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Journal of Experimental Botany, Vol 49, 249-253, Copyright © 1998 by Oxford University Press

ARTICLES

Short communication. Early changes in gene expression during direct somatic embryogenesis in alfalfa revealed by RAP-PCR

M Fowler, L Ong, E Russinova, A Atanassov, N Scott, A Slater and M Elliott
The Norman Borlaug Institute for Plant Science Research, De Montfort University, Scraptoft, Leicester LE7 9SU, UK; Present address: Palm Oil Research Institute of Malaysia (PORIM), PO Box 10620, 50720 Kuala Lumpur, Malaysia; Corresponding author e-mail: ads@dmu.ac.uk

A procedure for direct production of somatic embryos from alfalfa leaf explants has been developed. Wounding followed by 2,4-D treatment induces competent cells in the leaf to produce embryos. Changes in gene expression following induction of direct somatic embryogenesis were investigated by RNA arbitrarily primed PCR (RAP-PCR) with a view to identifying genes involved in controlling the onset of somatic embryo development. Amongst the induced sequences identified were a calnexin-like sequence (which was induced within 2 d), and a novel plant homologue of the yeast SN12 multi-drug resistance protein (which did not appear until 10 d after induction). RT-PCR amplification of these sequences confirmed the expression patterns revealed by RAP-PCR and was used to show that the calnexin-like gene is induced by 2,4-D, but that the level of expression is enhanced by wounding. The technique of RAP-PCR has the potential to isolate genes of interest by identifying specific expression patterns during complex developmental processes.Key words: ADP/ATP translocase, alfalfa, ATP binding cassette (ABC) protein, calnexin, differential display, Midicago falcata, SNQ2, somatic embryogenesis.
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