𝔖 Bobbio Scriptorium
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Gibberellin perception at the plasma membrane ofAvena fatuaaleurone protoplasts

✍ Scribed by Richard Hooley; Michael H. Beale; Sally J. Smith


Book ID
104659851
Publisher
Springer-Verlag
Year
1991
Tongue
English
Weight
998 KB
Volume
183
Category
Article
ISSN
0032-0935

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✦ Synopsis


A functional assay for gibberellin (GA) receptors is described based on the induction of a-amylase gene expression in isolated aleurone protoplasts of Avenafatua L. by GA4 immobilised to Sepharose beads.

A 17-thiol derivative of GA4, shown to be biologically active with aleurone protoplasts, has been coupled to epoxy-activated Sepharose 6B. This GA4-17-Sepharose induces high levels of a-amylase when incubated with isolated aleurone protoplasts, while cells of the intact aleurone layer do not respond appreciably to the immobilised GA~. In order to eliminate the possibility that GA4 may be released from the Sepharose when incubated with protoplasts, aleurone layers and isolated aleurone protoplasts have been co-incubated, and their responses to GA4, GA4-17-Sepharose and control Sepharose estimated by determining the relative amounts of a-amylase mRNA induced in each tissue. Evidence from these experiments is consistent with the view that GA4-17-Sepharose induces a-amylase gene expression in aleutone protoplasts by interacting with the protoplast surface. This indicates that GA receptors may be located at, or near, the external face of the aleurone plasma membrane.


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## Summary Plasma membrane ion channels of protoplasts from tobacco cell suspensions were characterized by patch‐clamp experiments. In the whole‐cell configuration, a voltage‐dependent current with a current maximum around −90 mV was observed that displayed a reversal potential close to the Nernst