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ABSTRACT: Transcriptome sequencing of Gloeostereum incarnatum

PROVIDER: PRJNA510218 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR8325745_1.fastq.gz	Fastqsanger.gz
	SRR8325745_2.fastq.gz	Fastqsanger.gz
	SRR8325746_1.fastq.gz	Fastqsanger.gz
	SRR8325746_2.fastq.gz	Fastqsanger.gz

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Project description:The Fusarium incarnatum strain K23, originally isolated from a habit-adapted wild plant Thapsia species, colonized the roots and shoots of tomato seedlings and protected them against salt stress. Comparison of expression and metabolite profile changes uncovered that the fungus completely reprogramed the tomato response to salt stress. Barely any overlap was observed among the genes and metabolites which are regulated by salt stress in uncolonized and colonized tomato seedlings. In colonized seedlings exposed to salt stress, less stress- related genes are activated than in un-colonized seedlings. Furthermore, K23 produced gibberellin and gibberellin-responsive genes were detected in all RNA samples. Our analysis demonstrates that K23 colonisation completely alters the salt-responsive gene and metabolite profiles in tomato seedlings.

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Project description:Objectives: To perform long-read transcriptome and proteome profiling of pathogen-stimulated peripheral blood mononuclear cells (PBMCs) from healthy donors. We aim to discover new transcripts and protein isoforms expressed during immune responses to diverse pathogens. Methods: PBMCs were exposed to four microbial stimuli for 24 hours: the TLR4 ligand lipopolysaccharide (LPS), the TLR3 ligand Poly(I:C), heat-inactivated Staphylococcus aureus, Candida albicans, and RPMI medium as negative controls. Long-read sequencing (PacBio) of one donor and secretome proteomics and short-read sequencing of five donors were performed. IsoQuant was used for transcriptome construction, Metamorpheus/FlashLFQ for proteome analysis, and Illumina short-read 3’-end mRNA sequencing for transcript quantification. Results: Long-read transcriptome profiling reveals the expression of novel sequences and isoform switching induced upon pathogen stimulation, including transcripts that are difficult to detect using traditional short-read sequencing. We observe widespread loss of intron retention as a common result of all pathogen stimulations. We highlight novel transcripts of NFKB1 and CASP1 that may indicate novel immunological mechanisms. In general, RNA expression differences did not result in differences in the amounts of secreted proteins. Interindividual differences in the proteome were larger than the differences between stimulated and unstimulated PBMCs. Clustering analysis of secreted proteins revealed a correlation between chemokine (receptor) expression on the RNA and protein levels in C. albicans- and Poly(I:C)-stimulated PBMCs. Conclusion: Isoform aware long-read sequencing of pathogen-stimulated immune cells highlights the potential of these methods to identify novel transcripts, revealing a more complex transcriptome landscape than previously appreciated.

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