Encroachment of shrubs into subalpine grasslands in the Pyrenees modifies the structure of soil fungal communities and soil properties.

The encroachment of shrubs into grasslands is common in terrestrial ecosystems dominated by grass. Land abandonment and favourable climatic trends in recent decades have favoured the expansion of shrubs into subalpine grasslands in many mountainous regions across Europe. The advance of the succession from grassland to shrubland is expected to have a major impact on ecosystem functioning. We used DNA metabarcoding to assess whether the structure of soil fungal communities varied along the succession from subalpine grassland to shrubland in the Pyrenees, and investigated whether shrub encroachment was associated with changes in soil properties. The expansion of shrubs increased the soil C:N ratio and/or reduced the N, P or K contents. Plant-driven changes in soil properties were strongly associated with the compositional turnover of fungi, including arbuscular mycorrhizal, ectomycorrhizal, ericoid, root endophytic, saprotrophic, lichenised and pathogenic fungi. Total richness and the richness of most functional groups were correlated with soil P, N and the C:N or N:P ratios. We show that the interplay between abiotic factors (changes in soil properties) and biotic factors (occurrence and identity of shrubs) played a key role in the structure and uniqueness of soil fungal communities along the succession.

Association of prevalent vaginal microbiome of mother with occurrence of type I diabetes in child.

Type I diabetes (T1D) is a rapidly increasing autoimmune disease especially in the Western countries and poses a serious global health problem. Incidence of T1D cannot be fully explained by genetic background, and environmental factors have been assumed to play a role. Environmental conditions and composition of human microbiome have been found to correlate with the incidence of T1D. We asked whether mothers' prevalent vaginal microbiome could correlate with the incidence of T1D in child. To test this hypothesis, we collected samples of vaginal microbiomes from eight mothers that had at least one child with T1D (child age maximum of 11 years at the time of sampling), born with a vaginal delivery. Eight control mothers had child/children with vaginal delivery and no diabetic child/children. The microbiomes were studied by using 16S rRNA Ion Torrent high throughput sequencing. We found that composition of total and Lactobacillus microbiome was altered, and saw an indication that diversity of vaginal microbiomes of the mothers with a diabetic child could be higher. Based on these pilot observations, we strongly encourage a larger population study to verify whether mother vaginal microbiome diversity and composition are linked to the prevalence of T1D in children.

Microbial diversity along a gradient in peatlands treating mining-affected waters.

Peatlands are used for the purification of mining-affected waters in Northern Finland. In Northern climate, microorganisms in treatment peatlands (TPs) are affected by long and cold winters, but studies about those microorganisms are scarce. Thus, the bacterial, archaeal and fungal communities along gradients of mine water influence in two TPs were investigated. The TPs receive waters rich in contaminants, including arsenic (As), sulfate (SO42-) and nitrate (NO3-). Microbial diversity was high in both TPs, and microbial community composition differed between the studied TPs. Bacterial communities were dominated by Proteobacteria, Actinobacteria, Chloroflexi and Acidobacteria, archaeal communities were dominated by Methanomicrobia and the Candidate phylum Bathyarchaeota, and fungal communities were dominated by Ascomycota (Leotiomycetes, Dothideomycetes, Sordariomycetes). The functional potential of the bacterial and archaeal communities in TPs was predicted using PICRUSt. Sampling points affected by high concentrations of As showed higher relative abundance of predicted functions related to As resistance. Functions potentially involved in nitrogen and SO42- turnover in TPs were predicted for both TPs. The results obtained in this study indicate that (i) diverse microbial communities exist in Northern TPs, (ii) the functional potential of the peatland microorganisms is beneficial for contaminant removal in TPs and (iii) microorganisms in TPs are likely well-adapted to high contaminant concentrations as well as to the Northern climate.

Bacterial and fungal communities in boreal forest soil are insensitive to changes in snow cover conditions.

