Does warming by open-top chambers induce change in the root-associated fungal community of the arctic dwarf shrub Cassiope tetragona (Ericaceae)?

Climate change may alter mycorrhizal communities, which impact ecosystem characteristics such as carbon sequestration processes. These impacts occur at a greater magnitude in Arctic ecosystems, where the climate is warming faster than in lower latitudes. Cassiope tetragona (L.) D. Don is an Arctic plant species in the Ericaceae family with a circumpolar range. C. tetragona has been reported to form ericoid mycorrhizal (ErM) as well as ectomycorrhizal (ECM) symbioses. In this study, the fungal taxa present within roots of C. tetragona plants collected from Svalbard were investigated using DNA metabarcoding. In light of ongoing climate change in the Arctic, the effects of artificial warming by open-top chambers (OTCs) on the fungal root community of C. tetragona were evaluated. We detected only a weak effect of warming by OTCs on the root-associated fungal communities that was masked by the spatial variation between sampling sites. The root fungal community of C. tetragona was dominated by fungal groups in the Basidiomycota traditionally classified as either saprotrophic or ECM symbionts, including the orders Sebacinales and Agaricales and the genera Clavaria, Cortinarius, and Mycena. Only a minor proportion of the operational taxonomic units (OTUs) could be annotated as ErM-forming fungi. This indicates that C. tetragona may be forming mycorrhizal symbioses with typically ECM-forming fungi, although no characteristic ECM root tips were observed. Previous studies have indicated that some saprophytic fungi may also be involved in biotrophic associations, but whether the saprotrophic fungi in the roots of C. tetragona are involved in biotrophic associations remains unclear. The need for more experimental and microscopy-based studies to reveal the nature of the fungal associations in C. tetragona roots is emphasized.

No divergence in Cassiope tetragona: persistence of growth response along a latitudinal temperature gradient and under multi-year experimental warming.

BACKGROUND AND AIMS: The dwarf shrub Cassiope tetragona (Arctic bell-heather) is increasingly used for arctic climate reconstructions, the reliability of which depends on the existence of a linear climate-growth relationship. This relationship was examined over a high-arctic to sub-arctic temperature gradient and under multi-year artificial warming at a high-arctic site. METHODS: Growth chronologies of annual shoot length, as well as total leaf length, number of leaves and average leaf length per year, were constructed for three sites. Cassiope tetragona was sampled near its cold tolerance limit at Ny-Ålesund, Svalbard, at its assumed climatic optimum in Endalen, Svalbard, and near its European southern limit at Abisko, Sweden. Together these sites represent the entire temperature gradient of this species. Leaf life span was also determined. Each growing season from 2004 to 2010, 17 open top chambers (OTCs) were placed near Ny-Ålesund, thus increasing the daily mean temperatures by 1·23°C. At the end of the 2010 growing season, shoots were harvested from OTCs and control plots, and growth parameters were measured. KEY RESULTS: All growth parameters, except average leaf length, exhibited a linear positive response (R(2) between 0·63 and 0·91) to mean July temperature over the temperature gradient. Average leaf life span was 1·4 years shorter in sub-arctic Sweden compared with arctic Svalbard. All growth parameters increased in response to the experimental warming; the leaf life span was, however, not significantly affected by OTC warming. CONCLUSIONS: The linear July temperature-growth relationships, as well as the 7 year effect of experimental warming, confirm that the growth parameters annual shoot length, total leaf length and number of leaves per year can reliably be used for monitoring and reconstructing temperature changes. Furthermore, reconstructing July temperature from these parameters is not hampered by divergence.

Ecophysiological response of Crambe maritima to airborne and soil-borne salinity.

