Reindeer grazing affects plants and mycelia in alpine treeline and subarctic ecosystems Frida Lindwall Uppsats för avläggande av naturvetenskaplig masterexamen i Miljövetenskap 60 hp Institutionen för biologi och miljövetenskaper, Göteborgs universitet Januari 2013
51
Embed
Reindeer grazing affects plants and mycelia in alpine ... · 2 Table of Content Reindeer!grazingaffect!plantsand!mycelia!in!alpine!and!arctic!ecosystems! 1! ... För att få djupare
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
Reindeer grazing affects plants
and mycelia in alpine treeline
and subarctic ecosystems
Frida Lindwall
Uppsats för avläggande av naturvetenskaplig masterexamen i Miljövetenskap 60 hp Institutionen för biologi och miljövetenskaper, Göteborgs universitet Januari 2013
Reindeer grazing affect plants and mycelia in alpine treeline and
subarctic ecosystems
This Master thesis consist of an introductory chapter followed by two papers:
Reindeer grazing has contrasting effect on species traits in Vaccinium vitis-idaea L. and Bistorta vivipara (L.) Gray. Frida Lindwall, Tage Vowles, Alf Ekblad and Robert G. Björk. (Submitted to Acta Oecologica)
Reindeer grazing affect the production of ectomycorrhizal mycelia in alpine tree-line ecosystems. Frida Lindwall, Tage Vowles, Alf Ekblad and Robert G. Björk (manuscript)
The native grazers on the tundra play a central role in structuring the vegetation (Oksanen,
1990) by shaping the tree line (Van Bogaert et al., 2011) and influencing the density of plants,
species composition, and species richness (Eskelinen and Oksanen, 2006; Manseau et al.,
1996; Ravolainen et al., 2011; Suominen and Olofsson, 2000; van der Wal et al., 2001;
Virtanen, 2000). Since tundra species also are adapted to exceptionally cold and harsh
conditions, rapid climate warming is likely to cause dramatic changes in the distribution of
plants and ecosystems (Callaghan et al., 2007; Elmendorf et al., 2012; Gottfried et al., 2012).
In order to gain a deeper insight into the processes that may shape our future landscapes it is
therefore important to understand how individual species respond to changes in different
environmental drivers.
There are three mechanisms by which plants are affected by grazing – defoliation,
fertilization and trampling (Sørensen et al., 2009) – and there is ample evidence to suggest
that these mechanisms, on their own or in combination, can have a profound impact on
vegetation (van der Wal, 2006). Reindeer (Rangifer tarandus L.) feed on different plant
species depending on season. Grasses, forbs and leaves of deciduous shrubs and trees
dominate their diet during the summer while during winter they mostly eat lichens (Cetraria
spp., Sterocaulon spp., and Cladonia sp.) but also tissue from evergreen dwarf shrubs such as
Empetrum nigrum ssp. hermaphroditum (Hagerup) Böcher and Vaccinium spp. (Eriksson et
al., 2007). Many tundra ecosystems are nitrogen (N) limited due to suppressed N
mineralization in the cold environment. The direct impact on vegetation caused by trampling
and browsing by herbivores may have immediate effects on the ecosystem N dynamics, but
today’s research is not conclusive on the grazing effects on N pools and availability (Cargill
and Jefferies, 1984; Gauthier et al., 1995; Johnson and Matchett, 2001; McNaughton, 1979;
Mulder, 1999; Olofsson et al., 2004; Pastor et al., 1993; Ritchie et al., 1998; Stark and
Grellman, 2002; Zhu et al., 2008). To determine plants’ internal allocation patterns of N and
C, stable isotopes analyses have frequently been used (Brooks et al., 1997; Brüggemann et al.,
2011). The heavier stable isotopes of C and N (13C and 15N) have almost exactly the same
chemical and physical properties as the lighter ones (12C and 14N), except when considering
mass. Diffusion rate is slower and chemical bonds are stronger for the heavier isotope, and
therefore heavier isotopes are discriminated against in transportation and chemical reactions;
an isotopic fractionation occurs (Brand and Coplen, 2012). Isotopes can give indications on
14
the origin of nutrients (Michelsen et al., 1996), the availability of N in soil (Craine et al.,
2009) and how nutrients and C are transported and allocated in plants (Brüggemann et al.,
2011). The C to N (C:N) ratio of the plant tissue determines the palatability of the plants
(White, 1978). It has been found that the C:N ratio in plant tissue tends to be lower when
herbivores are present, suggesting that herbivores improve plant tissue quality (Sjögersten et
al., 2011). It has also been found that plants with a high R:S ratio increase in abundance with
increased grazing pressure (Evju et al., 2009), suggesting that high R:S ratio is a tolerance
strategy against herbivores. However, no study exists on how reindeer grazing affects
individual plant species’ internal allocation of C and N in the arctic and sub-arctic region,
which is N limited (Shaver et al., 1992).
Therefore, the aim of this study was to investigate how traits of two plant species in a
sub-arctic dry heath were affected by the exclusion of reindeer. Utilising seventeen-year old
reindeer exclosures, the perennial forb Bistorta vivipara (L.) Gray and the perennial evergreen
dwarf shrub Vaccinium vitis-idaea L. were examined for responses to grazing. The traits
investigated were biomass allocation (above- and below ground), C and N content of fine
roots and leaves and their isotopic composition (δ15N and δ13C). Due to the chemical defence
in evergreen plants (Bryant et al., 1991), the higher digestibility of forbs than evergreens
(Cornelissen et al., 2004) and that reindeer seem to prefer forbs in favour of other plants due
to higher N content (Trudell and White, 1981) we hypothesise that the effect of exclosure
differs between the two species. Furthermore, we hypothesise that B. vivipara increases its
belowground biomass allocation when grazed, thus increasing the R:S ratio, whereas V. vitis-
idaea does not respond to grazing since it should be a less preferred food resource. The C and
N isotope signature in plants is hypothesised to differ between exclosures and ambient plots
due to the changes in carbon allocation patterns and nitrogen availability brought forth by
reindeer.
Materials and methods
Study site
The study site is a dry heath located at 840 m above sea level in northern Sweden (N67°46.5’
E17°32.1’). The site is situated in an all year grazing area for reindeer with an annual mean air
temperature of -1.0 °C and a precipitation of 460 mm per year. There is no information
available on the exact number of reindeer visiting the site each year. However, reindeer
droppings were counted at the site in 1997 and again in 2011. The number of droppings found
15
in 1997 (± standard deviation) was 2 076 ± 1 415 per hectare (Eriksson et al., 2007), and in
2011 it was 1667 ± 1414 (T Vowles, unpublished data). During this time the number of
reindeer owned by the adjacent Sami village, Unna Tjerusj, has fluctuated between about
4000 and 6000 (around 6000 at the time of sampling) and was even higher prior to 1997.
Thus, the grazing pressure has been fairly high and fairly constant over at least the past 15
years. Dominating plant species in the field layer are the dwarf shrubs Empetrum nigrum ssp.
hermaphroditum, Vaccinium myrtillus L. and V. vitis-idaea, the graminoids Deshampsia
flexuosa L., Calamagrostis lapponica Wahlenb. and Carex bigelowii Torr. ex Schwein.
Dominating forbs were B. vivipara, Solidago virgaurea L. and Hieracium alpina L. The
bottom layer consisted of several moss species of the genera Dicranum and Polytricum and
lichens of the genera Cladonia, Cetraria and Stereocaulon.
Sampling and processing
Six plots, 25×25 m, were established in 1996 (Eriksson et al., 2007). To exclude reindeer
three plots were surrounded by 1.7 m high fences (hereafter called exclosures) and three were
left unprotected (ambient plots). The sampling was done in mid-August 2011, where three B.
vivipara and three V. vitis-idaea were randomly collected from each plot. To standardize the
sample volume, a 125 cm3 soil cube (5×5×5 cm) were cut out from the soil and stored
separately in plastic bags and frozen (within a few hours of collection) before further
processing. After thawing, the root system was cleaned from soil (see Björk et al. (2007))
after which the above- and belowground parts of the plant were separated. The dry mass was
determined after drying for 24 h at 70°C. After weighing the above- and belowground
biomass, the leaves and the fine roots (diameter < 2 mm) were collected and weighed
separately. To get a sufficient amount of sample, the fine roots from the three samples from
each plot were put together, and ground for C and N analyses. The C and N concentrations
and isotopic composition (δ13C or δ15N = ((Rsample – Rstandard)/Rstandard) × 1000 (‰), where R is
the molar ratio 13C/12C or 15N/14N) were determined from the milled materials as described by
Boström et al. (2007).
