Characterisation of methylotrophs in the rhizosphere

Macey, Michael (2017) Characterisation of methylotrophs in the rhizosphere. Doctoral thesis, University of East Anglia.

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Methanol is the second most abundant volatile organic compound in the atmosphere, with the majority of this methanol being produced as a waste metabolic by-product of the growth and decay of plants. There is a large disparity between the amount of methanol estimated as being produced and that which enters the atmosphere. This disparity is believed to be due to the utilisation of methanol by plant associated methylotrophs. The diversity and activity of methylotrophs associated with the root and rhizosphere of pea and wheat plants was assessed through a range of cultivation independent and dependent approaches.
Enrichments performed with a range of environmental samples supplemented with methanol resulted in the isolation of several strains of methylotrophic bacteria, including two novel species of methylotroph belonging to the family Methylophilaceae, whose genomes were sequenced and their physiological capabilities assessed.
The diversity and abundance of methanol dehydrogenase encoding genes in bulk soil and the pea and wheat rhizosphere was assessed through 454 sequencing and qPCR respectively. Sequencing showed high levels of diversity of methylotrophic bacteria within the bulk soil and also showed a shift in this diversity between the bulk soil and the plant associated soils, in spite of no shift in the abundance of these genes occurring.
Active methylotrophs present in the bulk and plant associated soils were identified by DNA stable isotope probing using 13C labelled methanol. Next generation sequencing of the 16S rRNA genes and construction of metagenomes from the 13C labelled DNA revealed members of the Methylophilaceae as highly abundant in all of the soils. A greater diversity of the Methylophilaceae and the genus Methylobacterium were identified as active in the plant associated soils relative to the bulk soil.
A 13CO2 stable isotope probing experiment identified methylotrophs as utilising plant exudates in the pea and wheat root and rhizosphere communities. Several methylotrophic genera were identified as exudate utilising, in addition to heterotrophic genera and Actinomycetes. The specific 13C labelled genera were shown to vary between both the wheat and pea plants and between the rhizosphere and root communities.

Item Type: Thesis (Doctoral)
Faculty \ School: Faculty of Science > School of Environmental Sciences
Depositing User: Jackie Webb
Date Deposited: 30 Apr 2018 10:54
Last Modified: 30 Apr 2018 10:54

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