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Cell Biology International (1993) 17, 10131022 (Printed in Great Britain)
A methacrylate embedding procedure developed for immunolocalization on plant tissues is also compatible with in situ hybridization.
J. Kronenberger, T. Desprez, H. Höfte, M. Caboche and J. Traas
INRA, Laboratoire de Biologie Cellulaire, Route de Saint Cyr, 78026 Versailles cedex, France. fax: 33 1 30 83 30 99; tel. 33 1 30 83 30 41.
A recently described method that uses methacrylate embedding of aldehyde fixed plant tissues allows the immunolabelling of a range of antigens (Baskin et al. 1992). We have tested whether the same embedding procedure is also compatible with in situ hybridization. For this purpose we have used 2- 5 μm sections of methacrylate embedded plantlets of Arabidopsis thaliana. After removal of the resin the sections were prepared for in situ hybridization following standard procedures. Three different digoxygenin (dig)-labelled probes were used, recognizing RNAs coding for the chlorophyll a/b binding protein cab-140, the β-tubulin tub5 and meri a member of the meri-5 family. Each of the probes shows the labelling pattern expected from the literature. Moreover, the method allows a good structural preservation of very fragile tissues, in contrast to paraffin embedding. We conclude that methacrylate embedding, allowing both immunolabelling and in situ hybridization with high resolution and structural preservation, offers a high potential for the functional analysis of genes and proteins in plant development. This is especially true for Arabidopsis thaliana, a widely used model species where it seems to be the method of choice.
Key words: Arabidopsis thaliana, In situ hybridization, Immunofluorescence.