Plant Physiol. Drug Metab Dispos
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Processing, Targeting, and Antifungal Activity of Stinging Nettle Agglutinin in Transgenic Tobacco

Mirjam P. Does*, Petra M. Houterman, Henk L. Dekker, and Ben J.C. Cornelissen

Section for Plant Pathology, Institute for Molecular Cell Biology, BioCentrum Amsterdam, Kruislaan 318, 1098 SM Amsterdam, The Netherlands (M.P.D., P.M.H., B.J.C.C.); and Department of Biochemistry, E.C. Slater Institute, BioCentrum Amsterdam, Plantage Muidergracht 12, 1018 TV Amsterdam, The Netherlands (H.L.D.)

The gene encoding the precursor to stinging nettle (Urtica dioica L.) isolectin I was introduced into tobacco (Nicotiana tabacum). In transgenic plants this precursor was processed to mature-sized lectin. The mature isolectin is deposited intracellularly, most likely in the vacuoles. A gene construct lacking the C-terminal 25 amino acids was also introduced in tobacco to study the role of the C terminus in subcellular trafficking. In tobacco plants that expressed this construct, the mutant precursor was correctly processed and the mature isolectin was targeted to the intercellular space. These results indicate the presence of a C-terminal signal for intracellular retention of stinging nettle lectin and most likely for sorting of the lectin to the vacuoles. In addition, correct processing of this lectin did not depend on vacuolar deposition. Isolectin I purified from tobacco displayed identical biological activities as isolectin I isolated from stinging nettle. In vitro antifungal assays on germinated spores of the fungi Botrytis cinerea, Trichoderma viride, and Colletotrichum lindemuthianum revealed that growth inhibition by stinging nettle isolectin I occurs at a specific phase of fungal growth and is temporal, suggesting that the fungi had an adaptation mechanism.

*   Corresponding author; e-mail does{at}bio.uva.nl; fax 31-20-525-7934.

Plant Physiol. (1999) 120: 421-432
Copyright Clearance Center:   0032-0889/99/120//12
© 1999 American Society of Plant Physiologists




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