The northern regions are experiencing considerable changes in winter climate leading to more frequent warm periods, rain-on-snow events and reduced snow pack diminishing the insulation properties of snow cover and increasing soil frost and freeze-thaw cycles. In this study, we investigated how the lack of snow cover, formation of ice encasement and snow compaction affect the size, structure and activities of soil bacterial and fungal communities. Contrary to our hypotheses, snow manipulation treatments over one winter had limited influence on microbial community structure, bacterial or fungal copy numbers or enzyme activities. However, microbial community structure and activities shifted seasonally among soils sampled before snow melt, in early and late growing season and seemed driven by substrate availability. Bacterial and fungal communities were dominated by stress-resistant taxa such as the orders Acidobacteriales, Chaetothyriales and Helotiales that are likely adapted to adverse winter conditions. This study indicated that microbial communities in acidic northern boreal forest soil may be insensitive to direct effects of changing snow cover. However, in long term, the detrimental effects of increased ice and frost to plant roots may alter plant derived carbon and nutrient pools to the soil likely leading to stronger microbial responses.

Different endophyte communities colonize buds of sprouts compared with mature trees of mountain birch recovered from moth herbivory.

Plant meristems were previously thought to be sterile. Today, meristem-associated shoot endophytes are mainly reported as contaminants from plant tissue cultures, the number of observed species being very low. However, the few strains characterized have the capacity for infecting host cells and affecting plant growth and development. Here we studied the communities of endophytic bacteria in the buds of mountain birch (Betula pubescens ssp. czerepanovii (N. I. Orlova) Hämet-Ahti) exposed to winter moth (Operophtera brumata L.) herbivory, to identify differences between sprouts and branches of mature birch trees. Mountain birch of the high subarctic is cyclically exposed to winter moth and produces sprouts to generate new trees as a survival mechanism. The majority (54%) of operational taxonomic units belonged to Xanthomonadaceae and Pseudomonales of Proteobacteria. Most of the observed species were classified as Xanthomonas (28%). Sprout buds had the highest diversity, containing approximately three times more species, and significantly more (43%) Pseudomonas species than the mature trees (14%). Our results demonstrate that endophytic communities of buds are richer than previously thought. We suggest that the meristem-associated endophytes should be studied further for a possible role in sprouting and aiding regeneration of trees.

Competitive success of southern populations of Betula pendula and Sorbus aucuparia under simulated southern climate experiment in the subarctic.

Global warming has been commonly accepted to facilitate species' range shifts across latitudes. Cross-latitudinal transplantations support this; many tree species can well adapt to new geographical areas. However, these studies fail to capture species' adaptations to new light environment because the experiments were not designed to explicitly separate species' responses to light and temperature. Here we tested reaction norms of tree seedlings in reciprocal transplantations 1,000 km apart from each other at two latitudes (60°N and 69°N). In contrast to past studies, we exposed our experimental plants to same temperature in both sites (temperature of 60°N growing site is recorded to adjust temperature of 69°N site in real time via Internet connection) while light environment (photoperiod, light quality) remained ambient. Shoot elongation and autumn coloration were studied in seedlings of two deciduous trees (Betula pendula and Sorbus aucuparia), which were expected to respond differently to day length. Sorbus as a member of Rosaceae family was assumed to be indifferent to photoperiod, while Betula responds strongly to day length. We hypothesized that (1) southern and northern populations of both species perform differently; (2) southern populations perform better in both sites; (3) autumn phenology of southern populations may delay in the northern site; (4) and Sorbus aucuparia is less dependent on light environment. According to the hypotheses, shoot elongation of northern population was inherently low in both species. An evolutionary consequence of this may be a competitive success of southern populations under warming climate. Southern population of B. pendula was delayed in autumn coloration, but not in growth cessation. Sorbus aucuparia was less responsive to light environment. The results suggest that light provides selection pressure in range shifts, but the response is species dependent.

Moth outbreaks alter root-associated fungal communities in subarctic mountain birch forests.

