Os05g0343400

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Insert non-formatted text here==Annotated Information==

   OsWRKY53 is a member of WRKY Gene Family.

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Function

OsWRKY53 is a common transcription factor in the signal transduction pathways triggered by both chitin oligosaccharide and fungal cerebroside elicitors. OsWRKY53 localizes to the nucleus and activates the expression of defense-related genes. Involving in basal defense responses to pathogen infection in rice.

Expression

WRKY proteins form a large family of plant-specific transcription factors that specifically bind to the W-box elements (T)TGAC(C/T) and appear to play a regulatory role in a variety of stress responses.WRKY proteins have been investigated extensively for their possible involvement in defense responses against attacks by pathogens, mainly in dicotyledonous plants such as Arabidopsis and tobacco . The rice genome is predicted to contain over 100 Oryza sativa L.WRKY (OsWRKY)genes.

Fig. 1

1.1 cDNA cloning and expression analysis of OsWRKY53 A full-length cDNA of OsWRKY53 (AK121190) was isolated from elicited suspension-cultured rice cells by RT-PCR using primers that were designed based on information from a rice genome database. The nucleotide sequence of the OsWRKY53 cDNA reported in this paper will appear in the DNA Data Bank of Japan (DDBJ) nucleotide sequence database under the accession number AB190436.

OsWRKY53 belongs to group I, which is characterized by two WRKY domains that contain the Cys2His2 zinc finger-like motif.

To investigate the involvement of OsWRKY53 in defense responses to pathogen infection, examining its expression using Northern hybridization. The elicitor-induced changes in the steady-state levels of the OsWRKY53 mRNA in suspensioncultured rice cells are shown in Fig. 1a. The mRNA level peaked 0.5 h after the addition of each elicitor (chitin oligosaccharide and fungal cerebroside elicitors), and decreased gradually thereafter. analyzing the expression pattern of OsWRKY53 in rice plants (cv. Nipponbare) inoculated with the fungal pathogen M. grisea race 007, which is compatible with cv. Nipponbare.

Northern hybridization revealed a significant increase in the level of OsWRKY53 mRNA at 6 h after inoculation, after which it decreased gradually (Fig. 1b).

1.2 Comparison of the OsWRKY53 sequence with other

homologous WRKY proteins and subcellular localization The OsWRKY53 amino acid sequence was compared with those of other known group-IWRKYproteins involved in defense responses (Fig. 2). OsWRKY53 showed similarity to PcWRKY1 from parsley (38% identity) [31] and AtWRKY33 from Arabidopsis (43% identity). A putative nuclear localization signal was identified in the OsWRKY53 amino acid sequence, as in PcWRKY1 and AtWRKY33, suggesting that OsWRKY53 localizes to the nucleus.

To investigate the subcellular localization of the OsWRKY53 protein, OsWRKY53 was fused in-frame to the sGFP reporter gene and subcloned into an expression vector under the control of the CaMV 35S promoter. This construct and a similar construct encoding sGFP alone were introduced into onion epidermal cells via particle bombardment. Onion cells expressing sGFP alone fluoresced throughout the cell. In contrast, cells transformed with a plasmid expressing the sGFP:OsWRKY53 fusion protein fluoresced mainly in the nuclei.

The TGAC core sequence(W-box elements) is essential for the sequence-specific binding activity

1.3 Transactivation of gene transcription by OsWRKY53

To investigate the function of OsWRKY53, we constructed an effector plasmid that contains the CaMV 35S promoter driving a gene that encodes a fusion protein of the DNA-binding domain of the yeast transcriptional activator GAL4 (GAL4 DB) and the full-length OsWRKY53. This plasmid, or a control plasmid encoding GAL4 DB alone, were cotransfected by particle bombardment into rice cells along with the reporter plasmid GAL4-LUC, which contains four copies of a GAL4 binding site fused to LUC. Compared with rice cells expressing GAL4 DB alone, those expressing the GAL4-OsWRKY53 fusion protein showed approximately 25-fold greater luciferase reporter activity.

Fig. 6

1.4 Activation of defense-related genes in OsWRKY53-overexpressing transgenic rice

attempting to comprehensively identify genes upregulated in OsWRKY53-overexpressing transgenic rice cells using the Agilent Rice Oligo Microarray, which contains more than 21,000 rice genes. Total RNA samples were prepared from three independent lines of OsWRKY53-overexpressing transgenic rice cells (lines A, C, and E in Fig. 6) and used for microarray analysis. A total of 221 genes were upregulated in the OsWRKY53-overexpressing transgenic rice cells. One third of the upregulated genes are of unknown function, and the other two thirds include genes likely to be related to metabolism,regulation of gene expression, defense, transport, proteolysis,and signal transduction. Another microarray analysis revealed that 102 of the 221 genes were upregulated by chitin oligosaccharide elicitor treatment (data not shown). Table 1 shows the nine most-upregulated defenserelated genes in the OsWRKY53-overexpressing transgenic rice cells. Of these nine genes, six (PBZ1 (AK071613), a PR-14 gene encoding a putative lipid transfer protein (AK058896), Chitinase1 (AK073267), PR-5 (AK111104), Chitinase 2 (AK102505), and Peroxidase (AK102307)) are chitin oligosaccharide-inducible. The upregulation of these six genes in five independent OsWRKY53-overexpressing transgenic rice lines was demonstrated using real time RT-PCR.