BACKGROUND AND AIMS: There is a need to evaluate the salt tolerance of plant species that can be cultivated as crops under saline conditions. Crambe maritima is a coastal plant, usually occurring on the driftline, with potential use as a vegetable crop. The aim of this experiment was to determine the growth response of Crambe maritima to various levels of airborne and soil-borne salinity and the ecophysiological mechanisms underlying these responses. METHODS: In the greenhouse, plants were exposed to salt spray (400 mM NaCl) as well as to various levels of root-zone salinity (RZS) of 0, 50, 100, 200 and 300 mM NaCl during 40 d. The salt tolerance of Crambe maritima was assessed by the relative growth rate (RGR) and its components. To study possible salinity effects on the tissue and cellular level, the leaf succulence, tissue Na(+) concentrations, Na(+) : K(+) ratio, net K(+)/Na(+) selectivity, N, P, K(+), Ca(2+), Mg(2+), proline, soluble sugar concentrations, osmotic potential, total phenolics and antioxidant capacity were measured. KEY RESULTS: Salt spray did not affect the RGR of Crambe maritima. However, leaf thickness and leaf succulence increased with salt spray. Root zone salinities up to 100 mM NaCl did not affect growth. However, at 200 mM NaCl RZS the RGR was reduced by 41 % compared with the control and by 56 % at 300 mM NaCl RZS. The reduced RGR with increasing RZS was largely due to the reduced specific leaf area, which was caused by increased leaf succulence as well as by increased leaf dry matter content. No changes in unit leaf rate were observed but increased RZS resulted in increased Na(+) and proline concentrations, reduced K(+), Ca(2+) and Mg(2+) concentrations, lower osmotic potential and increased antioxidant capacity. Proline concentrations of the leaves correlated strongly (r = 0.95) with RZS concentrations and not with plant growth. CONCLUSIONS: Based on its growth response, Crambe maritima can be classified as a salt spray tolerant plant that is sensitive to root zone salinities exceeding 100 mM NaCl.

Do high levels of diffuse and chronic metal pollution in sediments of Rhine and Meuse floodplains affect structure and functioning of terrestrial ecosystems?

This paper (re)considers the question if chronic and diffuse heavy metal pollution (cadmium, copper, lead and zinc) affects the structure and functioning of terrestrial ecosystems of Biesbosch National Park, the floodplain area of rivers Meuse and Rhine. To reach this aim, we integrated the results of three projects on: 1. the origin, transfer and effects of heavy metals in a soil-plant-snail food chain; 2. the impact of bioavailability on effects of heavy metals on the structure and functioning of detritivorous communities; 3. the risk assessment of heavy metals for an herbivorous and a carnivorous small mammal food chain. Metal pollution levels of the Biesbosch floodplain soils are high. The bioavailability of metals in the soils is low, causing low metal levels in plant leaves. Despite this, metal concentrations in soil dwelling detritivores and in land snails at polluted locations are elevated in comparison to animals from 'non-polluted' reference sites. However, no adverse effects on ecosystem structure (species richness, density, biomass) and functioning (litter decomposition, leaf consumption, reproduction) have been found. Sediment metal pollution may pose a risk to the carnivorous small mammal food chain, in which earthworms with elevated metal concentrations are eaten by the common shrew. Additional measurements near an active metal smelter, however, show reduced leaf consumption rates and reduced reproduction by terrestrial snails, reflecting elevated metal bioavailability at this site. Since future management may also comprise reintroduction of tidal action in the Biesbosch area, changes in metal bioavailability, and as a consequence future ecosystem effects, cannot be excluded.

Depletion of stratospheric ozone over the Antarctic and Arctic: responses of plants of polar terrestrial ecosystems to enhanced UV-B, an overview.

Depletion of stratospheric ozone over the Antarctic has been re-occurring yearly since 1974, leading to enhanced UV-B radiation. Arctic ozone depletion has been observed since 1990. Ozone recovery has been predicted by 2050, but no signs of recovery occur. Here we review responses of polar plants to experimentally varied UV-B through supplementation or exclusion. In supplementation studies comparing ambient and above ambient UV-B, no effect on growth occurred. UV-B-induced DNA damage, as measured in polar bryophytes, is repaired overnight by photoreactivation. With UV exclusion, growth at near ambient may be less than at below ambient UV-B levels, which relates to the UV response curve of polar plants. UV-B screening foils also alter PAR, humidity, and temperature and interactions of UV with environmental factors may occur. Plant phenolics induced by solar UV-B, as in pollen, spores and lignin, may serve as a climate proxy for past UV. Since the Antarctic and Arctic terrestrial ecosystems differ essentially, (e.g. higher species diversity and more trophic interactions in the Arctic), generalization of polar plant responses to UV-B needs caution.

