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: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 5´and 3´adaptor 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)
Project description:We report high-throughput RNA sequencing of Δcrp and five adaptively evolved mutants of Δcrp in the batch exponential phase, IG116-Δcrp in batch exponential phase and Δcrp and two adaptively evolved mutants of Δcrp in chemostat cultivations
Project description:Transcriptome analysis is an important approach to associate genotype with phenotype. The content and dynamics of eukaryotic transcriptome are far more complex than previously anticipated. Here we integrated high-throughput RNA-seq and paired-end method to conduct an unprecedentedly deep survey of transcription profile for cultivated rice, one of the oldest domesticated crops species and has since spread worldwide to become one of the major staple foods. Analysis of reads mapping revealed 4,244 previously uncharacterized transcripts, including a mass of protein-coding genes and putative functional non-coding RNA genes. Alignment of junction reads indicated over 42% of rice multiple-exon genes produce two or more distinct splicing isoforms. It’s intriguing that we identified 1,356 putative gene fusion events, indicating the 234 fusion gene produced by trans-splicing vastly increases the complexity of rice transcriptome, together with the pervasive alternative splicing events. Digital gene expression profiling revealed most rice duplicate genes were maintained by the selection constraint on gene dosages, which would increase the genetic robustness of rice to counteract deleterious mutations Keywords: Expression profiling by high throughput sequencing mRNA expression of 8 independent rice tissues was determined by method of RNA-Seq using short reads from high throughput sequencing technology. Meanwhile small RNA populations from mixture solution pooled from total RNA of each 8 tissues were also sequenced.
Project description:Rice roots grown in hydroponic culture were inoculated with rice blast fungus and gene expression profiles were analyzed by microarray Roots of two isogenic lines of rice cv Nipponbare (blast-resistance gene: Pia or pia) were inoculated with rice blast fungus, P91-15B, carrying avirulence gene, AvrPia. Total RNA was isolated from crown roots, labeled with cy3, and probed with agilent rice oligoarray (4x44).
Project description:Rice blast fungus was inoculated to susceptible and resistant lines of rice leaves and the invaded cells and their adjacent cells were collected by Laser Microdissection method. The extracted RNA was amplified and analyzed by two color microarray.
Project description:Fourth leaves of rice seedlings (4.5 leaf stage) grown in hydroponic culture were inoculated with rice blast fungus and gene expression profiles were analyzed by microarray. Fourth leaves of the two isogenic lines of rice cv Nipponbare (blast-resistance gene: Pia or its mutant, pia) were inoculated with rice blast fungus, P91-15B, carrying avirulence gene, AvrPia. Total RNA was isolated, labeled with cy3, and probed with agilent rice oligoarray (4x44).
Project description:Fourth leaves of rice seedlings (4.5 leaf stage) grown in hydroponic culture were inoculated with rice blast fungus and gene expression profiles were analyzed by microarray. Fourth leaves of the two isogenic lines of rice cv Nipponbare (blast-resistance gene: Pish or its mutant, pish) were inoculated with rice blast fungus, Kyu77-07A, carrying avirulence gene, AvrPish. Total RNA was isolated, labeled with cy3, and probed with agilent rice oligoarray (4x44).
Project description:To investigate plant-fungus interactions in early stage of infection, we analyzed response of rice against Magnaporthe grisea infection deficient mutants. In M. grisea, Mgb1 and Mst12 are essential for development of infection structures. Deletion of MGB1 results in defect in appresorium formation, and MST12, in penetration peg development. Analysis of gene expression profiles in rice by microarray revealed the mutant-specific and R gene dependent gene expression, strongly suggesting that gene-for-gene interaction commences before the penetration into rice cell. Keywords: disease state analysis