A synchronized, large-scale field experiment using Arabidopsis thaliana reveals the significance of the UV-B photoreceptor UVR8 under natural conditions.

This study determines the functional role of the plant ultraviolet-B radiation (UV-B) photoreceptor, UV RESISTANCE LOCUS 8 (UVR8) under natural conditions using a large-scale 'synchronized-genetic-perturbation-field-experiment'. Laboratory experiments have demonstrated a role for UVR8 in UV-B responses but do not reflect the complexity of outdoor conditions where 'genotype × environment' interactions can mask laboratory-observed responses. Arabidopsis thaliana knockout mutant, uvr8-7, and the corresponding Wassilewskija wild type, were sown outdoors on the same date at 21 locations across Europe, ranging from 39°N to 67°N latitude. Growth and climatic data were monitored until bolting. At the onset of bolting, rosette size, dry weight, and phenolics and glucosinolates were quantified. The uvr8-7 mutant developed a larger rosette and contained less kaempferol glycosides, quercetin glycosides and hydroxycinnamic acid derivatives than the wild type across all locations, demonstrating a role for UVR8 under field conditions. UV effects on rosette size and kaempferol glycoside content were UVR8 dependent, but independent of latitude. In contrast, differences between wild type and uvr8-7 in total quercetin glycosides, and the quercetin-to-kaempferol ratio decreased with increasing latitude, that is, a more variable UV response. Thus, the large-scale synchronized approach applied demonstrates a location-dependent functional role of UVR8 under natural conditions.

Legacies of invertebrate exclusion and tree secondary metabolites control fungal communities in dead wood.

During decomposition of organic matter, microbial communities may follow different successional trajectories depending on the initial environment and colonizers. The timing and order of the species arrival (assembly history) can lead to divergent communities through priority effects. We explored how assembly history and resource quality affected fungal communities and decay rate of decomposing wood, 1.5 and 4.5 years after tree felling. Additionally, we investigated the effect of invertebrate exclusion during the first two summers. We measured initial resource quality of bark and wood of aspen (Populus tremula) logs and surveyed the fungal communities by DNA metabarcoding at different times during succession. We found that gradients in fungal community composition were related to resource quality and we discuss how this may reflect different fungal life history strategies. As with previous studies, the initial amount of bark tannins was negatively correlated with wood decomposition rate over 4.5 years. The initial fungal community explained variation in community composition after 1.5, but not 4.5, years of succession. Although the assembly history of initial colonizers may cause alternative trajectories in successional communities, our results indicate that the communities may converge with the arrival of secondary colonizers. We also identified a strong legacy of invertebrate exclusion on fungal communities, even after 4.5 years of succession, thereby adding crucial knowledge on the importance of invertebrates in affecting fungal community development. By measuring and manipulating aspects of assembly history and resource quality that have rarely been studied, we expand our understanding of the complexity of fungal community dynamics.

Fungal community dynamics across a forest-alpine ecotone.

Climate change is causing upward shift of forest lines worldwide, with consequences for soil biota and carbon (C) sequestration. We here analyse compositional changes in the soil biota across the forest line ecotone, an important transition zone between different ecosystems. We collected soil samples along transects stretching from subalpine mountain birch forests to alpine heath. Soil fungi and micro-eukaryotes were surveyed using DNA metabarcoding of the ITS2 and 18S markers, while ergosterol was used to quantify fungal biomass. We observed a strong shift in the soil biota across the forest line ecotone: Below the forest line, there were higher proportions of basidiomycetes and mucoromycetes, including ectomycorrhizal and saprotrophic fungi. Above it, we observed relatively more root-associated ascomycetes, including Archaeorhizomycetes, ericoid mycorrhizal fungi and dark septate endophytes. Ergosterol and percentage C content in soil correlated strongly and positively with the abundance of root-associated ascomycetes. The predominance of ectomycorrhizal and saprotrophic fungi below the forest line probably promote high C turnover, while root-associated ascomycetes above the forest line may enhance C sequestration. With further rise in forest lines, there will be a corresponding shift in the below-ground biota, probably leading to enhanced release of soil C.

Elevated air humidity increases UV mediated leaf and DNA damage in pea (Pisum sativum) due to reduced flavonoid content and antioxidant power.