Statistic analyses
To investigate grazing effects on R:S ratios, aboveground-, belowground- and total biomass,
each species was analysed separately using a nested ANOVA. In this analysis, treatment was
a fixed factor and plot and replications were random factors within a hierarchical design.
Because of the limited amount of fine root and leaf biomass, the three replicates were pooled
16
together to plot samples to ensure a large enough sample for C and N analysis. Thus, to
investigate treatment effects on C and N traits in fine root and leaf biomass, C:N ratio, δ13C
and δ15N, a MANOVA was used with treatment as fix factor and plot as random factor.
Furthermore, the differences in C:N ratio, C and N content, δ13C, and δ15N between species
and fine roots and leaves were determined using a nested ANOVA with species and treatment
as fixed factors and plot as random factor. However, the limited amount of fine roots of V.
vitis-idaea made it impossible to test for differences between species and grazing. All data
were, after addition of a constant, log-transformed and concomitantly scaled to unit variance
to achieve a normal distribution and to eliminate skewness and ensure homogeneity of
variances according to (Økland et al., 2001). To increase the statistical power and to reduce
the risk of making Type II errors an α = 0.10 was used to test for significant differences.
Results and discussion
The exclusion of reindeer made the leaves of B. vivipara more depleted in δ15N (P = 0.044),
whereas the leaves of V. vitis-idaea were unaffected (Fig. 1). It also seems possible that the
fine roots of V. vitis idaea were more depleted when excluding reindeer. However, the fine
roots of V. vitis-idaea was not statistically tested since we just got one pooled sample from
each treatment, but the effect size of the treatment is in the same range as for B. vivipara
leaves, 1‰. There are three possible explanations for the overall pattern of 15N enrichment in
the grazed plots. First, reindeer faeces, which are only deposited in grazed plots, are likely to
be enriched in 15N (Finstad and Kielland, 2011). Enrichment of faeces has also been found in
other organisms: 1-4 ‰ in goose faeces (Sjögersten et al., 2010), about 6‰ in faeces from
small mammals (Hwang et al., 2007), about 3‰ in goat faeces (Codron et al., 2011) and 3‰
in sheep faeces (Wittmer et al., 2010). But while faeces are enriched in 15N urine is depleted
compared to the diet. Therefore, if urine and faeces are circulated back to the soil-plant
system, our data suggest that faeces would be a central N source and also much more
important than urine for the plants targeted in this study. However, this N source is allocated
differently in the two species, since when reindeer were excluded the greatest depletion of 15N
in B. vivipara was in the leaves whereas in V. vitis-ideae it was in the roots. A second
possibility may be differences in mycorrhiza association. Hobbie and Hobbie (2006) found
that plants were more 15N depleted in symbiosis with fungi, i.e. mycorrhiza, than if not. The
transport of N through hyphae towards the plant discriminates against 15N. As a consequence,
the N remaining in the fungus is enriched in 15N, while the N transferred to the plant is 15N
depleted. Due to the lower grazing pressure on plants in exclosures it is feasible to believe that
17
more C is available to fungi inside the exclosures. With more C available, mycorrhiza
colonization may increase (Gehring and Whitham, 1991) and, consequently, more 15N
depleted N would be transported to the plants in exclosures compared to plants in the ambient
plots. However, when comparing extramatrical mycelia production at other sites in
Fennoscandia this trend was not conclusive among vegetation types (F. Lindwall, unpublished
data) and the effect of aboveground herbivores on mycorrhizal colonization and mycelia
production is still unclear (Gehring and Whitham, 2002). Thirdly, Schulze et al. (1994) found
that the δ15N increases with the soil depth, and since the whole root system was not examined
in this study, it is possible that plants in the ambient plots had deeper root systems than in the
exclosures. However, the δ15N signature follows the same pattern between treatments for both
species but the belowground biomass does not. Furthermore, earlier studies (Björk et al.,
2007; Michelsen et al., 1996) have found that the majority of the root system of arctic plants
is in the organic layer, usually only a few centimetres thick in dry tundra heaths, making the
third explanation unlikely. Thus, we conclude that the most likely explanation for the 15N
enriched plants in grazed plots is the addition of 15N enriched droppings into the system, but
changes in ectomycorrhizal mycelia production or community change cannot be ruled out.
Furthermore, we found a higher (P = 0.099) amount of C in the leaves of B. vivipara in
exclosures than in ambient plots (Tab. 2), suggesting that the plants protected from grazers are
able to allocate more C to the leaves. The R:S ratio in B. vivipara tends (P = 0.16) to be
higher in ambient plots, and as Evju et al. (2009) argue, an increase of C to the roots is a
tolerance strategy against herbivory. If plants are protected against herbivory, C may instead
be allocated to the leaves. If there also is a sufficient amount of N, the productivity inside the
exclosures will increase (Larcher, 2003). However in contrast with earlier studies that showed
an increased abundance and height of forbs in exclosures (Bråthen and Oksanen, 2001;
Oksanen and Moen, 1994; Pajunen et al., 2008), we found no effect on total, above ground or
below ground biomass or on total abundance (T Vowles, unpublished data) in either of the
species (Tab. 1), and also, no significant grazing effect on total C and N content in V. vitis-
idaea leaves. The effect of reindeer on primary production and soil nutrient availability is not
clear and contrasting results from earlier studies show both negative (Stark and Grellman,
2002) and positive effects (Johnson and Matchett, 2001) of grazing on the available N. The
lemming population peak this year, was one of the largest for decades and had a large impact
on the alpine areas in Sweden. This may explain the lack of treatment effect on biomass, and
even though we found treatment effects in C and N traits in B. vivipara, the differences may
18
have been larger if lemmings also were excluded (Olofsson et al., 2012). Despite potential
interfering lemming effects, our results, as hypothesized, show that individual plant species
respond differently to reindeer grazing, which also will have implications on how species will
respond to environmental change in sub-arctic ecosystems.
Moreover, there was a clear difference in δ15N, δ13C, C:N ratio, and C, N and biomass
allocation patterns between the species, under ambient conditions (see Tab. 2 and Fig. 1). The
C:N ratio, which is about twice as high in V. vitis-idaea than in B. vivipara (P < 0.001),
indicates that B. vivipara is, as hypothesised, of higher quality as forage. The C:N ratio is in
general higher in evergreens than deciduous plants because evergreens have higher
concentrations of lignin and other secondary C substances (often used as defense substance
against grazing) than plants with shorter leaf lifespan (Aerts, 1995). The δ15N was
significantly lower (P < 0.001) for B. vivipara than for V. vitis-idaea, especially in the leaves.
The δ15N in leaves of both plants were more depleted than the fine roots, which was also
found by Emmerton et al. (2001), and may be explained by an internal discrimination of 15N
that occurs between roots and shoots (Brüggemann et al., 2011). Thus, above- and
belowground plant parts should both be taken into account when considering plant isotope
composition. There is also a difference in δ13C in the leaves between the two species, where
B. vivipara is more depleted (P = 0.057), which is consistent with results shown by Brooks et
al. (1997). They explained the differences between plant functional types as an effect of
different photosynthetic rates and different degrees of discrimination against 13C in the
assimilation of carbon dioxide (CO2). Differences in boundary layer and stomatal- and
internal conductance (Brüggemann et al., 2011; Warren, 2007) may occur between the two
species and there are also different responses between plant functional groups due to
environmental factors (Brooks et al., 1997; Warren, 2007). The difference in δ13C and δ15N
between fine roots and leaves is larger for B. vivipara than V. vitis-idaea. This pattern is
probably a result of differences in rate and amount of assimilated C between deciduous plants
and evergreens (Warren, 2007), but also species-specific internal fractioning of isotopes
between shoot and roots (Brüggemann et al., 2011; Dawson et al., 2002). Thus, the different
δ13C and δ 15N signatures between species highlight the species specificity in the fractionation
process occurring in both photosynthesis and N transfer between the plant and its fungal
symbiont. The lower C:N ratio found in B. vivipara supports the hypothesis that reindeer
should prefer to eat B. vivipara over V. vitis-idaea, although V. vitis-idaea is also affected by
the reindeers, which may indicate a rather high grazing pressure in this area.
19
Concluding remarks
The herbivore population has important effects on plant carbon and nitrogen dynamics. The
higher δ15N signatures in the ambient plots are probably an indication that faeces, which are 15N enriched, are an important N source for plants in tundra ecosystems. The C:N ratio
supports the idea that B. vivipara is a food resource of higher quality than V. vitis-idaea.
