title: Immunofluorescence detection of callose deposition around plasmodesmata sites.
abstract: Accumulation of callose (β-1,3 glucans) at the plasmodesmata (PD) neck region dynamically regulates symplastic intercellular transport. Here we describe a 2-3-day immuno-labelling protocol to determine callose levels in the cell wall region at PD. The method relies on exposure of internal cell walls by hand-sectioning of the sample and digestion of the cell wall with enzymes in order to improve antibody penetration to deep tissue layers. By using this protocol, combined with high-resolution confocal imaging, we successfully detected PD-associated callose in Arabidopsis root apical meristem, vascular tissue, and developing lateral root primordia.
authors.affiliation: John Innes Centre, Norwich Research Park, Norwich, Norfolk, NR4 7UH, UK.
authors.forename: Ali
authors.initials: A
authors.lastname: Pendle
authors.forename: Yoselin
authors.initials: Y
authors.lastname: Benitez-Alfonso
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D017360:Arabidopsis / Q000737:chemistry / Q000648:ultrastructure*; D001692:Biological Transport; D002473:Cell Wall / Q000737:chemistry / Q000648:ultrastructure*; D044602:Cellulases / Q000737:chemistry; D005404:Fixatives / Q000737:chemistry; D005936:Glucans / Q000032:analysis* / Q000096:biosynthesis; D007150:Immunohistochemistry / Q000379:methods*; D018613:Microscopy, Confocal; D008867:Microtomy; D010539:Permeability; D018517:Plant Roots / Q000737:chemistry / Q000648:ultrastructure*; D031425:Plasmodesmata / Q000737:chemistry / Q000648:ultrastructure*; D016707:Tissue Fixation
chemical_list: D000911:Antibodies, Monoclonal; D005404:Fixatives; D005936:Glucans; C048306:callose; D044602:Cellulases