Growth in high relative air humidity (RH, >85%) affects plant morphology and causes diminished response to stomatal closing signals. Many greenhouses are prone to high RH conditions, which may negatively affect production and post-harvest quality. UV radiation induces stomatal closure in several species, and facilitates disease control. We hypothesised that UV exposure may trigger stomatal closure in pea plants (Pisum sativum) grown in high RH, thereby restoring stomatal function. The effects of UV exposure were tested on plants grown in moderate (60%) or high (90%) RH. UV exposure occurred at night, according to a disease control protocol. Lower stomatal conductance rates were found in UV-exposed plants, though UV exposure did not improve the rate of response to closing stimuli or desiccation tolerance. UV-exposed plants showed leaf curling, chlorosis, necrosis, and DNA damage measured by the presence of cyclobutane pyrimidine dimers (CPD), all of which were significantly greater in high RH plants. These plants also had lower total flavonoid content than moderate RH plants, and UV-exposed plants had less than controls. Plants exposed to UV had a higher content of cuticular layer uronic compounds than control plants. However, high RH plants had a higher relative amount of cuticular waxes, but decreased proteins and uronic compounds. Plants grown in high RH had reduced foliar antioxidant power compared to moderate RH. These results indicate that high RH plants were more susceptible to UV-induced damage than moderate RH plants due to reduced flavonoid content and oxidative stress defence.

No evidence of a protective or cumulative negative effect of UV-B on growth inhibition induced by gamma radiation in Scots pine (Pinus sylvestris) seedlings.

Exposure to ambient UV-B radiation may prime protective responses towards various stressors in plants, though information about interactive effects of UV-B and gamma radiation is scarce. Here, we aimed to test whether UV-B exposure could prime acclimatisation mechanisms contributing to tolerance to low-moderate gamma radiation levels in Scots pine seedlings, and concurrently whether simultaneous UV-B and gamma exposure may have an additive adverse effect on seedlings that had previously not encountered either of these stressors. Responses to simultaneous UV-B (0.35 W m-2) and gamma radiation (10.2-125 mGy h-1) for 6 days with or without UV-B pre-exposure (0.35 W m-2, 4 days) were studied across various levels of organisation, as compared to effects of either radiation type. In contrast to UV-B, and regardless of UV-B presence, gamma radiation at ≥42.9 mGy h-1 caused increased formation of reactive oxygen species and reduced shoot length, and reduced root length at 125 mGy h-1. In all experiments there was a gamma dose rate-dependent increase in DNA damage at ≥10.8 mGy h-1, generally with additional UV-B-induced damage. Gamma-induced growth inhibition and gamma- and UV-B-induced DNA damage were still visible 44 days post-irradiation, even at 20.7 mGy h-1, probably due to genomic instability, but this was reversed after 8 months. In conclusion, there was no evidence of a protective effect of UV-B on gamma-induced growth inhibition and DNA damage in Scots pine, and no additive adverse effect of gamma and UV-B radiation on growth in spite of the additional UV-B-induced DNA damage.

A Seven-Year Study of Phenolic Concentrations of the Dioecious Salix myrsinifolia.

In boreal woody plants, concentrations of defensive phenolic compounds are expected to be at a high level during the juvenile phase and decrease in maturity, although there is variation between plant species. Females of dioecious species, like most of the Salicaceae, are expected to invest their resources in defense and reproduction, while males are expected to be more growth-oriented. We studied age- and sex-dependent changes in leaf and stem phenolics, and in height and diameter growth in a dioecious Salix myrsinifolia plants over a seven-year time period. In addition, we registered flowering as well as rust damage in the leaves. From the first year and throughout ontogenetic development from juvenile to adult phases, there was no significant change in the concentrations of any of the studied compounds in the leaves of S. myrsinifolia. In the stems, the concentrations of six out of 43 identified compounds decreased slightly with age, which may be partly explained by dilution caused by the increment in stem diameter with age. The fairly steady chemistry level over seven years, accompanied by moderate genotypic phenolic variation, indicates important roles of chemical defenses against herbivory for this early-successional species.

Compositional Changes in Foliage Phenolics with Plant Age, a Natural Experiment in Boreal Forests.

