Identical promoter elements are involved in regulation of the OPR1 gene by senescence and jasmonic acid in Arabidopsis

Abstract

Like other developmental processes, the terminal phase of leaf development, generally referred to as leaf senescence. regulates a subset of genes whose transcript abundances are increased during senescence. Jasmonic acid (JA), a plant growth regulator, also regulates the expression of subsets of genes in many aspects of plant growth and development, including leaf senescence. However, the underlying molecular mechanisms by which senescence and JA modulate gene expression are poorly understood. During an effort to isolate senescence-associated genes, we identified an Arabidopsis enhancer trap line in which the reporter gene GUS is up-regulated by both senescence and JA. The T-DNA tagged gene was subsequently cloned using thermal asymmetric interlaced PCR (TAIL-PCR). This gene encodes a 12-oxo-phytodienoic acid-10,11-reductase (OPR1). Consistent with the GUS expression data, RNA gel blot analysis showed that OPR1 was indeed up-regulated by both senescence and JA. Promoter deletion analysis and linker-scanning mutagenesis assays were employed to unveil the molecular bases of OPR1 regulation by senescence and JA. Two regulatory cis elements, namely JASE1 (5'-CGTCAATGAA-3') and JASE2 (5'-CATACGTCGTCAA-3'), in the promoter region of the gene, were identified. While JASE2 contains a mixed A/C box-like motif, JASE1 represents a new motif without any signature sequence so far reported. Both elements were required for the up-regulation of OPR1 by leaf senescence and JA. suggesting that leaf senescence and JA may share a common molecular mechanism for modulating OPR1.