Organ-dependent regulation of a plant promoter isolated from rice by 'promoter-trapping' in tobacco

Abstract

A vector containing a transcriptionally inactive neomycin phosphotransferase II gene was used to select promoter sequences from a pool of random genomic DNA fragments. This paper describes how one such sequence (P4.7) isolated from Oryza sativa acts as a hormonally regulated promoter in Nicotiana tabacum. Relative expression ratios in leaf, root, midrib, callus, and stem tissue of tobacco plants are 1:5:4:10:17. Histochemical assays show that P4.7 activates the uidA reporter gene throughout the phloem and cortex of tobacco stems. Transcription from the P4.7 fragment is inducible in leaf tissue by low levels of alpha-naphthalene acetic acid or 6-benzyl-aminopurine, even when cell proliferation is inhibited by colchicine or hydroxyurea. Conversely, 1% DMSO was found to inhibit activation of P4.7 without interfering with callus formation. The fragment contains TATA and CAAT sequences normally found at the 5' end of many plant genes, and an additional region homologous to sequences located in similar positions in a variety of similarly regulated promoters. Promoter deletion and fusion experiments have indicated the location of a stem enhancer element in P4.7. The promoter trap system we have described may potentially be used to characterize transcriptional factors common to monocot and dicot species.