Fractionation of Tomato Fruit Chromoplasts
Chromoplast differentiation involves an active synthesis of carotenoids associated with the remodeling of the preexisting plastid membrane systems to form specialized structures involved in the sequestration and storage of the synthesized carotenoids. These subplastidial structures show remarkable morphological differences and seem to be adapted to the accumulation of particular carotenoids in some plant species and organs. At present, very little is known about chromoplast biogenesis and the role of the different suborganellar structures in the synthesis and storage of carotenoids. The combination of classical fractionation methods with the use of biochemical and -omics techniques represents an attractive approach to unravel novel aspects related with the biochemical and cellular mechanisms underlying the biogenesis of the structures involved in the biosynthesis and storage of carotenoids during chromoplast differentiation. Here we describe a combined protocol for the isolation, lysis and fractionation of tomato fruit chromoplast. The fractions obtained are suitable for metabolomics and proteomics analysis.
Key wordsChromoplast Carotenoid Proteomics Fractionation Tomato fruit
We acknowledge the financial support of AGAUR-Generalitat de Catalunya (Grant 2017 SGR 710), the CERCA Programme of the Generalitat de Catalunya and the Severo Ochoa Programme for Centres of Excellence in R&D 2016–2019 to CRAG (SEV-2015-0533). AB is member of the Spanish Carotenoid Network (CaRed) funded by the Spanish Ministry of Economy and Competitiveness (Grants BIO2015-71703-REDT and BIO2017-90877-REDT).
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