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VIEW ARTICLE   |    DOI: 10.1094/MPMI-9-0039

Secretory Granule Proteins from the Subventral Esophageal Glands of the Potato Cyst Nematode Identified by Monoclonal Antibodies to a Protein Fraction from Second-Stage Juveniles. Jan M. de Boer. Department of Nematology, Wageningen Agricultural University, P. O. Box 8213, 6700 ES Wageningen, The Netherlands. Geert Smant (1), Aska Goverse (1), Eric L. Davis(3), Hein A. Overmars (1), H. (Rikus) Pomp (2), Marga van Gent-Pelzer(2), Jacoline F. Zilverentant(2), Jack P. W. G. Slokkermans(2), Richard S. Hussey (4), Fred J. Gommers(1), Jaap Bakker(1), and Arjen Schots (2). (1) Department of Nematology, Wageningen Agricultural University, P. O. Box 8213, 6700 ES Wageningen, The Netherlands; (2) Laboratory for Monoclonal Antibodies, P. O. Box 9060, 6700 GW Wageningen, The Netherlands; (3)Department of Plant Pathology, North Carolina State University, Box 7616, Raleigh, NC 27695-7616, U.S.A. (4) Department of Plant Pathology, University of Georgia, Athens, GA 30602-7274, U.S.A. MPMI 9:39-46. Accepted 20 September 1995, Copyright 1996 The American Phytopathological Society.

Sodium dodecyl sulfate-extracted proteins from second-stage juveniles (J2) of the potato cyst nematode Globodera roslochiensis were fractionated by preparative continuous flow electrophoresis, and monoclonal antibodies (MAbs) were raised against the 38- to 40.5-kDa protein fraction. Screening of the hybridoma culture fluids by immuno-fluorescence microscopy of J2 resulted in the identification of 12 MAbs that bound specifically to the subventral esophageal glands. On Western blots of J2 these MAbs identified four protein bands with apparent molecular masses of 30, 31, 39, and 49 kDa. Immunoelectron microscopy with one of these MAbs showed an intense labeling of the electron dense core of the secretory granules in the subventral gland cells of J2. It is concluded that one or more of these proteins are localized within these secretory granules. Immunofluorescence microscopy of J2 from other plant parasitic nematode species showed that most of these MAbs also bind to the subventral glands of G. pallida and G. tabacum but not of Heterodera schachtii, H. glycines, Meloidogyne incognita, or M. hapla.

Additional Keywords: host-parasite interaction, virulence factor, host plant transformation, plantibodies.