Abstract
Plastids, a wide family of plant specific organelles, exist in all plant cells in a number of different forms with different functions essential for plant life. Among them, chloroplasts are by far the more extensively studied owing to their central role in photosynthesis. However, other plastid family members, often referred to as nongreen plastids, play also major roles in the physiology of higher plants and could be better suited for studies of specific metabolic processes in heterotrophic plant cells. Unfortunately, serious technical problems are frequently encountered with separating intact, active nongreen plastids from contaminating membranes and mitochondria. Here, we provide detailed protocols suitable for the large scale preparation of intact and highly pure proplastids from cauliflower buds, as well as amyloplasts from sycamore cultured cells, and for the subsequent separation of their surrounding envelope membranes from the stroma and other plastid fractions. Both methods proved to be highly reliable and have been instrumental for in-depth investigations on biochemistry and physiology of nongreen plastids.
Key words
- Proplastids
- Amyloplasts
- Envelope membranes
- Organelle purity
- Protoplasts
- Cauliflower buds
- Sycamore cells
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References
Jarvis P, Lopez-Juez E (2013) Biogenesis and homeostasis of chloroplasts and other plastids. Nat Rev Mol Cell Biol 14(12):787–802
Plaxton WC (2017) Leucoplast isolation and subfractionation. In: Taylor NL, Millar AH (eds) Isolation of plant organelles and structures: methods and protocols, vol 1511. Springer, New York, pp 73–81
Journet E-P, Douce R (1985) Enzymic capacities of purified cauliflower bud plastids for lipid synthesis and carbohydrate metabolism. Plant Physiol 79(2):458–467
Journet E-P, Bligny R, Douce R (1986) Biochemical changes during sucrose deprivation in higher plant cells. J Biol Chem 261(7):3193–3199
ap Rees T, Macdonald FD (1987) Isolation of amyloplasts from suspension cultures of soybean. Methods Enzymol 148:218–226
Shannon JC, Echeverria E, Boyer C (1987) Isolation of amyloplasts from developing endosperm of maize (Zea mays L.). Methods Enzymol 148:226–234
Liedvogel B (1987) Isolation of membranous chromoplasts from daffodil flowers. Methods Enzymol 148:241–246
Ngernprasirtsiri J, Harinasut P, Macherel D et al (1988) Isolation and characterization of the amyloplast envelope-membrane from cultured white-wild cells of sycamore (Acer pseudoplatanus L.). Plant Physiol 87(2):371–378
Alban C, Joyard J, Douce R (1988) Preparation and characterization of envelope membranes from nongreen plastids. Plant Physiol 88(3):709–717
Miernyk J (1989) Leucoplast isolation. In: Boyer CT, Shannon JC, Hardison RC (eds) Physiology, biochemistry, and genetics of nongreen plastids. Current Topics in Plant Physiology, vol 2. The American Society of Plant Physiologists, Rockville, MD, pp 15–23
Gounaris I (1989) Chromoplasts of Capsicum annuum: isolation of proteins and nucleic acids. In: Boyer CD, Shannon JC, Hardison RC (eds) Physiology, biochemistry and genetics of nongreen plastids. Current Topics in Plant Physiology, vol 2. The American Society of Plant Physiologists, Rockville, MD, pp 24–36
Shannon JC (1989) Aqueous and nonaqueous methods for amyloplast isolation. In: Boyer CD, Shannon JC, Hardison RC (eds) Physiology, biochemistry and genetics of nongreen plastids. Current Topics in Plant Physiology, vol 2. The American Society of Plant Physiologists, Rockville, MD, pp 37–48
Douce R, Alban C, Journet E et al (1989) Biochemical properties of plastids isolated from cauliflower buds and sycamore cells and of their envelope membranes. In: Boyer CD, Shannon JC, Hardison RC (eds) Physiology, biochemistry and genetics of non-green plastids. Current Topics in Plant Physiology, vol 2. The American Society of Plant Physiologists, Rockville, MD, pp 99–119
Joyard J, Block MA, Alban C et al (1989) The plastid envelope membranes and their contribution to plastid membrane biogenesis. In: Briggs WR (ed) Plant biology (USA), vol 8. Photosynthesis. Alan R. Liss Inc, New York, pp 331–345
Joyard J, Teyssier E, Miège C et al (1998) The biochemical machinery of plastid envelope membranes. Plant Physiol 118(3):715–723
Alban C, Dorne A-J, Joyard J et al (1989) [14C] acetate incorporation into glycerolipids from cauliflower proplastids and sycamore amyloplasts. FEBS Lett 249(1):95–99
Alban C, Joyard J, Douce R (1989) Comparison of glycerolipid biosynthesis in non-green plastids from sycamore (Acer pseudoplatanus) cells and cauliflower (Brassica oleracea) buds. Biochem J 259(3):775–783
Barsan C, Kuntz M, Pech J-C (2017) Isolation of Chromoplasts and Suborganellar compartments from tomato and bell pepper fruit. In: Taylor NL, Millar AH (eds) Isolation of plant organelles and structures: methods and protocols, vol 1511. Springer, New York, pp 61–71
Bligny R, Leguay J-J (1987) Techniques of cell suspension culture. Methods Enzymol 148:3–16
Duncan O, Millar AH, Taylor NL (2017) Isolation of mitochondria, their sub-Organellar compartments, and membranes. In: Taylor NL, Millar AH (eds) Isolation of plant organelles and structures: methods and protocols, vol 1511. Springer, New York, pp 83–96
Block MA, Dorne A, Joyard J et al (1983) Preparation and characterization of membrane fractions enriched in outer and inner envelope membranes from spinach chloroplasts. I electrophoretic and immunochemical analyses. J Biol Chem 258(21):13273–13280
Journet E-P (1987) Isolation of plastids from buds of cauliflower (Brassica oleracea L.). Methods Enzymol 148:234–240
Brouquisse R, Nishimura M, Gaillard J et al (1987) Characterization of a cytosolic aconitase in higher plant cells. Plant Physiol 84(4):1402–1407
Acknowledgments
We would like to thank Dr. Jacques Joyard and Pr Roland Douce for their pioneer role in the characterization of plastid envelope membranes and more generally in the elucidation of plant organelles physiology and biochemistry. Jacqueline Martin-Laffon is acknowledged for her help in End-Note library creation and for critical reading of the manuscript.
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Alban, C., Journet, EP. (2018). Purification of Nongreen Plastids (Proplastids and Amyloplasts) from Angiosperms, and Isolation of Their Envelope Membranes. In: Maréchal, E. (eds) Plastids. Methods in Molecular Biology, vol 1829. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8654-5_10
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DOI: https://doi.org/10.1007/978-1-4939-8654-5_10
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