Secondary metabolites in Cladina stellaris in relation to reindeer grazing and thallus nutrient content

We analysed concentrations of phenols and key macro-nutrients in a dominant mat-forming lichen, Cladina stellaris, in dry heath forests subjected to heavy reindeer grazing in Finnish Lapland. Lichen samples were collected in the beginning of two growth seasons from six to eight sites with old reindeer exclosures that served as control plots within sites. The concentration of perlatolic acid in lichen apices (top 10 mm) was higher in grazed plots than in control ones, whereas usnic acid did not seem to respond unequivocally to grazing. Moreover, there was a strong negative relationship between nitrogen and phenolic content of Cladina stellaris in intact plots but this relationship was absent in grazed ones. Changes in nitrogen and phosphorus contents caused by reindeer grazing were not correlated with changes in the level of phenolics in lichen thalli. The present result demonstrated that the carbon-nutrient balance (CNB) hypothesis may well explain some of the variation in lichen phenolic content but is not sufficient for explaining changes caused by reindeer grazing. We hypothesized that physical changes in the lichen microenvironment induced by reindeer have more profound impact on lichen phenolic content than alterations in thallus nutrient content.     

Copper toxicity to five Parmelia lichens in vitro

Treatment of Parmelia caperata, P. perlata, P. subrudecta, P. sulcata and P. tiliacea with CuSO₄ resulted in a time- and copper-concentration-dependent decrease in the total and intracellular potassium concentrations of the thallus, indicating that copper damaged the cytoplasmic membrane. Treatment with copper also resulted in a time-dependent increase in the total copper concentration of the thallus. After 4 h of exposure to copper, the process of potassium efflux was essentially completed but the absorption of copper was still increasing; moreover, the amount of copper bound to the thallus exceeded twice the amount of potassium released from the thallus, suggesting that cupric ions reached intracellular sites in the thallus, and K⁺/Cu²⁺ exchange was not electroneutral. After 5 h of exposure to copper, the extent of decrease in the total and intracellular potassium concentrations of the thallus was positively correlated with copper absorption levels, but only at 0.05<P<0.10, suggesting that membrane damage was proportional to the amount of bound copper, but other factors could have been operative, namely binding of copper to the cell wall. Acetone extracts of untreated thalli contained low concentrations of amino acids, polyols, and sugars, but considerable amounts of lichen substances: atranorin, caperatic, constictic, lecanoric, menegazziaic, protocetraric, salazinic, stictic, and usnic acids. Titration of the extracts with copper and assay of the free Cu²⁺ concentration revealed the presence of copper-binding ligands, and several successive absorption cycles, most probably corresponding to the binding of Cu²⁺ to each of the lichen substances detected in the extracts. However, no significant correlation (P>0.10) was found between the Cu²⁺-complexing capacity of acetone extracts and copper-induced membrane damage. It was concluded that in the studied Parmelia species, and in the experimental conditions used in this work, copper toxicity was not a simple function of the Cu²⁺-binding properties of the lichen substances present in the thallus. Several hypotheses were formulated to interpret the results.     

A Note on the Relationship Between Some Saxicolous Lichens and Manganese Ore in North Wales,UK

Manganese oxyhydroxides have been found deposited on the surface of the lichen Catillaria chalybeia on an altered rhodochrosite. It is suggested that the Mn was mobilised from the ore surface via surface-weathering and redeposited in the thallus. The lichen Acarospora smaragdula also grew upon the ore but showed no sign of Mn deposition. Nine further lichens and two bryophytes were also found on the ore.     