There was a larger difference in the isotope signature between roots and shoots in B. vivipara
than V. vitis-idaea, which is probably a result of differences in internal isotopic fractionation
and photosynthetic rate between plant functional types. Clearly, reindeer do affect plant traits
either directly, by removing and destroying biomass, or indirectly, by addition of nitrogen via
faeces. For deeper insights into how reindeer are affecting nutrient dynamics in tundra
ecosystems, more mechanistic studies (e.g. isotopic labelling studies) are needed to separate
the processes behind the species-specific differences in allocation patterns found in this study.
Acknowledgements
The authors thank Mathias Molau and Paloma Alvarez Blanco for their assistance in the field.
The Swedish Research Council for Environment, Agricultural Sciences and Spatial Planning
(grant no 214-2010-1411 to RGB) supported this work and is gratefully acknowledged. The
work was also conducted with financial support from the strategic research area BECC
(Biodiversity and Ecosystems in a Changing Climate; www.cec.lu.se/research/becc).
20
References
Aerts, R., 1995. The advantages of being evergreen. Trends Ecol. Evol. 10, 402-407.
Björk, R.G., Majdi, H., Klemedtsson, L., Lewis-Jonsson, L., Molau, U., 2007. Long-term warming effects on root morphology, root mass distribution, and microbial activity in two dry tundra plant communities in northern Sweden. New Phytol. 176, 862-873.
Boström, B., Comstedt, D., Ekblad, A., 2007. Isotope fractionation and C-13 enrichment in soil profiles during the decomposition of soil organic matter. Oecologia 153, 89-98.
Brand, W.A., Coplen, T.B., 2012. Stable isotope deltas: tiny, yet robust signatures in nature. Isotopes Environ. Health Stud. 48, 393-409.
Brooks, J.R., Flanagan, L.B., Buchmann, N., Ehleringer, J.R., 1997. Carbon isotope composition of boreal plants: Functional grouping of life forms. Oecologia 110, 301-311.
Bryant, J.P., Provenza, F.D., Pastor, J., Reichardt, P.B., Clauser, T.P., du Toit, J.T., 1991. Interactions between woody plants and browsing mammals mediated by secondary metabolites. Annu. Rev. Ecol. Syst. 22, 431-446.
Brüggemann, N., Gessler, A., Kayler, Z., Keel, S.G., Badeck, F., Barthel, M., Boeckx, P., Buchmann, N., Brugnoli, E., Esperschütz, J., et al., 2011. Carbon allocation and carbon isotope fluxes in the plant-soil-atmosphere continuum: a review. Biogeosciences 8, 3457-3489.
Bråthen, K.A., Oksanen, J., 2001. Reindeer reduce biomass of preferred plant species. Journal of vegetation science 12, 473-480.
Callaghan, T.V., Björn, L., Chapin, F.S., Chemov, Y., Christensen, T.R., Huntley, B., Ims, R.A., Johansson, M., Oechel, W.C., Panikov, N., et al., 2007. Arctic tundra and polars desert ecosystems, Arctic Climate Impact Assessment scientific report. Cambridge University Press, University of Alaska Fairbanks, pp. 243-352.
Cargill, S.M., Jefferies, R.L., 1984. The Effects of Grazing by Lesser Snow Geese on the Vegetation of a Sub- Arctic Salt Marsh. J. Appl. Ecol. 21, 669-686.
Codron, D., Sponheimer, M., Codron, J., Hammer, S., Tschuor, A., Braun, U., Bernasconi, S.M., Clauss, M., 2011. Tracking the fate of digesta 13C and 15N compositions along the ruminant gastrointestinal tract: Does digestion influence the relationship between diet and faeces? European Journal of Wildlife Research 58, 303-313.
Cornelissen, J.H., Quested, H.M., Gwynn Jones, D., van Logtestijn, R.S., De Beus, M.A.H., Kondratchuk, A., Callaghan, T.V., Aerts, R., 2004. Leaf Digestibility and Litter Decomposability Are Related in a Wide Range of Subarctic Plant Species and Types. Funct. Ecol. 18, 779-786.
Craine, J.M., Elmore, A.J., Aidar, M.P., Bustamante, M., Dawson, T.E., Hobbie, E.A., Kahmen, A., Mack, M.C., McLauchlan, K.K., Michelsen, A., et al., 2009. Global patterns of foliar nitrogen isotopes and their relationships with climate, mycorrhizal fungi, foliar nutrient concentrations, and nitrogen availability. New Phytol. 183, 980-992.
Elmendorf, S.C., Henry, G.H., Hollister, R.D., Björk, R.G., Bjorkman, A.D., Callaghan, T.V., Collier, L.S., Cooper, E.J., Cornelissen, J.H., Day, T.A., et al., 2012. Global assessment of experimental climate warming on tundra vegetation: heterogeneity over space and time. Ecol. Lett. 15, 164-175.
Emmerton, K.S., Callaghan, T.V., Jones, H.E., Leake, J.R., Michelsen, A., Read, D.J., 2001. Assimilation and isotopic fractionation of nitrogen by mycorrhizal fungi. New Phytol. 151, 503-511.
Eriksson, O., Niva, M., Caruso, A., 2007. Use and abuse of reindeer range, Uppsala.
Eskelinen, A., Oksanen, J., 2006. Changes in the abundance, composition and species richness of mountain vegetation in relation to summer grazing by reindeer Journal of vegetation science 17, 245-254.
Evju, M., Austrheim, G., Halvorsen, R., Mysterud, A., 2009. Grazing responses in herbs in relation to herbivore selectivity and plant traits in an alpine ecosystem. Oecologia 161, 77-85.
Finstad, G.L., Kielland, K., 2011. Landscape Variation in the Diet and Productivity of Reindeer in Alaska Based on Stable Isotope Analyses. Arctic, Antarctic, and Alpine Research 43, 543-554.
Gauthier, G., Hugher, J.R., Reed, A., Beaulieu, J., PRochefort, L., 1995. Effects of grazing by greater snow geese on the production of graminoids at an arctic site (Bylot island, NWT, Canada). J. Ecol. 83, 653-664.
Gehring, C.A., Whitham, T.G., 2002. Mycorrhizae-Herbivore Interactions: Population and Community Consequences, in: Heijden, V.d. (Ed.), Mycorrhizal Ecology. Springer, Berlin Heidelberg.
Gottfried, M., Pauli, H., Futschik, A., Akhalkatsi, M., Barancok, P., Alonso, J.L.B., Coldea, G., Dick, J., Erschbamer, B., Calzado, M.R.F., et al., 2012. Continent-wide response of mountain vegetation to climate change. Nat. Clim. Chang. 2, 111-115.
Hobbie, J.E., Hobbie, E.A., 2006. 15N in symbiotic fungi and plants estimates nitrogen and carbon flux rates in arctic tundra. Ecology 87, 816-822.
Hwang, Y.T., Millar, J.S., Longstaffe, F.J., 2007. Do δ15N and δ13C values of feces reflect the isotopic composition of diets in small mammals? Can. J. Zool. 85, 388-396.
Larcher, W., 2003. Physiological Plant Ecology, 4 ed. Springer, New York.
Manseau, M., Huot, J., Crete, M., 1996. Effects of summer grazing by caribou on composition and productivity of vegetation: Community and landscape level. J. Ecol. 84, 503-513.
McNaughton, S.J., 1979. Grazing as an Optimization Process: Grass-Ungulate Relationships in the Serengeti. The American Naturalist 113, 691-703.
Michelsen, A., Schmidt, I., K., Jonasson, S., Quarmby, C., Sleep, D., 1996. Leaf 15N of subarctic plants provides field evidence that ericoid, ectomycorrhizal and non- and arbuscular mycorrhizal species access different sources of soil nitrogen. Oecologia 105, 53-63.
22
Mulder, C.P.H., 1999. Vertebrate herbivores and plants in the Arctic and subarctic: effects on individuals, populations, communities and ecosystems. Perspectives in Plant Ecology, Evolution and Systematics 2, 29-55.
Økland, R.H., Økland, T., Rydgren, K., 2001. Vegetation-environment relationships of boreal spruce swamp forest in Østmarka nature reserve, SE Norway. Somerfeltia 29, 1-190.
Oksanen, L., Moen, J., 1994. Species-specific plant responses to exclusion of grazers in three Fennoscandian tundra habitats. Ecoscience 1, 31-39.
Olofsson, J., Stark, S., Oksanen, L., 2004. Reindeer influence on ecosystem processes in the tundra. Oikos 105, 386-396.
