Nitric oxide is induced by wounding and influences jasmonic acid signaling in Arabidopsis thaliana
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Nitric oxide (NO) has been associated with plant defense responses during microbial attack, and with induction and/or regulation of programmed cell death. Here, we addressed whether NO participates in wound responses in Arabidopsis thaliana (L.) Heynh.. Real-time imaging by confocal laser-scanning microscopy in conjunction with the NO-selective fluorescence indicator 4,5-diaminofluorescein diacetate (DAF-2 DA) uncovered a strong NO burst after wounding or after treatment with JA. The NO burst was triggered within minutes, reminiscent of the oxidative burst during hypersensitive responses. Furthermore, we were able to detect NO in plants (here induced by wounding) by means of electron paramagnetic resonance measurements using diethyldithiocarbamate as a spin trap. When plants were treated with NO, Northern analyses revealed that NO strongly induces key enzymes of jasmonic acid (JA) biosynthesis such as allene oxide synthase (AOS) and lipoxygenase (LOX2). On the other hand, wound-induced AOS gene expression was independent of NO. Furthermore, JA-responsive genes such as defensin (PDF1.2) were not induced, and NO induction of JA-biosynthesis enzymes did not result in elevated levels of JA. However, treatment with NO resulted in accumulation of salicylic acid (SA). In transgenic NahG plants (impaired in SA accumulation and/or signaling), NO did induce JA production and expression of JA-responsive genes. Altogether, the presented data demonstrate that wounding in Arabidopsis induces a fast accumulation of NO, and that NO may be involved in JA-associated defense responses and adjustments.
KeywordsArabidopsis Jasmonic acid Nitric oxide Salicylic acid Wounding
Allene oxide synthase
- DAF-2 DA
Electron paramagnetic resonance
Inducible nitric oxide synthase
Jasmonic acid-induced protein
Reactive oxygen species
We thank E. Mattes and G. Fricke-Bode for excellent technical assistance, and Wolfgang Mayr for SA measurements. Furthermore, we thank Syngenta (San Diego, CA) for NahG lines (Col). This work was supported by Freistaat Bayern (BStMLU) and by grant DU 246/4-1 (Deutsche Forschungsgemeinschaft).
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