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Articles

Control of -Amylase mRNA Accumulation by Gibberellic Acid and Calcium in Barley Aleurone Layers ¹

Department of Botany, University of California, Berkeley, California 94720

Pulse-labeling of barley (Hordeum vulgare L. cv Himalaya) aleurone layers incubated for 13 hours in 2.5 micromolar gibberellic acid (GA₃) with or without 5 millimolar CaCl₂ shows that -amylase isozymes 3 and 4 are not synthesized in vivo in the absence of Ca²⁺. A cDNA clone for -amylase was isolated and used to measure -amylase mRNA levels in aleurone layers incubated in the presence and absence of Ca²⁺. No difference was observed in -amylase mRNA levels between layers incubated for 12 hours in 2.5 micromolar GA₃ with 5 millimolar CaCl₂ and layers incubated in GA₃ alone. RNA isolated from layers incubated for 12 hours in GA₃ with and without Ca²⁺ was translated in vitro and was found to produce the same complement of translation products regardless of the presence of Ca²⁺ in the incubation medium. Immunoprecipitation of translation products showed that the RNA for -amylase synthesized in Ca²⁺-deprived aleurone layers was translatable. Ca²⁺ is required for the synthesis of -amylase isozymes 3 and 4 at a step after mRNA accumulation and processing.

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