title: Detection of Autophagy in Plants by Fluorescence Microscopy.
abstract: Autophagy is a key process for degradation and recycling of proteins or organelles in eukaryotes. Autophagy in plants has been shown to function in stress responses, pathogen immunity, and senescence, while a basal level of autophagy plays a housekeeping role in cells. Upon activation of autophagy, vesicles termed autophagosomes are formed to deliver proteins or organelles to the vacuole for degradation. The number of autophagosomes can thus be used to indicate the level of autophagy. Here, we describe two common methods used for detection of autophagosomes, staining of autophagosomes with the fluorescent dye monodansylcadaverine, and expression of a fusion between GFP and the autophagosomal membrane protein ATG8.
authors.affiliation: Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA, 50011, USA.
authors.forename: Yunting
authors.initials: Y
authors.lastname: Pu
authors.affiliation: Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA, 50011, USA. bassham@iastate.edu.|Plant Sciences Institute, Iowa State University, Ames, IA, 50011, USA. bassham@iastate.edu.
authors.forename: Diane C
authors.initials: DC
authors.lastname: Bassham
mesh_terms: D017360:Arabidopsis / Q000737:chemistry / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000302:isolation & purification; D000071182:Autophagosomes / Q000737:chemistry*; D001343:Autophagy / Q000235:genetics*; D000071190:Autophagy-Related Protein 8 Family / Q000235:genetics* / Q000302:isolation & purification; D049452:Green Fluorescent Proteins; D008856:Microscopy, Fluorescence; D030821:Plants, Genetically Modified / Q000737:chemistry / Q000235:genetics; D059748:Proteolysis
chemical_list: C000597073:ATG8 protein, Arabidopsis; D029681:Arabidopsis Proteins; D000071190:Autophagy-Related Protein 8 Family; D049452:Green Fluorescent Proteins
keywords: Arabidopsis thaliana; Autophagosome; Autophagy; GFP-ATG8; Monodansylcadaverine; Vacuole