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1.
Sci Rep ; 9(1): 1776, 2019 02 11.
Article in English | MEDLINE | ID: mdl-30742038

ABSTRACT

Botryococcus braunii is a colonial microalga that appears early in the fossil record and is a sensitive proxy of environmental and hydroclimatic conditions. Palaeozoic Botryococcus fossils which contribute up to 90% of oil shales and approximately 1% of crude oil, co-localise with diagnostic geolipids from the degradation of source-signature hydrocarbons. However more recent Holocene sediments demonstrate no such association. Consequently, Botryococcus are identified in younger sediments by morphology alone, where potential misclassifications could lead to inaccurate paleoenvironmental reconstructions. Here we show that a combination of flow cytometry and ancient DNA (aDNA) sequencing can unambiguously identify Botryococcus microfossils in Holocene sediments with hitherto unparalleled accuracy and rapidity. The application of aDNA sequencing to microfossils offers a far-reaching opportunity for understanding environmental change in the recent geological record. When allied with other high-resolution palaeoenvironmental information such as aDNA sequencing of humans and megafauna, aDNA from microfossils may allow a deeper and more precise understanding of past environments, ecologies and migrations.


Subject(s)
Fossils , Genome, Plant , Hydrocarbons/metabolism , Microalgae/genetics , Microalgae/metabolism , DNA, Plant/genetics
2.
Sci Data ; 4: 170190, 2017 12 19.
Article in English | MEDLINE | ID: mdl-29257137

ABSTRACT

European common ash, Fraxinus excelsior, is currently threatened by Ash dieback (ADB) caused by the fungus, Hymenoscyphus fraxineus. To detect and identify metabolites that may be products of pathways important in contributing to resistance against H. fraxineus, we performed untargeted metabolomic profiling on leaves from five high-susceptibility and five low-susceptibility F. excelsior individuals identified during Danish field trials. We describe in this study, two datasets. The first is untargeted LC-MS metabolomics raw data from ash leaves with high-susceptibility and low-susceptibility to ADB in positive and negative mode. These data allow the application of peak picking, alignment, gap-filling and retention-time correlation analyses to be performed in alternative ways. The second, a processed dataset containing abundances of aligned features across all samples enables further mining of the data. Here we illustrate the utility of this dataset which has previously been used to identify putative iridoid glycosides, well known anti-herbivory terpenoid derivatives, and show differential abundance in tolerant and susceptible ash samples.


Subject(s)
Fraxinus , Metabolome , Plant Diseases , Plant Leaves/metabolism
3.
J Vis Exp ; (119)2017 01 13.
Article in English | MEDLINE | ID: mdl-28117801

ABSTRACT

Metagenomics is defined as the direct analysis of deoxyribonucleic acid (DNA) purified from environmental samples and enables taxonomic identification of the microbial communities present within them. Two main metagenomic approaches exist; sequencing the 16S rRNA gene coding region, which exhibits sufficient variation between taxa for identification, and shotgun sequencing, in which genomes of the organisms that are present in the sample are analyzed and ascribed to "operational taxonomic units"; species, genera or families depending on the extent of sequencing coverage. In this study, shotgun sequencing was used to analyze the microbial community present in cattle silage and, coupled with a range of bioinformatics tools to quality check and filter the DNA sequence reads, perform taxonomic classification of the microbial populations present within the sampled silage, and achieve functional annotation of the sequences. These methods were employed to identify potentially harmful bacteria that existed within the silage, an indication of silage spoilage. If spoiled silage is not remediated, then upon ingestion it could be potentially fatal to the livestock.


Subject(s)
Metagenomics/methods , Silage/microbiology , Animals , Bacteria/genetics , Cattle , Computational Biology , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA/methods
4.
Nature ; 541(7636): 212-216, 2017 01 12.
Article in English | MEDLINE | ID: mdl-28024298

ABSTRACT

Ash trees (genus Fraxinus, family Oleaceae) are widespread throughout the Northern Hemisphere, but are being devastated in Europe by the fungus Hymenoscyphus fraxineus, causing ash dieback, and in North America by the herbivorous beetle Agrilus planipennis. Here we sequence the genome of a low-heterozygosity Fraxinus excelsior tree from Gloucestershire, UK, annotating 38,852 protein-coding genes of which 25% appear ash specific when compared with the genomes of ten other plant species. Analyses of paralogous genes suggest a whole-genome duplication shared with olive (Olea europaea, Oleaceae). We also re-sequence 37 F. excelsior trees from Europe, finding evidence for apparent long-term decline in effective population size. Using our reference sequence, we re-analyse association transcriptomic data, yielding improved markers for reduced susceptibility to ash dieback. Surveys of these markers in British populations suggest that reduced susceptibility to ash dieback may be more widespread in Great Britain than in Denmark. We also present evidence that susceptibility of trees to H. fraxineus is associated with their iridoid glycoside levels. This rapid, integrated, multidisciplinary research response to an emerging health threat in a non-model organism opens the way for mitigation of the epidemic.


Subject(s)
Fraxinus/genetics , Genetic Predisposition to Disease/genetics , Genetic Variation , Genome, Plant/genetics , Plant Diseases/genetics , Trees/genetics , Ascomycota/pathogenicity , Conserved Sequence/genetics , Denmark , Fraxinus/microbiology , Genes, Plant/genetics , Genomics , Iridoid Glycosides/metabolism , Plant Diseases/microbiology , Plant Diseases/prevention & control , Plant Proteins/genetics , Population Density , Sequence Analysis, DNA , Species Specificity , Transcriptome , Trees/microbiology , United Kingdom
5.
BMC Microbiol ; 14: 262, 2014 Oct 28.
Article in English | MEDLINE | ID: mdl-25348335

ABSTRACT

BACKGROUND: Campylobacter jejuni and C. coli are human intestinal pathogens that are the most frequent causes of bacterial foodborne gastroenteritis in humans in the UK. In this study, we aimed to characterise the metabolic diversity of both C. jejuni and C. coli using a diverse panel of clinical strains isolated from the UK, Pakistan and Thailand, thereby representing both the developed and developing world. Our aim was to apply multi genome analysis and Biolog phenotyping to determine differences in carbon source utilisation by C. jejuni and C. coli strains. RESULTS: We have identified a core set of carbon sources (utilised by all strains tested) and a set that are differentially utilised for a diverse panel of thirteen C. jejuni and two C. coli strains. This study used multi genome analysis to show that propionic acid is utilised only by C. coli strains tested. A broader PCR screen of 16 C. coli strains and 42 C. jejuni confirmed the absence of the genes needed for propanoate metabolism. CONCLUSIONS: From our analysis we have identified a phenotypic method and two genotypic methods based on propionic utilisation that might be applicable for distinguishing between C. jejuni and C. coli.


Subject(s)
Campylobacter coli/classification , Campylobacter coli/metabolism , Campylobacter jejuni/classification , Campylobacter jejuni/metabolism , Carbon/metabolism , Propionates/metabolism , Bacterial Typing Techniques/methods , Campylobacter Infections/microbiology , Campylobacter coli/isolation & purification , Campylobacter jejuni/isolation & purification , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Humans , Metabolic Networks and Pathways/genetics , Molecular Diagnostic Techniques/methods , Molecular Sequence Data , Pakistan , Polymerase Chain Reaction , Sequence Analysis, DNA , Thailand , United Kingdom
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