Center for Research in Agricultural Genomics (CSIC-IRTA-UAB-UB), Bellaterra, Barcelona, Spain.
Department of Biochemistry and Molecular Biomedicine, Faculty of Biology, University of Barcelona, Barcelona, Spain.
Methods Mol Biol. 2020;2083:189-197. doi: 10.1007/978-1-4939-9952-1_14.
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.
质体分化涉及类胡萝卜素的活跃合成，这与重塑预先存在的质体膜系统以形成专门的结构有关，这些结构参与了合成类胡萝卜素的隔离和储存。这些亚质体结构表现出显著的形态差异，似乎适应于某些植物物种和器官中特定类胡萝卜素的积累。目前，关于质体生物发生以及不同亚细胞器结构在类胡萝卜素合成和储存中的作用知之甚少。将经典的分级分离方法与生物化学和组学技术的应用相结合，是一种很有吸引力的方法，可以揭示与质体生物发生相关的生化和细胞机制的新方面，这些机制涉及到类胡萝卜素生物合成和储存过程中质体分化涉及的结构。在这里，我们描述了一种用于分离、裂解和分级分离番茄果实质体的组合方案。获得的级分适用于代谢组学和蛋白质组学分析。
