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Authors :
Thierry Jouenne
Michel Boulay
Jean Guern
Pierre Coutos-Thévenot
Olivier Maes
Source :
Plant Cell, Tissue and Organ Culture. 50:97-105
Publication Year :
1997
Publisher :
Springer Science and Business Media LLC, 1997.

Abstract

An embryogenic grapevine rootstock cell suspension, continuously grown in the presence of auxin, was predominantly composed of proembryogenic masses. When transferred to an auxin-free medium, grapevine somatic embryos developed but were rapidly blocked at the heart stage. This inhibition has been related to the presence of extracellular macromolecules (Coutos-Thevenot et al., 1992a). In this study, the initial cell population density has been found to influence markedly embryo development. Inoculations below 5·103 cells per ml were required to obtain fully grown cotyledonary embryos. Interestingly, extracellular proteins of molecular weights of 32, 34, 48 and 52 kDa accumulated in cultures grown at high population cell densities and disappeared in cultures inoculated at densities below 5·103 cells per ml. Protein fractions partially purified by ion exchange chromatography caused both an early inhibition of embryogenesis and a stimulation of secondary embryogenesis. Moreover, to test for the possibility of modulating embryo development through alterations of extracellular proteins, cultures were supplemented with proteases and protease inhibitors. The addition of trypsin increased the rate of embryo development only in cultures inoculated at a low cell population density. Conversely, the protease inhibitor aprotinin inhibited development, arresting embryos at globular and heart stages. Together, these results provide evidence that extracellular proteins modulate somatic embryogenesis and suggest that an extracellular proteolitic mechanism could be implicated in development.

Details

ISSN :
01676857
Volume :
50
Database :
OpenAIRE
Journal :
Plant Cell, Tissue and Organ Culture
Accession number :
edsair.doi...........6a6e3264e5a48b9018c86cde0d600951