Involvement of lipoxygenase in elicitor-stimulated sanguinarine accumulation in Papaver somniferum suspension cultures.
The involvement of lipoxygenase (LOX, EC 1.13.11.12) in elicitor-induced opium poppy defense response was investigated. Papaver somniferum L. suspension cultures were treated with abiotic elicitor methyl jasmonate (MJ), fungal elicitor (Botrytis cinerea homogenate) and phenidone (specific inhibitor of LOX) to determine the involvement of this enzyme in production of sanguinarine, the major secondary metabolite of opium poppy cultures. P. somniferum suspension cultures responded to elicitor treatment with strong and transient increase of LOX activity followed by sanguinarine accumulation. LOX activity increased in elicited cultures, reaching 9.8 times of the initial value at 10 h after MJ application and 2.9 times after B. cinerea application. Sanguinarine accumulated to maximal levels of 169.5 ± 12.5 μg g⁻¹ dry cell weight in MJ-elicited cultures and 288.0 ± 10.0 μg g⁻¹ dry cell weight in B. cinerea-elicited cultures. The treatment of cells with phenidone before elicitor addition, significantly reduced sanguinarine production. The relative molecular weight of P. somniferum LOX (83 kDa) was estimated by using immunobloting and its pH optimum was shown to be pH 6.5.
Holková
Ivana
I
Department of Cell and Molecular Biology of Drugs, Faculty of Pharmacy, Comenius University, Kalinčiakova 8, SK-832 32, Bratislava, Slovakia. holkova@fpharm.uniba.sk
Bezáková
Lýdia
L
Bilka
František
F
Balažová
Andrea
A
Vanko
Marián
M
Blanáriková
Vítazoslava
V
D000085:Acetates; D053119:Benzophenanthridines / Q000096:biosynthesis*; D020171:Botrytis; D018929:Cell Culture Techniques; D003517:Cyclopentanes; D006863:Hydrogen-Ion Concentration; D007546:Isoquinolines; D008084:Lipoxygenase / Q000378:metabolism*; D054883:Oxylipins; D010207:Papaver / Q000166:cytology / Q000378:metabolism*; D010935:Plant Diseases / Q000382:microbiology; D011720:Pyrazoles
D000085:Acetates; D053119:Benzophenanthridines; D003517:Cyclopentanes; D007546:Isoquinolines; D054883:Oxylipins; D011720:Pyrazoles; C072239:methyl jasmonate; C005705:sanguinarine; D008084:Lipoxygenase; C015429:phenidone