Resistance of soil bacterial communities from montane heathland ecosystems in the Cantabrian mountains (NW Spain) to a gradient of experimental nitrogen deposition.

Elevated atmospheric nitrogen (N) deposition on terrestrial ecosystems has become one of the most important drivers of microbial diversity loss on a global scale, and has been reported to alter the soil function of nutrient-poor, montane Calluna vulgaris heathlands in the context of global change. In this work we analyze for the first time the shifts of bacterial communities in response to experimental addition of N in Calluna heathlands as a simulation of atmospheric deposition. Specifically, we evaluated the effects of five N addition treatments (0, 10, 20, and 50 kg N ha-1 yr-1 for 3-years; and 56 kg N ha-1 yr-1 for 10-years) on the resistance of soil bacterial communities as determined by changes in their composition and alpha and beta diversities. The study was conducted in montane Calluna heathlands at different development stages (young and mature phases) in the southern side of the Cantabrian Mountains (NW Spain). Our results evidenced a substantial increase of long-term (10-years) N inputs on soil extractable N-NH4+, particularly in young Calluna stands. The alpha diversity of soil bacterial communities in mature Calluna stands did not show a significant response to experimental N addition, whereas it was significantly higher under long-term chronic N addition (56 kg N ha-1 yr-1 for 10-years) in young Calluna stands. These bacterial community shifts are mainly attributable to a decrease in the dominance of Acidobacteria phylum, the most representative in montane Calluna ecosystems, in favor of copiotrophic taxa such as Actinobacteria or Proteobacteria phyla, favored under increased N availability. Future research should investigate what specific ecosystem functions performed by soil bacterial communities may be sensitive to increased nitrogen depositions, which may have substantial implications for the understanding of montane Calluna ecosystems' stability.

Plant and vegetation functional responses to cumulative high nitrogen deposition in rear-edge heathlands.

Elevated atmospheric nitrogen (N) deposition is a major driver of change, altering the structure/functioning of nutrient-poor Calluna vulgaris-heathlands over Europe. These effects amply proven for north-western/central heathlands may, however, vary across the ecosystem's distribution, especially at the range limits, as heathlands are highly vulnerable to land-use changes combined with present climate change. This is an often overlooked and greatly understudied aspect of the ecology of heathlands facing global change. We investigated the effects of five N-fertilisation treatments simulating a range of N deposition rates (0, 10, 20, and 50 kg N ha-1 yr-1 for 1 year; and 56 kg N ha-1 yr-1 for 9 years) on the Calluna-plants, the plant functional groups, species composition and richness of two life-cycle stages (building/young- and mature-phase) of Calluna-heathlands at their rear-edge limit. Our findings revealed a dose-related response of the shoot length and number of flowers of young and mature Calluna-plants to the addition of N, adhering to the findings from other heathland locations. However, cumulative high-N loading reduced the annual growth and flowering of young plants, showing early signs of N saturation. The different plant functional groups showed contrasting responses to the cumulative addition of N: annual/perennial forbs and annual graminoids increased with quite low values; perennial graminoids were rather abundant in young heathlands but only slightly augmented in mature ones; while bryophytes and lichens strongly declined at the two heathland life-cycle stages. Meanwhile there were no significant N-driven changes in plant species composition and richness. Our results demonstrated that Calluna-heathlands at their low-latitude distribution limit are moderately resistant to cumulative high-N loading. As north-western/central European heathlands under high-N inputs broadly experienced the loss of plant diversity and pronounced changes in plant species dominance, rear-edge locations may be of critical importance to unravel the mechanisms of heathland resilience to future global change.

Persistent Supercooling of Reproductive Shoots Is Enabled by Structural Ice Barriers Being Active Despite an Intact Xylem Connection.

Extracellular ice nucleation usually occurs at mild subzero temperatures in most plants. For persistent supercooling of certain plant parts ice barriers are necessary to prevent the entry of ice from already frozen tissues. The reproductive shoot of Calluna vulgaris is able to supercool down to below -22°C throughout all developmental stages (shoot elongation, flowering, fruiting) despite an established xylem conductivity. After localization of the persistent ice barrier between the reproductive and vegetative shoot at the base of the pedicel by infrared differential thermal analysis, the currently unknown structural features of the ice barrier tissue were anatomically analyzed on cross and longitudinal sections. The ice barrier tissue was recognized as a 250 µm long constriction zone at the base of the pedicel that lacked pith tissue and intercellular spaces. Most cell walls in this region were thickened and contained hydrophobic substances (lignin, suberin, and cutin). A few cell walls had what appeared to be thicker cellulose inclusions. In the ice barrier tissue, the area of the xylem was as much as 5.7 times smaller than in vegetative shoots and consisted of tracheids only. The mean number of conducting units in the xylem per cross section was reduced to 3.5% of that in vegetative shoots. Diameter of conducting units and tracheid length were 70% and 60% (respectively) of that in vegetative shoots. From vegetative shoots water transport into the ice barrier must pass pit membranes that are likely impermeable to ice. Pit apertures were about 1.9 µm x 0.7 µm, which was significantly smaller than in the vegetative shoot. The peculiar anatomical features of the xylem at the base of the pedicel suggest that the diameter of pores in pit membranes could be the critical constriction for ice propagation into the persistently supercooled reproductive shoots of C. vulgaris.

