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Confocal Laser Scanning Microscopy Detection of Chlorophylls and Carotenoids in Chloroplasts and Chromoplasts of Tomato Fruit

  • Lucio D’Andrea
  • Montse Amenós
  • Manuel Rodríguez-ConcepciónEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1153)

Abstract

Plant cells are unique among eukaryotic cells because of the presence of plastids, including chloroplasts and chromoplasts. Chloroplasts are found in green tissues and harbor the photosynthetic machinery (including chlorophyll molecules), while chromoplasts are present in non-photosynthetic tissues and accumulate large amounts of carotenoids. During tomato fruit development, chloroplasts are converted into chromoplasts that accumulate high levels of lycopene, a linear carotenoid responsible for the characteristic red color of ripe fruit. Here, we describe a simple and fast method to detect both types of fully differentiated plastids (chloroplasts and chromoplasts), as well as intermediate stages, in fresh tomato fruits. The method is based on the differential autofluorescence of chlorophylls and carotenoids (lycopene) detected by Confocal Laser Scanning Microscopy.

Keywords

Chloroplast Chlorophylls Chromoplast Carotenoids Lycopene Confocal microscopy Tomato fruit Fluorescence 

Notes

Acknowledgements

Our work is funded by grants from the Catalan AGAUR (2009SGR-26 and XRB), Spanish DGI (BIO2011-23680 and PIM2010IPO-00660), and European Union FP7 (TiMet, contract 245143). We are members of the IBERCAROT network funded by CYTED (112RT0445). L.D. received a predoctoral fellowship of the Spanish Ministerio de Educación FPU program.

References

  1. 1.
    López-Juez E, Pike K (2005) Plastids unleashed: their development and their integration in plant development. Int J Dev Biol 410:557–577CrossRefGoogle Scholar
  2. 2.
    Neuhaus HE, Emes MJ (2000) Nonphoto-synthetic metabolism in plastids. Annu Rev Plant Physiol Plant Mol Biol 51:111–140PubMedCrossRefGoogle Scholar
  3. 3.
    López-Juez E (2007) Plastid biogenesis, between light and shadows. J Exp Bot 58:11–26PubMedCrossRefGoogle Scholar
  4. 4.
    Lu S, Li L (2008) Carotenoid metabolism: biosynthesis, regulation, and beyond. J Integ Plant Biol 50:778–785CrossRefGoogle Scholar
  5. 5.
    Cazzonelli C, Pogson B (2010) Source to sink: regulation of carotenoid biosynthesis in plants. Trends Plant Sci 15:1360–1385CrossRefGoogle Scholar
  6. 6.
    Ruiz-Sola MA, Rodriguez-Concepción M (2012) Carotenoid biosynthesis in arabidopsis: a colorful pathway. Arabidopsis Book 10:e0158PubMedCentralPubMedCrossRefGoogle Scholar
  7. 7.
    Waters M, Pyke K (2005) Plastid development and differentiation. Annu Plant Rev Plastids 13:30–59Google Scholar
  8. 8.
    Bian W, Barsan C, Egea I, Purgatto E, Chervin C, Zouine M, Latché A, Bouzayen M, Pech JC (2011) Metabolic and molecular events occurring during chromoplast biogenesis. J Bot 2011:289859Google Scholar
  9. 9.
    Egea I, Barsan C, Bian W, Purgatto E, Latche A, Chervin C, Bouzayen M, Pech JC (2010) Chromoplast differentiation: current status and perspectives. Plant Cell Physiol 51:1601–1611PubMedCrossRefGoogle Scholar
  10. 10.
    Egea I, Bian W, Barsan C, Jauneau A, Pech JC, Latche A, Li Z, Chervin C (2011) Chloroplast to chromoplast transition in tomato fruit spectral confocal microscopy analyses of carotenoid and chlorophylls in isolated plastids and time lapse recording on intact live tissue. Ann Bot 108:2101–2107CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Lucio D’Andrea
    • 1
  • Montse Amenós
    • 1
  • Manuel Rodríguez-Concepción
    • 1
    Email author
  1. 1.Centre for Research in Agricultural Genomics (CRAG) CSIC-IRTA-UAB-UBBarcelonaSpain

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