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Proteome analysis of embryogenic cell suspensions of cowpea (Vigna unguiculata)

  • Physiology and Biochemistry
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Abstract

Using a combination of two-dimensional gel electrophoresis protein mapping and mass spectrometry analysis, we have established proteome reference maps of embryogenic cell suspensions of cowpea (Vigna unguiculata). The cell suspensions were generated from young primary leaves and contained basically pro-embryogenic masses, which enabled us to dissect their proteome composition while eliminating the complexity of too many cell types. Over 550 proteins could reproducibly be resolved over a pI range of 3–10. A total of 128 of the most abundant protein spots were excised, digested in-gel with trypsin and analyzed by tandem mass spectrometry. This enabled the identification of 67 protein spots. Two of the most abundant proteins were identified as a chitinase and as a ribonuclease belonging to the family of PR-4 and PR-10 proteins, respectively. The expression of the respective genes was confirmed by RT-PCR and the pattern of deposition of the PR-10 protein in cell suspensions as well as in developing cowpea seeds, roots, shoots and flowers were determined by Western blot experiments, using synthetic antibodies raised against a 14-amino acid synthetic peptide located close to the C-terminal region of the PR-10 protein.

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Abbreviations

FEC:

Friable embryogenic calli

ECS:

Embryogenic cell suspension

PR:

Pathogenesis related

2,4-d :

2,4-Dichlorophenoxyacetic acid

2-DE:

Two-dimensional electrophoresis

MALDI-TOF-TOF-MS/MS:

Matrix assisted laser desorption ionization-time of flight-tandem mass spectrometry

EST:

Expressed sequence tag

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Federal University of Ceará, P.O. Box 6039, Fortaleza, Ceará, Brazil

    F. C. S. Nogueira, E. F. Gonçalves, E. S. Jereissati, J. H. Costa & F. A. P. Campos

  2. Department of Biology, Federal University of Ceará, Fortaleza, Ceará, Brazil

    A. A. Soares

  3. CENARGEN/EMBRAPA, Brasilia, DF, Brazil

    O. B. Oliveira-Neto

  4. Chemistry Institute, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil

    M. Santos & G. B. Domont

Authors
  1. F. C. S. Nogueira
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  2. E. F. Gonçalves
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  3. E. S. Jereissati
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  4. M. Santos
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  5. J. H. Costa
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  6. O. B. Oliveira-Neto
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  7. A. A. Soares
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  8. G. B. Domont
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  9. F. A. P. Campos
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Corresponding author

Correspondence to F. A. P. Campos.

Additional information

Communicated by P. Lakshmanan.

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Nogueira, F.C.S., Gonçalves, E.F., Jereissati, E.S. et al. Proteome analysis of embryogenic cell suspensions of cowpea (Vigna unguiculata). Plant Cell Rep 26, 1333–1343 (2007). https://doi.org/10.1007/s00299-007-0327-6

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  • DOI: https://doi.org/10.1007/s00299-007-0327-6

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