Table 1

Fig.7

1.5 Overexpression of OsWRKY53 enhances resistance to rice blast fungus To investigate the function of OsWRKY53 during defense responses to pathogen infection, 35S-OsWRKY53 transgenic rice plants was generated , obtaining 11 transgenic lines. RNA gel blot analysis revealed that OsWRKY53 is substantially elevated in at least six independent 35S-OsWRKY53 lines (Fig. 7a). The growth and morphology of the transgenic rice plants overexpressing OsWRKY53 were similar to those of normal-type rice plants (data not shown).To determine the contribution of OsWRKY53 to disease resistance in rice, the resistance of the transgenic plants to the fungal pathogen M. grisea race 007 was examined . As control plants, using both the wild type and 35S-OsWRKY53 transgenic rice plants inwhich the level of OsWRKY53mRNAwas not elevated. Infected leaves of the control plants developed typical disease symptoms. In contrast, most OsWRKY53-overexpressing plants showed greatly reduced symptom levels (Fig. 7b).

Evolution

figure 2.

figure 2.(continued)

WRKY proteins can possess one or more WRKY domains and, as with other transcription factors, the nature of the DNA-binding domain forms a basis for categorizing different members of the WRKY family. WRKY proteins containing a single WRKY domain are group II proteins; those with two are group I. The group II WRKY proteins are further subdivided intosubgroups a–e based on the presence of short conserved structural motifs . A third group, group III, contains a single WRKY domain with a variant zinc-finger CX7CX23HXC. Compared with domains from groups Ia and II proteins, the terminal histidine of the zinc-finger of groups Ib and III is replaced with cysteine and the spacing is altered within group III. Group IV WRKY proteins contain the WRKY motif, but lack a complete zinc-finger.

numerous WRKY gene duplications occurred after the divergence of the monocotyledons from dicotyledons some 50–80 million years ago. Early origin for the WRKY family in primitive eukaryotes, before the emergence of the plant phyla, and its gross expansion during the course of plant evolutionary radiation, likely because of selective pressures favoring greater adaptability. Group I WRKY genes represent the ancestral form, with other groups arising later through losses and gains of a WRKY domain, and that this family originated some 1.8–2 billion years ago. the C-terminal domains are required for DNA-binding activity, thus are constrained in their ability to mutate without losing function, whereas the N-terminal domains mediate protein-protein interactions and may be less functionally constrained . group Ia genes arose from a fusion of two group II WRKY genes evolved at an early time.

Labs working on this gene

 1. Rice Functional Genomics Group, Temasek Life Sciences Laboratory
 2. Key Laboratory of Horticultural Plant Biology of Ministry of Education, National Indoor Conservation Center of Virus-free Gemplasms of Fruit Crops
 3. Biotechnology Research Center, The University of Tokyo

References

1. Tetsuya Chujo;Ryota Takai;Chiharu Akimoto-Tomiyama;Sugihiro Ando;Eiichi Minami;Yoshiaki Nagamura;Hanae Kaku;Naoto Shibuya;Michiko Yasuda;Hideo Nakashita;Kenji Umemura;Atsushi Okada;Kazunori Okada;Hideaki Nojiri;Hisakazu Yamane(2007). "Involvement of the elicitor-induced gene OsWRKY53 in the expression of defense-related genes in rice." Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression 1769(7–8): 497-505.

2. Christian A. Ross;Yue Liu;Qingxi J. Shen (2007). "The WRKY Gene Family in Rice (Oryza sativa)." Journal of Integrative Plant Biology 49(6): 827-842.

3. Tetsuya Chujo;Naho Sugioka;Yuka Masuda;Naoto Shibuya;Tetsuo Takemura;Kazunori Okada;Hideaki Nojiri;Hisakazu Yamane(2009). "Promoter analysis of the elicitor-induced WRKY gene OsWRKY53, which is involved in defense responses in rice." Biosci Biotechnol Biochem 73(8): 1901-1904.

4. Zhen Xie;Zhong-Lin Zhang;Xiaolu Zou;Jie Huang;Paul Ruas;Daniel Thompson;Qingxi J. Shen (2005). "Annotations and Functional Analyses of the Rice WRKY Gene Superfamily Reveal Positive and Negative Regulators of Abscisic Acid Signaling in Aleurone Cells." Plant Physiology 137(1): 176-189.