Natural and Experimentally-induced Zinc and Copper Resistance in the Lichen Genus Peltigera

Using a tolerance index based on metal-induced reductions inphotosynthesis, members of the lichen genus Peltigera, sampledfrom a range of sites with contrasting metal status, were demonstratedto possess Zn tolerance directly proportional to the Zn contentof the thallus. Tolerance was not related to the concentrationof other cations present in the thallus or weight of thallusper unit surface area. Although differences in Cu tolerancewere found, these were not related to the Cu content of thethallus but rather to the Zn content. In the laboratory, pretreatmentof P. membranacea with low concentrations of Zn significantlydecreased the inhibition of photosynthesis caused when subsequentlysupplied with high levels of Zn compared to water pretreatedplants. Increased resistance to Cd but not Cu was also found,although it was not possible to increase either Cu or Zn resistanceby pretreatment with Cu. Peltigera membranacea, lichen, heavy metal tolerance, zinc, copper, cadmium     

五种南极地衣的Co、Cr、Pb和Cu元素富集能力的差异

随着人类活动对全球环境的影响,环境重金属污染负荷日益增加,重金属元素向南极地区的迁移和富集已成为该地区面临的严重问题之一。地衣是研究重金属元素大气沉降特征的良好材料,但对南极地衣的重金属元素大气沉降富集能力的比较研究甚少。作者以网袋法实验比较采自西南极乔治王岛菲尔德斯半岛的5种南极优势地衣,即:王橙衣Caloplaca regalis、夹心果衣Himantormia lugubris、孔树花Ramalina terebrata、球粉衣Sphaerophorus globosus和簇花石萝Usnea aurantiacoatra对钴(Co)、铬(Cr)、铅(Pb)和铜(Cu)的富集能力。2个月的网袋法实验结果表明:1)5种地衣对Pb元素均表现出富集能力,其中夹心果衣富集能力最强,簇花石萝和孔树花次之;2)簇花石萝和夹心果衣对Cu元素的富集能力相近,球粉衣对Cu元素的富集能力较低,其他2种未表现出对Cu的富集作用;3)Co和Cr元素的富集仅分别在夹心果衣和簇花石萝中检测出来。因此,在综合检测Co、Cr、Cu和Pb的大气沉降时,可考虑簇花石萝和夹心果衣的组合;在单独检测Co、Pb和Cu的大气沉降时,夹心果衣均为最佳的物种。     

Ultrastructural Responses of the LichenBryoria fuscescensto Simulated Acid Rain and Heavy Metal Deposition

Effects of simulated acid rain and heavy metal deposition onthe ultrastructure of the lichenBryoria fuscescens(Gyeln.) Brodoand Hawksw. were examined in a field study conducted in northernFinland. Lichens were exposed to simulated rain containing twolevels of a mixture of copper (Cu²⁺) and nickel (Ni²⁺) ionsalone or in combination with acid rain (H₂SO₄) at pH 3 over2 months in addition to ambient rainfall. The algal and fungalcomponents responded differently to pH and there was an interactionwith metal toxicity. The algal partner was the most sensitiveto acid rain and heavy metal combinations and had more degeneratecells than the fungal partner. Damage was apparent in chloroplastsand mitochondria, where thylakoid and mitochondrial cristaewere swollen. The fungal partner was the more sensitive to highconcentrations of metal ions in the absence of acidity, suggestingan ameliorating interaction between the metals and acidity.For algae, critical metal concentrations in the thallus were>50 µg g^-1for Cu and >7 µg g^-1for Ni in thepresence of acidity, and >20 µ g g^-1for Ni in the absenceof acidity. Detrimental effects of heavy metals on fungal ultrastructurewere seen when thallus metal concentrations exceeded 400 µgg^-1for Cu and 100 µg g^-1d. wt for Ni. The results suggestthat acid wet deposition containing metal ions may reduce survivalof lichens in northern environments.Copyright 1998 Annals ofBotany Company. Copper, nickel, sulphuric acid, ultrastructure,Bryoria fuscescens(Gyeln.) Brodo and Hawksw., epiphytic lichen, air pollution.     

