Overexpression of Phosphomimic Mutated OsWRKY53 Leads to Enhanced Blast Resistance in Rice

WRKY transcription factors and mitogen-activated protein kinase (MAPK) cascades have been shown to play pivotal roles in the regulation of plant defense responses. We previously reported that OsWRKY53-overexpressing rice plants showed enhanced resistance to the rice blast fungus. In this study, we i...

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Main Authors:	Chujo, Tetsuya, Miyamoto, Koji, Ogawa, Satoshi, Masuda, Yuka, Shimizu, Takafumi, Kishi-Kaboshi, Mitsuko, Takahashi, Akira, Nishizawa, Yoko, Minami, Eiichi, Nojiri, Hideaki, Yamane, Hisakazu, Okada, Kazunori
Format:	Online
Language:	English
Published:	Public Library of Science 2014
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4043820/

id	pubmed-4043820
recordtype	oai_dc
spelling	pubmed-40438202014-06-09 Overexpression of Phosphomimic Mutated OsWRKY53 Leads to Enhanced Blast Resistance in Rice Chujo, Tetsuya Miyamoto, Koji Ogawa, Satoshi Masuda, Yuka Shimizu, Takafumi Kishi-Kaboshi, Mitsuko Takahashi, Akira Nishizawa, Yoko Minami, Eiichi Nojiri, Hideaki Yamane, Hisakazu Okada, Kazunori Research Article WRKY transcription factors and mitogen-activated protein kinase (MAPK) cascades have been shown to play pivotal roles in the regulation of plant defense responses. We previously reported that OsWRKY53-overexpressing rice plants showed enhanced resistance to the rice blast fungus. In this study, we identified OsWRKY53 as a substrate of OsMPK3/OsMPK6, components of a fungal PAMP-responsive MAPK cascade in rice, and analyzed the effect of OsWRKY53 phosphorylation on the regulation of basal defense responses to a virulence race of rice blast fungus Magnaporthe oryzae strain Ina86-137. An in vitro phosphorylation assay revealed that the OsMPK3/OsMPK6 activated by OsMKK4 phosphorylated OsWRKY53 recombinant protein at its multiple clustered serine-proline residues (SP cluster). When OsWRKY53 was coexpressed with a constitutively active mutant of OsMKK4 in a transient reporter gene assay, the enhanced transactivation activity of OsWRKY53 was found to be dependent on phosphorylation of the SP cluster. Transgenic rice plants overexpressing a phospho-mimic mutant of OsWRKY53 (OsWRKY53SD) showed further-enhanced disease resistance to the blast fungus compared to native OsWRKY53-overexpressing rice plants, and a substantial number of defense-related genes, including pathogenesis-related protein genes, were more upregulated in the OsWRKY53SD-overexpressing plants compared to the OsWRKY53-overexpressing plants. These results strongly suggest that the OsMKK4-OsMPK3/OsMPK6 cascade regulates transactivation activity of OsWRKY53, and overexpression of the phospho-mimic mutant of OsWRKY53 results in a major change to the rice transcriptome at steady state that leads to activation of a defense response against the blast fungus in rice plants. Public Library of Science 2014-06-03 /pmc/articles/PMC4043820/ /pubmed/24892523 http://dx.doi.org/10.1371/journal.pone.0098737 Text en © 2014 Chujo et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Chujo, Tetsuya Miyamoto, Koji Ogawa, Satoshi Masuda, Yuka Shimizu, Takafumi Kishi-Kaboshi, Mitsuko Takahashi, Akira Nishizawa, Yoko Minami, Eiichi Nojiri, Hideaki Yamane, Hisakazu Okada, Kazunori
spellingShingle	Chujo, Tetsuya Miyamoto, Koji Ogawa, Satoshi Masuda, Yuka Shimizu, Takafumi Kishi-Kaboshi, Mitsuko Takahashi, Akira Nishizawa, Yoko Minami, Eiichi Nojiri, Hideaki Yamane, Hisakazu Okada, Kazunori Overexpression of Phosphomimic Mutated OsWRKY53 Leads to Enhanced Blast Resistance in Rice
author_facet	Chujo, Tetsuya Miyamoto, Koji Ogawa, Satoshi Masuda, Yuka Shimizu, Takafumi Kishi-Kaboshi, Mitsuko Takahashi, Akira Nishizawa, Yoko Minami, Eiichi Nojiri, Hideaki Yamane, Hisakazu Okada, Kazunori
author_sort	Chujo, Tetsuya
title	Overexpression of Phosphomimic Mutated OsWRKY53 Leads to Enhanced Blast Resistance in Rice
title_short	Overexpression of Phosphomimic Mutated OsWRKY53 Leads to Enhanced Blast Resistance in Rice
title_full	Overexpression of Phosphomimic Mutated OsWRKY53 Leads to Enhanced Blast Resistance in Rice
title_fullStr	Overexpression of Phosphomimic Mutated OsWRKY53 Leads to Enhanced Blast Resistance in Rice
title_full_unstemmed	Overexpression of Phosphomimic Mutated OsWRKY53 Leads to Enhanced Blast Resistance in Rice
title_sort	overexpression of phosphomimic mutated oswrky53 leads to enhanced blast resistance in rice
description	WRKY transcription factors and mitogen-activated protein kinase (MAPK) cascades have been shown to play pivotal roles in the regulation of plant defense responses. We previously reported that OsWRKY53-overexpressing rice plants showed enhanced resistance to the rice blast fungus. In this study, we identified OsWRKY53 as a substrate of OsMPK3/OsMPK6, components of a fungal PAMP-responsive MAPK cascade in rice, and analyzed the effect of OsWRKY53 phosphorylation on the regulation of basal defense responses to a virulence race of rice blast fungus Magnaporthe oryzae strain Ina86-137. An in vitro phosphorylation assay revealed that the OsMPK3/OsMPK6 activated by OsMKK4 phosphorylated OsWRKY53 recombinant protein at its multiple clustered serine-proline residues (SP cluster). When OsWRKY53 was coexpressed with a constitutively active mutant of OsMKK4 in a transient reporter gene assay, the enhanced transactivation activity of OsWRKY53 was found to be dependent on phosphorylation of the SP cluster. Transgenic rice plants overexpressing a phospho-mimic mutant of OsWRKY53 (OsWRKY53SD) showed further-enhanced disease resistance to the blast fungus compared to native OsWRKY53-overexpressing rice plants, and a substantial number of defense-related genes, including pathogenesis-related protein genes, were more upregulated in the OsWRKY53SD-overexpressing plants compared to the OsWRKY53-overexpressing plants. These results strongly suggest that the OsMKK4-OsMPK3/OsMPK6 cascade regulates transactivation activity of OsWRKY53, and overexpression of the phospho-mimic mutant of OsWRKY53 results in a major change to the rice transcriptome at steady state that leads to activation of a defense response against the blast fungus in rice plants.
publisher	Public Library of Science
publishDate	2014
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4043820/
_version_	1612096141278576640

Overexpression of Phosphomimic Mutated OsWRKY53 Leads to Enhanced Blast Resistance in Rice

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