Fungal depsidones – an inducible or constitutive defence against herbivores in the lichen Lobaria pulmonaria?

Herbivore-deterrent depsidones in the epiphytic lichen Lobaria pulmonaria were quantified after a 104-day exposure to indigenous lichen-feeding mollusc communities in broadleaved deciduous forests in southeastern Norway. Controls and acetone-rinsed living thalli were transplanted under open and shaded tree canopies. Rinsed thalli had their depsidone concentration reduced to 36% of the pre-rinsing level, which is below the level needed to deter grazing molluscs. Grazing did not raise the concentration of depsidones beyond the level occurring in control to which molluscs had no access. Inducible responses were not detected in controls nor in acetone-rinsed thalli. Depsidone resynthesis was negligible in acetone-rinsed thalli regardless of grazing and/or light regimes. Our results suggest that C-based depsidones represent a constitutive type of herbivore defence in L. pulmonaria. A constitutive defence is probably an advantage for stress-tolerant slow-growing lichens inhabiting habitats with a constant presence of generalist invertebrate herbivores.     

Mollusc grazing limits growth and early development of the old forest lichen Lobaria pulmonaria in broadleaved deciduous forests

This study aims: (1) to quantify mollusc grazing on juvenile and mature thalli of the foliose epiphytic lichen Lobaria pulmonaria, and (2) to test the hypothesis inferring a herbivore defensive role of lichen depsidones in forests with indigenous populations of lichen-feeding molluscs. Lichens were transplanted in shaded and less shaded positions in each of two calcareous broadleaved deciduous forests, one poor in lichens, one with a rich Lobarion community. Preventing the access of molluscs significantly reduced the loss of juvenile L. pulmonaria, particularly in the naturally lichen-poor forest. Molluscs also severely grazed mature thalli in the lichen-poor forest, especially thalli placed under the more shading canopies. Furthermore, reducing the natural concentration of depsidones by pre-rinsing with acetone increased subsequent grazing significantly, showing that lichen depsidones function as herbivore defence in natural habitats. Our results suggest that mollusc grazing may play important roles in shaping the epiphytic vegetation in calcareous deciduous forests, and that recently established juvenile L. pulmonaria thalli seem to be particularly vulnerable.     

Removal of secondary compounds increases invertebrate abundance in lichens

We investigated how lichen carbon-based secondary compounds (CBSCs) affect abundance of invertebrates in five lichen species growing on the forest floor (Cladonia rangiferina, Cladonia stellaris) or on tree trunks (Evernia prunastri, Hypogymnia physodes, Pseudevernia furfuracea). To do this, CBSCs were removed by rinsing lichen thalli in acetone (which has no adverse effects on the lichens) and the lichens were re-transplanted in their natural habitat. After 4 months there was higher abundance of mites, springtails and spiders in the three epiphytic lichens that had their CBSC concentrations reduced. The increase in predatory spiders following CBSC reduction suggests that the compounds have multitrophic consequences. The acetone treatment reduced the number of nematodes in four of the lichen species. Given that lichens serve as important habitats for a diverse range of invertebrates, increased knowledge of how lichen CBSCs may regulate their abundance helps us to better understand the role that lichens and their defence compounds play in structuring forest food webs.     

Ultrastructure of snail grazing damage to calcicolous lichens

The identification of damaged lichens is often difficult due to changes in the morphology of regenerating specimens. We examined the Ultrastructure of grazing damages to four species of calcicolous lichens ( Aspicilia calcarea, Physcia adscendens, Tephromela atra and Xanthoria parietina ) and free-living cyanobacteria (family Chroococcaceae) caused by individuals of four species of land snails ( Chondrina clienta, Balea perversa, Clausilia bidentata and Helicigona lapicida ). We also investigated the radular structure of the four lichen-feeding snails to examine whether differences in radular morphology result in species-specific grazing damages. Individuals of all four snail species removed the cyanobacteria layer covering the limestone or lichen surfaces. The four lichen species were grazed to a different extent by the different snail species. SEM-images showed that B. perversa left distinct depressions on the thalli of A. calcarea , whereas H. lapicida grazed off the thalli of this lichen rather evenly. Both snail species left visible radular traces on the lichen surface. In contrast, Ch. clienta left shallow depressions without radular traces on the thalli of A. calcarea. In Tephromela atra , grazing damages were observed on both thallus and ascocarp. Ascocarps of T. atra were partly grazed by B. perversa. Helicigona lapicida grazing on T. atra removed more or less evenly the entire lichen tissue including the ascocarps. In foliose lichens, grazing by Ch. clienta, B. perversa and Cl. bidentata resulted in depressions of different depths, while H. lapicida removed entire pieces of the thalli. In general, radular traces were less distinct in foliose lichens than in crustose lichens.     