Diversity of the Insect Visitors on Calluna vulgaris (Ericaceae) in Southern France Heathlands.

As part of an ongoing research project on the pollination networks in European heathlands, the objective of this study was to assess the insect visitor guild on Calluna vulgaris (L.) Hull (Ericaceae). We focused the study on a region renowned for its largely well-preserved heathlands, the Cévennes National Park, Southern France. In 2013, flower visitors were observed over 3 d per site, in four heathland sites at mont Lozère. Honeybees (Apis mellifera L.) were the main visitors (62-88% of total visitors). Besides honeybees, a high diversity of visitors was detected with 57 different species identified (42 Diptera and 15 Hymenoptera). Hoverflies (Syrphidae, Diptera) visitors were abundant and diverse, especially individuals belonging to the genera Eristalis and Episyrphus. The reported diversity of visitors was probably due to the preservation of large heathland areas at mont Lozère and to the generalist pollination system of C. vulgaris.

Management-driven evolution in a domesticated ecosystem.

Millennia of human land-use have resulted in the widespread occurrence of what have been coined 'domesticated ecosystems'. The anthropogenic imprints on diversity, composition, structure and functioning of such systems are well documented. However, evolutionary consequences of human activities in these ecosystems are enigmatic. Calluna vulgaris (L.) is a keystone species of coastal heathlands in northwest Europe, an ancient semi-natural landscape of considerable conservation interest. Like many species from naturally fire-prone ecosystems, Calluna shows smoke-adapted germination, but it is unclear whether this trait arose prior to the development of these semi-natural landscapes or is an evolutionary response to the anthropogenic fire regime. We show that smoke-induced germination in Calluna is found in populations from traditionally burnt coastal heathlands but is lacking in naturally occurring populations from other habitats with infrequent natural fires. Our study thus demonstrates evolutionary imprints of human land-use in semi-natural ecosystems. Evolutionary consequences of historic anthropogenic impacts on wildlife have been understudied, but understanding these consequences is necessary for informed conservation and ecosystem management.

AFLP-based genetic mapping of the "bud-flowering" trait in heather (Calluna vulgaris).

BACKGROUND: Calluna vulgaris is one of the most important landscaping plants produced in Germany. Its enormous economic success is due to the prolonged flower attractiveness of mutants in flower morphology, the so-called bud-bloomers. In this study, we present the first genetic linkage map of C. vulgaris in which we mapped a locus of the economically highly desired trait "flower type". RESULTS: The map was constructed in JoinMap 4.1. using 535 AFLP markers from a single mapping population. A large fraction (40%) of markers showed distorted segregation. To test the effect of segregation distortion on linkage estimation, these markers were sorted regarding their segregation ratio and added in groups to the data set. The plausibility of group formation was evaluated by comparison of the "two-way pseudo-testcross" and the "integrated" mapping approach. Furthermore, regression mapping was compared to the multipoint-likelihood algorithm. The majority of maps constructed by different combinations of these methods consisted of eight linkage groups corresponding to the chromosome number of C. vulgaris. CONCLUSIONS: All maps confirmed the independent inheritance of the most important horticultural traits "flower type", "flower colour", and "leaf colour". An AFLP marker for the most important breeding target "flower type" was identified. The presented genetic map of C. vulgaris can now serve as a basis for further molecular marker selection and map-based cloning of the candidate gene encoding the unique flower architecture of C. vulgaris bud-bloomers.

Conditional cold avoidance drives between-population variation in germination behaviour in Calluna vulgaris.