The composition of plant secondary metabolites (PSMs) extensively impacts ecosystem functioning. It is vital that we understand temporal patterns in the plants' allocation of resources to PSMs, particularly those influenced by human activity. Existing data are insufficient in the long-term perspective of perennial plants (age or ontogeny). We analysed phenolic concentrations in foliage from birch (Betula pubescens Ehr.) considered to be undamaged and growing on 5, 10 and 15 years old clear-cuts in two boreal forest landscapes in Norway, sampled at the peak of the growing season. In sum, low molecular weight phenolic concentrations decreased with age. Apart from one apigenin glycoside, the low molecular weight phenolics co-varied similarly at all ages, suggesting a lack of temporal compound-specific prioritisation of this group. In contrast, the concentration of MeOH-soluble condensed tannins increased with age. The compositional shift fits well with several hypotheses that may provide proximate explanations for age patterns in PSM allocations, including both resource constraints and external pressures. Regardless of these explanations, our study adds an important perennial perspective (plant age) to temporal PSM patterns already well-known in boreal plant phenology (foliage age).

Correction to: Compositional Changes in Foliage Phenolics with Plant Age, a Natural Experiment in Boreal Forests.

The article Compositional Changes in Foliage Phenolics with Plant Age, a Natural Experiment in Boreal Forests, written by Hilde Karine Wam, Caroline Stolter and Line Nybakken, was originally published electronically on the publisher's internet portal (currently SpringerLink) on August 29, 2017 without open access.

Gender differences in Salix myrsinifolia at the pre-reproductive stage are little affected by simulated climatic change.

Females of dioecious species are known often to prioritize defense, while males grow faster. As climatic change is known to influence both growth and defense in plants, it would be important to know whether it affects the sexes of dioecious species differently. This could have impacts on future sex ratios in nature. We grew four clones of each sex of Salix myrsinifolia in greenhouse chambers under ambient conditions, enhanced temperature, enhanced CO2 or enhanced temperature + enhanced CO2 . The females had the greatest growth and also the highest levels of phenolic compounds in twigs, while in leaves some compounds were higher in males, some in females. Enhanced CO2 increased growth equally in both sexes, while growth was not affected by elevated temperature. Phenolic compounds in twigs were, however, lowered under elevated temperature. The gender differences were not strongly affected by the simulated climatic changes, but the effects seen on some highly concentrated compounds may be important. We interpret the intensive growth at pre-reproductive phase as a strategy in females to get an initial advantage before later periods with fewer resources available for growth.

Combination treatment of elevated UVB radiation, CO2 and temperature has little effect on silver birch (Betula pendula) growth and phytochemistry.

Elevations of carbon dioxide, temperature and ultraviolet-B (UBV) radiation in the growth environment may have a high impact on the accumulation of carbon in plants, and the different factors may work in opposite directions or induce additive effects. To detect the changes in the growth and phytochemistry of silver birch (Betula pendula) seedlings, six genotypes were exposed to combinations of ambient or elevated levels of CO2 , temperature and UVB radiation in top-closed chambers for 7 weeks. The genotypes were relatively similar in their responses, and no significant interactive effects of three-level climate factors on the measured parameters were observed. Elevated UVB had no effect on growth, nor did it alter plant responses to CO2 and/or temperature in combined treatments. Growth in all plant parts increased under elevated CO2 , and height and stem biomass increased under elevated temperature. Increased carbon distribution to biomass did not reduce its allocation to phytochemicals: condensed tannins, most flavonols and phenolic acids accumulated under elevated CO2 and elevated UVB, but this effect disappeared under elevated temperature. Leaf nitrogen content decreased under elevated CO2 . We conclude that, as a result of high genetic variability in phytochemicals, B. pendula seedlings have potential to adapt to the tested environmental changes. The induction in protective flavonoids under UVB radiation together with the positive impact of elevated CO2 and temperature mitigates possible UVB stress effects, and thus atmospheric CO2 concentration and temperature are the climate change factors that will dictate the establishment and success of birch at higher altitudes in the future.

Long term effects of ionising radiation in the Chernobyl Exclusion zone on DNA integrity and chemical defence systems of Scots pine (Pinus sylvestris).