Climate change has important implications on the abundance and range of insect pests in forest ecosystems. We studied responses of root-associated fungal communities to defoliation of mountain birch hosts by a massive geometrid moth outbreak through 454 pyrosequencing of tagged amplicons of the ITS2 rDNA region. We compared fungal diversity and community composition at three levels of moth defoliation (intact control, full defoliation in one season, full defoliation in two or more seasons), replicated in three localities. Defoliation caused dramatic shifts in functional and taxonomic community composition of root-associated fungi. Differentially defoliated mountain birch roots harbored distinct fungal communities, which correlated with increasing soil nutrients and decreasing amount of host trees with green foliar mass. Ectomycorrhizal fungi (EMF) abundance and richness declined by 70-80 % with increasing defoliation intensity, while saprotrophic and endophytic fungi seemed to benefit from defoliation. Moth herbivory also reduced dominance of Basidiomycota in the roots due to loss of basidiomycete EMF and increases in functionally unknown Ascomycota. Our results demonstrate the top-down control of belowground fungal communities by aboveground herbivory and suggest a marked reduction in the carbon flow from plants to soil fungi following defoliation. These results are among the first to provide evidence on cascading effects of natural herbivory on tree root-associated fungi at an ecosystem scale.

Short-term impacts of energy wood harvesting on ectomycorrhizal fungal communities of Norway spruce saplings.

The increased demand for harvesting energy wood raises questions about its effects on the functioning of the forest ecosystems, soil processes and biodiversity. Impacts of tree stump removal on ectomycorrhizal fungal (EMF) communities of Norway spruce saplings were studied with 454-pyrosequencing in a 3-year field experiment replicated in 3 geographical areas. This is possibly the most thorough investigation of EMF communities associated with saplings grown on sites subjected to energy wood harvesting. To separate impacts of tree stump and logging residue removal on EMF and plant variables, we used three harvesting treatments with increasing complexity from patch mounding alone (P) to patch mounding combined with logging residue removal (RP), and patch mounding combined with both logging residue and stump removal (SRP). Saplings grown in uncut forests (F) served as references for harvesting treatments. A majority of sequences (>92%) and operational taxonomic units (OTUs, 55%) were assigned as EMF. EMF OTU richness, fungal community composition or sapling growth did not differ between harvesting treatments (P, RP and SRP), while EMF OTU richness, diversity and evenness were highest and sapling growth lowest in the undisturbed reference forests (F). The short study period may partially explain the similarities in fungal and sapling variables in different harvesting treatments. In conclusion, our results indicate that neither stump removal nor logging residue removal have significant additional negative impacts on EMF communities or growth of Norway spruce saplings in the short-term compared with the impacts of more conventional harvesting methods, including clear cutting and patch mounding.

Do elevations in temperature, CO2, and nutrient availability modify belowground carbon gain and root morphology in artificially defoliated silver birch seedlings?

Climate warming increases the risk of insect defoliation in boreal forests. Losses in photosynthetically active surfaces cause reduction in net primary productivity and often compromise carbon reserves of trees. The concurrent effects of climate change and removal of foliage on root growth responses and carbohydrate dynamics are poorly understood, especially in tree seedlings. We investigated if exposures to different combinations of elevated temperature, CO2, and nutrient availability modify belowground carbon gain and root morphology in artificially defoliated 1-year-old silver birches (Betula pendula). We quantified nonstructural carbohydrates (insoluble starch as a storage compound; soluble sucrose, fructose, and glucose) singly and in combination in fine roots of plants under winter dormancy. Also the total mass, fine root proportion, water content, and length of roots were defined. We hypothesized that the measured properties are lower in defoliated birch seedlings that grow with ample resources than with scarce resources. On average, fertilization markedly decreased both the proportion and the carbohydrate concentrations of fine roots in all seedlings, whereas the effect of fertilization on root water content and dry mass was the opposite. However, defoliation mitigated the effect of fertilization on the root water content, as well as on the proportion of fine roots and their carbohydrate concentrations by reversing the outcomes. Elevation in temperature decreased and elevation in CO2 increased the absolute contents of total nonstructural carbohydrates, whereas fertilization alleviated both these effects. Also the root length and mass increased by CO2 elevation. This confirms that surplus carbon in birch tissues is used as a substrate for storage compounds and for cell wall synthesis. To conclude, our results indicate that some, but not all elements of climate change alter belowground carbon gain and root morphology in defoliated silver birch seedlings.

