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Journal of Experimental Botany, Vol 49, 839-845, Copyright © 1998 by Oxford University Press

ARTICLES

In vitro development of maize immature embryos: a tool for embryogenesis analysis

E Matthys-Rochon, F Piola, E Le Deunff, , R l and C Dumas
Laboratoire de Reproduction et Developpement des Plantes, UMR 9938 (CNRS-INRA-ENS) Ecole Normale Superieure de Lyon, 46 Allee d'Italie, F-69364 Lyon Cedex 07, France; Laboratory of General Botany, Faculty of Biology, Adam Mickiewicz University, PL-61713 Poznan, Poland; Corresponding author; e-mail: Elisabeth.Matthys-Rochon@ens-lyon.fr

During monocot embryo development, the zygote goes through a proembryo stage characterized by a radial symmetry and later becomes a true embryo with a bilateral symmetry. In order to determine culture conditions for immature embryonic stages, proembryos and embryos were isolated from controlled pollinated maize plants and developed in vitro. Precise culture conditions were determined for each type of explant: a monolayer system for embryos using NBM medium enriched with maltose (0.25 M) but without hormones, and a bilayer system for proembryo stages using N6 medium supplemented with maltose (0.35 M) and zeatin (3 mM). Morphological, cytological, and in situ hybridization analysis have shown that the resulting embryos (stages 1-2), developed in vitro, were similar to those formed in vivo and subsequently gave rise to fertile plants. This work demonstrates that successful embryo differentiation is dependent on specific parameters including the genotype, the nature of the carbon source, the type and concentration of hormones used and orientation of the embryos on the medium. The potential use of these results for embryo rescue and mutant analysis are discussed.Keywords: Maize, proembryo in vitro culture, plant regeneration, ISH-Ltp2.
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