Isolation of Arabidopsis mutants altered in the light-regulation of chalcone synthase gene expression using a transgenic screening approach.

Transgenic Arabidopsis expressing beta-glucuronidase (GUS) driven by a chalcone synthase gene (CHS) promoter were produced. GUS activity in the leaves increased with increasing fluence rates of white light in parallel with endogenous CHS transcript levels. An isogenic line homozygous for the transgene was obtained and mutagenized seedlings of this line were screened for altered light-induction of the transgene. Putative mutants with low GUS activity were not altered in the light-induction of endogenous CHS transcripts and are therefore not regulatory mutants. Two mutant lines (A12 and C10) with elevated levels of GUS activity in the light show a corresponding increase in CHS transcript levels. The A12 mutant was focussed upon and designated icx1 (increased chalcone synthase expression). This mutant has enhanced light-stimulation of CHS expression since CHS transcript levels in darkness in icx1 are very low, as in the wild-type. The transcript levels of two other genes involved in flavonoid biosynthesis are elevated in the light in icx1 as is anthocyanin formation. However, there is no alteration in LHCII chlorophyll a/b-binding protein gene (CAB) transcript levels under the same conditions. The altered gene expression phenotype of icx1 co-segregates with several other phenotypic characteristics, including fewer leaf trichomes and alterations to the seed coat. On the basis of these data and comparison with the Arabidopsis ttg (transparent testa glabra) mutant, it is suggested that the ICX1 gene product may be concerned both with the light-regulation of gene expression and with developmental processes occurring in the epidermis.