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Genetic snapshots of the
Background: In nitrate-poor soils, many leguminous plants form nitrogen-fixing symbioses with members of the bacterial family Rhizobiaceae. We selected Rhizobium sp. NGR234 for its exceptionally broad host range, which includes more than I 12 genera of legumes. Unlike the genome of Bradyrhizobium japonicum, which is composed of a single 8.7 Mb chromosome, that of NGR234 is partitioned into three replicons: a chromosome of about 3.5 Mb, a megaplasmid of more than 2 Mb (pNGR234b) and pNGR234a, a 536,165 bp plasmid that carries most of the genes required for symbioses with legumes. Symbiotic loci represent only a small portion of all the genes coded by rhizobial genomes, however. To rapidly characterize the two largest replicons of NGR234, the genome of strain ANU265 (a derivative strain cured of pNGR234a) was analyzed by shotgun sequencing. Results: Homology searches of public databases with 2,275 random sequences of strain ANU265 resulted in the identification of 1,130 putative protein-coding sequences, of which 922 (41%) could be classified into functional groups. In contrast to the 18% of insertion-like sequences (ISs) found on the symbiotic plasmid pNGR234a, only 2.2% of the shotgun sequences represent known ISs, suggesting that pNGR234a is enriched in such elements. Hybridization data also indicate that the density of known transposable elements is higher in pNGR234b (the megaplasmid) than on the chromosome. Rhizobium-specific intergenic mosaic elements (RIMEs) were found in 35 shotgun sequences, 6 of which carry RIME2 repeats previously thought to be present only in Rhizobium meliloti. As non-overlapping shotgun sequences together represent approximately 10% of ANU265 genome, the chromosome and megaplasmid may carry a total of over 200 RIMEs. Conclusions: 'Skimming' the genome of Rhizobium sp. NGR234 sheds new light on the fine structure and evolution of its replicons, as well as on the integration of symbiotic functions in the genome of a soil bacterium. Although most putative coding sequences could be distributed into functional classes similar to those in Bacillus subtilis, functions related to transposable elements were more abundant in NGR234. In contrast to ISs that accumulated in pNGR234a and pNGR234b, the hundreds of RIME elements seem mostly attributes of the chromosome.
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The molecular genetic linkage map of the model legume
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Background The legume Medicago truncatula has emerged as a model plant for the molecular and genetic dissection of various plant processes involved in rhizobial, mycorrhizal and pathogenic plant-microbe interactions. Aiming to develop essential tools for such genetic approaches, we have established the first genetic map of this species. Two parental homozygous lines were selected from the cultivar Jemalong and from the Algerian natural population (DZA315) on the basis of their molecular and phenotypic polymorphism. Results An F2 segregating population of 124 individuals between these two lines was obtained using an efficient manual crossing technique established for M. truncatula and was used to construct a genetic map. This map spans 1225 cM (average 470 kb/cM) and comprises 289 markers including RAPD, AFLP, known genes and isoenzymes arranged in 8 linkage groups (2n = 16). Markers are uniformly distributed throughout the map and segregation distortion is limited to only 3 linkage groups. By mapping a number of common markers, the eight linkage groups are shown to be homologous to those of diploid alfalfa (M. sativa), implying a good level of macrosynteny between the two genomes. Using this M. truncatula map and the derived F3 populations, we were able to map the Mtsym6 symbiotic gene on linkage group 8 and the SPC gene, responsible for the direction of pod coiling, on linkage group 7. Conclusions These results demonstrate that Medicago truncatula is amenable to diploid genetic analysis and they open the way to map-based cloning of symbiotic or other agronomically-important genes using this model plant.
Data and analysis scripts for: Prolonged soybean absence in the field selects for rhizobia that accumulate more polyhydroxybutyrate during symbiosis
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,During symbiosis, C that rhizobia respire to power N fixation can be stored as polyhydroxybutyrate (PHB), shown to support rhizobia survival under laboratory starvation. We collected soil in 2015 from four replicate plots per treatment in two long-term experiments at Waseca, Minnesota. Treatments differed in the intervals between soybean (Glycine max (L.) Merr.) hosts. We measured PHB accumulation in eight nodules per plant from four soybean (cv. ‘MN0095’) trap plants per soil sample. Trap plants were arranged in a greenhouse, common-garden experiment, and PHB accumulation was measured using flow cytometry. Treatments sampled after long intervals without soybean (greater than two years) showed a greater relative abundance of high-PHB strains. Treatments sampled after the first year of soybean following five years of a non-host crop showed a decreased relative abundance of high-PHB strains, compared to treatments sampled after long intervals without soybean. The latter result is consistent with the hypothesis (not tested directly here) that some high-PHB strains were “sanctioned” by plants as less-beneficial. Our results suggest that rhizobia strains with the tendency to allocate more C to N fixation at the expense of PHB accumulation may be less likely to persist where soybean is grown infrequently or where soil conditions make PHB particularly valuable. However, with typical two-year rotations in Minnesota, differences in PHB storage are unlikely to have a major effect on the relative survival of strains.,See README.md for a detailed description of data files and scripts.,
Sequencing Data from: Environment, plant genetics, and their interaction shape important aspects of sunflower rhizosphere microbial communities [NCBI BioProject]
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,NCBI BioProject,,
Jana Sperschneider - Rhizoctonia AG8-1 and AG8-3 genome data
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Genome assemblies and sequencing data for two Rhizoctonia isolates, AG8-1 and AG8-3.
