Developmental Regulation of (1→3, 1→4)-β-Glucanase Gene Expression in Barley

Abstract

Two genes encode (1-->3,1-->4)-beta-d-glucan 4-glucanohydrolase (EC 3.2.1.73) isoenzymes EI and EII in barley (Hordeum vulgare L.). Specific DNA probes have been used in Northern analyses to examine the developmental regulation of individual (1-->3,1-->4)-beta-glucanase genes in the aleurone and scutellum of germinated grain and in young leaves and young roots. In aleurone and scutella excised from germinated grain, mRNAs encoding both isoenzymes are present but developmental patterns differ between the two tissues. Thus, levels of both isoenzyme EI and EII mRNA increase significantly in the aleurone between 1 and 3 days after the initiation of germination. In the scutellum, isoenzyme EI mRNA predominates and decreases as germination proceeds. Isoenzyme EI mRNA appears in young leaves approximately 8 days after the initiation of germination and levels rise until about 20 days. Enzyme activity in leaf extracts parallels the development of isoenzyme EI mRNA. No isoenzyme EII mRNA is detected in the leaves in this period. Analysis of RNA from different leaf segments indicates that the isoenzyme EI mRNA is distributed relatively evenly along the length of the leaf. In young roots, mRNA encoding (1-->3,1-->4)-beta-glucanase isoenzyme EI is detected at high levels 3 to 6 days after the initiation of germination; again, little or no isoenzyme EII mRNA is found. Overall, transcription of the (1-->3,1-->4)-beta-glucanase isoenzyme EII gene appears to be restricted to the germinating grain, whereas isoenzyme EI is expressed in a wider range of tissues during seedling development.