Moth herbivory enhances resource turnover in subarctic mountain birch forests?

Massive moth outbreaks cause large-scale damage in subarctic mountain birch forests with a concomitant decrease in carbon flux to mycorrhizal fungi and an increased deposition of dissolved carbon and nutrients as moth frass into soil. We investigated impacts of moth herbivory along three replicated gradients with three levels of moth herbivory (undamaged, once damaged, repeatedly damaged) on soil nutrient levels and biological parameters. We found an increase in soil nutrients and in the biomass of enchytraeid worms, which are key faunal decomposers. Fungi bacteria ratio and C:N ratio decreased in humus with increasing severity of herbivory. Our findings suggest enhanced resource turnover in mountain birch forests due to massive moth herbivory. This may provide a shortcut for carbon and nutrient input to subarctic soils, which largely bypasses the main routes of carbon from plants to soil via mycorrhizal and litter-decomposing fungi. Moreover, a temporal shift occurs in carbon allocation to soil, providing decomposers an opportunity to use an early-season peak in resource availability. Our results suggest a hitherto unappreciated role of massive insect herbivore attacks on resource dynamics in subarctic ecosystems.

Induced accumulation of phenolics and sawfly performance in Scots pine in response to previous defoliation.

Phenolic compounds often accumulate in foliar tissues of deciduous woody plants in response to previous insect defoliation, but similar responses have been observed infrequently in evergreen conifers. We studied the effects of defoliation on the foliar chemistry of Scots pine (Pinus sylvestris L.) and cocoon mass, and survival of the pine sawfly (Diprion pini L.). In two successive years, needles were excised early in the season leaving only the current-year shoot intact (defoliated trees); untreated entire shoots served as controls (control trees). A year after the second defoliation, pine sawfly larvae were transferred to the trees. Delayed induced resistance in Scots pine in response to defoliation was indicated by (1) reduced cocoon mass in defoliated trees and (2) increased concentrations of phenolics and soluble condensed tannins in the foliage of defoliated trees compared with controls. Myricetin-3-galactoside, which showed the strongest induced response (104% and 71% increase in current-year (C) and previous-year (C+1) needles) of the compounds analyzed, also entered the regression model explaining variation in sawfly performance. Other compounds that entered the model, e.g., (+)-catechin, showed weaker responses to defoliation than myricetin-3-galactoside. Hyperin, condensed tannins and quercitrin showed strong induced responses in C or C+1 needles, or both, but these compounds did not explain the variation in sawfly performance. Accumulation of phenolics is sometimes associated with the reduced foliage nitrogen (N) concentrations in deciduous trees, and our results suggest that this may also be the case in evergreen conifers. Based on the earlier findings that defoliation reduces needle N concentration and N deficiency results in the accumulation of the same phenolic compounds, i.e., myricetin and quercetin glycosides, and soluble condensed tannins, we suggest that the accumulation of phenolics in defoliated trees occurred in response to the reduced foliar N concentration.

Defoliation causes parallel temporal responses in a host tree and its fungal symbionts.

Growth of the host and its symbiont is often closely linked and so host damage may negatively affect the symbiont. While negative effects of aboveground herbivory on belowground fungal symbionts have been reported in several woody and herbaceous plants, here we report, for the first time, on differential effects of the timing of foliar damage on ectomycorrhizal (ECM) fungal symbionts. The phenologies of host trees and their ECM symbionts differ; the growth of the latter mainly occurs later in the season than that of the host. By removing Scots pine foliage on three occasions during the growing season (early, middle and late season defoliation) in one, two or three successive years, we demonstrate that, despite the differences in the seasonal growth dynamics of the tree and the symbionts, ECM fungi follow the host's response patterns to defoliation. Early season defoliation was most detrimental to the host and resulted in an increased proportion of low-biomass ectomycorrhizae which are presumed to require less carbon from the host tree. This may improve the recovery of the host, as most roots remained mycorrhizal in spite of the defoliation treatments repeated in successive years.