BACKGROUND AND AIMS: Across their range, widely distributed species are exposed to a variety of climatic and other environmental conditions, and accordingly may display variation in life history strategies. For seed germination in cold climates, two contrasting responses to variation in winter temperature have been documented: first, an increased ability to germinate at low temperatures (cold tolerance) as winter temperatures decrease, and secondly a reduced ability to germinate at low temperatures (cold avoidance) that concentrates germination towards the warmer parts of the season. METHODS: Germination responses were tested for Calluna vulgaris, the dominant species of European heathlands, from ten populations collected along broad-scale bioclimatic gradients (latitude, altitude) in Norway, covering a substantial fraction of the species' climatic range. Incubation treatments varied from 10 to 25 °C, and germination performance across populations was analysed in relation to temperature conditions at the seed collection locations. KEY RESULTS: Seeds from all populations germinated rapidly and to high final percentages under the warmer incubation temperatures. Under low incubation temperatures, cold-climate populations had significantly lower germination rates and percentages than warm-climate populations. While germination rates and percentages also increased with seed mass, seed mass did not vary along the climatic gradients, and therefore did not explain the variation in germination responses. CONCLUSIONS: Variation in germination responses among Calluna populations was consistent with increased temperature requirements for germination towards colder climates, indicating a cold-avoidance germination strategy conditional on the temperature at the seeds' origin. Along a gradient of increasing temperatures this suggests a shift in selection pressures on germination from climatic adversity (i.e. low temperatures and potential frost risk in early or late season) to competitive performance and better exploitation of the entire growing season.

The legacy of nitrogen pollution in heather moorlands: ecosystem response to simulated decline in nitrogen deposition over seven years.

Eutrophication and acidification of heather moorlands by chronic atmospheric nitrogen (N) pollution, is of major concern within these internationally important ecosystems. However, in the UK and Western Europe generally emissions of NO(y) and NH(x) peaked during the 20th century. Due to the history and scale of atmospheric N pollution, the legacy of these high levels of N deposition, through accumulation in soil, may hinder or prevent ecosystem recovery. Effects of N pollution on heather moorland were investigated throughout the ecosystem including; the dominant plant species, Calluna vulgaris, the bryophyte and lichen community and the soil system using a long-term experiment simulating wet N deposition. We observed an increase in C. vulgaris height, shoot extension and canopy density, litter mineral N, total N concentration, N:P and C:N ratios in response to N addition. Bryophyte species diversity, bryophyte and lichen frequency and the frequency of two individual bryophyte species (Lophozia ventricosa and Campylopus flexuosus) were significantly reduced by N addition. We developed an N recovery experiment, using a split-plot design, on the long-term N treatment plots to investigate ecosystem response to a simulated decline in N deposition. Two years after cessation of N treatment the only ecosystem component that responded to the recovery experiment was C. vulgaris shoot extension, however after seven years of recovery there were significant declines in litter total N concentration and mineral N and an increase in litter C:N ratio. Although bryophytes and lichens form a close relationship with atmospheric N deposition these organisms did not show a significant response to the N recovery experiment, two years after cessation of N treatment. These data indicate that low nutrient ecosystems, such as moorlands, have the capacity to respond to declines in N deposition however the accumulation of pollution may hinder recovery of sensitive organisms, such as bryophytes and lichens.

Malformation of gynoecia impedes fertilisation in bud-flowering Calluna vulgaris.

In Calluna vulgaris, a common bedding plant during autumn in the northern hemisphere, the bud-blooming mutation of flower morphology is of high economic importance. Breeding of new bud-blooming cultivars suffers from poor seed set in some of the desirable bud-flowering crossing partners. In the current study, fertilisation and seed development in genotypes with good or poor seed set were monitored in detail in order to examine pre- and post-zygotic cross breeding incompatibilities. Whereas no distinct differences were detected in seed development, pollen tube growth was impeded in the pistils of genotypes characterised by poor seed set. Detailed microscopic analysis revealed malformations of the gynoecia due to imperfect fusion of carpels. Hence, a pre-zygotic mechanism hindering pollen tube growth due to malformation of gynoecia was deduced. An interaction of putative candidate genes involved in malformation of gynoecia with floral organ identity genes controlling the flower architecture is discussed.

Recurring weather extremes alter the flowering phenology of two common temperate shrubs.

The aim of this study is to explore the effects of heavy rain and drought on the flowering phenology of two shrub species Genista tinctoria and Calluna vulgaris. We conducted a field experiment over five consecutive years in Central Europe, applying annually recurring extreme drought and heavy rain events on constructed shrubland communities and recorded the flowering status. Further, we correlated spring temperature and precipitation with the onset of flowering. Both species showed a response to extreme weather events: drought delayed the mid flowering date of Genista tinctoria in 3 of 5 years by about 1 month and in 1 year advanced the mid flowering date by 10 days, but did not affect the length of flowering. Mid flowering date of Calluna vulgaris was not affected by drought, but the length of flowering was extended in 2 years by 6 and 10 days. For C. vulgaris the closer the drought occurred to the time of flowering, the larger the impact on the flowering length. Heavy rainfall advanced mid flowering date and reduced the length of flowering of Genista tinctoria by about 2 months in 1 year. Mid flowering date of Calluna vulgaris was not affected by heavy rain, but the length of flowering was reduced in 1 year by 4 days. Our data suggest that extreme weather events, including alterations to the precipitation regime, induce phenological shifts of plant species of a substantial magnitude. Thus, the impacts of climate extremes on plant life cycles may be as influential as gradual warming. Particularly, the variability in the timing of precipitation events appears to have a greater influence on flowering dynamics than the magnitude of the precipitation.

