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Comparison of transcript abundance in mycelium from the rice blast fungus Magnaporthe oryzae grown in rich medium and in minimal medium using HT-SuperSAGE.

ABSTRACT: Mycelium from the rice blast fungus Magnaporthe oryzae was grown in both rich medium and under nutrient limiting conditions. Genes were identified that were more highly expressed in one condition as compared to the other. Samples were taken from mycelium grown in both complete medium and in glucose minimal medium. Three replicates were taken for each condition.

ORGANISM(S): Magnaporthe oryzae

SUBMITTER: Darren Soanes

PROVIDER: E-GEOD-30256 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Genome-wide transcriptional profiling of appressorium development by the rice blast fungus Magnaporthe oryzae.

Soanes Darren M DM Chakrabarti Apratim A Paszkiewicz Konrad H KH Dawe Angus L AL Talbot Nicholas J NJ

PLoS pathogens 20120209 2

The rice blast fungus Magnaporthe oryzae is one of the most significant pathogens affecting global food security. To cause rice blast disease the fungus elaborates a specialised infection structure called an appressorium. Here, we report genome wide transcriptional profile analysis of appressorium development using next generation sequencing (NGS). We performed both RNA-Seq and High-Throughput SuperSAGE analysis to compare the utility of these procedures for identifying differential gene express ...[more]

PMID: 22346750

Similar Datasets

Analysis of transcript abundance during appressorium formation in the rice blast fungus Magnaporthe oryzae using HT-SuperSAGE.

Project description:The rice-blast fungus Magnaporthe oryzae is disseminated using a three-celled spore or conidium. Upon landing on the surface of a rice leaf, the conidium germinates and forms a specialised structure for entry into the host plant - the appressorium. In this study we have followed gene expression thoughout germination of the conidium and formation of the appressorium to identify genes that may be important for appressorium function. We have also compared gene expression in the wild-type germinating conidium at four hours to a mutant strain deleted for the MAP kinase pmk1. The pmk1 mutant is unable to form appressoria. Samples were taken from five time-points in wild-type germinating conidia and one time-point in the pmk1 mutant. Two replicates were taken at each time-point.

2012-02-13 | E-GEOD-30069 | biostudies-arrayexpress

Comparison of transcript abundance in mycelium from the rice blast fungus Magnaporthe oryzae grown in rich medium and in minimal medium using RNA-Seq

Project description:Mycelium from the rice blast fungus Magnaporthe oryzae was grown in both rich medium and under nutrient limiting conditions. Genes were identified that were more highly expressed in one condition as compared to the other. Samples were taken from mycelium grown in both complete medium and in glucose minimal medium. One replicate for each sample.

2012-02-13 | E-GEOD-30327 | biostudies-arrayexpress

Small RNA sequencing of transgenes silencing by paramutation in C. elegans

Project description:In the nematode Caenorhabditis elegans, different small RNA-dependent gene silencing mechanisms act in the germline to initiate transgenerational gene silencing. Piwi-interacting RNAs (piRNAs) can initiate transposon and gene silencing by acting upstream of endogenous short interfering RNAs (siRNAs), which engage a nuclear RNA interference (RNAi) pathway to trigger transcriptional gene silencing. Once gene silencing has been established, it can be stably maintained over multiple generations without the requirement of the initial trigger and is also referred to as RNAe or paramutation. This heritable silencing depends on the integrity of the nuclear RNAi pathway. However, the exact mechanism by which silencing is maintained across generations is not understood.Here we demonstrate that silencing of piRNA targets involves the production of two distinct classes of small RNAs with different genetic requirements. The first class, secondary siRNAs, are localized close to the direct target site for piRNAs. Nuclear import of the secondary siRNAs by the Argonaute HRDE-1 leads to the production of a distinct class of small RNAs that map throughout the transcript, which we term tertiary siRNAs. Both classes of small RNAs are necessary for full repression of the target gene and can be maintained independently of the initial piRNA trigger. Consistently, we observed a form of paramutation associated with tertiary siRNAs. Once paramutated, a tertiary siRNA generating allele confers dominant silencing in the progeny regardless of its own transmission, suggesting germline-transmitted siRNAs are sufficient for multigenerational silencing. C. elegans strains containing transgenes silenced by piRNAs were crossed to strains with transgenes with similar sequences but without piRNA target sites, to investigate the spreading of silencing between transgenes mediated by small RNAs. Mutant backgrounds were used to investigate the genetic requirements for this process.

