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Plant Cell. 2020 Feb 7. pii: tpc.00525.2019. doi: 10.1105/tpc.19.00525. [Epub ahead of print]

CRK2 and C-terminal Phosphorylation of NADPH Oxidase RBOHD Regulate Reactive Oxygen Species Production in Arabidopsis.

Author information

1
Ritsumeikan University CITY: Shiga Japan [JP].
2
Norwegian University of Science and Technology CITY: Trondheim Norway [NO].
3
University of Helsinki.
4
University of Helsinki CITY: Helsinki STATE: Uusimaa Finland [FI].
5
University of Helsinki CITY: Helsinki Finland [FI].
6
University of Turku and Åbo Akademi University CITY: Turku Finland [FI].
7
Max-Planck Institute for Plant Breeding Research; CITY: Cologne POSTAL_CODE: 50829 Germany [DE].
8
Max-Planck Institute for Plant Breeding Research; CITY: Cologne Germany [DE].
9
Institute of Molecular Biology CITY: Mainz Germany [DE].
10
Norwegian University of Science and Technology CITY: Trondheim POSTAL_CODE: 7491 Norway [NO].
11
Saitama University CITY: Saitama Japan [JP].
12
Max Planck Institute for Plant Breeding Research CITY: Cologne Germany [DE].
13
University of Helsinki CITY: Helsinki STATE: Uusimaa POSTAL_CODE: 00014 Finland [FI] michael.wrzaczek@helsinki.fi.

Abstract

Reactive oxygen species (ROS) are important messengers in eukaryotic organisms and their production is tightly controlled. Active extracellular ROS production by NADPH oxidases in plants is triggered by receptor-like protein kinase (RLK)-dependent signaling networks. Here we show that the cysteine-rich RLK CRK2 kinase activity is required for plant growth and CRK2 exists in a preformed complex with the NADPH oxidase RBOHD in Arabidopsis. Functional CRK2 is required for the full elicitor-induced ROS burst and consequently the crk2 mutant is impaired in defense against the bacterial pathogen Pseudomonas syringae pv. tomato DC3000. Our work demonstrates that CRK2 regulates plant innate immunity. We identified in vitro CRK2-dependent phosphorylation sites in the C-terminal region of RBOHD. Phosphorylation of S703 RBOHD is enhanced upon flg22 treatment and substitution of S703 with alanine reduced ROS production in Arabidopsis. Phylogenetic analysis suggests that phospho-sites in C-terminal region of RBOHD are conserved throughout the plant lineage and between animals and plants. We propose that regulation of NADPH oxidase activity by phosphorylation of the C-terminal region might be an ancient mechanism and that CRK2 is an important element in regulating MAMP-triggered ROS production.

PMID:
32034035
DOI:
10.1105/tpc.19.00525
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