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Annals of Botany 78: 223-232, 1996
© 1996 Annals of Botany Company

There is No Somatic Meiosis in Embryogenic Leaves of Cichorium

CAROLINE RAMBAUD+, ANNE-SOPHIE BLERVACQ, PIERRE DEVAUX, THÉRÈSE DUBOIS, JEAN DUBOIS, FRANÇOIS LAMMIN and JACQUES VASSEUR

Laboratoire de Physiologie cellulaire et Morphogenèse végétales, Université des Sciences et Technologies de Lille, SN2, 59655 Villeneuve d'Ascq, Cedex Florimond-Desprez, Laboratoire de Biotechnologie, BP 41, Cappelle en Pévèle, 59242, France

October 2, 1995 ; March 5, 1996

Several papers dealing with carrot cell cultures describe meiosis-like divisions and haploid cells prior to somatic embryogenesis. We have studied the first division in embryogenic mesophyll cells of a diploidCichorium intybus L. and of a tetraploid hybridC. intybus L.xC. endivia L. which undergo direct somatic embryogenesis from single cells when leaf fragments are placed in a liquid agitated inductive medium (modified MS with 1x10-7M NAA and 2.5x10-6M 2-iP), in darkness, at 35°C. Microsporogenesis inC. intybus provided aspects of meiosis for comparison. In leaves incubated in inductive conditions, DAPI staining of nuclei showed normal mitosis on days 3–6; about 0.6% cells in prophase had undergone spontaneous endoreduplication leading to a tetraploid somatic embryo. Immunocytochemistry of tubulin revealed the constant presence of a preprophase band, as in a normal mitosis. The first pluricellular somatic embryos became visible on day 5 of culture. Flow cytometric determination of nuclear DNA on days 4, 5 and 6 did not show any peak corresponding to the 1C DNA level for the diploid plant or to the 2C DNA level for the tetraploid. Instead there was a weak but constant peak at the 4C and 8C levels. We conclude that inCichorium leaves, the first division of somatic embryogenesis is a normal mitosis, with a small shift to endoreduplication. In our opinion, somatic meiosis is not a prerequisite during direct somatic embryogenesis.

Cichorium ; chicory; somatic embryogenesis; cell division; flow cytometry; tubulin


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