Abstract
Penetration resistance to powdery mildew fungi, conferred by localized cell wall appositions (papillae), is one of the best-studied processes in plant innate immunity. The syntaxin PENETRATION (PEN)1 is required for timely appearance of papillae, which contain callose and extracellular membrane material, as well as PEN1 itself. Appearance of membrane material in papillae suggests secretion of exosomes. These are potentially derived from multivesicular bodies (MVBs), supported by our observation that ARA6-labeled organelles assemble at the fungal attack site. However, the trafficking components that mediate delivery of extracellular membrane material are unknown. Here, we show that the delivery is independent of PEN1 function. Instead, we find that application of brefeldin (BF)A blocks the papillary accumulation of GFP-PEN1-labeled extracellular membrane and callose, while impeding penetration resistance. We subsequently provide evidence indicating that the responsible BFA-sensitive ADP ribosylation factor-GTP exchange factor (ARF-GEF) is GNOM. Firstly, analysis of the transheterozygote gnom(B4049/emb30-1) (gnom(B)(/E)) mutant revealed a delay in papilla formation and reduced penetration resistance. Furthermore, a BFA-resistant version of GNOM restored the BFA-sensitive papillary accumulation of GFP-PEN1 and callose. Our data, therefore, provide a link between GNOM and disease resistance. We suggest that papilla formation requires rapid reorganization of material from the plasma membrane mediated by GNOM. The papilla material is subsequently presumed to be sorted into MVBs and directed to the site of fungal attack, rendering the epidermal plant cell inaccessible for the invading powdery mildew fungus.
MeSH Terms
Arabidopsis/genetics
Arabidopsis Proteins/metabolism,physiology
Biological Transport
Brefeldin A/pharmacology
Exosomes/metabolism
GTP Phosphohydrolases/metabolism
Gene Expression Regulation, Fungal
Glucans/chemistry
Guanine Nucleotide Exchange Factors/physiology
Heterozygote
Immunity, Innate
Microscopy, Confocal/methods
Microscopy, Fluorescence/methods
Models, Biological
Qa-SNARE Proteins/metabolism
Chemicals
Arabidopsis Proteins
GNOM protein, Arabidopsis
Glucans
Guanine Nucleotide Exchange Factors
PEN1 protein, Arabidopsis
Qa-SNARE Proteins
Brefeldin A
callose
GTP Phosphohydrolases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Nielsen Mads Eggert
Department of Agriculture and Ecology, Faculty of Science, University of Copenhagen, DK-1871 Frederiksberg C, Copenhagen, Denmark. maen@life.ku.dk
Feechan Angela
Böhlenius Henrik
Ueda Takashi
Thordal-Christensen Hans
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