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:Magnaporthe oryzae is the causative agent of the rice blast, the most relevant rice disease worldwide. To date expression analysis on rice infected with Magnaporthe oryzae have been carried out only with the strains FR13 (leaf) and Guy 11 (root). However different strains of Magnaporthe are present in the environment leading to different rice responses at molecular level. To gain more insight on the unknown molecular mechanisms activated by different Magnaporthe strains during rice defense, a global expression analysis was performed by using the GeneChip® Rice Genome Array. To identify rice genes differentially regulated upon infection by Magnaporthe isolates, inoculation with different strains were performed and samples were collected 24 hours post infection.
Project description:Magnaporthe oryzae is the causative agent of the rice blast, the most relevant rice disease worldwide. To date expression analysis on rice infected with Magnaporthe oryzae have been carried out only with the strains FR13 (leaf) and Guy 11 (root). However different strains of Magnaporthe are present in the environment leading to different rice responses at molecular level. To gain more insight on the unknown molecular mechanisms activated by different Magnaporthe strains during rice defense, a global expression analysis was performed by using the GeneChip® Rice Genome Array. To identify rice genes differentially regulated upon infection by Magnaporthe isolates, inoculation with different strains were performed and samples were collected 24 hours post infection. RNA were obtained from leaf samples after inoculation of rice 2 week-old plantlets with the following strains: rice isolates Magnaporthe oryzae FR13 and CL367, non-adapted strain BR32, isolated from wheat, and Magnaporthe grisea BR29 isolated from crabgrass. Treated and control (mock) rice leaves (cv. Nipponbare) were collected 24 hours post inoculation. Three biological replicates for each interaction type and the corresponding mock were extracted and analysed independently with the GeneChip® Rice Genome Array.
Project description:Rice blast disease caused by Magnaporthe oryzae is one of the most damaging diseases affecting rice productivity. Previously, we reported a novel M. oryzae- secreted protein MSP1, which triggers cell death and pathogen-associated molecular pattern (PAMP)-triggered immune (PTI) responses in rice. To investigate the MSP1 induced defense response in rice at the protein level, we employed a label-free quantitative proteomic approach, in parallel with the flg22, which is a wellknown elicitor. Proteomics analysis using the MaxQuant-Perseus platform led to the identification of 4087 proteins of which 417 showed significant differences (multiple sample test, ANOVA p<0.05) in response to MSP1 and/or flg22 treatments. Functional annotation of the differential proteins showed that proteins related to the primary metabolism, secondary metabolism and lipid metabolism were strongly down-regulated, while elevated proteins were mainly associated with the stress response, chromatin remodeling, post-translational modification of proteins and signaling.
Project description:To investigate the role of iron excess in rice immune responses to Magnaporthe oryzae infection. Gene expression profiling analysis were performed using data obtained from RNA-seq of rice plants grown in differential iron supply and challenged with Magnaporthe oryzae spores.
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 control and 2 hour were mixed and hybridized with chip besides control and twenty four hours were mixed and hybridized with another chip. Both chips were performed in duplicate
Project description:High-throughput sequencing of small RNAs from rice was used to identify distinct miRNAs that are responsive to elicitors from the fungal pathogen Magnaporthe oryzae. [Expression profiling by array] We used microarrays to determine the expression behaviour of target genes for elicitor-regulated miRNAs. [High throughput sequencing] High-throughput sequencing of rice small RNAs was performed in two different tissues, leaves and roots, and two different time point of elicitor treatment, 30' and 2h Amplicons were prepared by 5M-BM-4and 3M-BM-4adaptor ligation in which the 5'-adaptor contained a 'barcode' consisting of a 4-nucleotide identifier sequence for each sample. The libraries containing unique barcodes were combined and subjected to pyrosequencing (454 Life SciencesTM, Roche) [Expression profiling by array] Leaves from rice plants were harvested at two time points after the onset of treatment (30' and 2h) with elicitors of Magnaporthe oryzae 18.1 and used for RNA extraction and hybridization on Affymetrix microarrays. Mock inoculations were performed with sterile water for control experiments. Three biological replicates were analyzed. Each sample represented a pool of approximately 150 rice plants. [High throughput sequencing] 8 samples examined: leaves and roots, treated or not with elicitors at two different time points, 30' and 2h (2x2x2)
Project description:Rice blast disease is a major threat to rice production worldwide, but the mechanisms underlying rice resistance to the causal agent Magnaporthe oryzae remain elusive. In this whole-genome transcriptome study of rice early defense response to M. oryzae, we applied Affymetrix Rice Genome Genechip to compare the compatible and incompatible rice-M. oryzae interactions in 24 hours post-inoculation. Leaf samples were harvested from three biological replicates of fungal- and mock-inoculated seedlings at 24 hours post-inoculation, from which RNA were extracted and analyzed with Genechip Rice Genome Array.
Project description:Rice is a staple food crop worldwide, and its production is severely threatened by phloem-feeding insect herbivores, particularly the brown planthopper (BPH, Nilaparvata lugens), and destructive pathogens. Despite the identification of many BPH resistance genes, the molecular basis of rice resistance to BPH remains largely unclear. Here, we report that the plant elicitor peptide (Pep) signalling confers rice resistance to BPH. Both rice PEP RECEPTORs (PEPRs) and PRECURSORs of PEP (PROPEPs), particularly OsPROPEP3, were transcriptionally induced in leaf sheaths upon BPH infestation. Knockout of OsPEPRs impaired rice resistance to BPH, whereas exogenous application of OsPep3 improved the resistance. Hormone measurement and co-profiling of transcriptomics and metabolomics in OsPep3-treated rice leaf sheaths suggested potential contributions of jasmonic acid biosynthesis, lipid metabolism and phenylpropanoid metabolism to OsPep3-induced rice immunity. Moreover, OsPep3 elicitation also strengthened rice resistance to the fungal pathogen Magnaporthe oryzae and bacterial pathogen Xanthamonas oryzae pv. oryzae and provoked immune responses in wheat. Collectively, this work demonstrates a previously unappreciated importance of the Pep signalling in plants for combating piercing-sucking insect herbivores and promises exogenous application of OsPep3 as an eco-friendly immune stimulator in agriculture for crop protection against a broad spectrum of insect pests and pathogens.
Project description:We created a mutant in the MAP kinase-encoding Pmk1 gene of the rice blast fungus Magnaporthe oryzae (pmk1AS) that renders the gene sensitive to inhibition by 1NA-PP1. Fungal gene expression was compared during infection of rice sheath by the M. oryzae pmk1AS mutant in the presence and absence of 1NA-PP1.