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Genome-resolved metatranscriptomics reveals conserved root colonization determinants in a synthetic microbiota

ABSTRACT: Genome-resolved metatranscriptomics reveals conserved root colonization determinants in a synthetic microbiota

PROVIDER: PRJNA968016 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24458160.fastq.gz	Fastqsanger.gz
	SRR24458161.fastq.gz	Fastqsanger.gz
	SRR24458162.fastq.gz	Fastqsanger.gz
	SRR24458163.fastq.gz	Fastqsanger.gz
	SRR24458164.fastq.gz	Fastqsanger.gz

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Genome-resolved metatranscriptomics reveals conserved root colonization determinants in a synthetic microbiota

Project description:The identification of processes activated by specific microbes during microbiota colonization of plant roots has been hampered by technical constraints in metatranscriptomics. These include lack of reference genomes, high representation of host or microbial rRNA sequences in datasets, or difficulty to experimentally validate gene functions. Here, we recolonized germ-free Arabidopsis thaliana with a synthetic, yet representative root microbiota comprising 106 genome-sequenced bacterial and fungal isolates. We used multi-kingdom rRNA depletion, deep RNA-sequencing and read mapping against reference microbial genomes to analyse the in-planta metatranscriptome of abundant colonizers. We identified over 3,000 microbial genes that were differentially regulated at the soil-root interface. Translation and energy production processes were consistently activated in planta, and their induction correlated with bacterial strains’ abundance in roots. Finally, we used targeted mutagenesis to show that several genes consistently induced by multiple bacteria are required for root colonization in one of the abundant bacterial strains (a genetically tractable Rhodanobacter). Our results indicate that microbiota members activate strain-specific processes but also common gene sets to colonize plant roots.

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Genome-resolved metatranscriptomics reveals conserved root colonization determinants in a synthetic microbiota.

Project description:The identification of processes activated by specific microbes during microbiota colonization of plant roots has been hampered by technical constraints in metatranscriptomics. These include lack of reference genomes, high representation of host or microbial rRNA sequences in datasets, or difficulty to experimentally validate gene functions. Here, we recolonized germ-free Arabidopsis thaliana with a synthetic, yet representative root microbiota comprising 106 genome-sequenced bacterial and fungal isolates. We used multi-kingdom rRNA depletion, deep RNA-sequencing and read mapping against reference microbial genomes to analyse the in planta metatranscriptome of abundant colonizers. We identified over 3,000 microbial genes that were differentially regulated at the soil-root interface. Translation and energy production processes were consistently activated in planta, and their induction correlated with bacterial strains' abundance in roots. Finally, we used targeted mutagenesis to show that several genes consistently induced by multiple bacteria are required for root colonization in one of the abundant bacterial strains (a genetically tractable Rhodanobacter). Our results indicate that microbiota members activate strain-specific processes but also common gene sets to colonize plant roots.

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Transcriptional responses to arbuscular mycorrhizal symbiosis development are conserved in the liverwort Marchantia paleacea

Project description:The mutualistic arbuscular mycorrhizal (AM) symbiosis arose in land plants more than 450 million years ago. This symbiosis is still widely found across major land plant lineages, including bryophytes. Despite its broad taxonomic distribution, little is known about the molecular components underpinning symbiosis outside of flowering plants. Here, we demonstrate that a broad AM genetic programme is conserved amongst land plants. In this study, we characterised the dynamic response of the liverwort Marchantia paleacea to Rhizophagus irregularis colonization by time-resolved transcriptomics across three stages of symbiosis. Comparative analysis of transcriptional responses to symbiosis in the liverwort M. paleacea and the legume Medicago truncatula further revealed evolutionarily conserved expression patterns for genes underpinning pre-symbiotic signalling, intracellular colonization and nutrient exchange. This study demonstrates that the genetic machinery regulating key aspects of symbiosis in plant hosts is largely conserved and coregulated across land plants.

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