Differential responses of lichen symbionts to enhanced nitrogen and phosphorus availability: an experiment with Cladina stellaris

BACKGROUND AND AIMS: Lichens can be both nitrogen- (N) and phosphorous- (P) limited and thus may be susceptible to nutrient enrichment. Nutrient enrichment with N and P may have differing impacts on the lichen structure because of different physiological responses of fungal and algal partners to these nutrients. The hypothesis was tested that the differential responses of lichen symbionts to enhanced availability of N and P is reflected in the lichen thallus structure and the wall-to-wall interface between the algal and fungal cells. METHODS: Lichen cushions of Cladonia stellaris were treated with one P and two N concentrations alone and in combination that yielded total depositions of approx. 300 (moderate) and 1000 (high) mg N m(-2) and 100 (high) mg P m(-2) over an experiment lasting 14 weeks. The effects of N and P inputs on the relative volumes of fungal and algal cell in the medullary tissue and on the thallus structure were studied using light microscopy. The interface between algal and fungal cell walls was examined using transmission electron microscopy. KEY RESULTS: The influence of excess P on the lichen thallus structure was stronger than that of additional N. Addition of P reduced the N : P ratio in podetia, the proportion of the medullary layer volume occupied by the algal cells, the thallus volume occupied by the internal lumen, and the algal cell-wall area covered by fungal hyphae. CONCLUSIONS: Ecologically realistic changes in the availability of key macronutrients can alter the growth of symbionts. Reduction in the proportion of photobiont cells indicates that the application of P either stimulates fungal hyphal growth in the medullary tissue or impairs the cell division of the algal cells. The results suggest that both the N and P availability and thallus N : P ratio affect the growth rates of lichen symbionts.     

混合样品代表地衣元素积累的平均水平——来自元素含量不同的两种地衣的证据

地衣是大气元素沉降的良好监测生物,其元素积累能力具有物种间和个体间差异。地衣原位监测法常使用混合样品代表每个样点的地衣元素积累水平,但混合样品的代表性及其在不同地衣之间的差异尚需进一步研究。以电感耦合等离子体质谱法（inductively coupled plasma mass spectrometry,ICP-MS）测定了内蒙古多伦县的丽石黄衣Xanthoria elegans和皮果衣Dermatocarpon miniatum 52种元素的含量并比较了样点内变异和物种间差异。结果显示,丽石黄衣与相似生境中的地衣元素含量大致相似,证实了研究区域以沙尘沉降为主的大气沉降特点。两种地衣的元素排序基本一致,表明其元素来源相同。7种营养元素（Ca、K、Mo、P、Rb、S和Se）的含量在物种间差异不显著,与地衣的生理调节有关。Hg含量的物种间差异不显著,可能与Hg的挥发性有关。皮果衣中44种元素（Al、As、B、Ba、Be、Bi、Cd、Ce、Co、Cr、Cs、Cu、Dy、Er、Eu、Fe、Gd、Ge、Ho、La、Li、Lu、Mg、Mn、Na、Nb、Nd、Ni、Pb、Pr、Sb、Sc、Sm、Sr、Tb、Th、Ti、Tl、Tm、U、V、Y、Yb和Zn）的含量是丽石黄衣的1.32-2.05倍,表明皮果衣具有较高的、元素特异性的元素积累能力。样点内除Ca含量变异较大（CV>31%）之外,其他51种元素的含量样点内变异在皮果衣和丽石黄衣中均<27.5%,表明混合样品在两种地衣中均可较好地代表地衣元素积累的平均水平。     

Does the lichen mat alleviate the effects of wet deposited nickel on soil microorganisms and Scots pine (Pinus sylvestris L.) seedlings?

A field experiment was conducted in a dry heath forest dominated by Scots pine (Pinus sylvestris L.) and a mat-forming lichen (Cladina stellaris (Opiz) Brodo) to assess the effect of wet-deposited nickel (Ni) on pine seedlings and soil microorganisms, and to explore whether an intact lichen mat could act as a buffer against heavy metal deposits. Pine seedlings were planted in quadrats covered by a natural lichen layer and in quadrats from which the lichen layer had been completely removed. The quadrats were exposed to four levels of Ni deposition: 0 (i.e., distilled water), 10, 100 and 1000 mg m^–2 year^–1 in two growing seasons. Increasing Ni deposition led to an increase in the Ni content of the needles, roots and the soil organic layer. The lichen mat reduced Ni flow to the organic soil layer, but had no significant, reducing effect on needle or root Ni concentration. The most severe Ni treatment had detrimental effects on seedling growth and increased peroxidase activity in the previous years needles. Removal of the lichen layer did not increase susceptibility of seedlings to Ni. Values of maximal carbon use efficiency (Max) and metabolic quotient (qCO₂) of the soil microorganisms indicated protective value of the lichen mat to soil microorganisms at the highest Ni treatment. Skimming per se decreased basal respiration, qCO₂ and concentrations of potassium in the soil and also increased the lag period of the microorganisms as a response to in situ substrate addition.     

