Two negative cis-regulatory regions involved in fruit-specific promoter activity from watermelon (Citrullus vulgaris S.)

Tao Yin, Hanying Wu, Shanglong Zhang, Jingmei Liu, Hongyu Lu, Lingxiao Zhang, Yong Xu, Daming Chen

Research output: Contribution to journalArticlepeer-review

Abstract

A 1.8 kb 5′-flanking region of the large subunit of ADP-glucose pyrophosphorylase, isolated from watermelon (Citrullus vulgaris S.), has fruit-specific promoter activity in transgenic tomato plants. Two negative regulatory regions, from -986 to -959 and from -472 to -424, were identified in this promoter region by fine deletion analyses. Removal of both regions led to constitutive expression in epidermal cells. Gain-of-function experiments showed that these two regions were sufficient to inhibit RFP (red fluorescent protein) expression in transformed epidermal cells when fused to the cauliflower mosaic virus (CaMV) 35S minimal promoter. Gel mobility shift experiments demonstrated the presence of leaf nuclear factors that interact with these two elements. A TCCAAAA motif was identified in these two regions, as well as one in the reverse orientation, which was confirmed to be a novel specific cis-element. A quantitative β-glucuronidase (GUS) activity assay of stable transgenic tomato plants showed that the activities of chimeric promoters harbouring only one of the two cis-elements, or both, were ∼10-fold higher in fruits than in leaves. These data confirm that the TCCAAAA motif functions as a fruit-specific element by inhibiting gene expression in leaves.

Original languageEnglish (US)
Pages (from-to)169-185
Number of pages17
JournalJournal of Experimental Botany
Volume60
Issue number1
DOIs
StatePublished - Jan 2009

Keywords

  • ADP-glucose pyrophosphorylase
  • Cis-element
  • Fruit-specific promoter
  • Transcriptional factors
  • Transgenic tomato
  • Watermelon

ASJC Scopus subject areas

  • Physiology
  • Plant Science

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