Carbon-based secondary compounds in the lichen Hypogymnia physodes deter detrivorous woodlice

Woodlice are not widely recognized as lichen-feeding invertebrates. We sought to discover whether the woodlouse, Porcellio scaber, could feed on the lichen Hypogymnia physodes and if the lichen carbon-based secondary compounds CBSCs would reduce grazing. We cut lichen thalli in two pieces, one was non-destructively rinsed in acetone to remove CBSCs; the other served as a control. Both pieces were fed to woodlice in a choice experiment. The CBSC concentration of individual thalli ranged from 3 to 19%. The woodlice grazed all pieces, but preferred the acetone-rinsed pieces, depending on the amount of CBSCs present in the non-extracted counterpart. The woodlice were not deterred from feeding on samples with CBSC concentrations ≤5%, which corresponded to natural contents in shade-adapted thalli. This suggested that P. scaber tolerates this amount of compounds at least in the short-term. In conclusion, P. scaber can feed on H. physodes, but CBSCs deter them from feeding.     

To eat or not to eat—relationship of lichen herbivory by snails with secondary compounds and field frequency of lichens

Aims The biochemical defense of lichens against herbivores and its relationship to lichen frequency are poorly understood. Therefore, we tested whether chemical compounds in lichens act as feeding defense or rather as stimulus for snail herbivory among lichens and whether experimental feeding by snails is related to lichen frequency in the field.Methods In a no-choice feeding experiment, we fed 24 lichen species to snails of two taxa from the Clausilidae and Enidae families and compared untreated lichens and lichens with compounds removed by acetone rinsing. Then, we related experimental lichen consumption with the frequency of lichen species among 158 forest plots in the field (Schw?bische Alb, Germany), where we had also sampled snail and lichen species.Important findings In five lichen species, snails preferred treated samples over untreated controls, indicating chemical feeding defense, and vice versa in two species, indicating chemical feeding stimulus. Interestingly, compared with less frequent lichen species, snails consumed more of untreated and less of treated samples of more frequent lichen species. Removing one outlier species resulted in the loss of a significant positive relationship when untreated samples were analyzed separately. However, the interaction between treatment and lichen frequency remained significant when excluding single species or including snail genus instead of taxa, indicating that our results were robust and that lumping the species to two taxa was justified. Our results imply lichen-feeding snails to prefer frequent lichens and avoid less frequent ones because of secondary compound recognition. This supports the idea that consumers adapt to the most abundant food source.     

Differential herbivore damage to calcicolous lichens by snails

Abstract: A laboratory experiment was conducted to determine the damage caused by snail grazing to 35 calcicolous lichen species and cyanobacteria from the Baltic island of Öland, Sweden. Individuals of four species of land snails (Balea perversa, Chondrina clienta, Clausilia bidentata and Helicigona lapicida) were allowed to graze on identified lichen species growing on pieces of limestone. Snail grazing on thalli and ascocarps was classified into four categories ranging from no damage to completely eaten. In general, some lichen species were heavily grazed, whereas others were only slightly damaged or not injured. Aspicilia calcarea, Tephromela atra and Verrucaria nigrescens were preferred by all snail species except Chondrina clienta. In general, lichen thalli that were not immersed in the calcareous rock and cyanobacteria were preferred, whereas ascocarps were avoided by one of the snail species (C. clienta). Immersed perithecia with a carbonized outer layer were avoided by all snail species except C. bidentata. Possibilities of chemical and mechanical defence properties in calcicolous lichens are discussed.     

Do secondary substances in the thallus of a lichen promote CO2 diffusion and prevent depression of net photosynthesis at high water content?