Epiphytic lichens as biomonitors of airborne heavy metal pollution

The research aims to assess the performance of the lichen Parmotrema reticulatum as an air pollution biomonitor of four heavy metals, namely, chromium (Cr), copper (Cu), lead (Pb) and zinc (Zn). Four contrasting land use sites within the greater Auckland region of New Zealand are used. One site is located within the relatively clean air shed of conservation land, the others within residential, commercial, and industrial areas, characterised by increasingly polluted air sheds, respectively. Three groups of lichens are monitored over a two-year period using active and passive biomonitoring methods to assess ‘on-thallus’ and ‘in-thallus’ concentrations of heavy metals. Seasonal transplants are used to quantify heavy metals accumulated by the lichen during each season. Long-term transplants are used to measure how fast lichens accumulate heavy metals and to better understand how and when heavy metals within the lichen thallus achieve equilibrium with air pollutant concentrations over time. The results show that the lichens continuously accumulates pollutants from the air until equilibrium is reached, thus transplanted lichens are useful for monitoring air pollution concentrations over time. Since pollutant concentration in the transplanted lichen at equilibrium stabilises, at this point the lichen ceases to be useful for monitoring temporal trends in air pollution, but may useful for spatial air pollution monitoring. The industrial location has the highest total accumulation for all four heavy metals, followed by the commercial and residential locations, respectively. Overall, the results show that the lichen P. reticulatum may be successfully used to monitor spatial and temporal pollution patterns caused by even very low concentrations of Cr, Cu, Pb and Zn.     

Scots pine needle injuries at subarctic industrial sites

 Injuries to needles of Scots pines (Pinus sylvestris L.) growing in nutrient-poor soils on the Kola Peninsula collected in April 1991 were studied on a gradient of increasing distances (10 – 115 km) from the Monchegorsk nickel smelter, Russia, which emits SO₂, Ni and Cu. The condition of the mesophyll cells was quantified from needles of the two latest age classes using a light and an electron microscope. The damage to the ultrastructure consisted of multistress symptoms caused by excess sulphur, heavy metals, frost, acidic precipitation and ozone. Injuries were most commonly manifested in the form of dark, irregularly shaped chloroplasts with protrusions and light thylakoids and plastoglobuli. These symptoms gradually disappeared with increasing distance and decreasing deposition rate. Concentrations of sulphur, copper and nickel decreased towards more distant sites where normal levels of the latter two elements were reached. Sulphur concentrations remained above background throughout the distance gradient. In the closest plots to the smelter area, cell collapse under the stomata and epidermis related to acute SO₂ and heavy metal effects was found, whereas further away symptoms were more diverse, pointing towards the effects of ozone, acidic deposition and thereby decreased frost tolerance. The additive multistress symptoms were clearly seen in the area up to 40 km from the smelter where needle Cu concentration was above 110 ppm, Ni concentration above 39 ppm and S concentration above 1343 ppm. Received: 22 October 1995 / Accepted: 17 October 1996     

Assessment of the Reproduction Efficiency of the Atlantic salmon Salmo salar (Salmonidae) in the Kola River (Kola Peninsula) Based on Data on the Density of Juveniles

Journal of Ichthyology - In the Kola River (Kola Peninsula), on the basis of cluster analysis of long-term density data in the nursery areas of underyearlings, wild and wild + hatchery parr of...     

