Molecular Cloning and Characterization of a Brassinosteroid-Regulated Gene from Elongating Soybean (Glycine max L.) Epicotyls

Abstract

Brassinosteroids promote elongation and regulate gene expression in soybean (Glycine max L.) stems. We constructed a cDNA library from brassinosteroid-treated soybean epicotyls and used differential hybridization to isolate a cDNA (pBRU1) corresponding to a transcript whose abundance is increased by brassinosteroid treatment. Sequence analysis of pBRU1 revealed an open reading frame of 283 amino acids with a putative signal peptide of 29 amino acids. The sequence had extensive homology (77% identity, 89% similarity) over 114 contiguous amino acids to the meri-5 gene of Arabidopsis thaliana (J.I. Medford, J.S. Elmer, H.J. Klee [1991] Plant Cell 3: 359-370), and significant homology (48% identity, 62% similarity) to a xyloglucan endotransglycosylase localized in the cell walls of nasturtium (J. de Silva, C.D. Jarman, D.A. Arrowsmith, M.S. Stronach, S. Chengappa, C. Sidebottom, J.S. Reid [1993] Plant J 3: 701-711). RNase protection studies showed that BRU1 transcript levels are not increased by 1.0 microM auxins, cytokinins, abscisic acid, or gibberellic acid and that BRU1 expression is highest in stem tissue. Findings from studies with run-on transcripts from isolated soybean nuclei most likely indicate that the regulation of BRU1 by brassinosteroids is largely posttranscriptional. The elevated levels of BRU1 transcripts in elongating tissue and the homology with a xyloglucan endotransglycosylase suggest a possible role for the BRU1 protein in brassinosteroid-stimulated elongation.