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Plant Molecular Biology

, Volume 24, Issue 2, pp 327–340 | Cite as

A seed-specific Brassica napus oleosin promoter interacts with a G-box-specific protein and may be bi-directional

  • James S. Keddie
  • Miltos Tsiantis
  • Pietro Piffanelli
  • Rino Cella
  • Polydefkis Hatzopoulos
  • Denis J. Murphy
Research Article

Abstract

In Brassica napus, oleosins are expressed at high levels in the seed during the latter stages of embryo development. The cis-acting regulatory properties of an 872 bp promoter fragment of a B. napus oleosin gene were examined by analysis of β-glucuronidase (GUS) expression in transgenic tobacco plants containing an oleosin promoter-GUS transcriptional fusion. The reporter gene was expressed at high levels only in seeds, specifically in embryo and endosperm tissue and regulated throughout seed development. These data demonstrate that oleosin gene transcription is regulated in a tissue-specific and temporally regulated manner and clearly indicate that oleosin protein expression is co-ordinated primarily at the transcriptional level. Oleosin mRNA was shown to be abscisic acid (ABA) inducible and an ABA-response element in the oleosin promoter was shown to be bound by a protein factor in a sequence-specific manner. Sequence analysis of the oleosin promoter has identified several other putative cis-acting sequences which may direct oleosin gene expression. The presence of a large open reading frame in the bottom strand of the oleosin promoter (ORF2) which encodes a polypeptide similar to the ethylene-induced E4 gene of tomato is reported. A PCR-generated DNA probe containing the ORF2 sequence hybridised with a 1.4 kb transcript in total RNA extracts of a variety of tissues, including leaves and germinated seed cotyledons. This finding suggests that the oleosin gene promoter directs transcription in both directions. It is the first report of a bi-directional nuclear gene promoter in plants.

Key words

abscisic acid ABA-response element bi-directional promoter Brassica napus oleosin seed development 

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Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • James S. Keddie
    • 1
  • Miltos Tsiantis
    • 1
  • Pietro Piffanelli
    • 1
  • Rino Cella
    • 2
  • Polydefkis Hatzopoulos
    • 3
  • Denis J. Murphy
    • 1
  1. 1.Department of Brassica and Oilseeds ResearchJohn Innes CentreNorwichUK
  2. 2.Department of Genetics & MicrobiologyUniversity of PaviaPaviaItaly
  3. 3.Molecular Biology Laboratory, Department of Agricultural Biology & Biotechnology, AgriculturalUniversity of AthensGreece

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