2015-04-01 | E-GEOD-66344 | biostudies-arrayexpress

Small RNA sequencing study of the nematode phylum

Project description:Comparison of small RNA profiles across nematode evolution Sequencing of small RNAs (15-33nt)

2015-02-01 | E-GEOD-56651 | biostudies-arrayexpress

No evidence for a C. elegans small RNA response to feeding with OxyS-expressing bacteria

Project description:Small RNA sequencing shows that there are no Caenorhabditis elegans endo siRNAs made against the OxyS non-coding RNA. Sequencing of small RNAs from C. elegans fed bacteria overexpressing the non-coding RNA OxyS, control samples fed either standard E. coli or one without the OxyS RNA, an rde-4 deletion mutant deficient in endo siRNA production and the OxyS-overexpressing bacteria itself. 5' independent samples were treated with the enzyme RNA 5' polyphosphatase to remove 5' triphosphate; 5' dependent samples were not treated with 5' polyphosphatase.

2016-07-05 | E-GEOD-64990 | biostudies-arrayexpress

Analysis of small RNA response to viral infection in C.elegans

Project description:Dissection of the small RNA pathway required for generation of an antiviral small RNA response Small non-coding RNA (~15-30nt) was extracted from animals infected with the Orsay virus

2013-06-01 | E-GEOD-41693 | biostudies-arrayexpress

The loop position of shRNAs and pre-miRNAs is critical for the accuracy of Dicer processing in vivo

Project description:Short-hairpin RNA (shRNA)-induced RNAi is used for biological discovery and therapeutics. Dicer, whose normal role is to liberate endogenous miRNAs from their precursors, processes shRNAs into different biologically active siRNAs, affecting their efficacy and potential for off-targeting. We found that in cells, Dicer induced imprecise cleavage events around the expected sites based on the previously described 5'/3'-counting rules. These promiscuous non-canonical cleavages were abrogated when the cleavage site was positioned 2 nt from a bulge or loop. Interestingly, we observed that the ~1/3 of mammalian endogenous pre-miRNAs that contained such structures were more precisely processed by Dicer. Implementing a new "loop-counting rule", we designed potent anti-HCV shRNAs with substantially reduced off-target effects. Our results suggest that Dicer recognizes the loop/bulge structure in addition to the ends of shRNAs/pre-miRNAs for accurate processing. This has important implications for both miRNA processing and future design of shRNAs for RNAi-based genetic screens and therapies. Various shRNAs were expressed in Cell and processed by the RNase III enzyme Dicer. The profiles of the siRNA products were generated by deep sequencing with or without the Ago2-IP.

2012-12-25 | E-GEOD-41292 | biostudies-arrayexpress

EXPRSS: an Illumina based high-throughput expression-profiling method to reveal transcriptional dynamics - I

Project description:Background Next Generation Sequencing technologies have facilitated differential gene expression analysis through RNA-seq and Tag-seq methods. RNA-seq has biases associated with transcript lengths, lacks uniform coverage of regions in mRNA and requires 10–20 times more reads than a typical Tag-seq. Most existing Tag-seq methods either have biases or not high throughput due to use of restriction enzymes or enzymatic manipulation of 5’ ends of mRNA or use of RNA ligations. Results We have developed EXpression Profiling through Randomly Sheared cDNA tag Sequencing (EXPRSS) that employs acoustic waves to randomly shear cDNA and generate sequence tags at a relatively defined position (~150-200 bp) from the 3′ end of each mRNA. Implementation of the method was verified through comparative analysis of expression data generated from EXPRSS, NlaIII-DGE and Affymetrix microarray and through qPCR quantification of selected genes. EXPRSS is a strand specific and restriction enzyme independent tag sequencing method that does not require cDNA length-based data transformations. EXPRSS is highly reproducible, is high-throughput and it also reveals alternative polyadenylation and polyadenylated antisense transcripts. It is cost-effective using barcoded multiplexing, avoids the biases of existing SAGE and derivative methods and can reveal polyadenylation position from paired-end sequencing. Conclusions EXPRSS Tag-seq provides sensitive and reliable gene expression data and enables high-throughput expression profiling with relatively simple downstream analysis. Five weeks old Arabidopsis (Col-0) leaf discs treated with water or flg22 for 60min; mRNA profiles were generated by deep sequencing on Illumina GAIIx using EXPRSS and NlaIII-DGE protocols, in quadruplicate.

2014-06-20 | E-GEOD-45989 | biostudies-arrayexpress

Transgenerational antiviral RNAi does not occur in Orsay virus infection of C. elegans

Project description:Comparison of small RNAs between virus-infected animals and their progeny Sequencing of small RNAs (15-30nt)

2015-01-01 | E-GEOD-60020 | biostudies-arrayexpress

Small RNA sequencing study of C elegans piRNA biogenesis factors

Project description:Identification of defects in piRNAs and piRNA precursors Sequencing of small RNAs (18-30nt). Sample treatments: Secondary siRNAs in C. elegans carry a 5' triphosphate and this has to be removed prior to cloning, hence the use of 5' polyphosphatase treatment to reveal this population. TAP (tobacco acid pyrophosphatase) is another enzyme that removes 5' triphosphate but it also removes 5' CAP structures- we used this to look for piRNA precursor sequences in C. elegans that have a 5' cap.

2014-04-01 | E-GEOD-49220 | biostudies-arrayexpress

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