Requirement of divalent cations for fast exchange of actin monomers and actin filament subunits

The exchange of actin monomers and actin filament subunits was investigated in the presence of different cations. At high concentrations of calcium (1.2 mm) or magnesium (0.6 mm) the half-time of exchange was less than two hours. When the calcium ions were chelated by EGTA² and potassium (12 mm) was added the exchange was immediately stopped. In the presence of potassium ions (19 mm) at low concentrations of divalent cations (less than 40 μm) no exchange was observed within 15 hours. Fast exchange was initiated upon addition of calcium ions (1.2 mm).The results were interpreted by the model of head to tail polymerization in which filaments lengthen at one end and shorten simultaneously at the other. At high concentrations of divalent cations association and dissociation reactions occur mainly at different ends, whereas in the presence of potassium and at low concentrations of divalent cations these reactions take place at each end with similar frequencies. The efficiency of the head to tail polymerization depends strongly on the concentration and on the type of cations present.     

Exploring symbiont management in lichens

Lichens are unique among fungal symbioses in that their mycelial structures are compact and exposed to the light as thallus structures. The myriad intersections of unique fungal species with photosynthetic partner organisms (green algae in 90% of lichens) produce a wide variety of diverse shapes and colours of the fully synthesized lichen thallus when growing in nature. This characteristic complex morphology is, however, not achieved in the fungal axenic state. Even under ideal environmental conditions, the lichen life cycle faces considerable odds: first, meiotic spores are only produced on well-established thalli and often only after achieving considerable age in a stable environment, and second, even then in vivo resynthesis requires the presence of compatible algal strains where fungal spores germinate. Many lichen species have evolved a way around the resynthesis bottleneck by producing asexual propagules for joint propagation of symbionts. These different dispersal strategies ostensibly shape the population genetic structure of lichen symbioses, but the relative contributions of vertical (joint) and horizontal (independent) symbiont transmission have long eluded lichen evolutionary biologists. In this issue of Molecular Ecology, Dal Grande et al. (2012) close in on this question with the lung lichen, Lobaria pulmonaria, a flagship species in the conservation of old growth forests. By capitalizing on available microsatellite markers for both fungal and algal symbionts, they show that while vertical transmission is the predominant mode of reproduction, horizontal transmission is demonstrable and actively shapes population genetic structure. The resulting mixed propagation system is a highly successful balance of safe recruitment of symbiotic clones and endless possibilities for fungal recombination and symbiont shuffling.     

Lichenized species of Omphalina (Tricholomataceae) in the Iberian Peninsula

Abstract:A taxonomic revision of the lichenized species of the genus Omphalina in the Iberian Peninsula is presented, based upon fungal and lichen herbarium material, as well as on fresh collections. Relevant morphological and ecological features are discussed and a diagnostic key is provided. Four lichenized species, O. ericetorum, O. hudsoniana, O. meridionalis and O. velutina, are recognized. A squamulose vegetative thallus (Coriscium -type) is present only in O. hudsoniana, while the other three species form a thallus of hyphal globules (Botrydina -type). Anatomical features of the vegetative thallus cannot be used to distinguish between these last three species. Whilst O. hudsoniana is known only from the montane belt of the Eurosiberian Region and O. meridionalis seems to be restricted to the Mediterranean Region, living in more or less humid sites of the Supramediterranean belt, the other two species, O. ericetorum and O. velutina, are widespread.     

Paleoclimatic potential of the northernmost juniper trees in Europe

In late summer 2004 stem discs were collected from about 40 juniper trees (Juniperus Siberica Burgsd) growing at the remote central part of Kola Peninsula behind the polar circle at the northern timberline. Up to now these juniper trees are oldest ones found at Kola Peninsula. Data processing was difficult due to extremely small tree rings as well as the occurrence of missing and false rings. However, finally it was possible to build up a 676-year long chronology and retrieve information on the past climatic variations at Kola Peninsula that could partly be linked to extraterrestrial factors such as changes in solar activity and galactic cosmic ray activity. It was obtained that:

(1) There is a rather good agreement between long-term climatic variation in Europe and at Kola Peninsula.

