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Elicitor-mediated induction of anthraquinone biosynthesis and regulation of isopentenyl diphosphate isomerase and farnesyl diphosphate synthase activities in cell suspension cultures of Cinchona robusta How.
Authors:Ana C Ramos-Valdivia  Robert van der Heijden  Robert Verpoorte
Institution:(1) Division of Pharmacognosy, Leiden/Amsterdam Center for Drug Research, Gorlaeus Laboratories, P.O. Box 9502, NL-2300 RA Leiden, The Netherlands, NL;(2) Departamento de Biotecnología, Centro de Investigación y Estudios Avanzados (CINVESTAV), Apto Postal 14-740, CP 07000, México D.F., México,
Abstract:Treatment of a Cinchona robusta How. cell suspension culture with a homogenate of Phytophthora cinnamomi resulted in cessation of growth and a rapid induction of the biosynthesis of anthraquinone-type phytoalexins. The strongest induction of anthraquinone biosynthesis was obtained when the elicitor was added in the early growth phase of the growth cycle. The accumulation of anthraquinones was accompanied by a tri-phasic response in the activity of isopentenyl diphosphate (IPP) isomerase (EC 5.3.3.2): phase I was characterised by a rapid induction of activity, reaching a maximum at 12 h after elicitation. During phase II, IPP isomerase rapidly decreased to levels below those found in untreated cells. At phase III, IPP isomerase activity increased again, reaching a second maximum at about 72 h after elicitation. During phase I, the activity of farnesyl diphosphate synthase (EC 2.5.1.10) was found to be suppressed. Extraction and assay conditions were optimised for IPP isomerase. The presence of Mn2+ in the incubation buffer resulted in a marked increase in the activity of the enzymes obtained from cells in phase I. The induction of IPP isomerase in combination with a concomitant inhibition of farnesyl diphosphate synthase might result in an efficient channeling of C5-precursors into phytoalexin biosynthesis. Received: 23 August 1996 / Accepted: 20 March 1997
Keywords:: Anthraquinone  Cinchona  Farnesyl diphosphate synthase  Isopentenyl diphosphate isomerase  Phytoalexin  Phytophthora
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