AsA). More importantly, the networks led to the functional characterization of three transcription factors: an A. chinensis ethylene response factor 1, which negatively controls sugar accumulation and ethylene production by perceiving the ripening signal, a basic-leucine zipper 60 (AcbZIP60) transcription factor, which is involved in the biosynthesis of AsA as part of the L-galactose pathway, and a transcription factor related to apetala 2.4 (RAP2.4), which directly activates the expression of the kiwi fruit aroma terpene synthase gene AcTPS1b. Our findings provide insights into spatiotemporal changes in kiwifruit metabolism and generate a valuable resource for the study of metabolic regulatory processes in kiwifruit as well as other fruits.