Floral tissue ofPetunia hybrida(V30) expresses only one member of the chalcone synthase multigene family

Abstract

Twenty independent, petal-specific chalcone synthase (CHS) cDNA clones have been isolated from Petunia hybrida variety Violet 30 (V30). Sequence analysis shows that the largest of these clones contains the entire coding sequence. Using this clone in Southern blot analysis reveals the presence of multiple CHS gene copies in the genome of Petunia hybrida V30. Hybridization and sequence analysis of the CHS cDNA clones shows that they are all copied from a single mRNA species. This indicates the presence of only one transcriptionally active CHS gene in petals. Finally we report the identification, cloning and partial characterization of this gene.

Sugar-Dependent Expression of the CHS-A Gene for Chalcone Synthase from Petunia in Transgenic Arabidopsis

“…The CHS-A gene was initially isolated as a petal-specific gene from Petunia hybrida (18). The 1-kb fragment of the CHS-A gene, which is expressed in petals specifically (16,18), was introduced into Arabidopsis thaliana in the form of a CHS-A::GUS fusion in order to examine whether the organ specificity is conserved in other plant species.…”

“…The 1-kb fragment of the CHS-A gene, which is expressed in petals specifically (16,18), was introduced into Arabidopsis thaliana in the form of a CHS-A::GUS fusion in order to examine whether the organ specificity is conserved in other plant species. We chose this host because A. thaliana is an excellent system for plant molecular biology for the many reasons described elsewhere (21).…”

“…Mol of Free University, Amsterdam; ref. 18), extending from the EcoRI site at approximately position -820 to the BclI site at position + 170, as measured from the site of initiation of transcription, was ligated into the EcoRI-BamHI sites of plasmid pBluescript KS(+) (Stratagene, La Jolla, CA). The HindIII-XbaI fragment from the resultant plasmid, pHTS7, which included the subcloned 1-kb fragment, was ligated into the HindIII-XbaI site of plasmid pHTS6.2 (a derivative of plasmid pBI 101.2 [11] with the gene for hygromycin phosphotransferase in its T-DNA region).…”

“…Weiss and Halevy demonstrated the effect of sucrose, in the presence of gibberellin, on the appearance of anthocyanin pigment in stamendetached flowers (28). Koes et al (18) isolated the cDNA and a genomic clone that encode an isozyme of CHS2 (CHS-A) as a petal-specific gene from petunia (16,18). They also discovered that there are at least seven genes for isozymes of chalcone synthase in petunia, including CHS-A and CHS-J, which are expressed in petals ( 17,18).…”

Sugar-Dependent Gibberellin-Induced Chalcone Synthase Gene Expression in Petunia Corollas

“…Following electrophoresis, the formaldehyde gel was briefly stained with ethidium bromide and photographed before blotting to ensure that equal amounts of RNA had been used for each sample. The blots were hybridized with 32P-labeled, full-size petunia cDNA probes for chs (Koes et al, 1986). Following hybridization, the blots were washed twice with 0.3 M NaC1, 30 mM sodium citrate, pH 7.0 (2X SSC), and 0.1% (w/v) SDS at 60°C for 10 min and then autoradiographed.…”