Transition from chloroplasts to chromoplasts is an important biological process in fruit ripening. Many significant structural and metabolic shifts occur. Spectral confocal microscopy analyses of carotenoids and chlorophylls in isolated fruit plastids showed that chromoplasts arise from already existing chloroplasts, and that no further differentiation process occur. Real-time in-situ recording of intact live tissue showed that the process is synchronized within a cell, but with less synchrony within the whole tissue. Proteomic analyses gave better insight into metabolic shifts during chromoplastogenesis. Major shifts included decrease in abundance of proteins involved in light reactions, including photosynthesis, Calvin cycle and photorespiration. Proteins involved in carbohydrate metabolism also decrease in abundance. Proteins whose abundance increases during chromoplastogenesis include stress proteins, and proteins related to terpenoid biosynthesis. One interesting feature is that the large subunit of the RUBISCO enzyme, an enzyme typically abundant in photosynthetic plastids, although continuously decreasing in abundance, is present in fully developed chromoplasts. These metabolic shifts are preceded by the accumulation of acetyl CoA carboxylase, which accounts for the generation of a storage matrix that will accumulate carotenoids. Metabolic shifts are followed by structural changes. The biogenesis of the machinery for building up of thylakoid and photosystems is disrupted, coupled with elevated energy production components and loss of plastid division machinery.