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JXB Advance Access originally published online on April 28, 2003
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Journal of Experimental Botany, Vol. 54, No. 387, pp. 1553-1564, June 1, 2003
© 2003 Oxford University Press

In situ localization associates biologically active plant natriuretic peptide immuno-analogues with conductive tissue and stomata

Received 13 March 2003; Accepted 18 March 2003

M. M. Maryani4,1, M. V. Morse2, G. Bradley2, H. R. Irving3, D. M. Cahill1 and C. A. Gehring5,2

1 School of Biological and Chemical Sciences, Deakin University, Geelong, Victoria, 3217, Australia
2 Department of Biotechnology, University of the Western Cape, Private Bag X17, Bellville 7535, South Africa
3 Department of Pharmaceutical Biology and Pharmacology, Victorian College of Pharmacy, Monash University, 381 Royal Parade, Parkville, Melbourne, Victoria 3052, Australia

4 Present address: Faculty of Biology, Gadjah Mada University, Yogyakarta, Indonesia 55281.
5 To whom correspondence should be addressed. Fax: +27 21 959 2266. E-mail:cgehring{at}uwc.ac.za
Abbreviations: ANP, atrial natriuretic peptide; PNP, plant natriuretic peptides.

Plant natriuretic peptide immuno-analogues (irPNP) have previously been shown to affect a number of biological processes including stomatal guard cell movements, ion fluxes and osmoticum-dependent water transport. Tissue printing and immunofluorescent labelling techniques have been used here to study the tissue and cellular localization of irPNP in ivy (Hedera helix L.) and potato (Solanum tuberosum L.). Polyclonal antibodies active against human atrial natriuretic peptide (anti-hANP) and antibodies against irPNP from potato (anti-StPNP) were used for immunolabelling. Tissue prints revealed that immunoreactants are concentrated in vascular tissues of leaves, petioles and stems. Phloem-associated cells, xylem cells and parenchymatic xylem cells showed the strongest immunoreaction. Immunofluorescent microscopy with fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG supported this finding and, furthermore, revealed strong labelling to stomatal guard cells and the adjacent apoplastic space as well. Biologically active immunoreactants were also detected in xylem exudates of a soft South African perennial forest sage (Plectranthus ciliatus E. Mey ex Benth.) thus strengthening the evidence for a systemic role of the protein. In summary, in situ cellular localization is consistent with physiological responses elicited by irPNPs reported previously and is indicative of a systemic role in plant homeostasis.

Key words: Conductive tissue, expansins, Hedera helix L., homeostasis, immuno-analogues, plant natriuretic peptides, Plectranthus ciliatus E. Mey ex Benth., Solanum tuberosum L., tissue printing.


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