Olofsson, J., Tømmervik, H., Callaghan, T.V., 2012. Vole and lemming activity observed from space. Nat. Clim. Chang. DOI: 10.1038/nclimate1537.
Pajunen, A., Virtanen, R., Roininen, H., 2008. The effects of reindeer grazing on the composition and species richness of vegetation in forest–tundra ecotone. Polar Biol. 31, 1233-1244.
Pastor, J., Dewey, B., Naiman, R.J., McInnes, P.F., Cohen, Y., 1993. Moose Browsing and Soil Fertility in the Boreal Forests of Isle Royale National Park. Ecology 74, 467-480.
Ravolainen, V.T., Brathen, K.A., Ims, R.A., Yoccoz, N.G., Henden, J.-A., Killengreen, S.T., 2011. Rapid, landscape scale responses in riparian tundra vegetation to exclusion of small and large mammalian herbivores. Basic Appl. Ecol. 12, 643-653.
Ritchie, M.E., Tilman, D., Knops, J.M.H., 1998. Herbivore Effects on Plant and Nitrogen Dynamics in Oak Savanna. Ecology 79, 165-177.
Schulze, E.D., Chapin, F.S., Gebauer, G., 1994. Nitrogen nutrition and isotope differences among life forms at the northern treeline of Alaska. Oecologia 100, 406-412.
Shaver, G.R., Billings, W.D., Chapin, F.S., Giblin, A.E., Nadelhoffer, K.J., Oechel, W.C., Rastetter, E.B., 1992. Global change and the carbon balance of arctic ecosystems.
Sjögersten, S., Kuijper, D.P.J., Wal, R., Loonen, M.J.J.E., Huiskes, A.H.L., Woodin, S.J., 2010. Nitrogen transfer between herbivores and their forage species. Polar Biol. 33, 1195-1203.
Sjögersten, S., Wal, R., Loonen, M.J.J.E., Woodin, S.J., 2011. Recovery of ecosystem carbon fluxes and storage from herbivory. Biogeochemistry 106, 357-370.
Stark, S., Grellman, D., 2002. Soil microbial responses to herbivory in an arctic tundra heath at two levels of nutrient availability. Ecology 83, 2736-2744.
Suominen, O., Olofsson, J., 2000. Impacts of semi-domesticated reindeer on structure of tundra and forest communities in Fennoscandia: a review. Ann. Zool. Fenn. 37, 233-249.
Sørensen, L.I., Mikola, J., Kytöviita, M.-M., Olofsson, J., 2009. Trampling and Spatial Heterogeneity Explain Decomposer Abundances in a Sub-Arctic Grassland Subjected to Simulated Reindeer Grazing. Ecosystems 12, 830-842.
Trudell, J., White, R.G., 1981. The effect of forage structure and availability on food-intake, biting rate, bite size and daily eating time of reindeer. J. Appl. Ecol. 18, 63-81.
23
Van Bogaert, R., Haneca, K., Hoogesteger, J., Jonasson, C., De Dapper, M., Callaghan, T.V., 2011. A century of tree line changes in sub-Arctic Sweden shows local and regional variability and only a minor influence of 20th century climate warming. J. Biogeogr. 38, 907-921.
van der Wal, R., 2006. Do herbivores cause habitat degradation or vegetation state transition? Evidence from the tundra. Oikos 114, 177-186.
van der Wal, R., van Lieshout, S.M.J., Loonen, M., 2001. Herbivore impact on moss depth, soil temperature and arctic plant growth. Polar Biol. 24, 29-32.
Virtanen, R., 2000. Effects of grazing on above-ground biomass on a mountain snowbed, NW Finland. Oikos 90, 295-300.
Warren, C.R., 2007. Stand aside stomata, another actor deserves centre stage: the forgotten role of the internal conductance to CO2 transfer. J. Exp. Bot. 59, 1475-1487.
White, T.C.R., 1978. <white 1978.pdfThe importance of a relative shortage of food in animal ecology. Oecologia 33, 71-86.
Wittmer, M.H.O.M., Auerswald, K., Schönbach, P., Bai, Y., Schnyder, H., 2010. 15N fractionation between vegetation, soil, faeces and wool is not influenced by stocking rate. Plant Soil 340, 25-33.
Zhu, Z.H., Lundholm, J., Li, Y., Wang, X., 2008. Response of Polygonum viviparum species and community level to long-term livestock grazing in alpine shrub meadow in Qinghai-Tibet Plateau. J Integr Plant Biol 50, 659-672.
24
Figure 1. δ13C (a) and δ15N (b) in leaves and fine roots in Vaccinium vitis-idaea and Bistorta vivipara (n = 3)
exposed (Ambient) or not exposed (Exclosures) to grazing by reindeer. The error bars represent standard error.
** P < 0.05 indicates significant difference between treatments.
25
Table 1. Total biomass, above- and belowground biomass and root: shoot (R:S) ratio for the species Bistorta
vivipara and Vaccinium vitis-idaea in ambient plots (exposed to grazing by reindeer) and exclosures (not
exposed reindeer). Values show mean and standard error (±SE) for n = 3 plots
a. The fine root samples of V. vitis-idaea were, because of the limited amount of sample, merged into one
pooled sample, making it impossible to test for differences between species and grazing.
27
Manuscript
Text pages: 18, Tables: 2, Figures: 3
Reindeer grazing affects the production of ectomycorrhizal mycelia in
alpine treeline ecosystems
Frida Lindwalla, Tage Vowlesa, Alf Ekbladb, Robert G. Björkc,2
aDepartment of Biological and Environmental Sciences, University of Gothenburg, P.O. Box 461, SE-405 30
Gothenburg, Sweden. bSchool of Science and Technology, Örebro University, SE-701 82 Örebro, Sweden. cDepartment of Earth Sciences, University of Gothenburg, Gothenburg, Sweden
2 Corresponding author: Department of Earth Sciences, University of Gothenburg, P.O. Box
Deschampsia flexuosa L., Hieracium alpine L. and Trientalis europaea L. and the bottom
layer is dominated by Cladonia spp and Cetraria islandica (L.) Ach., The birch forest heath
type with mosses is a low forest dominated by B. pubescens L. with undergrowth of dwarf
shrubs, Vaccinium sp., and E. nigrum and grasses, D. flexuosa and Nardus stricta as well as
herbs, T. europea, Linnea borealis L. and Solidago virgaurea L. In the bottom layer mosses,
such as Pleurozium schreberi L. and Barbilophozia lycopodioides (Wall.) Loeske, are more
common than lichens. Since there are no Sami villages nearby Fulufjället, reindeer from
Sweden never visit this site (Eriksson et al., 2007). However, on rare occasion small groups
of reindeers from Norway cross the border and visit the site, but the grazing pressure from
reindeer is negligible. Nevertheless, the moose population on Fulufjället is, according to the
Swedish Environmental Protection Agency, very large (Naturvårdsverket, 2002), and the
grazing pressure in the birch forest, a preferred forage species (Wam & Hjeljord, 2010) is
therefore assumed to be large.
Långfjället is also located in the southern part of the mountain range, just 400 km north of
Fulufjället. The vegetation is similar to Fulufjället although the birch forest also contains
Pinus sylvestris L. In summer time reindeer frequently graze this site. The abundance of
Betula nana, the utterly dominant ectomycorrhizal plant in the field at this site, was on the dry
heath higher in exclosures compared to ambient plots. The dry heath is located on an east-
facing slope.
Poullanvare is the northernmost sampling site located about 50 km north of the town
Kiruna. The dry heath is situated in a north-westerly slope and the vegetation is similar to the
other two mountains with the main difference that C. vulgaris does not grow here but the
grass Calamagrostis lapponica (Wahlenb.) Hartm. is common. Unfortunately, due to sabotage
of the fences, reindeer were not properly excluded at this site. Instead the herd was enclosed
and the grazing pressure inside the exclosures was extremely large early in the season.
In each plot, four ingrowths bags (mesh size 50 µm; length 10 cm and diameter 2 cm)
(also used and explained in detail by Wallander et al. (2001)), were, in early season (see table
1 for details) placed in the soil (0-10 cm soil depth). The bags contained 40 g pure sand (with
no nutrients), burned in 550°C for 24 hours to ensure that no carbon remained, allowing only
33
mycorrhizal mycelia to grow. In fall 2011 the bags were collected and frozen within a few
hours before further processing.
