Stable Vindoline Production in Transformed Cell Cultures of Catharanthus roseus

• Published in: Journal of natural products (Washington, D.C.) Vol. 60; no. 3; pp. 261 - 264
• Main Authors: O'Keefe, Barry R, Mahady, Gail B, Gills, Joell J, Beecher, Christopher W. W, Schilling, Alex B
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 01.03.1997; Glendale, AZ American Society of Pharmacognosy
• Subjects: ACETYL-COENZYME-A-DEACETYLVINDOLINE-O-ACETYLTRANSFERASE; ACILTRANSFERASA; ACTIVIDAD ENZIMATICA; ACTIVITE ENZYMATIQUE; ACYLTRANSFERASE; ACYLTRANSFERASES; AGROBACTERIUM RHIZOGENES; AGROBACTERIUM TUMEFACIENS; AJMALICINE; ALCALOIDE; ALCALOIDES; ALKALOIDS; ANALISIS CUANTITATIVO; ANALYSE QUANTITATIVE; BIOCHEMICAL PATHWAYS; Biological and medical sciences; BIOLOGICAL DIFFERENCES; BIOSINTESIS; BIOSYNTHESE; BIOSYNTHESIS; Biotechnology; CATHARANTHINE; CATHARANTHUS ROSEUS; CELL CULTURE; CELL LINES; CELL SUSPENSIONS; CHEMICAL COMPOSITION; CHEMICAL REACTIONS; COMPOSICION QUIMICA; COMPOSITION CHIMIQUE; CULTIVO DE CELULAS; CULTURE DE CELLULE; DIFERENCIAS BIOLOGICAS; DIFFERENCE BIOLOGIQUE; ENZYMIC ACTIVITY; Eukaryotic cell cultures; EXPLANT; EXPLANTES; EXPLANTS; FEUILLE; Fundamental and applied biological sciences. Psychology; General pharmacology; GENETIC TRANSFORMATION; HOJAS; INDOL; INDOLES; LEAVES; Medical sciences; Methods. Procedures. Technologies; Miscellaneous; MONOTERPENE; MONOTERPENES; MONOTERPENOS; Pharmacognosy. Homeopathy. Health food; Pharmacology. Drug treatments; Plant cells and fungal cells; PLANTAS TRANSGENICAS; PLANTE TRANSGENIQUE; QUANTITATIVE ANALYSIS; REACCIONES QUIMICAS; REACTION CHIMIQUE; STEMS; STRAIN DIFFERENCES; TALLO; TIGE; TRANSFORMACION GENETICA; TRANSFORMATION GENETIQUE; TRANSGENIC PLANTS; VIA BIOQUIMICA DEL METABOLISMO; VOIE BIOCHIMIQUE DU METABOLISME; Pentacyclic compound; Cell culture; Vegetals; Genetic transformation; Angular nitrogen heterocycle; Pharmacognosy; Transformed cell; Agrobacterium tumefaciens; Catharanthus roseus; Medicinal plant; Apocynaceae; Alkaloid; Dicotyledones; Angiospermae; Bacteria; Isolation; Spermatophyta; Aromatic compound; Rhizobiaceae; Agrobacterium rhizogenes
• ISSN: 0163-3864; 1520-6025
• DOI: 10.1021/np960703n

Catharanthus roseus L. (G. Don) leaf disks were transformed by cocultivation with seven strains of Agrobacterium tumefaciens and Agrobacterium rhizogenes. Four distinct transformed cell lines were obtained with A. tumefaciens strains A281, BO542, and A. rhizogenes strain K599, and selected on hormone-free medium. Transformation of the cell lines was confirmed by Southern blot analysis, and preliminary data on the alkaloid profiles was determined by 2D TLC and HPLC/MS. Stable vindoline production was demonstrated in the undifferentiated suspension cell lines strains, CR-BO542, CR-K599, and strain CR-A281S (shooty teratomas), by HPLC/MS analysis of the alkaloid fractions. Active vindoline biosynthesis in the suspension-cell lines was further confirmed by measuring the activity of acetyl-coenzyme-A-deacetylvindoline-O-acetyltransferase, one of the final enzymes in the biosynthetic pathway of vindoline, in cell-free extracts.