26. Activation of Tomato Cellulases by the Potato Cyst Nematode Globodera Rostochiensis During Feeding Site Induction and Development

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26. ACTIVATION OF TOMATO CELLULASES BY THE POTATO CYST NEMATODE GLOBODERA ROSTOCHIENSIS DURING FEEDING SITE INDUCTION AND DEVELOPMENT A. Karczmarek1, W. Kurek2, L. Malgorzata2, S. Fudali2, M. Sobczak2, S. Janakowski2, W. Golinowski2, J. Helder1 and A. Goverse1 1Laboratory of Nematology, Department of Plant Sciences, Binnenhaven 5,6709 PD Wageningen, The Netherlands 2Department of Botany, Faculty of Agriculture and Biology, Warsaw Agricultural University, Nowoursynowska 159, Building 30, Warsaw, Poland [email protected] In the root of a host plant, cyst nematodes induce the formation of private food source, a syncytium. The establishment of such a group of plant cells in which cytoplasmic continuity is maintained starts with the selection of an initial syncytial cell by a pre-parasitic second stage juvenile. For syncytium proliferation, the nematode recruits plant cell wall-degrading enzymes. Endo-1,4-ß-glucanases (cellulases) reside among plant enzymes that are responsible for progressive cell wall dissolution resulting in syncytium expansion towards and within the vascular cylinder. Tomato is harbouring at least 8 different cellulases, and RT-PCR experiments revealed that only two isoforms, Le-Cel7 and Le-Cel8, are consistently up-regulated in young, Globodera rostochiensis-induced syncytia. In order to localise Le-Cel7 and Le-Cel8 expression, in situ RT-PCR experiments were performed. Careful hybridisation experiments in longitudinal sections revealed slightly different syncytium-specific and temporal expression patterns for these two cellulase isoforms. Antibodies were raised against characteristic fragments of Le-Cel7 and Le-Cel8, and subsequent immunolocalisation studies confirmed the presence of Le-Cel7 and Le-Cel8 proteins in developing syncytia. Transgenic tomato, tobacco and potato plants carrying hpRNA-silencing constructs for Le-Cel7 and Le- Cel8, respectively, are currently being tested to assess the importance of Le-Cel7 and Le-Cel8 in the development of cyst nematode-induced feeding sites. .

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