Biomass and stable isotope analyses
To obtain an even distribution the sand from the four ingrowths bags was mixed and 80 g, of
totally 160 g, was randomly collected. The sand was extracted in water and mycelia and
rhizomorphs floating in the water were collected with tweezers and freeze-dried. To get the
water content of the sand, the remaining sand was freeze-dried and then weighed for
compensation in the biomass estimation (see below). When dry, the extracted mycelia were
further rinsed from sand grains before analysed for C concentrations and C and N isotopic
composition as described by Boström et al. (2007). The isotopic composition is expressed in
the standard notation (δ13C or δ15N) in parts per thousand (‰) relative to the international
standards, N in air or C in Vienna Pee Dee Belemnite (δ13C or δ15N = ((Rsample –
Rstandard)/Rstandard) × 1000 (‰), where R is the molar ratio 13C/12C or 15N/14N) (Högberg et al.,
1999). The production (g C m-2) of extramatrical mycelia (EMM) was calculated as
𝐸𝑀𝑀 =!!"" × !% ×
!!"!!!"#! !!"#∗!%
! (Eq. 1)
where mEMM is the EMM dry mass in g, C% is the carbon content in % of EMM dry weight,
mtot is the total weight in g of the sampled sand (160g in this study), mext is the weight of the
extracted wet sand (i.e. 80 g in this study), w% is the sand water content in % and A is the
tube area (i.e. 0.00063 m2 in this study).
PLFA, NLFA and ergosterol
Ten grams of freeze-dried sand from each sample was used for PLFAs analysis. To compare
the PLFA and ergosterol content in the mycelia extracted sand with the sand not extracted
from mycelia, both types were analysed. Also totally new, unused sand was examined. Thirty-
six mycelia extracted samples, 36 non-extracted samples and 5 pure sand samples were
analysed, making a total of 77 samples. Lipids were extracted using the method described by
Frostegård et al. (1991) with some small modifications. In each 10 g sample di-nonadecanoyl
phosphatidylcholine was added as an internal standard. Extracted lipids were fractionated
using a silicic column. The first, chloroform, fraction, was further analysed for NLFA 16:1ω5
(biomarker for arbuscular fungi) and ergosterol while the third, methanol fraction was further
34
analysed for PLFA 18:1ω9 and 18:2ω6,9 (biomarker for fungi). The phospholipids were
transesterified with 0.5 M sodium methoxide, which was analysed on a gas chromatograph
(GC) with a 30 m DB-5 column and a flame ionization detector.
The ergosterol was extracted from the NLFA fraction, by adding pyridine and BSTFA
and incubated for 10 min in 60 °C. The NLFA fraction was further processed; sodium-
metoxide and heptan was added and the samples were sonicated for 30 min. HAc and water
was added and the upper phase was dried and dissolved in acetonitrile. All samples were
analysed in a GC with a 30 m DB-5 column and a flame ionization detector.
Data analyses
Significant differences between exclosures and ambient plots were tested with two-sample t-
test using R (R version 2.15.1). To achieve a normal distribution all data were log transformed
and to increase the statistical power and to reduce the risk of making Type II errors (i.e. to
accept the null hypothesis, even though it is false) an α = 0.10 was used to test for significant
differences.
Results
EMM production
In general, the EMM production was significantly higher (P = 0.042) in the birch forests (2.0
± 0.4 SE g C m-2) compared to the dry heaths (1.6 ± 0.3 g C m-2). The EMM production in the
birch forest on Fulufjället and Långfjället was significantly higher (P < 0.05) in the exclosures
compared to the ambient plots, 4 and 2 times, respectively. No treatment effect was found in
the birch forest on Poullanvare. On the dry heaths, only Långfjället showed a significant
difference (P = 0.065) in EMM production between the treatments. However, in contrast to
the birch forest the ambient plots had 3 times higher EMM production than exclosures. No
treatment effect was found on the dry heath on Fulufjället or Poullanvare.
Ergosterol
Not all samples were successfully extracted, resulting in a limited amount of data of
ergosterol content, and no PLFA or NLFA data. Since no replications were extracted from the
sites no statistical analysis was done. However, the results of ergosterol are shown in Fig. 3
35
and the average conversion factor between ergosterol and mycelia was 5.5 ± 1.25 SE mg/g C,
calculated from the extracted ergosterol in 10 g of sand and the weighed mycelia in 80 g sand.
Isotopic compositions
In general the EMM sampled in the ambient plots in the birch forests were 1.2 ‰ more 13C
depleted compared to the ambient plots on the dry heaths (P = 0.042). Furthermore, the EMM
were significantly 1.0‰ (P = 0.078) and 1.2‰ (P = 0.025) more 13C depleted in exclosures in
the birch forest on Fulufjället and Poullanvare dry heath, respectively, compared to the
ambient plots (Tab. 2). No treatment effect on 13C in EMM was found in the birch forest or
the dry heath at Långfjället.
There was no general difference in 15N-signature between the vegetation types and the
treatment effect varied between sites and vegetation types (Tab. 2). The EMM in dry heath
exclosures on Fulufjället were significantly 1.7‰ (P = 0.032) more 15N-depleted in exclosures
than ambient plots while the opposite was found on Långfjället where EMM in the dry heath
exclosures were significantly 1.5‰ (P = 0.080) more enriched compared to the ambient plots.
No treatment effect on 15N in EMM was found in the birch forest or the dry heath at
Poullanvare.
Discussion
EMM production
This study demonstrates, for the first time, that aboveground grazing has an effect on EMM
production in an alpine treeline ecosystem. The EMM production is overall very low, as
expected in this low productive environment, with a maximum EMM production of 6.1 g C
m-2 and an average of 2.0 g C m-2 in the birch forests and 1.6 g C m-2 on the dry heaths,
respectively. Wallander et al. (2001) report an hyphal production of 29.5 and 21 g C m-2 in a
Norway spruce forest and a mixed oak–spruce forest respectively and EMM production is
reported to vary between 8 and 50 g m-2 in a review by Ekblad et al. (2012). These findings
show a much higher production than seen in the alpine treeline ecosystem in the present
study. However, our results are in line with Clemmensen et al. (2006), who showed a biomass
production of 1.3 g m-2 on a dry heath in subarctic Sweden. The larger production of EMM in
the birch forest than on the dry heaths is most likely due to the different dominating plant
species and their ability to support the fungi with C. The ectomycorrhizal B. pubescens
36
dominate the forest while B. nana is the dominating ectomycorrhizal species in the dry heath.
B. pubescens can because of the higher production in forests than dry heaths (Hartley et al.,
2012) and because of lower reindeer grazing pressure due to the higher stature of trees than
shrubs, transport more C to the fungi than B. nana, which explains the higher production.
Within the birch forest an effect of reindeer exclusion was found on both Långfjället and
Fulufjället, where the EMM production was higher in exclosures. Since plants in exclosures
are protected from heavy grazing pressure, more of the assimilated C can be transported to the
fungi, and thereby increase the production of EMM and, thus, the nitrogen intake to the plant.
The loss of aboveground biomass due to grazing may reduce the amount of assimilated C
transported to the roots and fungi (Trent et al., 1988), which may explain the lower mycelia
production in ambient plots in the birch forest on Långfjället. However, the effect seen on
Fulufjället is, most likely, not an effect of reindeer grazing since reindeer rarely visit this site.
Instead, the pattern is probably due to a difference in the abundance of birch trees, with more
trees in exclosures, which simply is due to an unluckily plot selection and not an effect of
grazing. Also the effect may partly be due to grazing by moose (Alces alces) which
population has been reported, by the Swedish Environmental Protection Agency, to be very
large on Fulufjället (Naturvårdsverket, 2002). The EMM production did not differ between
treatments in the birch forest at Poullanvare, which probably is an effect of an autumnal moth
(Epirrita autumnata L.) outbreak in 1995 (Karlsson & Weih, 2003) and 2004 (Klemola et al.,
2008). The population of E. autumnata fluctuates with periods of 9-10 years and does serious
damage on the foliage of mountain birch in northern Fennoscandia (Tanhuanpää et al., 2002).
This outbreak often occurs in mature forests, leaving young trees and stands less damaged
(Ruohomaki et al., 1997; Tenow et al., 2004), changing the forest structure, making the effect
of reindeer grazing negligible. Between 1995 and 2011 the amount of mountain birch stems
increased in both exclosures and ambient plots in the birch forest on Poullanvare. However,
the average height of the trees decreased (Eriksson et al., 2007; T. Vowles, unpublished data),
suggesting a rejuvenation of the birch forest. Younger trees allow higher EMM production
than old trees (Wallander et al., 2010). Thus, the rejuvenation of the forest may explain the
somewhat higher EMM production in Poullanvare than Långfjället and Fulufjället (Tab. 2).
