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Enolases: storage compounds in seeds? Evidence from a proteomic comparison of zygotic and somatic embryos of Cyclamen persicum Mill.

Plant Molecular Biology volume 75, pages 305–319 (2011)Cite this article

Abstract

Somatic embryogenesis is well established for the economic relevant ornamental crop Cyclamen and thus could supplement the elaborate propagation via seeds. However, the use of somatic embryogenesis for commercial large scale propagation is still limited due to physiological disorders and asynchronous development within emerged embryos. To overcome these problems, profound knowledge of the physiological processes in Cyclamen embryogenesis is essential. Thus, the proteomes of somatic and zygotic embryos were characterised in a comparative approach. Protein separation via two dimensional IEF-SDS PAGE led to a resolution of more than 1,000 protein spots/gel. Overall, 246 proteins were of differential abundance in the two tissues compared. Mass spectrometry analysis of the 300 most abundant protein spots resulted in the identification of 247 proteins, which represent 90 distinct protein species. Fifty-five percent of the 247 proteins belong to only three physiological categories: glycolysis, protein folding and stress response. The latter physiological process was especially predominant in the somatic embryos. Remarkably, the glycolytic enzyme enolase was the protein most frequently detected and thus is supposed to play an important role in Cyclamen embryogenesis. Data are presented that indicate involvement of “small enolases” as storage proteins in Cyclamen. A digital reference map was established via a novel software tool for the web-based presentation of proteome data linked to KEGG and ExPasy protein-databases and both were made publicly available online.

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Abbreviations

AA:

Amino Acids

CBB-G 250:

Coomassie brilliant blue-G 250

2D:

Two-dimensional

2iP:

6-(γ,γdimethylallylamino)purine

2,4-D:

2,4-Dichlorophenoxyacetic acid

DIGE:

Differential gel electrophoresis

EST:

Expressed sequence tag

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

Glu:

Glutamine

His:

Histidine

HSP:

Heat shock protein

IEF-SDS PAGE:

Isoelectric focusing—sodium dodecyl sulphate polyacrylamide gel electrophoresis

LC:

Liquid chromatography

Lys:

Lysine

MS:

Mass spectrometry

SE:

Somatic embryos,

Ser:

Serine

ZE:

Zygotic embryos

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Acknowledgments

The authors would like to thank Michael Senkler for programming GelMap and for computer-related assistance, Kathrin Lindhorst for her excellent technical assistance in the lab, Dr. Frank Colditz and Jenniffer W. Mwangi for critically reading the manuscript and Varinova for providing Cyclamen plants. The project was funded by the DFG (Deutsche Forschungsgemeinschaft-German Research Foundation).

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Affiliations

  1. Institute of Plant Genetics, Leibniz Universitaet Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany

    Christina Rode & Hans-Peter Braun

  2. Laboratoire de Spectrométrie de Masse Bio-Organique, IPHC-DSA, Université de Strasbourg, CNRS, UMR7178, 25 rue Becquerel, 67087, Strasbourg, France

    Sébastien Gallien & Alain Van Dorsselaer

  3. Institut de Biologie Moléculaire des Plantes (IBMP), Université de Strasbourg, CNRS-UPR2357, 67083, Strasbourg, France

    Dimitri Heintz

  4. Institute of Floriculture and Woody Plant Science, Leibniz Universitaet Hannover, Herrenhaeuser Str. 2, 30419, Hannover, Germany

    Traud Winkelmann

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  1. Christina Rode
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  2. Sébastien Gallien
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  3. Dimitri Heintz
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  4. Alain Van Dorsselaer
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  5. Hans-Peter Braun
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Correspondence to Christina Rode.

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Rode, C., Gallien, S., Heintz, D. et al. Enolases: storage compounds in seeds? Evidence from a proteomic comparison of zygotic and somatic embryos of Cyclamen persicum Mill.. Plant Mol Biol 75, 305–319 (2011). https://doi.org/10.1007/s11103-010-9729-x

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  • DOI: https://doi.org/10.1007/s11103-010-9729-x

Keywords

  • Differential gel electrophoresis
  • Embryogenesis
  • Mass spectrometry
  • Proteome reference maps
  • Protein identification
  • Two-dimensional polyacrylamide gel electrophoresis