The Chernobyl Nuclear Power Plant (ChNPP) accident in 1986 resulted in extremely high levels of acute ionising radiation, that killed or damaged Scots pine (Pinus sylvestris) trees in the surrounding areas. Dead trees were cleared and buried, and new plantations established a few years later. Today, more than three decades later, gamma and beta-radiation near the ChNPP is still elevated compared with ambient levels but have decreased by a factor of 300 and 100, respectively. In the present work, Scots pine-trees growing at High (220 µGy h-1), Medium (11 µGy h-1), and Low (0.2 µGy h-1) total (internal + external) dose rates of chronically elevated ionising radiation in the Chernobyl Exclusion zone were investigated with respect to possible damage to DNA, cells and organelles, as well as potentially increased levels of phenolic and terpenoid antioxidants. Scots pine from the High and Medium radiation sites had elevated levels of DNA damage in shoot tips and needles as shown by the COMET assay, as well as increased numbers of resin ducts and subcellular abnormalities in needles. Needles from the High radiation site showed elevated levels of monoterpenes and condensed tannins compared with those from the other sites. In conclusion, more than three decades after the ChNPP accident substantial DNA damage and (sub)cellular effects, but also mobilisation of stress-protective substances possessing antioxidant activity were observed in Scots pine trees growing at elevated levels of ionising radiation. This demonstrates that the radiation levels in the Red Forest still significantly impact the plant community.

Combined enhancements of temperature and UVB influence growth and phenolics in clones of the sexually dimorphic Salix myrsinifolia.

Although several climatic factors are expected to change simultaneously in the future, the effect of such combined changes on plants have seldom been tested under field conditions. We report on a field experiment with dark-leaved willow, Salix myrsinifolia, subjected to enhancements in ultraviolet-A (UVA), UVB radiation and temperature, setup in Joensuu, Eastern Finland. S. myrsinifolia is a dioecious species, known as an important food plant for many herbivores. Cuttings of eight clones, four of each sex, of dark-leaved willow were planted in the field in spring 2009. In both 2009 and 2010, the total biomass increased significantly with temperature, and in 2010 there was an additive effect of UVB radiation. Both height and diameter increased with temperature in 2009, while the effect on height growth ceased in 2010. Males had greater diameter growth than females in 2010. Most phenolic compounds in the leaves decreased under enhanced temperature in both growing seasons. In 2010, four of six salicylates increased in response to enhanced temperature. Some quercetin derivatives increased under enhanced UVB radiation. Females had higher concentrations of chlorogenic acids than males, and while enhanced temperature reduced chlorogenic acid in females only, luteolins were reduced only in males. In summary, the combined enhancements gave no effects in addition to those that appeared under the single-factor treatments, except for the additive effect of UVB on temperature-increased biomass. The few gender-related differences found in response to climate change do not allow any marked expectations of future climate-induced changes in sex ratios.

Dark-leaved willow (Salix myrsinifolia) is resistant to three-factor (elevated CO2, temperature and UV-B-radiation) climate change.

Elevated carbon dioxide (CO2 ), temperature (T) and ultraviolet-B (UV-B) radiation may affect plant growth and secondary chemistry in different directions, but the effect of the combination of the three factors has seldom been tested. Here, we grew four dark-leaved willow (Salix myrsinifolia) clones under combinations of ambient or elevated CO2, T and UV-B radiation in top-closed chambers for 7 wk. Elevated UV-B had no effects on growth or phenolic compounds, and there were no significant interactions between UV-B, CO2 and T. CO2 alone increased most growth parameters, but the magnitude of the effect varied among the clones. Total phenolics increased at elevated CO2 , whereas they decreased at elevated T. The responses varied between the clones. The results imply that dark-leaved willow are fairly resistant to the applied three-factor climate change, probably because of high constitutive defense. However, the interactions between clone and climate change factors implies that some clones are more susceptible than the species as a whole.

Establishment of spruce plantations in native birch forests reduces soil fungal diversity.

Plantations of Norway spruce have been established well beyond its natural range in many parts of the world, potentially impacting native microbial ecosystems and the processes they mediate. In this study, we investigate how the establishment of spruce plantations in a landscape dominated by native birch forests in western Norway impacts soil properties and belowground fungal communities. Soil cores were collected from neighboring stands of planted spruce and native birch forests. We used DNA metabarcoding of the rDNA internal transcribed spacer 2 region and ergosterol measurements to survey the fungal community composition and its biomass, respectively. In the two investigated soil layers (litter and humus), fungal community composition, diversity and biomass were strongly affected by the tree species shift. Native birch stands hosted markedly richer fungal communities, including numerous fungi not present in planted spruce stands. In contrast, the spruce stands included higher relative abundance of ectomycorrhizal fungi as well as higher fungal biomass. Hence, establishing plantations of Norway spruce in native birch forests leads to significant losses in diversity, but increase in biomass of ectomycorrhizal fungi, which could potentially impact carbon sequestration processes and ecosystem functioning.

