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

被引:101
作者
Chujo, Tetsuya [1 ]
Miyamoto, Koji [1 ,3 ]
Ogawa, Satoshi [1 ]
Masuda, Yuka [1 ]
Shimizu, Takafumi [1 ]
Kishi-Kaboshi, Mitsuko [2 ]
Takahashi, Akira [2 ]
Nishizawa, Yoko [2 ]
Minami, Eiichi [2 ]
Nojiri, Hideaki [1 ]
Yamane, Hisakazu [1 ,3 ]
Okada, Kazunori [1 ]
机构
[1] Univ Tokyo, Biotechnol Res Ctr, Bunkyo Ku, Tokyo, Japan
[2] Natl Inst Agrobiol Sci, Genetically Modified Organism Res Ctr, Tsukuba, Ibaraki, Japan
[3] Teikyo Univ, Dept Biosci, Utsunomiya, Tochigi, Japan
来源
PLOS ONE | 2014年 / 9卷 / 06期
关键词
ACTIVATED PROTEIN-KINASE; WRKY TRANSCRIPTION FACTORS; MAP KINASE; DEFENSE RESPONSES; PLANT DEFENSE; PHYTOALEXIN BIOSYNTHESIS; BINDING-PROTEINS; CELL-DEATH; GENE; ARABIDOPSIS;
D O I
10.1371/journal.pone.0098737
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
收藏
页数:14
相关论文
共 52 条
[12]   Early nuclear events in plant defence signalling: rapid gene activation by WRKY transcription factors [J].
Eulgem, T ;
Rushton, PJ ;
Schmelzer, E ;
Hahlbrock, K ;
Somssich, IE .
EMBO JOURNAL, 1999, 18 (17) :4689-4699
[13]   The WRKY superfamily of plant transcription factors [J].
Eulgem, T ;
Rushton, PJ ;
Robatzek, S ;
Somssich, IE .
TRENDS IN PLANT SCIENCE, 2000, 5 (05) :199-206
[14]   Networks of WRKY transcription factors in defense signaling [J].
Eulgem, Thomas ;
Somssich, Imre E. .
CURRENT OPINION IN PLANT BIOLOGY, 2007, 10 (04) :366-371
[15]   Identification of the minimal repression domain of SUPERMAN shows that the DLELRL hexapeptide is both necessary and sufficient for repression of transcription in Arabidopsis [J].
Hiratsu, K ;
Mitsuda, N ;
Matsui, K ;
Ohme-Takagi, M .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2004, 321 (01) :172-178
[16]  
Ichimura K, 2002, TRENDS PLANT SCI, V7, P301, DOI 10.1016/S1360-1385(02)02302-6
[17]   Post-translational regulation of WRKY transcription factors in plant immunity [J].
Ishihama, Nobuaki ;
Yoshioka, Hirofumi .
CURRENT OPINION IN PLANT BIOLOGY, 2012, 15 (04) :431-437
[18]   Phosphorylation of the Nicotiana benthamiana WRKY8 Transcription Factor by MAPK Functions in the Defense Response [J].
Ishihama, Nobuaki ;
Yamada, Reiko ;
Yoshioka, Miki ;
Katou, Shinpei ;
Yoshioka, Hirofumi .
PLANT CELL, 2011, 23 (03) :1153-1170
[19]   Function of a mitogen-activated protein kinase pathway in N gene-mediated resistance in tobacco [J].
Jin, HL ;
Liu, YD ;
Yang, KY ;
Kim, CY ;
Baker, B ;
Zhang, SQ .
PLANT JOURNAL, 2003, 33 (04) :719-731
[20]   The plant immune system [J].
Jones, Jonathan D. G. ;
Dangl, Jeffery L. .
NATURE, 2006, 444 (7117) :323-329