Growth and nitrogen fixation of legumes at increased salinity under field conditions: implications for the use of green manures in saline environments.

The use of legumes as green manure can potentially increase crop productivity in saline environments and thus contribute to the sustainability of agricultural systems. Here, we present results from a field experiment conducted in the Netherlands that addressed the efficiency of nitrogen (N) fixation by a legume at varying salinities. We grew Melilotus officinalis in an agricultural field using drip irrigation with water salinity varying in electrical conductivity between 1.7 and 20 dS m(-1). In the experiment, nearly 100 % of total plant N in M. officinalis was derived from symbiotic fixation at all but the highest salinity level (20 dS m(-1)). Our results indicated that this species derived substantial amounts of N via symbiotic fixation, the N becoming available in the soil (and thus available to crops) when cultivated legumes senesce and decompose. Based on the growth performance of M. officinalis and its ability to fix N at moderate soil salinity in our field experiments, we identified this species as a promising source for green manure in saline agriculture in temperate regions.

Physiology and ecologic relevance of salt secretion by the salt gland of Glaux maritima L.

The effects of salinity in the root medium, time, and relative humidity on the salt secretion of Glaux maritima were investigated. Both in the greenhouse and in the field increasing salinity stimulated sodium and chloride secretion, whereas the essential elements potassium, calcium, and magnesium remained at low secretion levels, which might be interpreted as efficient mineral economy. The low secretion level of potassium is remarkable, because growing on a nutrient solution containing 6 mM potassium, the concentration of the plant sap increases to 150 mM K+ and the secreted quantity amounts to only 2 m mol l-1 plant sap 24 h-1.Attempts were made to establish the secretion rate. The maximum secretion rate calculated may be 80 pEq NaCl cm-2 s-1, but for long periods (days) the secretion rate will be lower. Measurement of salt secretion unavoidably leads to removal of secreted salt. Salt was removed by rinsing with distilled water, which artificially accelerates the secretion process or parts of it by diffusion of salt from the cuticle cavity or secretory cells. At increasing salinities the amount of secreted ions showed a fivefold increase, whereas the osmotic potential of the plant sap was raised only twofold, indicating the importance of secretion as a rapid regulation mechanism with regard to the salt economy.

Differences in proton pumping and Na/H exchange at the leaf cell tonoplast between a halophyte and a glycophyte.

The tonoplast Na(+)/H(+) antiporter and tonoplast H(+) pumps are essential components of salt tolerance in plants. The objective of this study was to investigate the transport activity of the tonoplast Na(+)/H(+) antiporter and the tonoplast V-H(+)-ATPase and V-H(+)-PPase in a highly tolerant salt-accumulating halophyte, Salicornia dolichostachya, and to compare these transport activities with activities in the related glycophyte Spinacia oleracea. Vacuolar membrane vesicles were isolated by density gradient centrifugation, and the proton transport and hydrolytic activity of both H(+) pumps were studied. Furthermore, the Na(+)/H(+)-exchange capacity of the vesicles was investigated by 9-amino-6-chloro-2-methoxyacridine fluorescence. Salt treatment induced V-H(+)-ATPase and V-H(+)-PPase activity in vesicles derived from S. oleracea, whereas V-H(+)-ATPase and V-H(+)-PPase activity in S. dolichostachya was not affected by salt treatment. Na(+)/H(+)-exchange capacity followed the same pattern, i.e. induced in response to salt treatment (0 and 200 mM NaCl) in S. oleracea and not influenced by salt treatment (10 and 200 mM NaCl) in S. dolichostachya. Our results suggest that S. dolichostachya already generates a high tonoplast H(+) gradient at low external salinities, which is likely to contribute to the high cellular salt accumulation of this species at low external salinities. At high external salinities, S. dolichostachya showed improved growth compared with S. oleracea, but V-H(+)-ATPase, V-H(+)-PPase and Na(+)/H(+)-exchange activities were comparable between the species, which might imply that S. dolichostachya more efficiently retains Na(+) in the vacuole.