5. Ramamoorthy Rengasamy;Jiang Shu-YeKumar Nadimuthu;Venkatesh Prasanna Nori;Ramachandran Srinivasan (2008). "A Comprehensive Transcriptional Profiling of the WRKY Gene Family in Rice Under Various Abiotic and Phytohormone Treatments." Plant and Cell Physiology 49(6): 865-879.

Structured Information

Gene Name	Os05g0343400
Description	WRKY transcription factor 53 (Transcription factor WRKY12)
Version	NM_001061787.1 GI:115463304 GeneID:4338474
Length	2368 bp
Definition	Oryza sativa Japonica Group Os05g0343400, complete gene.
Source	Oryza sativa Japonica Group ORGANISM Oryza sativa Japonica Group Eukaryota; Viridiplantae; Streptophyta; Embryophyta; Tracheophyta; Spermatophyta; Magnoliophyta; Liliopsida; Poales; Poaceae; BEP clade; Ehrhartoideae; Oryzeae; Oryza.
Chromosome	Chromosome 5
Location	Chromosome 5:16128079..16130446
Sequence Coding Region	16128193..16128504,16128618..16128782,16128881..16129341,16129441..16129617,16129715..16130063
Expression	GEO Profiles:Os05g0343400
Genome Context	<gbrowseImage1> name=NC_008398:16128079..16130446 source=RiceChromosome05 preset=GeneLocation </gbrowseImage1>
Gene Structure	<gbrowseImage2> name=NC_008398:16128079..16130446 source=RiceChromosome05 preset=GeneLocation </gbrowseImage2>
Coding Sequence	<cdnaseq>atggcgtcctcgacgggggggttggaccacgggttcacgttcacgccgccgccgttcatcacgtcgttcaccgagctgctgtcggggggcggtggggacctgctcggcgccggcggtgaggagcgctcgccgagggggttctccagaggcggagcgagggtgggcggcggggtgcccaagttcaagtccgcgcagccgccgagcctgccgctctcgccgccgccggtgtcgccgtcgtcctacttcgccatcccgccggggctcagccccaccgagctgctcgactcccccgtcctcctcagctcctcccatatcttggcgtccccgaccaccggtgcaatcccggctcagaggtacgactggaaggccagcgccgatctcatcgcttctcagcaagatgacagccgcggcgacttctccttccacaccaactccgacgccatggccgcgcaaccggcctctttcccttccttcaaggagcaagagcagcaagtggtcgagtcgagcaagaacggcgccgccgccgcgtcgagcaacaagagcggcggcggcgggaacaacaagctggaggacgggtacaactggaggaagtacgggcagaagcaggtgaaggggagcgagaacccgaggagctactacaagtgcacctacaacggctgctccatgaagaagaaggtggagcgctcgctcgccgacggccgcatcacccagatcgtctacaagggcgcacacaaccaccccaagccgctctccacccgccgcaacgcctcctcctgcgccaccgccgccgcctgcgccgacgacctcgcggcgcccggcgcgggcgcggaccagtactccgccgcgacgcccgagaactcctccgtcacgttcggcgacgacgaggccgacaacgcatcgcaccgcagcgagggcgacgagcccgaagccaagcgctggaaggaggatgctgacaacgagggcagctccggcggcatgggcggcggcgccggcggcaagccggtgcgcgagccgaggcttgtggtgcagacgctgagcgacatcgacatcctcgacgacggcttccggtggaggaagtacggccagaaggtcgtcaagggcaaccccaacccaaggagctactacaagtgcacgacggtgggctgcccggtgcggaagcacgtggagcgggcgtcgcacgacacgcgcgccgtgatcaccacctacgagggcaagcacaaccacgacgtcccggtcggccgcggcggcggcggcggacgcgccccggcgccggcgccgccgacgtcgggggcgatccggccgtcggccgtcgccgccgcccagcaggggccctacaccctcgagatgctccccaaccccgccggcctctacggcggctacggcgccggcgccggcggcgccgcgttcccgcgcaccaaggacgagcggcgggacgacctgttcgtcgagtcgctcctctgctag</cdnaseq>