Also, no damage on vegetation due to reindeer grazing was observed and the dropping count
was low at this site (T. Vowles, unpublished data). Thus, the lack of effect by excluding
reindeer may partly be explained by a fairly low grazing pressure.
37
The situation is the opposite on the dry heath, were the dominated EcM plant species is B.
nana. The EMM production was, as seen in Långfjället, positively affected by grazing. There
is a trend that B. nana increases in abundance and height when protected from grazing, (Fig. 1
and 2), which is in line with other exclosure studies (Eskelinen & Oksanen, 2006; Oksanen &
Moen, 1994; Pajunen et al., 2008). In ambient plots, where grazers suppress B. nana, C is
allocated to the roots as a tolerance strategy against herbivory (Evju et al., 2009) allowing
higher EMM production (Treseder & Allen, 2000). Hence, less EMM production in
exclosures is probably due to an investment of C in aboveground tissue, due to the release of
stress from grazing, and less to roots. On the other hand, on Fulufjället, where no reindeer
graze, the EMM production seems to increase with height of B. nana. Thus, our data suggest
that B. nana invest in roots when affected by herbivores, and when released from the stress C
is allocated to aboveground tissue until it reaches a certain height, where the investment is
directed to both above- and belowground plant parts.
Furthermore, the input of N, derived from faeces, in ambient plots may also affect the
production of EMM. For example, Nilsson and Wallander (2003) and Kjøller et al. (2012)
found that EMM production decreased with fertilization. This pattern was explained by a
change in EcM community structure but also as a decrease of fine root production when
plants are fertilized, and thereby a reduction in the mycorrhiza colonization and also the
production of mycelia. It has also been shown that nitrogen fertilization had a small effect on
the belowground EcM community (Deslippe et al., 2011; Treseder et al., 2007; Wallenda &
Kottke, 1998) and Clemmensen et al. (2006) found an increase of EMM production during
fertilization. Lilleskov et al. (2002) found that some EcM fungal species increased linearly
with net nitrification rate in soil, some species were found only at sites with high nitrification
rate and some species were nitrophobic, i.e. their abundance increased with decreasing
nitrification rate. Hence, it is very likely that the fungal taxa and abundance differ between the
treatments and a fungal species analysis is required for accurate conclusions about the effect
of added N.
To conclude the above-discussed results this study has shown that EMM production has
responded differently to grazing at the different sites. This variance comes from different host
plant species inhabiting these areas that respond differently to grazing and with different
supporting abilities. Our results, as hypothesised, show that grazing has an impact on the
production of mycelia, because it affects the allocation patterns of C in plants and the N
38
availability. The patterns are not consistent throughout the different vegetation types and we
conclude that the surrounding environment and the host plant species is important for the
response in fungi. Also, there are 5000-6000 different EcM fungal species (Agerer, 2006) and
it is not very likely that all of those species respond in the same way to changes in their
surroundings (Deslippe et al., 2011). Thus, studies covering the response of treatment effects
on fungal species composition are needed to truly understand the link between aboveground
herbivores and the fungi.
Isotopic signature
In general the EMM in the dry heaths were more 13C-enriched than in the birch forests, which
most likely is due to drier soils at the dry heaths. Drier conditions impacts the stomatal
conductance, as stomata closes, changing the isotope signature toward the 13C value of air; i.e.
drought is making the assimilated C less depleted (Brüggemann et al., 2011). Drier conditions
may also decrease photosynthesis, which leads to more internal carbon being used and a
change in the isotopic signature to less 13C depleted occurring. It is also possible that the more 13C-depleted EMM found in exclosures in the birch forest on Fulufjället is an effect of grazing
by moose (Alces alces L.), which prefer birch as forage (Wam & Hjeljord, 2010). The moose
population on Fulufjället is very large (Naturvårdsverket, 2002) and the grazing pressure in
the birch forest is assumed to be high. As the photosynthetic capacity in each individual birch
tree declines with an increased grazing pressure, the isotopic signature of C in the tree may
change. When the input of C highly enriched in 12C decreases, due to grazing, the plant will
become less depleted in 13C with time. Thus, mycelia forming mycorrhiza with plants affected
by grazing will receive less depleted C, as in the ambient plots (Brüggemann et al., 2011).
This pattern was not found in the forest on Långfjället, suggesting that moose grazing may be
influential on the isotopic signature in mountain birch forests.
Furthermore, excluding grazing did not consistently alter the 15N signature of the mycelia
across sites. In the dry heaths, the EMM was more 15N-depleted in exclosures on Fulufjället,
while on Långfjället exclosures were more 15N-enriched. Several factors can affect the isotope
signature; the fungal species composition (Clemmensen et al., 2006; Emmerton et al., 2001;
Hobbie & Colpaert, 2003), soil microbial activity (Clemmensen et al., 2008), availability of
different N-forms in the soil (Emmerton et al., 2001; Hobbie & Colpaert, 2003), degree of
mycorrhizal colonization and magnitude of C flux to fungi (Hobbie & Colpaert, 2003). Since
39
none of them can solely explain the 15N pattern in this study it is likely that several of them
are interacting in different ways to create the pattern found in our study.
Ergosterol
Varying ergosterol content was found at the different sites, and it does not correspond very
well to the weighed mycelia biomass (Fig. 3). The conversion factor of 5.5 mg/g C in EMM is
slightly lower than found and used elsewhere (Clemmensen et al., 2006; Salmanowicz &
Nylund, 1988; Wallander et al., 2001) and there is a very large variance in the data. Thus, the
interpretation of these results should be made with caution. It has been shown that different
results are obtained if using a GC-MS or a high performance liquid chromatographer (HPLC)
(Olsson et al., 2003). But also, if using the same instrument the results may vary a lot, even
within the same species; Frey et al. (1994) found an ergosterol content of 0.063 mg/g mycelia
of Glomus intraradices, while Fontaine et al. (2001) studying the same species did not find
any ergosterol at all. Also, as Olsson et al. (2003) concludes; there are always taxonomic
differences in the substances used as biomarkers. Since it is very likely that the fungal species
compositions will change with shifting environmental conditions, and if the variation in
ergosterol content between species is not addressed, it may not be trustful to use as a
biomarker for comparison between treatments. Furthermore, some samples where the mycelia
had already been extracted, contained fairly high amounts of ergosterol indicating that some
mycelia was left behind when handpicked from sand. Hence, both methods have
shortcomings.
Concluding remarks
This study confirms that grazing affects the production of EMM and, consequently, has an
impact on the carbon budget in soil. As reported by Hartley et al. (2012) there is a potential
loss of C associated with an altered amount of trees and shrubs in the arctic, due to less carbon
storage and fast turnover. Our data suggest that reindeer restrain the otherwise expanding
cover of shrubs (Sturm et al., 2001), as also reported by Olofsson et al. (2009), but also affect
the internal C allocation patterns leading to an increase of the EMM production in the dry
heaths. Thus, as the mycelium is the dominant pathway of C to the SOM pool (Godbold et al.,
2006), reindeer grazing could, due to an increased EMM production, mitigate the increased
CO2 concentration in the atmosphere. This study highlights the importance of taking
herbivores into account when trying to address the impact of climate change on alpine treeline
40
ecosystem. Questions that remain unanswered are how the fungal species composition is
affected by aboveground grazing, if the fungal species composition affects the efficiency of
the symbiosis and also how different fungal species respond to enhance nitrogen availability
due to grazing. These questions are fundamental when trying to understand the carbon and
nitrogen dynamics and transports between herbivores, plants and fungi, and require further
research. A DNA analyses together with more mechanistic studies (i.e. isotope labeling
studies) are needed for an accurate conclusion on the response to grazing by the fungi.
Acknowledgment
The authors thank Kjell Vowles for his assistance in the field. The Swedish Research Council
for Environment, Agricultural Sciences and Spatial Planning (grant no 214-2010-1411 to
RGB) supported this work and is gratefully acknowledged. The work was also conducted with
financial support from the strategic research area BECC (Biodiversity and Ecosystems in a
Changing Climate; www.cec.lu.se/research/becc).
41
References
Agerer, R. (2006). Fungal relationships and structural identity of their ectomycorrhizae. Mycological Progress, 5(2), 67-107. doi: 10.1007/s11557-006-0505-x
Allen, M. F. (1991). The Ecology of Mycorrhizae (Vol. 6): Cambridge University press.
