Immunolocalization of chitin synthases in the phytopathogenic dimorphic fungusUstilago maydis

• DOI: 10.1111/j.1567-1364.2006.00133.x
• Publication Date: 2006-07-17T15:12:50Z
• Authors: José Ruiz-Herrera; Beatriz Xoconostl...; Cristina G. Reyna...; Claudia León-Ram...; Alfonso Cárabez-...
• Keywords: Ustilago; Chitin synthase; Immunoelectron microscopy; Dimorphic fungus; Polyclonal antibodies; Immunofluorescence

Conserved polypeptides of the chitin synthase genes UmCHS3 and UmCHS6 from phytopathogenic fungus Ustilago maydis were utilized as immunogens to obtain polyclonal antibodies that purified by affinity procedures. Because their similarities at regions encoded either polypeptide, it was concluded anti-Chs3 recognized both Chs3 Chs4 synthases, whereas anti-Chs6 Chs6 Chs8 polypeptides. These used analyze localization corresponding synthases in U. cells, using indirect immunofluorescence microscopy immunoelectron with colloidal-gold-labeled secondary antibodies. It observed proteins accumulated surface cytoplasm fungal cells. Electron images revealed accumulation clusters gold particles vesicles, providing evidence for possible origin destination