Processing of NODULE INCEPTION controls transition to nitrogen fixation in root nodules
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA737413 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR14804665_1.fastq.gz | Fastqsanger.gz | |||
| SRR14804665_2.fastq.gz | Fastqsanger.gz | |||
| SRR14804666_1.fastq.gz | Fastqsanger.gz | |||
| SRR14804666_2.fastq.gz | Fastqsanger.gz | |||
| SRR14804667_1.fastq.gz | Fastqsanger.gz |
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Project description:Legumes establish endosymbiotic associations with nitrogen-fixing rhizobia, which they host inside root nodules. Here, specific physiological and morphological adaptations, such as the production of oxygen-binding leghemoglobin proteins and the formation of an oxygen diffusion barrier in the nodule periphery, are essential to protect the oxygen-labile bacterial nitrogenase enzyme. The molecular basis of the latter process remains elusive, as the identification of required genes is limited by the epistatic effect of nodule organogenesis over nodule infection and rhizobia accommodation. We overcame this by exploring the phenotypic diversity of Lotus japonicus accessions that uncouple nodule organogenesis from nodule infection when inoculated with a sub-compatible Rhizobium strain. Using comparative transcriptomics, we identified genes with functions associated with oxygen homeostasis and deposition of lipid polyesters on cell walls to be specifically upregulated in infected compared to uninfected nodules. As such hydrophobic modifications on cell walls are pivotal for creating diffusion barriers like the root endodermis, we focused on two Fatty acyl-CoA reductase genes that were specifically activated in the nodule or in the root endodermis. Mutant lines in a Fatty acyl-CoA reductase gene expressed exclusively in the nodule endodermis showed had decreased suberization of this cell layer and increased nodule permeability compared to wild type plants. Oxygen concentrations were significantly increased in the inner cortex of mutant nodules, which correlated with reduced nitrogen fixation rates, and impaired shoot growth. These results provide the first genetic evidence for the formation of the nodule oxygen diffusion barrier, a key adaptation enabling nitrogen fixation in legume nodules.
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Project description:We used laser-capture microdissection (LCM) to isolate specific cells from the Medicago truncatula nodule meristem (M), the distal infection (DIZ), the proximal infection zone (PIZ), infected cells (IC) and uninfected cells (UIC) from the fixation zone. Based on Medicago GeneChips, we identified the cell- and tissue-specific programm of gene expression in Medicago truncatula root nodules. Nodules were harvested three weeks after inoculation of Medicago truncatula (genotype Jemalong A17) plants with Sinorhizobium meliloti strain 2011. Nodules were fixed in Farmer's fixative and subsequently embedded in paraffin. 8 M-BM-5m de-paraffined sections were used to capture cells from the nodule meristem, distal infection zone, proximal infection zone, infected cells and uninfected cells from the fixation zone, using an Arcturus Pixcell II laser capture microscope. 3 biological replicates were used for each cell-/tissue type. After RNA extraction, the RNA was amplified and used for Medicago Gene Chip hybridization.
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Project description:Legume plants can establish symbiotic nitrogen fixation (SNF) with rhizobia mostly in root nodules, where rhizobia-infected cells are accompanied with uninfected cells in a mosaic pattern. Inside the mature nodules of legume, carbon and nitrogen nutrients between host plant cells and their resident bacteria are actively exchanged. To elucidate the metabolite dynamics relevant for SNF in nodules, three cell-types from nodule tissues of a model legume, Lotus japonicus, were isolated using laser microdissesction, and transcriptome analysis was done by an oligoarray with 60-mer length representing 21,495 genes. In our cell-type-specific profiling, many genes were identified as being expressed in nodules with spatial-specific manners. Among them, genes coding for metabolic enzymes were classified according to their function, and detailed data analysis figured out that secondary metabolic pathway was highly activated in nodule cortex. In particular, a number of metabolic genes for phenyl propanoid pathway were found as highly expressed genes accompanied with those encoding putative transporters of secondary metabolites. These data suggest the involvement of novel physiological function of phenylpropanoids in SNF. Gene expression in three different cell-types of Lotus japonicus nodule was measured. Three independent experiments were performed at each cell-types.
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