Hutchison Katie A, O'Connell Richard J, Pain Naomi A, Green Jonathan R
School of Biological Sciences, University of Birmingham, P.O. Box 363, Birmingham B15 2TT, UK.
IACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, Long Ashton, Bristol BS18 9AF, UK.
New Phytol. 1996 Dec;134(4):579-585. doi: 10.1111/j.1469-8137.1996.tb04923.x.
Monoclonal antibodies were raised to Colletotrichum lindemuthianum (Sacc. & Magn.) Briosi & Cav. infection structures which had been isolated from leaves of Phaseolus vulgaris L. by isopycnic centrifugation and immunomagnetic separation. One of the antibodies, UB29, recognized a carbohydrate epitope in a 200 kDa glycoprotein present on the surface of conidia, germ-tubes and appressoria when they developed on host tissue. Intracellular hyphae were not labelled. Immunogold labelling showed that the antigen was confined to the extracellular matrices around such structures. No such antigen was detected by immunofluorescence or Western blotting when the fungus developed in vitro. UB29 recognized a glycoprotein of identical M located in the epicuticular wax layer of uninoculated bean hypocotyls. The results suggest that a host epicuticular glycoprotein is present in the extracellular matrices of fungal structures during growth on the plant surface.
通过等密度离心和免疫磁珠分离技术,从菜豆叶片中分离出炭疽菌(Colletotrichum lindemuthianum (Sacc. & Magn.) Briosi & Cav.)的侵染结构,并以此制备单克隆抗体。其中一种抗体UB29能识别一种200 kDa糖蛋白中的碳水化合物表位,该糖蛋白存在于分生孢子、芽管和附着胞在宿主组织上发育时的表面。细胞内菌丝未被标记。免疫金标记显示,抗原局限于这些结构周围的细胞外基质中。当真菌在体外生长时,通过免疫荧光或蛋白质免疫印迹法未检测到此类抗原。UB29识别位于未接种菜豆下胚轴表皮蜡层中相同分子量的糖蛋白。结果表明,在植物表面生长期间,宿主表皮糖蛋白存在于真菌结构的细胞外基质中。