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Plant and Cell Physiology Advance Access originally published online on January 13, 2006
Plant and Cell Physiology 2006 47(3):346-354; doi:10.1093/pcp/pci252
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Polyamines Induce Rapid Biosynthesis of Nitric Oxide (NO) in Arabidopsis thaliana Seedlings

Ni Ni Tun1,3, Claudete Santa-Catarina2,3, Tahmina Begum1, Vanildo Silveira2, Walter Handro2, Eny Iochevet Segal Floh2 and Günther F. E. Scherer1,*

1 Universität Hannover, Institut für Zierpflanzenbau, AG Molekulare Ertragsphysiologie, Herrenhäuser Str. 2, D-30419 Hannover, Germany
2 Plant Cell Biology Laboratory, Department of Botany, IB-University of São Paulo, São Paulo, CP 11461, 05422-970, Brazil

* Corresponding author: E-mail, scherer{at}zier.uni-hannover.de; Fax, +49-511-762-3608.

In this study, we examined the regulation by putrescine, spermidine and spermine of nitric oxide (NO) biosynthesis in Arabidopsis thaliana seedlings. Using a fluorimetric method employing the cell-impermeable NO-binding dye diaminorhodamine-4M (DAR-4M), we observed that the polyamines (PAs) spermidine and spermine greatly increased NO release in the seedlings, whereas arginine and putrescine had little or no effect. Spermine, the most active PA, stimulated NO release with no apparent lag phase. The response was quenched by addition of 2-aminoethyl-2-thiopseudourea (AET), an inhibitor of the animal nitric oxide synthase (NOS) and plant NO biosynthesis, and by 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-1-oxy-3-oxide (PTIO), an NO scavenger. By fluorescence microscopy, using the cell-permeable NO-binding dye diaminorhodamine-4M acetoxymethyl ester (DAR-4M AM), we observed that PAs induced NO biosynthesis in specific tissues in Arabidopsis seedlings. Spermine and spermidine increased NO biosynthesis in the elongation zone of the Arabidopsis root tip and in primary leaves, especially in the veins and trichomes, while in cotyledons little or no effect of PAs beyond the endogenous levels of NO-induced fluorescence was observed. We conclude that PAs induce NO biosynthesis in plants.

3 These authors contributed equally to this work.

(Received November 17, 2005; Accepted December 26, 2005)
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