Accumulation of phenolics and growth of dioecious Populus tremula (L.) seedlings over three growing seasons under elevated temperature and UVB radiation.

Accumulation of secondary metabolites may exhibit developmentally regulated variation in different plant organs. Moreover, prevailing environmental conditions may interact with development-related variations in plant traits. In this study, we examined developmentally regulated variation in phenolic accumulation in the twigs of dioecious Populus tremula (L.) and how the effects of elevated temperature and ultraviolet B (UVB) radiation on growth and phenolics accumulation varied as the plants get older. In an open-field experiment, six female and six male genotypes were exposed to single and combined elevated temperature and UVB radiation treatments for three consecutive growing seasons. The concentrations of low molecular weight phenolics and condensed tannins did not show age-dependent variation in the twigs. In temperature-treated plants, diameter growth rate decreased, and concentration of condensed tannins increased as plants aged; there were no cumulative effects of elevated UVB radiation on growth and phenolic accumulation. Females maintained a higher concentration of low molecular weight phenolics throughout the experimental period; however, growth and phenolic concentration did not vary over time in females and males. Our results suggest that phenolic accumulation in perennial plants may not necessarily always exhibit age-dependent variation and the effects of elevated temperature on growth and phenolic may diminish as plants get older.

Lichen compounds restrain lichen feeding by bank voles (Myodes glareolus).

Some lichen compounds are known to deter feeding by invertebrate herbivores. We attempted to quantify the deterring efficiency of lichen compounds against a generalist vertebrate, the bank vole (Myodes glareolus). In two separate experiments, caged bank voles had the choice to feed on lichens with natural or reduced concentrations of secondary compounds. We rinsed air-dry intact lichens in 100% acetone to remove extracellular compounds non-destructively. In the first experiment, pairs of control and rinsed lichen thalli were hydrated and offered to the bank voles. Because the lichens desiccated fast, we ran a second experiment with pairs of ground control and compound-deficient thalli, each mixed with water to porridge. Eight and six lichen species were tested in the first and second experiment, respectively. In the first, bank voles preferred compound-deficient thalli of Cladonia stellaris and Lobaria pulmonaria, but did not discriminate between the other thallus pairs. This was likely a result of deterring levels of usnic and stictic acid in the control thalli. When lichens were served as porridge, significant preference was found for acetone-rinsed pieces of Cladonia arbuscula, C. rangiferina, Platismatia glauca, and Evernia prunastri. The increased preference was caused mainly by lower consumption of control thalli. Grinding and mixing of thallus structures prevented bank voles from selecting thallus parts with lower concentration of secondary compounds and/or strengthened their deterring capacity. We conclude that some lichen secondary compounds deter feeding by bank voles.

Responses in growth and phenolics accumulation to lateral bud removal in male and female saplings of Populus tremula (L.) under simulated climate change.

The responses in growth and defense after tissue damage are highly variable in plants depending on species, damaged-tissue type and the intensity of damage. The prevailing abiotic conditions can also influence these responses. In this study, our aim was to examine how the removal of lateral vegetative buds affects the growth and accumulation of phenolics in saplings of the dioecious Populus tremula grown under simulated climate change. For three growing seasons, the saplings were grown under ambient conditions (control), elevated temperature (+2°C) and elevated UV radiation (30%) (UVB and UVA as its control), or a combination of these. In the fourth growing season, all saplings were grown under ambient conditions. The bud removal was performed twice - in summer and autumn - in the third year. The responses of growth and the accumulation of phenolics to the bud removal were measured at the end of the fourth growing season. Removal of 5% of the lateral buds resulted in higher leaf, stem and total plant biomass in both sexes of P. tremula saplings, compared to intact plants. The effects were greater in the temperature-treated plants, especially in the temperature-treated females. The concentrations of flavonoids and condensed tannins were higher in the bud-removed individuals. The concentration of condensed tannins was also higher in the males than in the females, opposite to the concentration of phenolic acids. There was no significant interaction between bud removal and UVB treatment on either growth or phenolics. Our results suggest that plants can allocate resources to both growth and defense simultaneously in response to tissue loss, and that global warming can modify the responses to some extent.

