TitleCloning and functional characterization of a gene for capsanthin-capsorubin synthase from tiger lily (Lilium lancifolium Thunb. 'Splendens').
Publication TypeJournal Article
Year of Publication2012
AuthorsJeknić, Z, Morré, JT, Jeknić, S, Jevremović, S, Subotić, A, Chen, THH
JournalPlant Cell Physiol
Volume53
Issue11
Pagination1899-912
Date Published2012 Nov
ISSN1471-9053
KeywordsAmino Acid Sequence, Chromatography, High Pressure Liquid, Cloning, Molecular, Color, DNA, Complementary, Flowers, Gene Expression Regulation, Enzymologic, Gene Expression Regulation, Plant, Genes, Plant, Intramolecular Lyases, Iris Plant, Lilium, Molecular Sequence Data, Open Reading Frames, Oxidoreductases, Phylogeny, Plant Proteins, Plants, Genetically Modified, Reverse Transcriptase Polymerase Chain Reaction, Sequence Homology, Amino Acid, Tandem Mass Spectrometry, Xanthophylls
Abstract

The orange color of tiger lily (Lolium lancifolium 'Splendens') flowers is due, primarily, to the accumulation of two κ-xanthophylls, capsanthin and capsorubin. An enzyme, known as capsanthin-capsorubin synthase (CCS), catalyzes the conversion of antheraxanthin and violaxanthin into capsanthin and capsorubin, respectively. We cloned the gene for capsanthin-capsorubin synthase (Llccs) from flower tepals of L. lancifolium by the rapid amplification of cDNA ends (RACE) with a heterologous non-degenerate primer that was based on the sequence of a gene for lycopene β-cyclase (lcyB). The full-length cDNA of Llccs was 1,785 bp long and contained an open reading frame of 1,425 bp that encoded a polypeptide of 474 amino acids with a predicted N-terminal plastid-targeting sequence. Analysis by reverse transcription-PCR (RT-PCR) revealed that expression of Llccs was spatially and temporally regulated, with expression in flower buds and flowers of L. lancifolium but not in vegetative tissues. Stable overexpression of the Llccs gene in callus tissue of Iris germanica, which accumulates several xanthophylls including violaxanthin, the precursor of capsorubin, resulted in transgenic callus whose color had changed from its normal yellow to red-orange. This novel red-orange coloration was due to the accumulation of two non-native κ-xanthophylls, capsanthin and capsorubin, as confirmed by HPLC and ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis with authentic standards. Cloning of the Llccs gene should advance our understanding of the molecular and genetic mechanisms of the biosynthesis of κ-carotenoids in general and in the genus Lilium in particular, and will facilitate transgenic alterations of the colors of flowers and fruits of many plant species.

DOI10.1093/pcp/pcs128
Alternate JournalPlant Cell Physiol.
PubMed ID23008421
PubMed Central IDPMC3494009
Grant ListP30ES000210 / ES / NIEHS NIH HHS / United States
S10RR027878 / RR / NCRR NIH HHS / United States