Many lichens show seriously depressed net photosynthesis (NP) at high thallus water contents due to increased carbon dioxide diffusion resistance through blockage of diffusion pathways by water. The soil lichen Diploschistes muscorum, however, shows no depression and NP is close to maximal even at the highest thallus water content. We investigated whether lichen substances (lecanoric and diploschistesic acids) in the cortex and medulla contributed to this ability to maintain high NP. Dry thalli were extracted with water-free acetone and, after this treatment, were found to be fully viable to the extent of continued growth after replanting in the field. No differences were found in the response of NP to thallus water content between the normal and extracted thalli, in fact the response curves were often nearly identical. Thus, in this species it seems that lichen substances did not maintain the water-free diffusion pathways and some other explanation, possibly structural, needs to be sought. Received: 5 April 1997 / Accepted: 26 April 1997     

UV triggers the synthesis of the widely distributed secondary lichen compound usnic acid

Synthesis of the cortical dibenzofuran derivative usnic acid and the medullary depsidone salazinic acid was studied in Xanthoparmelia stenophylla thalli from which the compounds had been removed by acetone rinsing prior to a 21-day field experiment with UV absorbing and transmitting screens. Natural levels of ultraviolet radiation clearly induced the re-synthesis of usnic acid. The re-synthesis was boosted by the addition of ribitol, the carbohydrate delivered from the Trebouxia photobiont to the mycobiont. Salazinic acid was also weakly induced by UV. Re-synthesis was relatively low, up to 2.5 and 3.1% of start values for usnic and salazinic acid, respectively. However, given that the natural content of both compounds was high, constituting 12% of thallus dry weight, the absolute amounts of lichen compounds re-synthesised were not so small. We also studied the extractability of nine extracellular lichen compounds in three species X. stenophylla, Hypogymnia tubulosa, and Vulpicida pinastri, and found two distinct fractions of cortical compounds, one major that was completely extractable from living lichens and one minor that was only extractable with grinding. Medullary compounds were completely extracted without grinding. These findings did not influence the relative differences between treatments in our experiment, but may be of importance for future assessments of, e.g., quantitative studies of extracellular lichen compounds.     

Surface hydrophobicity causes SO2 tolerance in lichens

BACKGROUND AND AIMS: The superhydrophobicity of the thallus surface in one of the most SO(2)-tolerant lichen species, Lecanora conizaeoides, suggests that surface hydrophobicity could be a general feature of lichen symbioses controlling their tolerance to SO(2). The study described here tests this hypothesis. METHODS: Water droplets of the size of a raindrop were placed on the surface of air-dry thalli in 50 lichen species of known SO(2) tolerance and contact angles were measured to quantify hydrophobicity. KEY RESULTS: The wettability of lichen thalli ranges from strongly hydrophobic to strongly hydrophilic. SO(2) tolerance of the studied lichen species increased with increasing hydrophobicity of the thallus surface. Extraction of extracellular lichen secondary metabolites with acetone reduced, but did not abolish the hydrophobicity of lichen thalli. CONCLUSIONS: Surface hydrophobicity is the main factor controlling SO(2) tolerance in lichens. It presumably originally evolved as an adaptation to wet habitats preventing the depression of net photosynthesis due to supersaturation of the thallus with water. Hydrophilicity of lichen thalli is an adaptation to dry or humid, but not directly rain-exposed habitats. The crucial role of surface hydrophobicity in SO(2) also explains why many markedly SO(2)-tolerant species are additionally tolerant to other (chemically unrelated) toxic substances including heavy metals.     

Herbivory on calcicolous lichens: different food preferences and growth rates in two co-existing land snails

A total of 32 calcicolous lichen species, one alga and one bryophyte were recorded on a limestone wall in the grassland Great Alvar on the Baltic island of Öland, Sweden. Fourteen (41%) of these 34 species and free-living cyanobacteria showed herbivore damage, most probably due to grazing by the land snails Chondrina clienta and Balea perversa which inhabited the limestone wall. Three laboratory experiments were conducted to examine the food preferences of individuals of C. clienta and B. perversa collected at this site and to evaluate any association between their preference and the net food quality of the lichens to the snails. Chondrina clienta and B. perversa exhibited food preferences, which differed significantly between species. Within species, variation in food choice was similar among individuals. This indicates that snail populations may be composed of polyphagous individuals with similar food preferences. Different lichen species were of different net food quality to the snails as indicated by growth rate differences. In both snail species the most preferred lichen species of the choice experiment caused the largest weight increase in juveniles, viz. Caloplaca flavovirescens for C. clienta and Aspicilia calcarea for B. perversa. This suggest that the snail species studied differ in their abilities to deal with secondary compounds and physical characteristics of certain lichens or that they can utilize the energy and nutrients of these lichens to a different extent. It is suggested that differential food preferences might reduce the intensity of interspecific competition for resources (lichens) between C. clienta and B. perversa.     

