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Identification of Magnaporthe oryzae elicited small RNAs and expression profiles in resistant and susceptible rice cultivars

ABSTRACT: To identify blast pathogen elicited miRNAs, four sRNA libraries were constructed: 8749036, A library, susceptible rice cultivar Zhonger-Ruanzhan; 12157795, B library, mock-treated blast resistant line H4; 9937360, C library, blast-treated susceptible rice cultivar Zhonger-Ruanzhan; and 11138187 clean reads, D library, blast-treated blast resistant line H4. The Short Oligonucleotide Analysis Package (SOAP; Beijing Genomics Institute) matched 85.46% (A, 7476714), 79.35% (B, 9647324), 77.70% (C, 7721715) and 76.10% (D, 8476386) sRNA reads to the genome. After removing other RNA categories matched to NCBI Genbank, Rfam database, known rice miRNA precursor, repeat associated RNA and siRNA, the remaining reads: 1681359(A); 3829741(B); 3182403(C); and 3970132(D), were used for further novel miRNA prediction. Since some miRNAs were tissue-specific, time-specific or stress-induced, only 291, 210, 164 and 220 registered miRNAs were identified in libraries A, B, C and D respectively. Thirty-one (A, 9385 reads), 399(B, 41024 read), 351(C, 36622 reads) and 333(D, 38064 reads) unique sRNAs were projected to be candidate miRNAs.

ORGANISM(S): Oryza sativa Indica Group

PROVIDER: GSE36205 | GEO | 2016/01/01

SECONDARY ACCESSION(S): PRJNA152877

REPOSITORIES: GEO

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Project description:The rice blast fungus, Magnaporthe oryzae is a destructive pathogen of rice and other related crops, causing significant yield losses worldwide. Endogenous small RNAs (sRNAs), including small interfering RNAs (siRNAs) and microRNAs (miRNAs) are critical components of gene regulation in many eukaryotic organisms. Recently several new species of sRNAs have been identified in fungi. This fact along with the availability of genome sequence makes M. oryzae a compelling target for sRNA profiling. We have examined sRNA species and their biosynthetic genes in M. oryzae, and the degree to which these elements regulate fungal stress responses. To this end, we have characterized sRNAs under different physiological stress conditions, which had not yet been examined in this fungus. The resulting libraries are composed of more than 37 million total genome matched reads mapping to intergenic regions, coding sequences, retrotransposons, inverted, tandem, and other repeated regions of the genome with more than half of the small RNAs arising from intergenic regions. The 24 nucleotide (nt) size class of sRNAs was predominant. A comparison to transcriptional data of M. oryzae undergoing the same physiological stresses indicates that sRNAs play a role in transcriptional regulation for a small subset of genes. Support for this idea comes from generation and characterization of mutants putatively involved in sRNAs biogenesis; our results indicate that the deletion of Dicer-like genes and an RNA-Dependent RNA Polymerase gene increases the transcriptional regulation of this subset of genes, including one involved in virulence. Various physiological stressors and in planta conditions alter the small RNA profile of the rice blast fungus. Characterization of sRNA biosynthetic mutants helps to clarify the role of sRNAs in transcriptional control. Examination of small RNA populations of eight mycelial as well as In planta tissues of M. oryzae strain 70-15. Mycelia grown in complete medium (CM), minimal medium (MM), carbon starved medium (CS: MM lacking carbon source), nitrogen starved medium (NS: MM lacking nitrogen source) and a reactive oxygen species-rich environment generated by amendment of Paraquat (PQ) to the CM. In planta tissues include mock inoculated rice leaves (LMg0), 72 hours post-inoculation in rice leaves (LMg72) and 96 hours post-inoculation in rice leaves (LMg96).

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Identification of Magnaporthe oryzae elicited small RNAs and expression profiles in resistant and susceptible rice cultivars

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