Interactions between ectomycorrhizal associations and bacteria

Boreal forest podzol soils have vertically stratified horizons with different physico-chemical characteristics and high microbial diversity. Ectomycorrhizal fungi play key roles in accessing nutrients from both organic and mineral substrates. The role of associated bacteria in these processes is still poorly understood. The aim of the studies described in this thesis was to improve understanding of the distribution, diversity and community structure of fungi and bacteria on roots and in soil and their responses to environmental perturbations such as N-fertilisation. In two microcosm studies a single-root tip microbiome method was used to sample bacteria associated with different ectomycorrhizal roots at defined time-points, using high throughput sequencing of both fungi and bacteria The first study revealed highly dynamic patterns of assembly of bacterial communities associated with ectomycorrhizal roots at different time points in organic soil. Bacterial community structure differed between roots colonised by different species of ectomycorrhizal fungi from different genera. The second study extended these results to include both organic and mineral horizons, demonstrating significant differences between fungal and bacterial communities colonising soil from different horizons. Responses of both fungi and bacteria to short-term N additions were context dependent, influenced by both soil horizon and the dominant ectomycorrhizal fungi colonising the roots. Bacterial communities associated with roots colonised by pairs of closely related fungal species within the same fungal genera were also shown to be statistically distinct. Field studies of fungi and bacteria in a forest fertilised with 150 kg N ha⁻¹ 15 months previously, revealed 1017 unique fungal OTUs, (877 in the soil, 652 in the roots). N increased fungal diversity slightly in the O horizon soil but decreased it in the roots, particularly in the B horizon. Fungal community structure varied significantly between horizons and within each soil horizon the community structure of fungi colonising ectomycorrhizal roots was significantly different from that associated with the soil, suggesting that analyses of both soil and roots are necessary for accurate monitoring of environmental perturbations. 10925 unique bacterial OTUs were distinguished in total (8560 in the soil, 5512 in the roots). Bacteria displayed similar trends to the fungi but were less strongly influenced by N. These studies pave the way for more detailed functional studies of specific combinations of fungi and bacteria

Changes in the root fungal microbiome of strawberry following application of residues of the biofumigant oilseed radish

Biofumigation has been proposed as an environmentally friendly method of plant protection against soil-borne pathogens, but its effects on microbial communities are still incompletely understood. Using high throughput DNA sequencing, we investigated the effects of oilseed radish residues on the root fungal microbiome of strawberry in the presence of a soil-borne fungal pathogen, Verticillium dahliae. Results of our greenhouse study show that early flowering occurred in response to residue addition, suggesting a plant stress-response and there was a significant decrease in berry yield. The fungal microbiome of roots was significantly restructured by both biofumigation and inoculation with Verticillium. In particular, the abundance of root endophyte- and arbuscular mycorrhizal functional guilds was reduced significantly as a result of biofumigant and V. dahliae addition, whereas the abundance of saprotrophs increased significantly when both treatments were applied together. Alpha diversity analyses of fungi associated with roots indicated a significant increase in species richness following Verticillium inoculation, whereas the biofumigant alone or in the presence of V. dahliae resulted in no significant effect, suggesting that apparently some rare taxa may have been enriched/stimulated in the presence of the pathogen. Further investigations should reveal whether negative effects of biofumigation on potentially beneficial root associated endophytes and arbuscular mycorrhizal fungi are host genotype- or soil-dependent

Global similarity, and some key differences, in the metagenomes of Swedish varroa-surviving and varroa-susceptible honeybees

There is increasing evidence that honeybees (Apis mellifera L.) can adapt naturally to survive Varroa destructor, the primary cause of colony mortality world-wide. Most of the adaptive traits of naturally varroa-surviving honeybees concern varroa reproduction. Here we investigate whether factors in the honeybee metagenome also contribute to this survival. The quantitative and qualitative composition of the bacterial and viral metagenome fluctuated greatly during the active season, but with little overall difference between varroa-surviving and varroa-susceptible colonies. The main exceptions were Bartonella apis and sacbrood virus, particularly during early spring and autumn. Bombella apis was also strongly associated with early and late season, though equally for all colonies. All three affect colony protein management and metabolism. Lake Sinai virus was more abundant in varroa-surviving colonies during the summer. Lake Sinai virus and deformed wing virus also showed a tendency towards seasonal genetic change, but without any distinction between varroa-surviving and varroa-susceptible colonies. Whether the changes in these taxa contribute to survival or reflect demographic differences between the colonies (or both) remains unclear

Global similarity, and some key differences, in the metagenomes of Swedish varroa-surviving and varroa-susceptible honeybees

There is increasing evidence that honeybees (Apis mellifera L.) can adapt naturally to survive Varroa destructor, the primary cause of colony mortality world-wide. Most of the adaptive traits of naturally varroa-surviving honeybees concern varroa reproduction. Here we investigate whether factors in the honeybee metagenome also contribute to this survival. The quantitative and qualitative composition of the bacterial and viral metagenome fluctuated greatly during the active season, but with little overall difference between varroa-surviving and varroa-susceptible colonies. The main exceptions were Bartonella apis and sacbrood virus, particularly during early spring and autumn. Bombella apis was also strongly associated with early and late season, though equally for all colonies. All three affect colony protein management and metabolism. Lake Sinai virus was more abundant in varroa-surviving colonies during the summer. Lake Sinai virus and deformed wing virus also showed a tendency towards seasonal genetic change, but without any distinction between varroa-surviving and varroa-susceptible colonies. Whether the changes in these taxa contribute to survival or reflect demographic differences between the colonies (or both) remains unclear

Caf1A usher possesses a Caf1 subunit-like domain that is crucial for Caf1 fibre secretion

The chaperone/usher pathway controls assembly of fibres of adhesive organelles of Gram-negative bacteria. The final steps of fibre assembly and fibre translocation to the cell surface are co-ordinated by the outer membrane proteins, ushers. Ushers consist of several soluble periplasmic domains and a single transmembrane beta-barrel. Here we report isolation and structural/functional characterization of a novel middle domain of the Caf1A usher from Yersinia pestis. The isolated UMD (usher middle domain) is a highly soluble monomeric protein capable of autonomous folding. A 2.8 angstrom (1 angstrom = 0.1 nm) resolution crystal structure of UMD revealed that this domain has an immunoglobulin-like fold similar to that of donor-strand-complemented Caf1 fibre subunit. Moreover, these proteins displayed significant structural similarity. Although UMD is in the middle of the predicted amphipathic beta-barrel of Caf1A, the usher still assembled in the membrane in the absence of this domain. UMD did not bind Caf1M-Caf1 complexes, but its presence was shown to be essential for Caf1 fibre secretion. The study suggests that UMD may play the role of a subunit-substituting protein (dummy subunit), plugging or priming secretion through the channel in the Caf1A usher. Comparison of isolated UMD with the recent strcture of the corresponding domain of PapC usher revealed high similarity of the core structures, suggesting a universal structural adaptation of FGL (F(1)G(1) long) and FGS (F(1)G(1) short) chaperone/usher pathways for the secretion of different types of fibres. The functional role of two topologically different states of this plug domain suggested by structural and biochemical results is discussed