Project description:WRKY45-dependent rice genes: Mock- and benzothiadiazole (BTH)-treated wild-type (Nipponbare NB) rice and mock- and BTH-treated WRKY45-knockdown (W45-kd) rice (line #15) were analyzed in four biological replicates at 12 and 24 h after the treatment. Mock- and benzothiadiazole (BTH)-treated wild-type (Nipponbare NB) rice and mock- and BTH-treated WRKY45-knockdown (W45-kd) rice (line #3) were analyzed in four biological replicates at 24 h after the treatment.
Project description:NPR1 is a central positive regulator of salicylic-acid (SA)-mediated defense signaling in Arabidopsis. Here, we report characterization of OsNPR1, an Oryzae sativa (rice) ortholog of NPR1, focusing on its role in blast disease resistance and identification of OsNPR1-regulated genes followed by their comparison with NPR1-regulated genes in Arabidopsis. Blast resistance tests using OsNPR1–knockdown and –overexpressing rice lines indicated that OsNPR1 plays an essential role in benzothiadiazole-induced blast resistance. Genome-wide transcript profiling using OsNPR1–knockdown lines revealed that 358 genes out of 1,228 BTH-upregulated genes and 724 genes out of 1,069 BTH-downregulated genes were OsNPR1 dependent with respect to their BTH responsiveness, indicating that OsNPR1 plays a major role in the downregulation. Inspection of OsNPR1-dependent genes revealed that many genes involved in photosynthesis and chloroplastic translation and transcription were downregulated by BTH in an OsNPR1 dependent manner, indicating that photosynthesis and chloroplast activities is coordinately suppressed by OsNPR1 in response to BTH-induced activation of SA-signaling pathway. ABA-responsive genes were also OsNPR1-dependently downregulated, suggesting antagonistic interaction of SA signaling on ABA signaling. None of 11 BTH-upregulated genes for WRKY transcription factors was OsNPR1 dependent, whereas most of those are NPR1-dependently upregulated in Arabidopsis, indicating that the role of OsNPR1 is distinct from that of NPR1 in Arabidopsis. We discuss the significance of OsNPR1-regulated gene expression in SA-regulated defense program and the role of OsNPR1 in rice SA-signaling pathway that is branched to OsNPR1- and rice WRKY45-dependent sub-pathways. mock-treated wild-type (Nipponbare) rice, benzothiadiazole (BTH)-treated wild-type rice, mock-treated WRKY45-knockdown rice (2 lines) and BTH-treated WRKY45-knockdown rice (2 lines) were analyzed in four biological replicates.
Project description:NPR1 is a central positive regulator of salicylic-acid (SA)-mediated defense signaling in Arabidopsis. Here, we report characterization of OsNPR1, an Oryzae sativa (rice) ortholog of NPR1, focusing on its role in blast disease resistance and identification of OsNPR1-regulated genes followed by their comparison with NPR1-regulated genes in Arabidopsis. Blast resistance tests using OsNPR1–knockdown and –overexpressing rice lines indicated that OsNPR1 plays an essential role in benzothiadiazole-induced blast resistance. Genome-wide transcript profiling using OsNPR1–knockdown lines revealed that 358 genes out of 1,228 BTH-upregulated genes and 724 genes out of 1,069 BTH-downregulated genes were OsNPR1 dependent with respect to their BTH responsiveness, indicating that OsNPR1 plays a major role in the downregulation. Inspection of OsNPR1-dependent genes revealed that many genes involved in photosynthesis and chloroplastic translation and transcription were downregulated by BTH in an OsNPR1 dependent manner, indicating that photosynthesis and chloroplast activities is coordinately suppressed by OsNPR1 in response to BTH-induced activation of SA-signaling pathway. ABA-responsive genes were also OsNPR1-dependently downregulated, suggesting antagonistic interaction of SA signaling on ABA signaling. None of 11 BTH-upregulated genes for WRKY transcription factors was OsNPR1 dependent, whereas most of those are NPR1-dependently upregulated in Arabidopsis, indicating that the role of OsNPR1 is distinct from that of NPR1 in Arabidopsis. We discuss the significance of OsNPR1-regulated gene expression in SA-regulated defense program and the role of OsNPR1 in rice SA-signaling pathway that is branched to OsNPR1- and rice WRKY45-dependent sub-pathways.
