The cantharidin-induced oxidative burst in tobacco BY-2 cell suspension cultures |
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Authors: | P Van Gestelen P Ledeganck I Wynant R J Caubergs H Asard |
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Institution: | (1) Department of Biology, University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp, Belgium |
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Abstract: | Summary The in vivo induction of H2O2 production was tested on tobacco cell suspension cultures (Nicotiana tabacum cv. Bright Yellow-2). The measurement of H2O2 was based on the oxidation of 3,5-dichloro-2-hydroxybenzensulfonic acid by endogenous peroxidases and spectrophotometric detection after reaction with 4-aminoanti-pyrine. The phosphatase inhibitor cantharidin induced a transient increase in H2O2 synthesis. The timing of the H2O2 production, the level of induction by cantharidin and the background H2O2 production were dependent on the tobacco cell concentration used. A concentration curve of cantharidin revealed saturating kinetics for the H2O2 detection (E50=46 to 70 M, Emax=101 to 128 mol/h·g fresh weight). An inhibitor study with the tobacco BY-2 cells showed high inhibitions of the H2O2 induction with the flavin analogues diphenylene iodonium (I50=1.26 M) and acridine orange and with membrane-permeative thiol reagents (N-ethyl maleimide, N-pyrene maleimide, iodoacetate); whereas the nonpermeative thiol reagentp-chloromercuribenzoic acid was ineffective. Therefore, the induction of H2O2 production with phosphatase inhibitors (cantharidin) showed comparable properties to the elicitor-induced oxidative-burst response in other plant cells.Abbreviations AcOr
acridine orange
- AOS
active-oxygen species
- BY-2
Bright Yellow-2
- pCMBS
p-chloromercuribenzoic acid
- DHBS
3,5-dichloro-2-hydroxybenzenesulfonic acid
- DMSO
dimethylsulfoxide
- DPI
diphenylene iodonium
- EtOH
ethanol
- H2O2
hydrogen peroxide
- HRP
horseradish peroxidase
- MS
Murashige and Skoog
- NEM
N-ethyl maleimide
- NPM
N-pyrene maleimide
- O
2
–
superoxide
- SOD
superoxide dismutase |
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Keywords: | Active-oxygen species Tobacco Bright Yellow-2 cells Defence Nicotiana tabacum Phosphatase Protein phosphorylation |
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