Allen, M. F., Swenson, W., Querejeta, J. I., Egerton-Warburton, L. M., & Treseder, K. K. (2003). Ecology of mycorrhizae: A conceptual framework for complex interactions among plants and fungi. [Review]. Annual Review of Phytopathology, 41, 271-303. doi: 10.1146/annurev.phyto.41.052002.095518
Boström, B., Comstedt, D., & Ekblad, A. (2007). Isotope fractionation and C-13 enrichment in soil profiles during the decomposition of soil organic matter. Oecologia, 153(1), 89-98. doi: 10.1007/s00442-007-0700-8
Brüggemann, N., Gessler, A., Kayler, Z., Keel, S. G., Badeck, F., Barthel, M., . . . Bahn, M. (2011). Carbon allocation and carbon isotope fluxes in the plant-soil-atmosphere continuum: a review. Biogeosciences, 8(11), 3457-3489. doi: 10.5194/bg-8-3457-2011
Bråthen, K. A., & Oksanen, J. (2001). Reindeer reduce biomass of preferred plant species. Journal of vegetation science, 12(4), 473-480. doi: 10.2307/3236999
Cairney, J. W. G. (1999). Gaining water and nutrients: root function. In B. J. Atwell, P. E. Kriedemann & C. G. N. Turnbull (Eds.), Plants in action- Adaption in Nature, Performance in cultivation (pp. 664). Australia: Macmillan Education Australia PTY Ltd.
Clemmensen, K. E., Michelsen, A., Jonasson, S., & Shaver, G. R. (2006). Increased ectomycorrhizal fungal abundance after long-term fertilization and warming of two arctic tundra ecosystems. [Article]. New Phytologist, 171(2), 391-404. doi: 10.1111/j.1469-8137.2006.01778.x
Clemmensen, K. E., Sorensen, P. L., Michelsen, A., Jonasson, S., & Strom, L. (2008). Site-dependent N uptake from N-form mixtures by arctic plants, soil microbes and ectomycorrhizal fungi. [Research Support, Non-U.S. Gov't]. Oecologia, 155(4), 771-783. doi: 10.1007/s00442-008-0962-9
Deslippe, J. R., Hartmann, M., Mohn, W. W., & Simard, S. W. (2011). Long-term experimental manipulation of climate alters the ectomycorrhizal community of Betula nana in Arctic tundra. Global Change Biology, 17(4), 1625-1636. doi: 10.1111/j.1365-2486.2010.02318.x
Djajakirana, G., Joergensen, R. G., & Meyer, B. (1996). Ergosterol and microbial biomass relationship in soil. Biology and Fertility of Soils, 22(4), 299-304. doi: 10.1007/bf00334573
Ekblad, A., Wallander, H., Godbold, D. L., Cruz, C., Baldrian, P., Björk, R. G., . . . Plassard, C. (2012). The production and turnover of extramatrical mycelium of ectomycorrhizal fungi in forest soils: role in carbon cycling
Emmerton, K. S., Callaghan, T. V., Jones, H. E., Leake, J. R., Michelsen, A., & Read, D. J. (2001). Assimilation and isotopic fractionation of nitrogen by mycorrhizal fungi. New Phytologist, 151, 503-511.
Eriksson, O., Niva, M., & Caruso, A. (2007). Use and abuse of reindeer range (Vol. 87). Uppsala.
Eskelinen, A., & Oksanen, J. (2006). Changes in the abundance, composition and species richness of mountain vegetation in relation to summer grazing by reindeer Journal of vegetation science, 17, 245-254.
42
Evju, M., Austrheim, G., Halvorsen, R., & Mysterud, A. (2009). Grazing responses in herbs in relation to herbivore selectivity and plant traits in an alpine ecosystem. [Research Support, Non-U.S. Gov't]. Oecologia, 161(1), 77-85. doi: 10.1007/s00442-009-1358-1
Finlay, R. D. (2008). Ecological aspects of mycorrhizal symbiosis: with special emphasis on the functional diversity of interactions involving the extraradical mycelium. [Research Support, Non-U.S. Gov't
Fontaine, J., Grandmougin-Ferjani, A., Hartmann, M. A., & Sancholle, M. (2001). Sterol biosynthesis by the arbuscular mycorrhizal fungus Glomus intraradices. [Article]. Lipids, 36(12), 1357-1363. doi: 10.1007/s11745-001-0852-z
Frey, B., Vilarino, A., Schuepp, H., & Arines, J. (1994). Chitin and ergosterol content of extraradical and intraradical mycelium of the vesicular-arbuscular mycorrhizal funguf Glomus-intraradices Soil Biology & Biochemistry, 26(6), 711-717. doi: 10.1016/0038-0717(94)90263-1
Frostegård, Å., Tunlid, A., & Bååth, E. (1991). Microbial biomass measured as total lipid phosphate in soils of different organic content Journal of Microbiological Methods, 14(3), 151-163. doi: 10.1016/0167-7012(91)90018-l
Gange, A. C., Bower, E., & Brown, V. K. (2002). Differential effects of insect herbivory on arbuscular mycorrhizal colonization. Oecologia, 131(1), 103-112. doi: 10.1007/s00442-001-0863-7
Gehring, C. A., & Whitham, T. G. (1994). Interactions between aboveground berbivores and the mycorrhizal mutualists of plants. TREE, 9(7), 251-255.
Gehring, C. A., & Whitham, T. G. (2002). Mycorrhizae-Herbivore Interactions: Population and Community Consequences. In V. d. Heijden (Ed.), Mycorrhizal Ecology (Vol. 157). Berlin Heidelberg: Springer.
Godbold, D. L., Hoosbeek, M. R., Lukac, M., Cotrufo, M. F., Janssens, I. A., Ceulemans, R., . . . Peressotti, A. (2006). Mycorrhizal hyphal turnover as a dominant process for carbon input into soil organic matter. [Article]. Plant and Soil, 281(1-2), 15-24. doi: 10.1007/s11104-005-3701-6
Hartley, A. E., Garnett, M. H., Sommerkorn, M., Hopkins, D. W., Fletcher, B. J., Sloan, V. L., . . . Wookey, P. A. (2012). A potentail loss of Carbon associated with greater plant growth in the European arctic. Nature Climate Change, 2(2), 875-879. doi: 10.1038
Hobbie, E. A., & Colpaert, J. V. (2003). Nitrogen availability and colonization by mycorrhizal fungi correlate with nitrogen isotope patterns in plants. New Phytologist, 157(1), 115-126. doi: 10.1046/j.1469-8137.2003.00657.x
Högberg, P., Plamboeck, A. H., Taylor, A. F. S., & Fransson, P. M. A. (1999). Natural C-13 abundance reveals trophic status of fungi and host-origin of carbon in mycorrhizal fungi in mixed forests. Proceedings of the National Academy of Sciences of the United States of America, 96(15), 8534-8539. doi: 10.1073/pnas.96.15.8534
Karlsson, P. S., & Weih, M. (2003). Long-term patterns of leaf, shoot and wood production after insect herbivory in the Mountain Birch. Functional Ecology, 17(6), 841-850. doi: 10.1111/j.1365-2435.2003.00792.x
Kjøller, R., Nilsson, L.-O., Hansen, K., Schmidt, I. K., Vesterdal, L., & Gundersen, P. (2012). Dramatic changes in ectomycorrhizal community composition, root tip abundance and mycelial production along a stand-scale nitrogen deposition gradient. New Phytologist, 194(1), 278-286. doi: 10.1111/j.1469-8137.2011.04041.x
43
Klemola, T., Andersson, T., & Ruohomaeki, K. (2008). Fecundity of the autumnal moth depends on pooled geometrid abundance without a time lag: implications for cyclic population dynamics. Journal of Animal Ecology, 77(3), 597-604. doi: 10.1111/j.1365-2656.2008.01369.x
Lilleskov, E. A., Fahey, T. J., Horton, T. R., & Lovett, G. M. (2002). Belowground ectomycorrhizal fungal community change over a nitrogen deposition gradient in Alaska. Ecology, 83(1), 104-115. doi: 10.2307/2680124
Manseau, M., Huot, J., & Crete, M. (1996). Effects of summer grazing by caribou on composition and productivity of vegetation: Community and landscape level. journal of ecology, 84(4), 503-513.
Montgomery, H. J., Monreal, C. M., Young, J. C., & Seifert, K. A. (2000). Determination of soil fungal biomass from soil ergosterol analyses. Soil Biology & Biochemistry, 32(8-9), 1207-1217. doi: 10.1016/s0038-0717(00)00037-7
Naturvårdsverket. (2002). Skötselplan Fulufjällets nationalpark. Uppsala: Lindblom & Co Retrieved from http://www.naturvardsverket.se/Documents/publikationer/620-5246-6.pdf.
