Promoter analysis of the chalcone synthase (chsA) gene of Petunia hybrida: a 67 bp promoter region directs flower-specific expression

Plant Mol Biol. 1990 Jul;15(1):95-109. doi: 10.1007/BF00017727.

Abstract

In order to scan the 5' flanking region of the chalcone synthase (chs A) gene for regulatory sequences involved in directing flower-specific and UV-inducible expression, a chimaeric gene was constructed containing the chs A promoter of Petunia hybrida (V30), the chloramphenicol acetyl transferase (cat) structural sequence as a reporter gene and the chs A terminator region of Petunia hybrida (V30). This chimaeric gene and 5' end deletions thereof were introduced into Petunia plants with the help of Ti plasmid-derived plant vectors and CAT activity was measured. A 220 bp chs A promoter fragment contains cis-acting elements conferring flower-specific and UV-inducible expression. A promoter fragment from -67 to +1, although at a low level, was still able to direct flower-specific expression but could not drive UV-inducible expression in transgenic Petunia seedlings. Molecular analysis of binding of flower nuclear proteins to chs A promoter fragments by gel retardation assays showed strong specific binding to the sequences from -142 to +81. Promoter sequence comparison of chs genes from other plant species, combined with the deletion analysis and gel retardation assays, strongly suggests the involvement of the TACPyAT repeats (-59 and -52) in the regulation of organ-specificity of the chs A gene in Petunia hybrida. We also describe an in vitro organ-specific transient expression system, in which flower or purple callus protoplasts are used, that enables us to pre-screen organ-specific expression of a chimaeric reporter gene.

MeSH terms

  • Acyltransferases / genetics*
  • Base Sequence
  • Chloramphenicol O-Acetyltransferase / genetics
  • Chromosome Deletion
  • Cloning, Molecular
  • DNA / genetics
  • Gene Expression Regulation, Enzymologic / radiation effects
  • Genetic Engineering
  • Molecular Sequence Data
  • Mutation
  • Organ Specificity
  • Plants / genetics*
  • Plants / metabolism
  • Promoter Regions, Genetic*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Repetitive Sequences, Nucleic Acid
  • Ultraviolet Rays

Substances

  • RNA, Messenger
  • DNA
  • Acyltransferases
  • Chloramphenicol O-Acetyltransferase
  • flavanone synthetase