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Hormonal Regulation of -Amylase Gene Transcription in Wild Oat (Avena fatua L.) Aleurone Protoplasts

AFRC Weed Research Organization, Begbroke Hill, Oxford OX5 1 PF, United Kingdom

The time of appearance and relative amounts of -amylase mRNA in wild oat (Avena fatua L.) aleurone protoplasts incubated with 1 micromolar gibberellin A₄ (GA₄) were closely correlated with the amounts of -amylase enzyme secreted by the protoplasts. In the absence of GA₄, or when protoplasts were incubated with 25 micromolar abscisic acid (ABA) together with 1 micromolar GA₄ no -amylase mRNA was detected and only very low levels of -amylase were secreted. Nuclei were isolated in high yields (65-71%) from aleurone protoplasts and in an in vitro transcription system displayed characteristics of a faithful DNA-dependent RNA synthesizing system. The time course of incorporation of [³H]-UTP suggested that the RNA synthesized was mainly `run off' transcription and therefore that the transcripts produced in vitro were those being synthesized in the protoplasts at the times when the nuclei were isolated. By hybridizing in vitro synthesized [³²P]RNA to barley -amylase cDNA and control filters we have estimated that 90 ± 10 ppm of the transcripts synthesized by nuclei isolated from GA₄ treated protoplasts can be attributed to -amylase sequences and that statistically insignificant amounts of these transcripts are obtained from control and GA₄ plus ABA treatments. The results suggest that GA₄ and ABA influence the transcription of -amylase genes in aleurone protoplasts of wild oat.

² Permanent address: University of Bristol, Department of Agricultural Sciences, Long Ashton Research Station, Weed Research Division, Begbroke Hill, Oxford OX5 1 PF, United Kingdom.

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