The induction of sensitivity to gibberellin in aleurone tissue of developing wheat triticum aestivum cultivar sappo grains 3. sensitization of isolated protoplasts

Norman, H.A.; Black, M.; Chapman, J.C.

Planta (Heidelberg) 158(3): 264-271


ISSN/ISBN: 0032-0935
Accession: 006694272

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A method for isolating viable protoplasts in high yield from the aleurone layers of developing wheat grains is described, and the techniques for their subsequent culture outlined. Protoplasts from untreated tissue do not produce .alpha.-amylase in response to GA3 if the incubation temperature is left at C. However, pre-treatment of the protoplast preparation at temperatures above C for at least 8 h followed by a short incubation at C induces sensitivity to the growth regulator such that .alpha.-amylase is produced. The requirements of the sensitization process are similar to those for intact aleurone tissue although additional adjustment to the Ca2+ is beneficial. Pre-treatment of aleurone layers with the sensitizing temperature regimes prior to protoplast isolation have the advantage of increasing protoplast viability. Once sensitized, the protoplasts respond to a GA3 concentration as low as 10-11 mol dm-3 with a maximal response at 10-9 mol dm-3. The successful isolation of wheat aleurone protoplasts whose sensitivity to GA3 can be manipulated represents a useful step towards investigating the role of cell membranes in growth regulator action.