title: The leaf epidermome of Catharanthus roseus reveals its biochemical specialization.
abstract: Catharanthus roseus is the sole commercial source of the monoterpenoid indole alkaloids (MIAs), vindoline and catharanthine, components of the commercially important anticancer dimers, vinblastine and vincristine. Carborundum abrasion technique was used to extract leaf epidermis-enriched mRNA, thus sampling the epidermome, or complement, of proteins expressed in the leaf epidermis. Random sequencing of the derived cDNA library established 3655 unique ESTs, composed of 1142 clusters and 2513 singletons. Virtually all known MIA pathway genes were found in this remarkable set of ESTs, while only four known genes were found in the publicly available Catharanthus EST data set. Several novel MIA pathway candidate genes were identified, as demonstrated by the cloning and functional characterization of loganic acid O-methyltransferase involved in secologanin biosynthesis. The pathways for triterpene biosynthesis were also identified, and metabolite analysis showed that oleanane-type triterpenes were localized exclusively to the cuticular wax layer. The pathways for flavonoid and very-long-chain fatty acid biosynthesis were also located in this cell type. The results illuminate the biochemical specialization of Catharanthus leaf epidermis for the production of multiple classes of metabolites. The value and versatility of this EST data set for biochemical and biological analysis of leaf epidermal cells is also discussed.
authors.affiliation: Department of Biological Sciences, Brock University, St. Catharines, Ontario L2S3A1 Canada.
authors.forename: Jun
authors.initials: J
authors.lastname: Murata
authors.forename: Jonathon
authors.initials: J
authors.lastname: Roepke
authors.forename: Heather
authors.initials: H
authors.lastname: Gordon
authors.forename: Vincenzo
authors.initials: V
authors.lastname: De Luca
mesh_terms: D000595:Amino Acid Sequence; D030008:Catharanthus / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D020224:Expressed Sequence Tags; D018506:Gene Expression Regulation, Plant; D015723:Gene Library; D057888:Iridoid Glucosides; D039823:Iridoids / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D008969:Molecular Sequence Data; D015394:Molecular Structure; D019441:Plant Epidermis / Q000235:genetics* / Q000378:metabolism; D018515:Plant Leaves / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014747:Vinblastine / Q000031:analogs & derivatives / Q000096:biosynthesis / Q000737:chemistry; D014748:Vinca Alkaloids / Q000096:biosynthesis / Q000737:chemistry
chemical_list: D018076:DNA, Complementary; D057888:Iridoid Glucosides; D039823:Iridoids; D014748:Vinca Alkaloids; C021086:secologanin; C002947:loganic acid; C009667:vindoline; D014747:Vinblastine; D008780:Methyltransferases; C017836:catharanthine