Nilsson, L. O., & Wallander, H. (2003). Production of external mycelium by ectomycorrhizal fungi in a Norway spruce forest was reduced in response to nitrogen fertilization. [Article]. New Phytologist, 158(2), 409-416. doi: 10.1046/j.1469-8137.2003.00728.x
Oksanen, L., & Moen, J. (1994). Species-specific plant responses to exclusion of grazers in three Fennoscandian tundra habitats. Ecoscience, 1(1), 31-39.
Olofsson, J., Oksanen, L., Callaghan, T., Hulme, P. E., Oksanen, T., & Suominen, O. (2009). Herbivores inhibit climate-driven shrub expansion on the tundra. Global Change Biology, 15(11), 2681-2693. doi: 10.1111/j.1365-2486.2009.01935.x
Olsson, P. A., Larsson, L., Bago, B., Wallander, H., & van Aarle, I. M. (2003). Ergosterol and fatty acids for biomass estimation of mycorrhizal fungi. New Phytologist, 159(1), 7-10. doi: 10.1046/j.1469-8137.2003.00810.x
Pajunen, A., Virtanen, R., & Roininen, H. (2008). The effects of reindeer grazing on the composition and species richness of vegetation in forest–tundra ecotone. Polar Biology, 31(10), 1233-1244. doi: 10.1007/s00300-008-0462-8
Parrent, J. L., Morris, W. F., & Vilgalys, R. (2006). CO2-enrichment and nutrient availability alter ectomycorrhizal fungal communities. ecology, 87(9), 2278-2287. doi: 10.1890/0012-9658(2006)87[2278:canaae]2.0.co;2
R version 2.15.1. (2012). R: A language and environment for statistical computing. Vienna, Austria: R foundation for statistical computing. Retrieved from www.R-project.org/
Ricklefs, R. E. (2008). The economy of nature (6 ed.). USA: W. H. Freeman and Company.
Rousseau, J. V. D., Sylvia, D. M., & Fox, A. J. (1994). Contribution of Ectomycorrhiza to the Potential Nutrient-Absorbing Surface of Pine. New Phytologist, 128(4), 639-644. doi: 10.1111/j.1469-8137.1994.tb04028.x
Ruohomaki, K., Virtanen, T., Kaitaniemi, P., & Tammaru, T. (1997). Old mountain birches at high altitudes are prone to outbreaks of Epirrita autumnata (Lepidoptera: Geometridae). Environmental Entomology, 26(5), 1096-1104.
44
Ruotsalainen, A. L., & Eskelinen, A. (2011). Root fungal symbionts interact with mammalian herbivory, soil nutrient availability and specific habitat conditions. Oecologia, 166(3), 807-817.
Salmanowicz, B., & Nylund, J. E. (1988). High performance liquid chromatography determination of ergosterol a measure of ectomycorrhiza infection in scots pine. European Journal of Forest Pathology, 18(5), 291-298.
Simard, S. W., Jones, W. S., & Durall, D. M. (2002). Carbon and nutrient fluxes within and between mycorrhizal plants. In M. G. A. van der Heijden & I. Sanders (Eds.), Mycorrhizal Ecology (Vol. 157). Berlin Heidenberg: Springer.
Smith, S., & Read, D. J. (2008). Ectomycorrhizas Mycorrhizal symbiosis (3 ed., pp. 189-349). San Diego: Academic press.
Sturm, M., Racine, C., & Tape, K. (2001). Climate change - Increasing shrub abundance in the Arctic. [Article]. Nature, 411(6837), 546-547. doi: 10.1038/35079180
Talbot, J. M., Allison, S. D., & Treseder, K. K. (2008). Decomposers in disguise: mycorrhizal fungi as regulators of soil C dynamics in ecosystems under global change. Functional Ecology, 22(6), 955-963. doi: 10.1111/j.1365-2435.2008.01402.x
Tanhuanpää, M., Ruohomäki, K., Turchin, P., Ayres, M. P., Bylund, H., Kaitaniemi, P., . . . Haukioja, E. (2002). Population cycles of the autumnal moth in Fennoscandia. In A. A. Berryman (Ed.), Population Cycles: The case of trophic interaction (pp. 192). New york, USA: Oxford Univerrsity press.
Tenow, O., Bylund, H., Karlsson, P. S., & Hoogesteger, J. (2004). Rejuvenation of a mountain birch forest by an Epirrita autumnata (Lepidoptera : Geometridae) outbreak. Acta Oecologica-International Journal of Ecology, 25(1-2), 43-52. doi: 10.1016/j.actao.2003.10.006
Trent, J. D., Wallace, L. L., Svejcar, T. J., & Christiansen, S. (1988). Effect of grazing on growth, carbohydrate pools, and mycorrhizae in winter-wheat Canadian Journal of Plant Science, 68(1), 115-120.
Treseder, K. K., & Allen, M. F. (2000). Mycorrhizal fungi have a potential role in soil carbon storage under elevated CO2 and nitrogen deposition. New Phytologist, 147(1), 189-200. doi: 10.1046/j.1469-8137.2000.00690.x
Treseder, K. K., Turner, K. M., & Mack, M. C. (2007). Mycorrhizal responses to nitrogen fertilization in boreal ecosystems: potential consequences for soil carbon storage. [Review]. Global Change Biology, 13(1), 78-88. doi: 10.1111/j.1365-2486.2006.01279.x
Wallander, H., Ekblad, A., Godbold, D. L., Johnson, D. A., Bahr, A., Baldrian, P., . . . Rudawska, M. (2012). Evaluation of methodes to estimate production, biomass and turnover of ectomycorrhizal mycelium i forest soils- A review. Soil Biology & Biochemistry, in press.
Wallander, H., Johansson, U., Sterkenburg, E., Brandstrom Durling, M., & Lindahl, B. D. (2010). Production of ectomycorrhizal mycelium peaks during canopy closure in Norway spruce forests. [Research Support, Non-U.S. Gov't]. New Phytol, 187(4), 1124-1134. doi: 10.1111/j.1469-8137.2010.03324.x
Wallander, H., Nilsson, L., Ola., Hagerberg, D., & Bååth, E. (2001). Estimation of the biomass and seasonal growth of external mycelium of ectomycorrhizal fungi in the field. New Phytologist, 151, 753-760.
Wallenda, T., & Kottke, I. (1998). Nitrogen deposition and ectomycorrhizas. [Article; Proceedings Paper]. New Phytologist, 139(1), 169-187. doi: 10.1046/j.1469-8137.1998.00176.x
45
Wam, H. K., & Hjeljord, O. (2010). Moose summer and winter diets along a large scale gradient of forage availability in southern Norway. European Journal of Wildlife Research, 56(5), 745-755. doi: 10.1007/s10344-010-0370-4
Zhao, X. R., Lin, Q., & Brookes, P. C. (2005). Does soil ergosterol concentration provide a reliable estimate of soil fungal biomass? [Article]. Soil Biology & Biochemistry, 37(2), 311-317. doi: 10.1016/j.soilbio.2004.07.041
46
Table 1. Coordinates, meters above sea level (m.a.s.l), Temperature measured at 2 cm depth during the time the
bags were in the soil, annual precipitation (Precip.) and days the sand bags were in the soil are given for the six
sites on three different mountains Fulufjället (FU), Långfjället (LO) and Poullanvare (PO) and two different
vegetation types, birch forest heath with mosses (BM) and dry heath (DH).
Site Coordinates m.a.s.l Temp (C°) Precip. (mm) Days in soil
FU-‐BM N 61°38'42.2" E012°35'24.5"
880 9.1 834 148
FU-‐DH N 61°38'08.1" E012°38'18.9"
930 8.5
LO-‐BM N 62°03'55.5" E012°14'45.9"
800 8.6 697 148
LO-‐DH N 62°06'49.8" E012°16'20.1"
840 8.4
PO-‐BM N 68°20'13.2" E021°19'15.6" 460 7.6
444 112 PO-‐DH
N 68°20'23.7" E021°10'41.0" 580 -
47
Table 2. The average EMM production (g C m-2), δ15N and δ13C in ambient plots (Amb.) and exclosures (Exc.)
on the three mountains, Fulufjället (FU), Långfjället (LO) and Poullanvare (PO), and two vegetation types, birch
forest heath type with mosses (BM) and dry heath (DH). The bold numbers show a significant difference
between treatments. Standard error is given in brackets. * P < 0.1 and ** P < 0.05