Abstract
That chloroplasts are green and therefore recognizable by eye should have been an advantage in developing methods for their isolation, but historically it has been a disadvantage. It was relatively easy to grind up cells, subject the brei to differential centrifugation, and obtain a green pellet. It was also easy to infer that the pellet contained chloroplasts when it actually consisted of the swollen relics of thylakoid membranes. Much was learned about the light reactions of photosynthesis from studying such suspensions of thylakoids, but they were useless for the study of CO2 fixation or other processes that we now know occur in chloroplasts. A fundamental principle of biochemistry is (or should be) that we can only claim that a process has been reconstructed in vitro when the isolated system is shown to mimic the process in vitro both qualitatively and quantitatively. Failure to do so should not be explained away by ad-hoc hypotheses.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Granick S (1938) Quantitative isolation of chloroplasts from higher plants. Am J Bot 25: 538–541.
Kahn A, von Wettstein D (1961) Macromolecular physiology of plastids. II. Structure of isolated spinach chloroplasts. J Ultrastruct Res 5: 557–574.
Gibbs M, Robinson JM (1974) Photosynthetic CO2 incorporation in isolated spinach chloroplasts. In: A San Pietro (ed) Experimental Plant Physiology, pp. 19–20. St. Louis: Mosby.
Walker DA (1980) Preparation of higher plant chloroplasts. Meth Enzymol 69: 94–104.
Morgenthaler J-J, Price CA, Robinson JM, Gibbs M (1974) Photosynthetic activity of spinach chloroplasts after isopycnic centrifugation in gradients of silica. Plant Physiol 54: 532–534.
Morgenthaler J-J, Marsden MPF, Price CA (1975) Factors affecting the separation of photosynthetically competent chloroplasts in gradients of silica sols. Arch Biochem Biophys 168: 289–301.
Price CA, Cushman JC, Mendiola-Morgenthaler LR, Reardon EM (1987) Isolation of plastids in density gradients of Percoll and other silica sols. In: L Packer and R Douce (eds) Methods in Enzymology, vol. 148: Plant Membranes, pp. 157–179. Academic Press, San Diego.
Hadjeb N, Gounaris I, Price CA (1988) Chromoplast-specific proteins in Capsicum annuum. Plant Physiol 88: 42–45.
Cushman JC, Price CA (1986) Synthesis and turnover of proteins in proplastids and chloroplasts of Euglena gracilis. Plant Physiol 82: 972–977.
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning, A Laboratory Manual, vol. 2. Cold Spring Harbor Laboratory Press.
Gray PW, Hallick RB (1979) Isolation of Euglena gracilis chloroplast 5S ribosomal RNA and mapping the 5S rRNA gene on chloroplast DNA. Biochemistry 18: 1820–1825.
Shigeoka SA, Yokota A, Nakano Y, Kitaoka S (1980) Preparation of physiologically intact chloroplasts from Euglena gracilis z. Bull Univ Osaka Pref Ser 3 32: 7–51
Ortiz W, Reardon EM, Price CA (1980) Preparation of chloroplasts from Euglena highly active in protein synthesis. Plant Physiol 66: 291–294.
Nishimura M, Douce R, Akazawa T (1982) Isolation and characterization of metabolically competent mitochondria from spinach leaf protoplasts. Plant Physiol 69: 916–920.
Price CA (1982) Centrifugation in Density Gradients, pp 430. Academic Press, New York.
Pertoft H, Laurent TC (1977) Isopycnic separation of cells and cell organelles by centrifugation in modified colloidal silica gradients. In: N Catsimpoolas (eds) Methods of Cell Separation 1, pp 25–65. Plenum, New York.
Schmitt JM, Herrmann RG (1977) Fraction of cell organelles in silica sol gradients. In: D Prescott (ed) Methods of Cell Biology 10, pp 177–200. Academic Press, New York.
Thorsteinson KE, Wolyn DJ (1994) Simultaneous purification of chloroplast and mitochon-drial DNA without isolation of intact organelles from green carrot tissue and suspension cultures. Plant Mol Biol Rep 12: 26–36.
Matthews BF, DeBonte LR (1985) Chloroplast and mitochondrial DNAs of the carrot and its wild relatives. Plant Mol Biol Rep 3: 12–16.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1994 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Price, C.A., Hadjeb, N., Newman, L., Reardon, E.M. (1994). Isolation of chloroplasts and chloroplast DNA. In: Gelvin, S.B., Schilperoort, R.A. (eds) Plant Molecular Biology Manual. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-0511-8_15
Download citation
DOI: https://doi.org/10.1007/978-94-011-0511-8_15
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-011-7654-5
Online ISBN: 978-94-011-0511-8
eBook Packages: Springer Book Archive