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. 1989 Jul;8(7):1907–1913. doi: 10.1002/j.1460-2075.1989.tb03593.x

A wheat α-Amy2 promoter is regulated by gibberellin in transformed oat aleurone protoplasts

A K Huttly 1, D C Baulcombe 2
PMCID: PMC401049  PMID: 16453890

Abstract

Gibberellin (GA3)-responsive aleurone protoplasts isolated from Avena sativa have been successfully used as a transient expression system to analyse promoter fusions between the wheat α-amylase gene α-Amy2/54 and the reporter gene GUS. Following PEG-mediated uptake of plasmid DNA, transient expression directed by the α-Amy2/54 promoter was found to be regulated in the same way as the endogenous oat α-amylase genes. Expression was thus dependent on the inclusion of GA3 in the protoplast incubation media, could not be detected before a lag phase of 2 days following transformation and was inhibited by simultaneous addition of abscisic acid (ABA) with GA3 to the media. In contrast, expression from the CaMV 35S promoter in the same system was not affected by GA3 or ABA and could be detected 1 day after transformation. Introduction of a further three different promoters into the aleurone protoplasts confirmed that GA3 specifically controlled transient expression from the α-Amy2/54 promoter only. Promoter deletions of the α-Amy2/54: GUS fusion demonstrated that sequences within 300 bp of the start of transcription of the gene were sufficient to direct high-level expression that was regulated by GA3 and ABA.

Keywords: GA3-regulated, transient expression, oat aleurone protoplasts, α-amylase, β-glucuronidase

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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