Ultraviolet radiation accelerates photodegradation under controlled conditions but slows the decomposition of senescent leaves from forest stands in southern Finland.

Depending on the environment, sunlight can positively or negatively affect litter decomposition, through the ensemble of direct and indirect processes constituting photodegradation. Which of these processes predominate depends on the ecosystem studied and on the spectral composition of sunlight received. To examine the relevance of photodegradation for litter decomposition in forest understoreys, we filtered ultraviolet radiation (UV) and blue light from leaves of Fagus sylvatica and Betula pendula at two different stages of senescence in both a controlled-environment experiment and outdoors in four different forest stands (Picea abies, Fagus sylvatica, Acer platanoides, Betula pendula). Controlling for leaf orientation and initial differences in leaf chlorophyll and flavonol concentrations; we measured mass loss at the end of each experiment and characterised the phenolic profile of the leaf litter following photodegradation. In most forest stands, less mass was lost from decomposing leaves that received solar UV radiation compared with those under UV-attenuating filters, while in the controlled environment UV-A radiation either slightly accelerated or had no significant effect on photodegradation, according to species identity. Only a few individual phenolic compounds were affected by our different filter treatments, but photodegradation did affect the phenolic profile. We can conclude that photodegradation has a small stand- and species-specific effect on the decomposition of surface leaf litter in forest understoreys during the winter following leaf fall in southern Finland. Photodegradation was wavelength-dependent and modulated by the canopy species filtering sunlight and likely creating different combinations of spectral composition, moisture, temperature and snowpack characteristics.

Size-dependent growth of two old-growth associated macrolichen species.

Relationships between thallus size and growth variables were analysed for the foliose Lobaria pulmonaria and the pendulous Usnea longissima with the aim of elucidating their morphogenesis and the factors determining thallus area (A) versus biomass (dry weight (DW) gain. Size and growth data originated from a factorial transplantation experiment that included three boreal climate zones (Atlantic, suboceanic and continental), each with three successional forest stands (clear-cut, young and old). When A was replaced by the estimated photobiont layer area in an area-DW scatterplot including all thalli (n = 1080), the two separate species clusters merged into one, suggesting similar allocation patterns between photobionts and mycobionts across growth forms. During transplantation, stand-specific water availability boosted area gain in foliose transplants, consistent with a positive role of water in fungal expansion. In pendulous lichens, A gain greatly exceeded DW gain, particularly in small transplants. The A gain in U. longissima increased with increasing DW:A ratio, consistent with a reallocation of carbon, presumably mobilized from the dense central chord. Pendulous lichens with cylindrical photobiont layers harvest light from all sides. Rapid and flexible three-dimensional A gain allows the colonization of spaces between canopy branches to utilize temporary windows of light in a growing canopy. Foliose lichens with a two-dimensional photobiont layer have more coupled A and DW gains.

Spruce and beech as local determinants of forest fungal community structure in litter, humus and mineral soil.

Beech forests reaches its native distribution limit in SE Norway, but is expected to expand substantially northwards due to climate warming. This may potentially result in a fundamental transformation of contemporary Northern European forests, with tentative effects on the associated belowground fungi. Fungal communities mediate vital ecosystem processes such as ecosystem productivity and carbon sequestration in boreal forests. To investigate how soil fungi is affected by the vegetation transition from spruce to beech forest, we sampled litter, humus and mineral soil in a forest landscape dominated by beech, spruce or a mixture of these. The fungal communities in the soil samples were analyzed by DNA metabarcoding of the rDNA ITS2 region. Although soil layers were the most important structuring gradient, we found clear differences in fungal species composition between spruce and beech plots. The differences in fungal community composition were most evident in the litter and least in the mineral soil. Decomposers, most notably Mycena, dominated the litter layer while various mycorrhizal fungi dominated the humus and mineral layers. Some ectomycorrhizal taxa, such as Cenoccocum and Russula, were more abundant in spruce forests. Differences in fungal community composition between forest types can potentially have large impacts on carbon sequestration rates.