OsWRKY53 - Revision history

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2014-06-04T13:33:16Z

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2014-06-02T15:19:02Z

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2014-06-02T12:45:11Z

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2014-06-02T12:23:07Z

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Ricelove: Created page with " == Function: == vvvOsWRKY53 is a transcription factor that plays important roles in chitin oligosaccharide elicitor-induced defense signaling pathways in rice.OsWRKY53 was a..."

2014-06-02T09:17:15Z

Created page with " == Function: == vvvOsWRKY53 is a transcription factor that plays important roles in chitin oligosaccharide elicitor-induced defense signaling pathways in rice.OsWRKY53 was a..."

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== Function: ==

vvvOsWRKY53 is a transcription factor that plays important roles in chitin oligosaccharide elicitor-induced defense signaling pathways in rice.OsWRKY53 was also induced in suspension-cultured rice cells by a fungal cerebroside elicitor and in rice plants by infection with the blast fungus Magnaporthe grisea. OsWRKY53 is a transcriptional activator and specifically bound to W-box elements,overexpression of OsWRKY53 in rice plants resulted in enhanced resistance to M. grisea.

← Older revision		Revision as of 13:33, 4 June 2014
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−	~~== Function: ==~~
−
−	OsWRKY53 is a transcription factor that plays important roles in chitin oligosaccharide elicitor-induced defense signaling pathways in rice.OsWRKY53 was also induced in suspension-cultured rice cells by a fungal cerebroside elicitor and in rice plants by infection with the blast fungus Magnaporthe grisea. OsWRKY53 is a transcriptional activator and specifically bound to W-box elements,overexpression of OsWRKY53 in rice plants resulted in enhanced resistance to M. grisea.
−
−	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.
−	~~1.1 cDNA cloning and expression analysis of OsWRKY53~~
−	A full-length cDNA of OsWRKY53 (AK121190; named by Xie et al.) 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.
−	All known WRKY proteins contain up to two WRKY domains and are classified based on both the number of WRKY domains and the features of their zinc finger-like motifs. 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). These results strongly suggest that OsWRKY53 is involved in the basal defense responses of rice against pathogen infection.
−
−	~~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.
−
−	~~1.3 Binding of OsWRKY53 to conserved W-box motifs~~
−	The DNA-binding activity of OsWRKY53 was investigated in vitro by a GMSA using partially purified Trx-OsWRKY53(Fig. 4). The DNA probe used was the double-stranded synthetic oligonucleotide BS65, which has been reported to represent a binding site of WRKY-family proteins in Arabidopsis and tobacco. When incubated with the recombinant OsWRKY53, BS65 produced retarded bands; in contrast, no retardation of the probe was observed with Trx alone. To verify the binding specificity of OsWRKY53 to the DNA probe, a parallel binding assay was performed with competitor DNA. A 250-fold molar excess of the cold target DNA was sufficient to compete with, and eliminate, the retardation of the labeled probe.
−	To investigate whether the TGAC core sequence is essential for the sequence-specific binding activity, testing a mutant probe in which the two TGAC sequences were changed to CCTA. This mutant probe, mBS65, was not recognized by OsWRKY53,indicating that the probe retardation was due to the specific
−	~~binding activity of OsWRKY53 to the W-box elements.~~
−
−	~~1.4 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.
−
−	~~1.5 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~~

@@ Line 5: / Line 5: @@
     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.
-.1. cDNA cloning and expression analysis of OsWRKY53
+.1 cDNA cloning and expression analysis of OsWRKY53
     A full-length cDNA of OsWRKY53 (AK121190; named by Xie et al.) 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.
     All known WRKY proteins contain up to two WRKY domains and are classified based on both the number of WRKY domains and the features of their zinc finger-like motifs. 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
 .5 h after the addition of each elicitor (chitin oligosaccharide and fungal cerebroside elicitors), and decreased gradually thereafter.
-We also analyzed 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.
+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.

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 .1. cDNA cloning and expression analysis of OsWRKY53
     A full-length cDNA of OsWRKY53 (AK121190; named by Xie et al.) 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.
-    All known WRKY proteins contain up to two WRKY domains and are classified based on both the number of WRKY domains and the features of their zinc finger-like motifs. 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, we examined 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
+    All known WRKY proteins contain up to two WRKY domains and are classified based on both the number of WRKY domains and the features of their zinc finger-like motifs. OsWRKY53 belongs to group I, which is characterized by two WRKY domains that contain the Cys2His2 zinc finger-like motif.
 .5 h after the addition of each elicitor (chitin oligosaccharide and fungal cerebroside elicitors), and decreased gradually thereafter.
 We also analyzed 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.

← Older revision		Revision as of 12:23, 2 June 2014
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	== Function: ==		== Function: ==

−	~~vvvOsWRKY53~~ is a transcription factor that plays important roles in chitin oligosaccharide elicitor-induced defense signaling pathways in rice.OsWRKY53 was also induced in suspension-cultured rice cells by a fungal cerebroside elicitor and in rice plants by infection with the blast fungus Magnaporthe grisea. OsWRKY53 is a transcriptional activator and specifically bound to W-box elements,overexpression of OsWRKY53 in rice plants resulted in enhanced resistance to M. grisea.	+	OsWRKY53 is a transcription factor that plays important roles in chitin oligosaccharide elicitor-induced defense signaling pathways in rice.OsWRKY53 was also induced in suspension-cultured rice cells by a fungal cerebroside elicitor and in rice plants by infection with the blast fungus Magnaporthe grisea. OsWRKY53 is a transcriptional activator and specifically bound to W-box elements,overexpression of OsWRKY53 in rice plants resulted in enhanced resistance to M. grisea.
		+
		+	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.
		+	3.1. cDNA cloning and expression analysis of OsWRKY53
		+	A full-length cDNA of OsWRKY53 (AK121190; named by Xie et al.) 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.
		+	All known WRKY proteins contain up to two WRKY domains and are classified based on both the number of WRKY domains and the features of their zinc finger-like motifs. 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, we examined 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.
		+	We also analyzed 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.