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Plastids pp 189–203Cite as

Isolation of Plastid Fractions from the Diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1829))

Abstract

The so-called “complex” plastids from diatoms possessing four envelope membranes are a typical feature of algae that arose from secondary endosymbiosis. Studying isolated plastids from these algae may allow answering a number of fundamental questions regarding diatom photosynthesis and plastid functionality. Due to their complex architecture and their integration into the cellular endoplasmic reticulum (ER) system, their isolation though is still challenging. In this work, we report a reliable isolation technique that is applicable for the two model diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum. The resulting plastid-enriched fractions are of homogenous quality, almost free from cellular contaminants, and feature structurally intact thylakoids that are capable to perform oxygenic photosynthesis, though in most cases they seem to lack most of the stromal components as well as plastid envelopes.

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References

  1. Walker DA (1964) Improved rates of carbon dioxide fixation by illuminated chloroplasts. Biochem J 92(3):22C–23C. https://doi.org/10.1042/bj0920022C

    Article  PubMed  CAS  Google Scholar 

  2. Jensen RG, Bassham JA (1966) Photosynthesis by isolated chloroplasts. Proc Natl Acad Sci U S A 56(4):1095–1101

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Stankovic ZS, Walker DA (1977) Photosynthesis by isolated pea chloroplasts: some effects of adenylates and inorganic pyrophosphate. Plant Physiol 59(3):428–432. https://doi.org/10.1104/pp.59.3.428

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  4. Lyttleton JW (1962) Isolation of ribosomes from spinach chloroplasts. Exp Cell Res 26(2):312–317. https://doi.org/10.1016/0014-4827(62)90183-0

    Article  PubMed  CAS  Google Scholar 

  5. Edwards GE, Robinson SP, Tyler NJ et al (1978) Photosynthesis by isolated protoplasts, protoplast extracts, and chloroplasts of wheat: influence of orthophosphate, pyrophosphate, and adenylates. Plant Physiol 62(2):313–319. https://doi.org/10.1104/pp.62.2.313

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  6. Pollock SV, Colombo SL, Prout DL Jr et al (2003) Rubisco activase is required for optimal photosynthesis in the green alga Chlamydomonas reinhardtii in a low-CO(2) atmosphere. Plant Physiol 133(4):1854–1861. https://doi.org/10.1104/pp.103.032078

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  7. Schnell RA, Lefebvre PA (1993) Isolation of the Chlamydomonas regulatory gene NIT2 by transposon tagging. Genetics 134(3):737–747

    PubMed  PubMed Central  CAS  Google Scholar 

  8. Mendiola-Morgenthaler L, Leu S, Boschetti A (1985) Isolation of biochemically active chloroplasts from chlamydomonas. Plant Sci 38(1):33–39. https://doi.org/10.1016/0168-9452(85)90076-7

    Article  CAS  Google Scholar 

  9. Mason CB, Bricker TM, Moroney JV (2006) A rapid method for chloroplast isolation from the green alga Chlamydomonas reinhardtii. Nat Protocols 1(5):2227–2230. https://doi.org/10.1038/nprot.2006.348

    Article  PubMed  CAS  Google Scholar 

  10. Round FE, Crawford RM, Mann DG (2007) Diatoms: biology and morphology of the genera. Plastids. Cambridge University Press, Cambridge, p 59

    Google Scholar 

  11. Wittpoth C, Kroth PG, Weyrauch K et al (1998) Functional characterization of isolated plastids from two marine diatoms. Planta 206(1):79–85. https://doi.org/10.1007/s004250050376

    Article  CAS  Google Scholar 

  12. Guillard RL (1975) Culture of phytoplankton for feeding marine invertebrates. In: Smith W, Chanley M (eds) Culture of marine invertebrate animals. Springer, New York, pp 29–60. https://doi.org/10.1007/978-1-4615-8714-9_3

    Chapter  Google Scholar 

  13. Flori S, Jouneau P-H, Bailleul B et al (2017) Plastid thylakoid architecture optimizes photosynthesis in diatoms. Nat Commun 8:15885. https://doi.org/10.1038/ncomms15885

    Article  PubMed  PubMed Central  CAS  Google Scholar 

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Acknowledgments

We would like to thank Doris Ballert for the help with the cultivation of T. pseudonana and P. tricornutum and the BioImaging Center at the University of Konstanz for the assistance during fluorescence microscopy. This work was supported by a stipend of the Graduate School Chemical Biology (KoRS-CB) and by the University of Konstanz to AFS. CRB further wishes to acknowledge the German Research Foundation (DFG, grant KR 1661/6-1) and the Gordon and Betty Moore Foundation GBMF 4966 (grant DiaEdit). SF and GF acknowledge funds from the Marie Curie Initial Training Network Accliphot (FP7-PEOPLE-2012-ITN, 316427) and from the DRF impulsion FIB-Bio program.

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Authors and Affiliations

  1. Plant Ecophysiology, Department of Biology, University of Konstanz, Konstanz, Germany

    Alexander F. Schober, Peter G. Kroth & Carolina Río Bártulos

  2. Department of Cell and Molecular Processes, Marine Biological Association of the United Kingdom, Plymouth, UK

    Serena Flori

  3. Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Institut National Recherche Agronomique, Commissariat à l’Energie Atomique et aux Energies Alternatives, CEA Grenoble, UMR5168, Université Grenoble Alpes, Grenoble, France

    Giovanni Finazzi

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  1. Alexander F. Schober
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  2. Serena Flori
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  3. Giovanni Finazzi
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  4. Peter G. Kroth
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  5. Carolina Río Bártulos
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Corresponding author

Correspondence to Carolina Río Bártulos .

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Editors and Affiliations

  1. Laboratoire de Physiologie Cellulaire et Végétale, Centre National de la Recherche Scientifique, Commissariat à l’Energie Atomique et aux Energies Alternatives, CEA Grenoble, Institut National Recherche Agronomique, UMR5168, Université Grenoble Alpes, Grenoble, France

    Eric Maréchal

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Schober, A.F., Flori, S., Finazzi, G., Kroth, P.G., Bártulos, C.R. (2018). Isolation of Plastid Fractions from the Diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum. 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_13

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  • DOI: https://doi.org/10.1007/978-1-4939-8654-5_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8653-8

  • Online ISBN: 978-1-4939-8654-5

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