ITS1 metabarcoding highlights low specificity of lichen mycobiomes at a local scale

As self‐supporting and long‐living symbiotic structures, lichens provide a habitat for many other organisms beside the traditionally considered lichen symbionts—the myco‐ and the photobionts. The lichen‐inhabiting fungi either develop diagnostic phenotypes or occur asymptomatically. Because the degree of specificity towards the lichen host is poorly known, we studied the diversity of these fungi among neighbouring lichens on rocks in an alpine habitat. Using a sequencing metabarcoding approach, we show that lichen mycobiomes clearly reflect the overlap of multiple ecological sets of taxa, which differ in their trophic association with lichen thalli. The lack of specificity to the lichen mycobiome is further supported by the lack of community structure observed using clustering and ordination methods. The communities encountered across samples largely result from the subsampling of a shared species pool, in which we identify three major ecological components: (i) a generalist environmental pool, (ii) a lichenicolous/endolichenic pool and (iii) a pool of transient species. These taxa majorly belong to the fungal classes Dothideomycetes, Eurotiomycetes and Tremellomycetes with close relatives in adjacent ecological niches. We found no significant evidence that the phenotypically recognized lichenicolous fungi influence the occurrence of the other asymptomatic fungi in the host thalli. We claim that lichens work as suboptimal habitats or as a complex spore and mycelium bank, which modulate and allow the regeneration of local fungal communities. By performing an approach that minimizes ambiguities in the taxonomic assignments of fungi, we present how lichen mycobiomes are also suitable targets for improving bioinformatic analyses of fungal metabarcoding.     

Lichen endozoochory by snails

Endozoochory plays a prominent role for the dispersal of seed plants. However, for most other plant taxa it is not known whether this mode of dispersal occurs at all. Among those other taxa, lichens as symbiotic associations of algae and fungi are peculiar as their successful dispersal requires movement of propagules that leaves the symbiosis functional. However, the potential for endozoochorous dispersal of lichen fragments has been completely overlooked. We fed sterile thalli of two foliose lichen species (Lobaria pulmonaria and Physcia adscendens) differing in habitat and air-quality requirements to nine snail species common in temperate Europe. We demonstrated morphologically that L. pulmonaria regenerated from 29.0% of all 379 fecal pellets, whereas P. adscendens regenerated from 40.9% of all 433 fecal pellets, showing that lichen fragments survived gut passage of all snail species. Moreover, molecular analysis of regenerated lichens confirmed the species identity for a subset of samples. Regeneration rates were higher for the generalist lichen species P. adscendens than for the specialist lichen species L. pulmonaria. Furthermore, lichen regeneration rates varied among snail species with higher rates after gut passage of heavier snail species. We suggest that gastropods generally grazing on lichen communities are important, but so far completely overlooked, as vectors for lichen dispersal. This opens new ecological perspectives and questions the traditional view of an entirely antagonistic relationship between gastropods and lichens.     

LICHEN HERBIVORE PREFERENCE: A TEST OF TWO HYPOTHESES

Field data from a saxicolous lichen community in Shenandoah National Park, Virginia, suggested that the slug Pallifera varia grazed more frequently on certain lichen species than would be expected if it fed randomly. Two hypotheses might explain this grazing pattern. The preference hypothesis, that slugs select lichens of highest quality, was tested by measuring the concentration of essential elements in lichens eliciting high and low preference by slugs. Lichens with the highest element contents were assumed to be of the highest quality to herbivores. The avoidance hypothesis, that preference by slugs actually results from the rejection of unpalatable species, was tested by offering slugs choices of baited filter-paper disks impregnated with secondary products of the lichen species in question. Results suggested that preferred lichens had significantly lower concentrations of N, P, and Ca, and that avoided lichens produced secondary products that effectively inhibited Pallifera grazing activity. These results suggested that the avoidance hypothesis was the better explanation for nonrandom Pallifera grazing patterns. Furthermore, they suggested that lichens with the highest concentrations of essential elements are most likely to produce defense compounds, an observation that supports predictions to explain patterns of chemical defense in vascular plants.     