Comparing salt tolerance of beet cultivars and their halophytic ancestor: consequences of domestication and breeding programmes.

Salt tolerance of higher plants is determined by a complex set of traits, the timing and rate of evolution of which are largely unknown. We compared the salt tolerance of cultivars of sugar beet and their ancestor, sea beet, in hydroponic studies and evaluated whether traditional domestication and more recent breeding have changed salt tolerance of the cultivars relative to their ancestor. Our comparison of salt tolerance of crop cultivars is based on values of the relative growth rate (RGR) of the entire plant at various salinity levels. We found considerable salt tolerance of the sea beet and slightly, but significantly, reduced salt tolerance of the sugar beet cultivars. This indicates that traditional domestication by selection for morphological traits such as leaf size, beet shape and size, enhanced productivity, sugar content and palatability slightly affected salt tolerance of sugar beet cultivars. Salt tolerance among four sugar beet cultivars, three of which have been claimed to be salt tolerant, did not differ. We analysed the components of RGR to understand the mechanism of salt tolerance at the whole-plant level. The growth rate reduction at higher salinity was linked with reduced leaf area at the whole-plant level (leaf area ratio) and at the individual leaf level (specific leaf area). The leaf weight fraction was not affected by increased salinity. On the other hand, succulence and leaf thickness and the net assimilation per unit of leaf area (unit leaf rate) increased in response to salt treatment, thus partially counteracting reduced capture of light by lower leaf area. This compensatory mechanism may form part of the salt tolerance mechanism of sea beet and the four studied sugar beet cultivars. Together, our results indicate that domestication of the halophytic ancestor sea beet slightly reduced salt tolerance and that breeding for improved salt tolerance of sugar beet cultivars has not been effective.

Heavy metal pollution affects consumption and reproduction of the landsnail Cepaea nemoralis fed on naturally polluted Urtica dioica leaves.

This study is one of the very first that investigates the effects of heavy metal pollution on food consumption and reproduction of terrestrial snails under semi-realistic field conditions. Two experiments were carried out using snails (Cepaea nemoralis) and food (Urtica dioica leaves) from different metal polluted locations and one reference location. The first experiment showed that both polluted and reference snails fed on high-metal leaves from a highly polluted location had significantly lower consumption rates than snails consuming leaves from the reference location. In the second experiment, snails from both locations used in the consumption experiment and from two low-polluted locations were kept on native soil and food in order to reproduce. No negative effect of heavy metal pollution on clutch size was found for the snails from the reference location and the low-polluted locations. Snails from the highly polluted location laid no eggs. This suggests that at high levels of metal pollution, reproduction is strongly negatively affected. We suggest that the absence of egg laying by snails from the highly polluted location results from a combination of decreased consumption and an increased demand of energy for the accumulation and detoxification of metals (decreased scope for growth).

Development of a proxy for past surface UV-B irradiation: a thermally assisted hydrolysis and methylation py-GC/MS method for the analysis of pollen and spores.

A method was developed for the analysis of the UV-absorbing sporopollenin monomers p-coumaric acid and ferulic acid in very low numbers of pollen. This enables the analysis of pollen or spores from cultured plants, from herbarium collections, and from sediment, soil, and peat cores. The method involves thermally assisted hydrolysis and methylation using tetramethylammonium hydroxide combined with gas chromatography and mass spectrometry. Pyrolysis, gas chromatographic, and mass spectrometric conditions were optimized for the analysis of minimal amounts of pollen. The method has a detection limit of approximately 60 fresh pollen of Alnus glutinosa and a relative standard deviation of approximately 10% between 100 and 600 pollen.