Interactive effects of drought, elevated CO2 and warming on photosynthetic capacity and photosystem performance in temperate heath plants.

Increased temperature, atmospheric CO(2) and change in precipitation patterns affect plant physiological and ecosystem processes. In combination, the interactions between these effects result in complex responses that challenge our current understanding. In a multi-factorial field experiment with elevated CO(2) (CO2, FACE), nighttime warming (T) and periodic drought (D), we investigated photosynthetic capacity and PSII performance in the evergreen dwarf shrub Calluna vulgaris and the grass Deschampsia flexuosa in a temperate heath ecosystem. Photosynthetic capacity was evaluated using A/C(i) curves, leaf nitrogen content and chlorophyll-a fluorescence OJIP induction curves. The PSII performance was evaluated via the total performance index PI(total), which integrates the function of antenna, reaction centers, electron transport and end-acceptor reduction according to the OJIP-test. The PSII performance was negatively influenced by high air temperature, low soil water content and high irradiance dose. The experimental treatments of elevated CO(2) and prolonged drought generally down-regulated J(max), V(cmax) and PI(total). Recovery from these depressions was found in the evergreen shrub after rewetting, while post-rewetting up-regulation of these parameters was observed in the grass. Warming effects acted indirectly to improve early season J(max), V(cmax) and PI(total). The responses in the multi-factorial experimental manipulations demonstrated complex interactive effects of T×CO2, D×CO2 and T×D×CO2 on photosynthetic capacity and PSII performance. The impact on the O-J, J-I and I-P phases which determine the response of PI(total) are discussed. The single factor effects on PSII performance and their interactions could be explained by parallel adjustments of V(cmax), J(max) and leaf nitrogen in combination. Despite the highly variable natural environment, the OJIP-test was very robust in detecting the impacts of T, D, CO2 and their interactions. This study demonstrates that future climate will affect fundamental plant physiological processes in a way that is not predictable from single factor treatments. The interaction effects that were observed depended upon both the growth strategy of the species considered, and their ability to adjust during drought and rewetting periods.

Effects of elevated CO2, warming and drought episodes on plant carbon uptake in a temperate heath ecosystem are controlled by soil water status.

The impact of elevated CO2, periodic drought and warming on photosynthesis and leaf characteristics of the evergreen dwarf shrub Calluna vulgaris in a temperate heath ecosystem was investigated. Photosynthesis was reduced by drought in midsummer and increased by elevated CO2 throughout the growing season, whereas warming only stimulated photosynthesis early in the year. At the beginning and end of the growing season, a T × CO2 interaction synergistically stimulated plant carbon uptake in the combination of warming and elevated CO2. At peak drought, the D × CO2 interaction antagonistically down-regulated photosynthesis, suggesting a limited ability of elevated CO2 to counteract the negative effect of drought. The response of photosynthesis in the full factorial combination (TDCO2) could be explained by the main effect of experimental treatments (T, D, CO2) and the two-factor interactions (D × CO2, T × CO2). The interactive responses in the experimental treatments including elevated CO2 seemed to be linked to the realized range of treatment variability, for example with negative effects following experimental drought or positive effects following the relatively higher impact of night-time warming during cold periods early and late in the year. Longer-term experiments are needed to evaluate whether photosynthetic down-regulation will dampen the stimulation of photosynthesis under prolonged exposure to elevated CO2.

Habitat management: a tool to modify ecosystem impacts of nitrogen deposition?

Atmospheric nitrogen deposition has been shown to affect both the structure and the function of heathland ecosystems. Heathlands are semi-natural habitats and, as such, undergo regular management by mowing or burning. Different forms of management remove more or less nutrients from the system, so habitat management has the potential to mitigate some of the effects of atmospheric deposition. Data from a dynamic vegetation model and two field experiments are presented. The first involves nitrogen addition following different forms of habitat management. The second tests the use of habitat management to promote heathland recovery after a reduction in nitrogen deposition. Both modelling and experimental approaches suggest that plant and microbial response to nitrogen is affected by management. Shoot growth and rates of decomposition were lowest in plots managed using more intensive techniques, including mowing with litter removal and a high temperature burn. Field data also indicate that ecosystem recovery from prolonged elevated inputs of nitrogen may take many years, or even decades, even after the removal of plant and litter nitrogen stores which accompanies the more intensive forms of habitat management.