Abstract
Measurement of the concentrations of bioactive gibberellin (GA) in germinating cereal seeds is important to grasp the molecular mechanisms of storage starch degradation following the activation of aleurone cells. We investigated a simple method for the estimation of bioactive GA in germinating rice seeds using α-amylase secreted by the aleurone. The α-amylase enzyme activities released from aleurone in response to GA application were plotted against the applied GA concentrations and the amount of α-amylase proteins were visualized on nitrocellulose filter using polyclonal antibodies to compare with those synthesized in germinating seeds. We could define the bioactive GA concentration in a rice seedling ranged from 0.1 to 1 nM and identify the 42 kDa α-amylase isoform, which appears only when the internal GA condition becomes higher than 1 nM. This isoform will be a good index for the analysis of signaling cascades mediated by high concentrations of GA.
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