Data and Code from: Environment, plant genetics, and their interaction shape important aspects of sunflower rhizosphere microbial communities [GitHub repository]
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,- Description: Contains data, processing and analysis code, initial exploratory figures, final publication figures, and final publication tables.,- Link: https://github.com/cliffbueno/Sunflower_AEM,- Note: A release of this repository has been archived on Zenodo with a stable DOI: https://zenodo.org/doi/10.5281/zenodo.12193724,
USDA ARS National Rhizobium Germplasm Collection
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,Our mission is to support application of low-input sustainable agriculture by: Providing, to the best of our ability, technical information about rhizobia, their preservation, and cultural and symbiotic characteristics; Acquiring and preserving the nitrogen-fixing bacterial symbionts of leguminous plants with the goal of maintaining widest possible genetic diversity; Maintaining quality control of new and existing germplasm by evaluation of microbiological purity and by examination of nodulation of the original trap host plant; Distributing cultures to the public and private sectors without charge for these services; Developing or adapting techniques in molecular biology for the determination of genetic diversity of rhizobia, to investigate interactions with their host plants and to identify novel characteristics; Acquiring, maintaining, evaluating quality, and distributing type strains for all the different taxa of nitrogen-fixing legume symbionts; Participating in the UNESCO program.,
Histochemical study of nitrogen-transfer endosymbiosis
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Plant roots have the best-understood mutualisms with microbes, but leaf and bract cell endosymbiosis have not been previously reported. Leaf and bract cells of more than 30 species in 18 families of seed plants were surveyed for the presence of intracellular bacteria and several experiments were designed to find and analyze nutrient exchanges between bacteria and plant cells. This dataset contains the results of 1) histochemical analyses to detect hormones, superoxide, and nitrogenous chemicals around bacteria within plant leaf and bract cells, 2) experiments to assess the differential absorption of isotopic nitrogen into plants, and 3) genetic analysis of bacteria isolated from plant material.
Histochemical study of nitrogen-transfer endosymbiosis
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Plant roots have the best-understood mutualisms with microbes, but leaf and bract cell endosymbiosis have not been previously reported. Leaf and bract cells of more than 30 species in 18 families of seed plants were surveyed for the presence of intracellular bacteria and several experiments were designed to find and analyze nutrient exchanges between bacteria and plant cells. This dataset contains the results of 1) histochemical analyses to detect hormones, superoxide, and nitrogenous chemicals around bacteria within plant leaf and bract cells, 2) experiments to assess the differential absorption of isotopic nitrogen into plants, and 3) genetic analysis of bacteria isolated from plant material.
Data from: Heritable differences in abundance of bacterial rhizosphere taxa are correlated with fungal necrotrophic pathogen resistance
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,This Ag Data Commons submission includes the 94 sunflower paired-end sequencing FASTQ files, the corresponding 16S bacterial FASTQ files, and other relevant data to the study described below:,Host-microbe interactions are increasingly recognized as important drivers of organismal health, growth, longevity, and community-scale ecological processes. However, less is known about how genetic variation affects hosts' associated microbiomes and downstream phenotypes. We demonstrate that sunflower Helianthus annuus harbors substantial, heritable variation in microbial communities under field conditions. We show that microbial communities co-vary with heritable variation in resistance to root infection caused by the necrotrophic pathogen Sclerotinia sclerotiorum, and that plants grown in autoclaved soil showed almost complete elimination of pathogen resistance. Association mapping suggests at least 59 genetic locations with effects on both microbial relative abundance and Sclerotinia resistance. Although the genetic architecture appears quantitative, we have elucidated previously unexplained genetic variation for resistance to this pathogen. We identify new targets for plant breeding and demonstrate the potential for heritable microbial associations to play important roles in defense in natural and human-altered environments.,See README for details of each table in the spreadsheet and related information.,
Amplicon sequencing data for: Heritable genetic differences affect sunflower rhizosphere archaeal, bacterial, and fungal communities across all taxonomic levels
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,Amplicon sequencing data (16S and ITS) was deposited to NCBI Genbank under BioProject ID PRJNA1200133 (https://www.ncbi.nlm.nih.gov/bioproject/PRJNA1200133).,