Protein patterns associated with Pisum sativum somatic embryogenesis

Abstract

Total protein patterns were studied in the course of development of pea somatic embryos using simple protocol of direct regeneration from shoot apical meristems on auxin supplemented medium. Protein content and total protein spectra (SDS-PAGE) of somatic embryos in particular developmental stages were analysed in Pisum sativum, P. arvense, P. elatius and P. jomardi. Expression of seed storage proteins in somatic embryos was compared with their accumulation in zygotic embryos of selected developmental stages. Pea vegetative tissues, namely leaf and root, were used as a negative control not expressing typical seed storage proteins. The biosynthesis and accumulation of seed storage proteins was observed during somatic embryo development (since globular stage), despite of the fact that no special maturation treatment was applied. Major storage proteins typical for pea seed (globulins legumin, vicilin, convicilin and their subunits) were detected in somatic embryos. In general, the biosynthesis of storage proteins in somatic embryos was lower as compared to mature dry seed. However, in some cases the cotyledonary somatic embryos exhibited comparatively high expression of vicilin, convicilin and pea seed lectin, which was even higher than those in immature but morphologically fully developed zygotic embryos. Desiccation treatments did not affect the protein content of somatic embryos. The transfer of desiccated somatic embryos on hormone-free germination medium led to progressive storage protein degradation. The expression of true seed storage proteins may serve as an explicit marker of somatic embryogenesis pathway of regeneration as well as a measure of maturation degree of somatic embryos in pea.

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Abbreviations

MSB-medium:

medium with mineral salts after Murashige and Skoog (1962) and vitamins after Gamborg et al. (1968)

SDS-PAGE:

sodium dodecylsulphate polyacrylamide gel electrophoresis

SE:

somatic embryo

ZE:

zygotic embryo

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Griga, M., Horáček, J. & Klenotičová, H. Protein patterns associated with Pisum sativum somatic embryogenesis. Biol Plant 51, 201–211 (2007). https://doi.org/10.1007/s10535-007-0041-9

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Additional key words

  • embryo development
  • pea
  • seed proteins
  • storage substances
  • zygotic embryogenesis