Insight into the pattern of heavy-metal accumulation in lichen thalli

BackgroundHeavy metals that pass through the plasmalemma are expected to influence on lichen metabolic processes; however, lichens may tolerate high concentrations of metals by sequestrating them extracellularly. Heavy metal accumulation level fundamentally determine the success of lichens in the colonisation of polluted sites; however, the proportions between extra- and intracellular metal concentrations in lichen thalli are still poorly recognized. In this study metal accumulation patterns of selected toxic trace elements, i.e. Pb, Cd, and micronutrients, i.e. Zn, Cu and Ni, in Cladonia cariosa thalli were recognised in relation to extra- and intracellular fractions.MethodsThe intracellular and total concentrations of Zn, Pb, Cd, Cu and Ni in lichen thalli collected from eleven variously polluted sites were determined by means of atomic absorption spectrometry. Additionally, organic carbon and total nitrogen contents as well as pH of soil substrate were measured.ResultsThe accumulation patterns differed between studied metal elements; the major part of Zn, Pb and Cd loads was accumulated extracellularly, whereas Cu and Ni accumulation was mostly intracellular. Like toxic trace elements, Zn was accumulated mainly extracellularly at high polluted sites. The non-linear models most reliably reflect relationships between intracellular and extracellular metal contents in C. cariosa thalli. The intracellular contents of Zn, Pb, Cd and Cu increased slower at higher than at lower extracellular concentrations. Moreover, at higher total concentrations of elements in the thalli, their extracellular proportions were markedly increased.ConclusionThe results suggest that in the face of extreme Zn-enrichment, lichens demonstrate the ability to accumulate the excess of Zn outside the cells. Therefore, it can be concluded that metal accumulation depend not only on the element but also on its abundance in the environment and direct availability for lichens. The studied species showed a defence against excessive intracellular accumulation when a given element is in excess. Such capability may facilitate the colonization of extremely polluted sites by certain pioneer lichens.     

肉疣衣属中国新记录种——乌普萨拉肉疣衣

采用形态解剖、化学、生态类型及地理成分分析等传统分类方法对新疆天山南部巴音布鲁克山区地衣外部形态和内部构造、地衣附属结构、地衣繁殖器官进行观察。利用显色反应法(CT)、微量化学结晶法(MCT)、薄层层析法(TLC)等生物化学方法,鉴定地衣次生代谢产物,发现了1个中国新记录种:乌普萨拉肉疣衣[Ochrolechia upsaliensis(L.)A.Massal]。文中描述了该新记录种地衣的形态解剖、化学特征和生境,并提供了地衣体、子囊及子囊孢子的彩色图片。     

Do lichen secondary compounds play a role in highly specific fungal parasitism?

Chemical interactions between highly host-specific lichenicolous fungi and their lichen hosts have been little studied. In an allometric study, we quantified carbon-based secondary compounds (CBSCs) in a mixed natural Lobarina scrobiculata population (N = 147) of the normal and the stictic acid-deficient chemotypes, both with and without galls of Plectocarpon scrobiculatae. We assessed the correlation between the presence/abundance of parasite galls and the lichen CBSCs contents, and quantified size-dependent contents of CBSCs. The parasite produced galls similarly in both chemotypes, indicating that the stictic acid complex does not deter Plectocarpon. Within both chemotypes, thalli with Plectocarpon had half the contents of all individual CBSCs than those without galls. There was a significant size-dependent increase in CBSC contents in thalli without galls, but not in those with. This study shows that lichen chemistry is involved in highly host-specific fungal parasitism, and widens our knowledge of specialized biotrophic fungal interactions.     

Heterogeneity of Chlorophyll Fluorescence over Thalli of Several Foliose Macrolichens Exposed to Adverse Environmental Factors: Interspecific Differences as Related to Thallus Hydration and High Irradiance