Project description:Here, we present OryzaPG-DB, a rice proteome database based on shotgun proteogenomics, which incorporates the genomic features of experimental shotgun proteomics data. This version of the database was created from the results of 27 nanoLC-MS/MS runs on a hybrid ion trap-orbitrap mass spectrometer, which offers high accuracy for analyzing tryptic digests from undifferentiated cultured rice cells. Peptides were identified by searching the product ion spectra against the protein, cDNA, transcript and genome databases from Michigan State University, and were mapped to the rice genome. Approximately 3200 genes were covered by these peptides and 40 of them contained novel genomic features. Users can search, download or navigate the database per chromosome, gene, protein, cDNA or transcript and download the updated annotations in standard GFF3 format, with visualization in PNG format. In addition, the database scheme of OryzaPG was designed to be generic and can be reused to host similar proteogenomic information for other species. OryzaPG is the first proteogenomics-based database of the rice proteome, providing peptide-based expression profiles, together with the corresponding genomic origin, including the annotation of novelty for each peptide.
Project description:WRKY45-dependent rice genes: Mock- and benzothiadiazole (BTH)-treated wild-type (Nipponbare NB) rice and mock- and BTH-treated WRKY45-knockdown (W45-kd) rice (line #15) were analyzed in four biological replicates at 12 and 24 h after the treatment. Mock- and benzothiadiazole (BTH)-treated wild-type (Nipponbare NB) rice and mock- and BTH-treated WRKY45-knockdown (W45-kd) rice (line #3) were analyzed in four biological replicates at 24 h after the treatment. Mock- and benzothiadiazole (BTH)-treated wild-type (Nipponbare NB) rice and mock- and BTH-treated WRKY45-knockdown (W45-kd) rice (line #15) were analyzed in four biological replicates at 12 and 24 h after the treatment. Mock- and benzothiadiazole (BTH)-treated wild-type (Nipponbare NB) rice and mock- and BTH-treated WRKY45-knockdown (W45-kd) rice (line #3) were analyzed in four biological replicates at 24 h after the treatment.
Project description:5 leaves old rice plantlets were infected with Magnaporthe grisea spores and zero, two hours and twenty four houres after infection samples were collected
Project description:RNA-dependent RNA polymerase 6 (RDR6) is a core component of the small RNA biogenesis pathway, but its function in meiosis is unclear. Herein we report a new allele of OsRDR6 (Osrdr6-mei), which causes meiosis-specific phenotype in rice. In Osrdr6-mei, meiotic DSB formation was blocked to some extent. We further created a biallelic mutant, Osrdr6-bi, by crossing Osrdr6-mei with a knock out mutant, Osrdr6-edit (Osrdr6-edi). In Osrdr6-bi, 24 univalents were observed at diakinesis, and no histone H2AX phosphorylation (γH2AX) foci was detected in meiocytes, indicating that OsRDR6 is crucial for meiotic DSB formation. Compared with the wild type, the number of 21-nt small RNAs was dramatically reduced, while the number of 24-nt small RNAs was significantly increased in Osrdr6-mei. And thousands of DMRs discovered in Osrdr6-mei, implying that OsRDR6 plays an essential role on DNA methylation, especially CHH methylation. Meanwhile, the transcriptome data showed that 457 genes were down-regulated in the mutant; including three genes related to DSB formation: OsSDS, P31comet and CRC1. Interestingly, the increased 24-nt small RNA level surrounding down-regulated genes was largely associated with the silencing of these genes, while DMRs were rarely related to them. In summary, we speculated that the small RNA levels are disordered in Osrdr6 mutants, which may lead to the defection of DSB formation.