The chalcone synthase multigene family of Petunia hybrida (V30): sequence homology, chromosomal localization and evolutionary aspects (original) (raw)
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Plant Molecular Biology, 1989
Chalcone synthase (CHS) genes in Petunia hybrida comprise a multigene family containing at least 7 complete members in the strain Violet 30 (V30). Based on a high sequence homology in both coding and non-coding sequence, a number of CHS genes can be placed into two subfamilies. By restriction fragment length polymorphism (RFLP) analysis it was shown that both chromosomes II and V carry one of these subfamilies, in addition to the other CHS genes identified so far. Members of a subfamily were found to be closely linked genetically. Analysis of the Petunia species that contributed to the hybrid nature of P. hybrida (P. axillaris, P. parodii, P inflata and P. violacea) shows that none of the CHS gene clusters is specific for either one of the parents and therefore did not arise as a consequence of the hybridization. The number of CHS genes within a subfamily varies considerably among these Petunia species. From this we infer that the CHS subfamilies arose from very recent gene duplications.
Nucleic Acids Research, 1986
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. Plarnt variety, plasmids, phages and bacterial strains are listed in
Chalcone isomerase in flowers of mutants of Petunia hybrida
Planta, 1983
The effect of the gene Po on the activity of chalcone isomerase was investigated in Petunia hybrida. Furthermore, isomerase activities isolated from petals were compared with those extracted from anthers. No effect of Po on the pH-dependence of the isomerase and its kinetic properties was observed. With respect to these criteria, the enzyme extracted from anthers behaved in an identical
Evolution of the chalcone synthase gene family in the genus Ipomoea
Proceedings of the National Academy of Sciences, 1995
The evolution of the chalcone synthase [CHS; malonyl-CoA:4-coumaroyl-CoA malonyltransferase (cyclizing), EC 2.3.1.74] multigene family in the genus Ipomoea is explored. Thirteen CHS genes from seven Ipomoea species (family Convolvulaceae) were sequenced-three from genomic clones and the remainder from PCR amplification with primers designed from the 5' flanking region and the end of the 3' coding region of Ipomoea purpurea Roth. Analysis of the data indicates a duplication of CHS that predates the divergence of the Ipomoea species in this study. The Ipomoea CHS genes are among the most rapidly evolving of the CHS genes sequenced to date. The CHS genes in this study are most closely related to the Petunia CHS-B gene, which is also rapidly evolving and highly divergent from the rest of the Petunia CHS sequences.
Plant Science, 2005
Chalcone synthase (CHS), the first committed enzyme in the flavonoid biosynthetic pathway, is commonly encoded by multi-gene families with select members of these families accounting for the majority of expression. We have examined the CHS gene family in Viola cornuta, a plant whose flowers undergo ontogenetic color change. Using both RNA and RNA/DNA samples isolated from floral tissues at different pigment stages, we obtained 14 unique sequences from 60 total clones of a 288 bp fragment from the catalytic region of CHS. The V. cornuta sequences were monophyletic when compared to CHS orthologs from other taxa. Substitution models generally indicated unequal rates of transition and transversion as well as significant rate variation among sites. With a Tamura-Nei correction, nucleotide divergence ranged from 0.3 to 10.6% with the vast majority as synonymous changes. The nucleotide divergence pattern suggests designation of three V. cornuta CHS clades; based on divergence of CHS orthologs, these clades are consistent with three CHS orthologs in V. cornuta. Sequences from only a single clade were found to be expressed in all three floral pigment stages.
The Plant Journal, 1996
DNA sequences homologous to single-copy genes were labelled with biotinylated dUTP or digoxygenin-labelled dUTP and hybridized to chromosome spreads, The hybridization signals were visualized with fluorescent avidin-or antibody-conjugates. This method allowed the detection of DNA targets on metaphase chromosomes as small as 1.4 kb. The hybridization signals were identified as fluorescent spots on both sister chromatids. Using an 18S rDNA probe as marker to identify chromosomes II and III it was possible to assign single-copy genes to these chromosomes. In the line V30 the endogenous chalcone synthase gene (chsA) was mapped at the distal end of the short arm of chromosome 5. The cDNA probe for this single-copy gene was 1.4 kb. In contrast, in the lines Mitchell and V26 chsA was localized at the distal end of the long arm of chromosome 3, suggesting that a chromosomal rearrangement had taken place. In a transformed Petunia uidA, transgenes were detected using a 2.7 kb probe. One transgene was mapped on one of the homologues of chromosome II proximal to the ribosomal genes. This homologue could be distinguished from the other by having the ribosomal genes at the distal end of the long arm. Using multicolour fluorescence in situ hybridization it was shown that it is possible to detect the endogenous chsA genes and both transgenes simultaneously.
Chalcone Synthase Promoters in Petunia Are Active in Pigmented and Unpigmented Cell Types
THE PLANT CELL ONLINE, 1990
Chalcone synthase (CHS) catalyzes the first step in the biosynthesis of flavonoids that function in flower pigmentation, protection against stress, and induction of nodulation. The petunia genome contains eight complete chs genes, of which four are differentially expressed in floral tissues and UV-light-induced seedlings. The 5'-flanking regions of these four chs genes were fused to the 8-glucuronidase (GUS) reporter gene and introduced into petunia plants by Agrobacterium-mediated transformation. We show that expression of each construct is identical to the expression of the authentic chs gene, implying that the differences in expression pattern between these chs genes are caused at least in part by their promoters. Histochemical analyses of GUS expression show that chs promoters are not only active in pigmented cell types (epidermal cells of the flower corolla and tube and [sub] epidermal cells of the flower stem) but also in a number of unpigmented cell types (mesophylic cells of the corolla, several cell types in the ovary and the seed coat). Comparison of chs-GUS expression and flavonoid accumulation patterns in anthers suggests that intercellular transport of flavonoids and enzymes occurs in this organ. Analysis of the flavonoids accumulated in tissues from mutant lines shows that only a subset of the genes that control flavonoid biosynthesis in the flower operates in the ovary and seed. This implies that (genetic) control of flavonoid biosynthesis is highly tissue specific.
Chalcone Synthase and Flavonol Accumulation in Stigmas and Anthers of Petunia hybrida
1993
Flavonol aglycones are required for pollen germination in pe- tunia (Petunia hybrida 1.). Mutant plants lacking chalcone synthase (CHS), which catalyzes the first committed step in flavonoid syn- thesis, do not accumulate flavonols and are self-sterile. lhe mutant pollen can be induced to germinate by supplementing it with kaempferol, a flavonol aglycone, either at the time of pollination or by