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Free Content Identification of a methyl jasmonate-responsive region in the promoter of a lipoxygenase 1 gene expressed in barley grain

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A genomic DNA fragment was isolated containing 5′ upstream sequences and part of the open reading frame corresponding to the lipoxygenase 1 cDNA (LoxA) expressed in barley grains during development and germination. Lox1 transcription was shown to be methyl jasmonate (MeJA)- and wound-inducible in leaves, but Lox1 transcripts were not detected in mildew-infected leaves, although this is a commonly observed response to pathogenic attack in various plants. Transient gene expression assays were used to identify a promoter region involved in MeJA-responsive expression. Analysis of 5′ and 3′ promoter deletions indicated that sequences between −363 and −294 conferred MeJA-responsive expression. Deletions/replacements covering this part of the promoter further defined a MeJA-responsive region between −331 and −291. Insertion of the region −328 to −293 into the constitutive CaMV 35S promoter conferred MeJA-responsive expression. The 36 bp fragment contains the motif TGACG as inverted repeats, which has been previously identified as a binding site for bZIP transactivating factors. Site-directed mutagenesis on these TGACG motifs abolished MeJA-responsive expression, clearly identifying them as MeJA-responsive elements. Sequence comparisons found no similar motif in other characterized promoters of MeJA-inducible genes, but suggested a common spatial structure which may serve as a binding site for transacting factors involved in the MeJA signal transduction pathway.
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Document Type: Research Article

Publication date: March 1, 1997

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