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Planta

, Volume 176, Issue 1, pp 83–90 | Cite as

Alkaloid N-oxides as transport and vacuolar storage compounds of pyrrolizidine alkaloids in Senecio vulgaris L

  • Adelheid Ehmke
  • Kirsten von Borstel
  • Thomas Hartmann
Article

Abstract

Cell-suspension cultures of pyrrolizidinealkaloid-producing species selectively take up and accumulate senecionine (sen) and its N-oxide (sen-Nox). Cultures established from non-alkaloid-producing species are unable to accumulate the alkaloids. The uptake and accumulation of 14C-labelled alkaloids was studied using a Senecio vulgaris cell-suspension culture as well as protoplasts and vacuoles derived from it. The alkaloid uptake exhibits all characteristics of a carrier-mediated transport. The uptake of sen-Nox follows a multiphasic saturation kinetics. The Km-values for sen Nox of 53 μM and 310 μM are evaluated. Senecionine competitively inhibits sen-Nox uptake, indicating that the tertiary alkaloid and its N-oxide share the same membrane carrier. The N-oxide of sen shows a pH optimum below 5.5, whereas sen is taken up over a range from pH 4 to 8. Activation energies of 90 and 53 kJ·mol-1 are calculated for sen-Nox and sen transport, respectively. At concentrations of 10 to 100 μM, sen-Nox is rapidly taken up by cells and protoplasts; within 2 h >90% of total N-oxide is within the cells. By contrast the uptake of sen is less efficient. Vacuoles isolated from protoplasts preloaded with sen-Nox totally retained the alkaloid N-oxide, whereas sen is rapidly lost during the procedure of vacuole preparation. N-oxidation converts the weak lipophilic tertiary base into a charged polar molecule which is excellently adapted to serve as the cellular transport and storage form of pyrrolizidine alkaloids.

Key words

Alkaloid N-oxide (transport, accumulation) Cell culture (alkaloids) Protoplast Pyrrolizidine alkaloid Senecio Vacuole (alkaloid storage) 

Abbreviations

CCCP

carbonylcyanide m-chlorophenylhydrazone

DCCD

N,N′-dicyclohexylcarbodiimide

DIDS

4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid

DNP

2,4-dinitrophenol

sen

senecionine

sen-Nox

senecionine N-oxide

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Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Adelheid Ehmke
    • 1
  • Kirsten von Borstel
    • 1
  • Thomas Hartmann
    • 1
  1. 1.Institut für Pharmazeutische Biologie der Technischen Universität BraunschweigBraunschweigFederal Republic of Germany

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