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ABSTRACT: Transcriptome profiling of Microdochium nivale grown in vitro and in planta

PROVIDER: PRJNA785089 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17081788.fastq.gz	Fastqsanger.gz
	SRR17081789.fastq.gz	Fastqsanger.gz
	SRR17081790.fastq.gz	Fastqsanger.gz
	SRR17081791.fastq.gz	Fastqsanger.gz
	SRR17081792.fastq.gz	Fastqsanger.gz

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Microdochium nivale strain:F00608

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Global transcriptome changes in perennial ryegrass during early infection by pink snow mould

Project description:Lack of resistance to pink snow mould (Microdochium nivale) is seen as a major constraint for adaptation of perennial ryegrass (Lolium perenne L.) at higher latitudes. Plants generally become more resistant to snow moulds after cold acclimation, and almost all investigations of genetic variation in resistance have been performed using cold acclimated plants. However, there may be variation in resistance mechanisms that are functioning independently of cold acclimation. In this study our aim was to identify candidate genes involved in such resistance mechanisms. We first characterized variation in resistance to M. nivale among non-acclimated plants of eight genotypes from the Norwegian cultivar Fagerlin and selected one resistant and one susceptible genotype for transcriptome analysis. Total RNA was extracted from leaf blade tissue of plants exposed to three different treatments: non-inoculated and non-incubated plants, non-inoculated plants after four days of incubation, and inoculated plants after four days of incubation. cDNA libraries were prepared and paired-end sequencing performed using Illumina Hiseq 2000. Transcriptome profiles, GO enrichment and KEGG pathway analysis indicate that defence response related genes are differentially expressed between incubated non-inoculated and incubated inoculated conditions both within resistant and susceptible genotypes. A significant up-regulation of defence related genes as well as genes involved in cell wall cellulose metabolic processes and aryl-alcohol dehydrogenase (NADP+) activity was observed in the resistant genotype. The candidate genes identified in this study might be potential molecular marker resources for breeding perennial ryegrass cultivars with improved resistance to pink snow mould.

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Project description:Elucidation of the involvement of MOR (morphogenesis-related NDR kinase network) pathway in the gene expression during appressorium development in planta, transcriptome analyses at the stage of appressorium formation (4 hours post inoculation) in vitro and in planta were performed and revealed that MOR is responsible for the expression of a subset of the plant signal induced genes with potential roles in pathogenicity.

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Project description:Pantoea stewartii subsp. stewartii is a bacterial phytopathogen that causes Stewart’s wilt disease in corn. It uses quorum sensing to regulate expression of some genes involved in virulence in a cell density-dependent manner as the bacterial population grows from small numbers at the initial infection site in the leaf apoplast to high cell numbers in the xylem where it forms a biofilm. There are also other genes important for pathogenesis not under quorum-sensing control such as a Type III secretion system. The purpose of this study was to compare gene expression during a high-density in planta infection versus a low-density pre-inoculum liquid culture and in a high-density culture grown on agar medium to identify genes specifically expressed in planta that may also be important for colonization and/or virulence. RNA was purified from each sample type to determine the transcriptome via RNA-Seq using Illumina sequencing of cDNA. Fold gene expression changes in the high-density in planta data set in comparison to the two in vitro grown samples were determined and a list of the most differentially expressed genes was generated to elucidate genes important for plant association. Quantitative reverse transcription PCR (qRT-PCR) was used to validate expression patterns for a select subset of genes. Analysis of the transcriptome data via gene ontology revealed that bacterial transporters and systems active under low oxygen tensions appear to play a critical role for P. stewartii as it colonizes and causes wilt disease in corn plants.

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