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Journal of Experimental Botany, Vol. 51, No. 345, pp. 695-701, April 2000
© 2000 Oxford University Press

Development of Fe-deficiency responses in cucumber (Cucumis sativus L.) roots: involvement of plasma membrane H+-ATPase activity

Marta Dell'Orto1, Simonetta Santi2, Patrizia De Nisi1, Stefano Cesco2, Zeno Varanini2, Graziano Zocchi1,3 and Roberto Pinton2

1 Dipartimento di Produzione Vegetale, University of Milan, Via Celoria 2, I-20133 Milano, Italy
2 Dipartimento di Produzione Vegetale e Tecnologie Agrarie, University of Udine, Via Delle Scienze 208, I-33100 Udine, Italy

One of the mechanisms through which some strategy I plants respond to Fe-deficiency is an enhanced acidification of the rhizosphere due to proton extrusion. It was previously demonstrated that under Fe-deficiency, a strong increase in the Hplus;-ATPase activity of plasma membrane (PM) vesicles isolated from cucumber roots occurred. This result was confirmed in the present work and supported by measurement of ATP-dependent proton pumping in inside-out plasma membrane vesicles. There was also an attempt to clarify the regulatory mechanism(s) which lead to the activation of the H+-ATPase under Fe-deficiency conditions. Plasma membrane proteins from Fe-deficient roots submitted to immunoblotting using polyclonal antibodies showed an increased level in the 100 kDa polypeptide. When the plasma membrane proteins were treated with trypsin a 90 kDa band appeared. This effect was accompanied by an increase in the enzyme activity, both in the Fe-deficient and in the Fe-sufficient extracts. These results suggest that the increase in the plasma membrane H+-ATPase activity seen under Fe-deficiency is due, at least in part, to an increased steady-state level of the 100 kDa polypeptide.

Key words: Fe-deficiency, H+-ATPase, plasma membrane, proton extrusion, rhizosphere.


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