Chapter 14
Fractionation of Tomato Fruit Chromoplasts
Karel De Pourcq and Albert Boronat
Abstract
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 words Chromoplast, Carotenoid, Proteomics, Fractionation, Tomato fruit
1 Introduction
Chromoplasts are plastids specialized in the synthesis and accumulation of carotenoids found in many fruits, flowers, seeds and roots.
Chromoplasts derive from preexisting plastids (such as proplastids,
chloroplasts, amyloplasts, or leucoplasts) in a process characterized
by a massive biosynthesis of carotenoids along with a remodeling of
the preexisting membrane systems and the formation of suborganellar structures involved in carotenoid sequestration and storage
[1–3]. Because of their hydrophobic nature, carotenoids are usually
stored in the form of lipoproteic complexes [4]. Based on the
pigment storage substructures, chromoplasts are classified into
five major types: globular, crystalline, membranous, fibrillar, and
tubular [1, 2]. This variety of suborganellar structures has been
associated with the large diversity of carotenoid profiles found in
the chromoplasts present in different plant species, tissues, organs,
or developmental stages [5]. Interestingly, different chromoplast
Manuel Rodrı ´guez-Concepcio ´ n and Ralf Welsch (eds.), Plant and Food Carotenoids: Methods and Protocols,
Methods in Molecular Biology, vol. 2083, https://doi.org/10.1007/978-1-4939-9952-1_14,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
189
Fractionation of Tomato Fruit Chromoplasts
Karel De Pourcq and Albert Boronat
Abstract
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 words Chromoplast, Carotenoid, Proteomics, Fractionation, Tomato fruit
1 Introduction
Chromoplasts are plastids specialized in the synthesis and accumulation of carotenoids found in many fruits, flowers, seeds and roots.
Chromoplasts derive from preexisting plastids (such as proplastids,
chloroplasts, amyloplasts, or leucoplasts) in a process characterized
by a massive biosynthesis of carotenoids along with a remodeling of
the preexisting membrane systems and the formation of suborganellar structures involved in carotenoid sequestration and storage
[1–3]. Because of their hydrophobic nature, carotenoids are usually
stored in the form of lipoproteic complexes [4]. Based on the
pigment storage substructures, chromoplasts are classified into
five major types: globular, crystalline, membranous, fibrillar, and
tubular [1, 2]. This variety of suborganellar structures has been
associated with the large diversity of carotenoid profiles found in
the chromoplasts present in different plant species, tissues, organs,
or developmental stages [5]. Interestingly, different chromoplast
Manuel Rodrı ´guez-Concepcio ´ n and Ralf Welsch (eds.), Plant and Food Carotenoids: Methods and Protocols,
Methods in Molecular Biology, vol. 2083, https://doi.org/10.1007/978-1-4939-9952-1_14,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
189