Diacylglycerol kinases activate tobacco NADPH oxidase-dependent oxidative burst in response to cryptogein.
Cryptogein is a 10 kDa protein secreted by the oomycete Phytophthora cryptogea that activates defence mechanisms in tobacco plants. Among early signalling events triggered by this microbial-associated molecular pattern is a transient apoplastic oxidative burst which is dependent on the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity of the RESPIRATORY BURST OXIDASE HOMOLOG isoform D (RBOHD). Using radioactive [33P]-orthophosphate labelling of tobacco Bright Yellow-2 suspension cells, we here provide in vivo evidence for a rapid accumulation of phosphatidic acid (PA) in response to cryptogein because of the coordinated onset of phosphoinositide-dependent phospholipase C and diacylglycerol kinase (DGK) activities. Both enzyme specific inhibitors and silencing of the phylogenetic cluster III of the tobacco DGK family were found to reduce PA production upon elicitation and to strongly decrease the RBOHD-mediated oxidative burst. Therefore, it appears that PA originating from DGK controls NADPH-oxidase activity. Amongst cluster III DGKs, the expression of DGK5-like was up-regulated in response to cryptogein. Besides DGK5-like is likely to be the main cluster III DGK isoform silenced in one of our mutant lines, making it a strong candidate for the observed response to cryptogein. The relevance of these results is discussed with regard to early signalling lipid-mediated events in plant immunity.
Agroécologie, AgroSup Dijon, CNRS, INRA, Univ. Bourgogne Franche-Comté, F-21000, Dijon, France.
Jean-Luc
JL
Cacas
Agroécologie, AgroSup Dijon, CNRS, INRA, Univ. Bourgogne Franche-Comté, F-21000, Dijon, France.
Patricia
P
Gerbeau-Pissot
Agroécologie, AgroSup Dijon, CNRS, INRA, Univ. Bourgogne Franche-Comté, F-21000, Dijon, France.
Jérôme
J
Fromentin
UPMC UnivParis06, UR5, Physiologie Cellulaire et Moléculaire des Plantes, 4 place Jussieu, 75252, Paris cedex 05, France.
Catherine
C
Cantrel
Agroécologie, AgroSup Dijon, CNRS, INRA, Univ. Bourgogne Franche-Comté, F-21000, Dijon, France.
Dominique
D
Thomas
UPMC UnivParis06, UR5, Physiologie Cellulaire et Moléculaire des Plantes, 4 place Jussieu, 75252, Paris cedex 05, France.
Emmanuelle
E
Jeannette
UPE, UPEC, Institut d'Ecologie et des Sciences de l'Environnement de Paris, 61 avenue du général de Gaulle, 94010, Créteil, France.|CNRS, UMR7618, Institut d'Ecologie et des Sciences de l'Environnement de Paris, 61 avenue du général de Gaulle, 94010, Créteil, France.
Tetiana
T
Kalachova
CNRS, UMR 5200 Laboratoire de Biogenèse Membranaire, INRA Bordeaux Aquitaine, BP81, F-33883, Villenave d'Ornon, France.
Sébastien
S
Mongrand
Agroécologie, AgroSup Dijon, CNRS, INRA, Univ. Bourgogne Franche-Comté, F-21000, Dijon, France.
Françoise
F
Simon-Plas
UPMC UnivParis06, UR5, Physiologie Cellulaire et Moléculaire des Plantes, 4 place Jussieu, 75252, Paris cedex 05, France.|UPE, UPEC, Institut d'Ecologie et des Sciences de l'Environnement de Paris, 61 avenue du général de Gaulle, 94010, Créteil, France.|CNRS, UMR7618, Institut d'Ecologie et des Sciences de l'Environnement de Paris, 61 avenue du général de Gaulle, 94010, Créteil, France.
Eric
E
Ruelland
D002460:Cell Line; D016000:Cluster Analysis; D019852:Diacylglycerol Kinase / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D005656:Fungal Proteins / Q000494:pharmacology*; D000073659:Gain of Function Mutation / Q000235:genetics; D020868:Gene Silencing; D035683:MicroRNAs / Q000378:metabolism; D019255:NADPH Oxidases / Q000378:metabolism*; D000069452:Pathogen-Associated Molecular Pattern Molecules / Q000378:metabolism; D010712:Phosphatidic Acids / Q000378:metabolism; D010802:Phylogeny; D010940:Plant Proteins / Q000378:metabolism; D030821:Plants, Genetically Modified; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D016897:Respiratory Burst / Q000187:drug effects; D014026:Nicotiana / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000378:metabolism
D005656:Fungal Proteins; D035683:MicroRNAs; D000069452:Pathogen-Associated Molecular Pattern Molecules; D010712:Phosphatidic Acids; D010940:Plant Proteins; D047428:Protein Kinase Inhibitors; C061032:cryptogein protein, Phytophthora cryptogea; D019255:NADPH Oxidases; D019852:Diacylglycerol Kinase; D010738:Type C Phospholipases
DGK; RBOHD; cryptogein; microbial-associated molecular pattern; phosphatidic acid; phospholipase C; reactive oxygen species