Spatial heterogeneity of chlorophyll (Chl) fluorescence over thalli of three foliose lichen species was studied using Chl fluorescence imaging (CFI) and slow Chl fluorescence kinetics supplemented with quenching analysis. CFI values indicated species-specific differences in location of the most physiologically active zones within fully hydrated thalli: marginal thallus parts (Hypogymnia physodes), central part and close-to-umbilicus spots (Lasallia pustulata), and irregulary-distributed zones within thallus (Umbilicaria hirsuta). During gradual desiccation of lichen thalli, decrease in Chl fluorescence parameters (F_O - minimum Chl fluorescence at point O, F_P - maximum Chl fluorescence at P point, ₂ - effective quantum yield of photochemical energy conversion in photosystem 2) was observed. Under severe desiccation (>85 % of water saturation deficit), substantial thalli parts lost their apparent physiological activity and the resting parts exhibited only a small Chl fluorescence. Distribution of these active patches was identical with the most active areas found under full hydration. Thus spatial heterogeneity of Chl fluorescence in foliose lichens may reflect location of growth zones (pseudomeristems) within thalli and adjacent newly produced biomass. When exposed to high irradiance, fully-hydrated thalli of L. pustulata and U. hirsuta showed either an increase or no change in F_O, and a decrease in F_P. Distribution of Chl fluorescence after the high irradiance treatment, however, remained the same as before the treatment. After 60 min of recovery in the dark, F_O and F_P did not recover to initial values, which may indicate that the lichen used underwent a photoinhibition. The CFI method is an effective tool in assessing spatial heterogeneity of physiological activity over lichen thalli exposed to a variety of environmental factors. It may be also used to select a representative area at a lichen thallus before application of single-spot fluorometric techniques in lichens.     

Co-occurrence of the multicopper oxidases tyrosinase and laccase in lichens in sub-order peltigerineae

BACKGROUND AND AIMS: Following previous findings of high extracellular redox activity in lichens and the presence of laccases in lichen cell walls, the work presented here additionally demonstrates the presence of tyrosinases. Tests were made for the presence of tyrosinases in 40 species of lichens, and from selected species their cellular location and molecular weights were determined. The effects of stress and inhibitors on enzyme activity were also studied. METHODS: Tyrosinase and laccase activities were assayed spectrophotometrically using a variety of substrates. The molecular mass of the enzymes was estimated using polyacrylamide gel electrophoresis. KEY RESULTS: Extracellular tyrosinase and laccase activity was measured in 40 species of lichens from different taxonomic groupings and contrasting habitats. Out of 20 species tested from the sub-order Peltigerineae, all displayed significant tyrosinase and laccase activity, while activity was low or absent in other species tested. Representatives from both groups of lichens displayed low peroxidase activities. Identification of the enzymes as tyrosinases was confirmed by the ability of lichen thalli or leachates derived by shaking lichens in distilled water to metabolize substrates such as L-dihydroxyphenylalanine (DOPA), tyrosine and epinephrine readily in the absence of hydrogen peroxide, the sensitivity of the enzymes to the inhibitors cyanide, azide and hexylresorcinol, activation by SDS and having typical tyrosinase molecular masses of approx. 60 kDa. Comparing different species within the Peltigerineae showed that the activities of tyrosinases and laccase were correlated to each other. Desiccation and wounding stimulated laccase activity, while only wounding stimulated tyrosinase activity. CONCLUSIONS: Cell walls of lichens in sub-order Peltigerineae have much higher activities and a greater diversity of cell wall redox enzymes compared with other lichens. Possible roles of tyrosinases include melanization, removal of toxic phenols or quinones, and production of herbivore deterrents.     

Growth of reindeer lichens and effects of reindeer grazing on ground cover vegetation in a Scots pine forest and a subarctic heathland in Finnish Lapland

Reindeer lichens are an important component of northern ecosystems. The aim of this study was to measure the growth rate of terricolous lichens as it is a key parameter involved in productivity of these ecosystems and an important part of lichen tolerance to reindeer grazing. Furthermore, the natural succession and the long-term effects of reindeer grazing on lichen community characteristics in two contrasting habitats were investigated as well as the interactions between lichen cover and mosses and vascular plants. Biomass and coverage measurements were conducted in a lichen woodland and in a subarctic heath with grazed and ungrazed areas in northern Finland. Measurements spanning over 13 yr of undisturbed development show that the growth rate of Cladina stellaris can be as high as >0.17 g g⁻¹ produced annually, although in average growth rates were much lower. During the succession of ground vegetation, C. stellaris , C. rangiferina , C. mitis and Cetraria nivalis increased in biomass in fenced areas and were reduced most in biomass by reindeer in unprotected areas. Reindeer grazing and trampling seem to change the vegetation towards a type that is dominated by small dwarf shrubs, bare soil and minute-cup lichens ( Cladonia spp.). Removing the lichen layer by reindeer may reduce natural regeneration of pine trees as implied by increasing numbers of pine seedlings with increasing lichen cover.