Gibberellic-acid-responsive protoplasts from mature aleurone of Himalaya barley

Gibberellic acid (GA3)-responsive protoplasts were prepared from mature aleurone layers of Himalaya barley. Protoplasts prepared in air (air-protoplasts) synthesized α-amylase (EC 3.2.1.1) in the presence of GA3 at a rate which was 4-5 times greater that in its absence. Protoplasts prepared in nitrogen (N2-protoplasts) took longer than air-protoplasts to respond to GA3 but α-amylase synthesis ultimately attained a rate which was similar to that for air-protoplasts and which was many times that occurring in the absence of the hormone. Many characteristics of the protoplast response were similar to those of intact aleurone layers. α-Amylase arose by new synthesis, its synthesis was inhibited by abscisic acid, it was isozymically similar to aleurone layer enzyme, most of it was secreted into the incubation medium and its synthesis was accompanied by accumulation of α-amylase mRNA. GA3-induced changes in protein synthesis and cell structure also resembled those of intact aleurone cells. We conclude that the response of the protoplasts to GA3 is normal and that they present a useful system for the study of GA3 action in barley aleurone.