Protein Sequence	<aaseq>MASSTGGLDHGFTFTPPPFITSFTELLSGGGGDLLGAGGEERSP RGFSRGGARVGGGVPKFKSAQPPSLPLSPPPVSPSSYFAIPPGLSPTELLDSPVLLSS SHILASPTTGAIPAQRYDWKASADLIASQQDDSRGDFSFHTNSDAMAAQPASFPSFKE QEQQVVESSKNGAAAASSNKSGGGGNNKLEDGYNWRKYGQKQVKGSENPRSYYKCTYN GCSMKKKVERSLADGRITQIVYKGAHNHPKPLSTRRNASSCATAAACADDLAAPGAGA DQYSAATPENSSVTFGDDEADNASHRSEGDEPEAKRWKEDADNEGSSGGMGGGAGGKP VREPRLVVQTLSDIDILDDGFRWRKYGQKVVKGNPNPRSYYKCTTVGCPVRKHVERAS HDTRAVITTYEGKHNHDVPVGRGGGGGRAPAPAPPTSGAIRPSAVAAAQQGPYTLEML PNPAGLYGGYGAGAGGAAFPRTKDERRDDLFVESLLC</aaseq>
Gene Sequence	<dnaseqindica>115..426#540..704#803..1263#1363..1539#1637..1985#actcccgtctcgtcgtctcgttctcgtctccgatcactctcctcctcatcttcgtcacggtctcctcgcttcgctagctcgcttgcttgctggctgagctgtggtacgctcgccatggcgtcctcgacgggggggttggaccacgggttcacgttcacgccgccgccgttcatcacgtcgttcaccgagctgctgtcggggggcggtggggacctgctcggcgccggcggtgaggagcgctcgccgagggggttctccagaggcggagcgagggtgggcggcggggtgcccaagttcaagtccgcgcagccgccgagcctgccgctctcgccgccgccggtgtcgccgtcgtcctacttcgccatcccgccggggctcagccccaccgagctgctcgactcccccgtcctcctcagctcctcccatgtgcgttccgcgtgacacaccttcatcttcttcttctacctttagtgcgcaattgctgtgtagtgtggtgctgatgatgatccatggcgtttgtgcgtgtttgtgcttttcagatcttggcgtccccgaccaccggtgcaatcccggctcagaggtacgactggaaggccagcgccgatctcatcgcttctcagcaagatgacagccgcggcgacttctccttccacaccaactccgacgccatggccgcgcaaccggcctctttcccttccttcaaggtacgtacaaatgcttcagctcatctagctcaacccaagctgaataaactgttgacagtctagctcaccttggttcatgtacgcttgtgctggagcaggagcaagagcagcaagtggtcgagtcgagcaagaacggcgccgccgccgcgtcgagcaacaagagcggcggcggcgggaacaacaagctggaggacgggtacaactggaggaagtacgggcagaagcaggtgaaggggagcgagaacccgaggagctactacaagtgcacctacaacggctgctccatgaagaagaaggtggagcgctcgctcgccgacggccgcatcacccagatcgtctacaagggcgcacacaaccaccccaagccgctctccacccgccgcaacgcctcctcctgcgccaccgccgccgcctgcgccgacgacctcgcggcgcccggcgcgggcgcggaccagtactccgccgcgacgcccgagaactcctccgtcacgttcggcgacgacgaggccgacaacgcatcgcaccgcagcgagggcgacgagcccgaagccaagcgctggtaagcctcacatcatcactcaaatcagcaatcgaagcgtaatatctccatgcgtgtgcgtgtgttcttcttgggtctcatggatgtcatggttttcaggaaggaggatgctgacaacgagggcagctccggcggcatgggcggcggcgccggcggcaagccggtgcgcgagccgaggcttgtggtgcagacgctgagcgacatcgacatcctcgacgacggcttccggtggaggaagtacggccagaaggtcgtcaagggcaaccccaacccaaggtaatcaatcaatcaagctgatcaaaatccgtcgattagtcgattaatcatatctgttcttgcgtctctaataatcaaatctgttcttgaatcgcaggagctactacaagtgcacgacggtgggctgcccggtgcggaagcacgtggagcgggcgtcgcacgacacgcgcgccgtgatcaccacctacgagggcaagcacaaccacgacgtcccggtcggccgcggcggcggcggcggacgcgccccggcgccggcgccgccgacgtcgggggcgatccggccgtcggccgtcgccgccgcccagcaggggccctacaccctcgagatgctccccaaccccgccggcctctacggcggctacggcgccggcgccggcggcgccgcgttcccgcgcaccaaggacgagcggcgggacgacctgttcgtcgagtcgctcctctgctagtcgagccgagccgagccgagctgagctgggccccacatccccctgctcgccacgtggcgtattttcgcctcgccgtatacgtacggccgtatagcgtacgtatacacgctcgcacgccctgcccaacacggcaatacacacatacatactctcgtacacacgtagtagcatacatatacagtatagtaggtggtagtggtagctagctagggagtgagatccaatttgttgattcgttgcaggccactgccacgtgggccacaccggaaacagtacacgcgtatacaccacacttgggatacgcgtacgtacgcacatgtacacgtagttttgtgcctttgtaactgctgagagacaggtcaaataagactgatgaatttttc</dnaseqindica>
External Link(s)	NCBI Gene:Os05g0343400, RefSeq:Os05g0343400