(2) The minima of solar activity Sporer (1416–1534 AD), Maunder (1645–1715 AD) and Dalton (1801–1816 AD) were accompanied by temperature decreases. Cooling during the end of the Wolf minimum (until 1350) is reflected in the juniper tree-ring series from Kola Peninsula whereas it is not reflected in the European temperature reconstructions.

(3) Some recent decreases in solar activity around 1900 and 1960 are linked to phases of reduced growth in juniper.

(4) The juniper chronologies from Kola Peninsula do not indicate a temperature rise at the end of the XX century.

(5) MTM spectral and wavelet analysis of juniper tree-ring records showed:

(a) more pronounced 22- and 80–100-year periodicities;

(b) the main cycle of solar activity, the 11-year Schwabe cycle, was not present;

(c) 20–22-year periodicity was not significant throughout the entire ca.700-year period, but during certain time intervals: 1328–1550, 1710–1800, 1985 to present.

High-light stress and photoprotection in Umbilicaria antarctica monitored by chlorophyll fluorescence imaging and changes in zeaxanthin and glutathione

The effect of high light on spatial distribution of chlorophyll (Chl) fluorescence parameters over a lichen thallus (Umbilicaria antarctica) was investigated by imaging of Chl fluorescence parameters before and after exposure to high light (1500 micro mol m (-2) s (-1), 30 min at 5 degrees C). False colour images of F (V)/F (M) and Phi (II) distribution, taken over thallus with 0.1 mm (2) resolution, showed that maximum F (V)/F (M) and Phi (II) values were located close to the thallus centre. Minimum values were typical for thallus margins. After exposure to high light, a differential response of F (V)/F (M) and Phi (II) was found. The marginal thallus part exhibited a loss of photosynthetic activity, manifested as a lack of Chl fluorescence signal, and close-to-centre parts showed a different extent of F (V)/F (M) and Phi (II) decrease. Subsequent recovery in the dark led to a gradual return of F (V)/F (M) and Phi (II) to their initial values. Fast (30 min) and slow (1 - 22 h) phase of recovery were distinguished, suggesting a sufficient capacity of photoprotective mechanisms in U. antarctica to cope with low-temperature photoinhibition. Glutathione and xanthophyll cycle pigments were analyzed by HPLC. High light led to an increase in oxidized glutathione (GSSG), and a conversion of violaxanthin to zeaxanthin, expressed as their de-epoxidation state (DEPS). The responses of GSSG and DEPS were reversible during subsequent recovery in the dark. GSSG and DEPS were highly correlated to non-photochemical quenching (NPQ), indicating involvement of these antioxidants in the resistance of U. antarctica to high-light stress. Heterogeneity of Chl fluorescence parameters over the thallus and differential response to high light are discussed in relation to thallus anatomy and intrathalline distribution of the symbiotic alga Trebouxia sp.     

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     

Lichen communities and the associated fauna on a rocky sea shore on Bornholm in the Baltic

Thirty-three sampling sites representing 11 lichen dominated plant communities on maritime rocks on Bornholm were analysed with respect to composition of the vegetation, number of animals present and gross taxonomic composition of the fauna. The specific fauna of one moss and seven lichen species was examined.
The lichen communities studied were largely the same as those found in similar studies in NW Europe, except for the recognition of a distinct Ramalina cuspidata community. The highest average number of animals per area was found in the Schistidium maritimum and the Ramalina siliquosa communities, whereas the highest average number of animals g⁻¹ dry plant biomass was recorded in the Parmelia saxatilis community. Among the species examined Anaptychia fusca contained the highest average number of animals g⁻¹ dry thallus and Ramalina cuspidata the lowest number, 114 and 7, respectively. Collemboles appeared to dominate in the moss Schistidium maritimum and in foliose lichens, whereas mites dominated in crustose and fruticose lichens.     

Modern pollen and stomate deposition in lake surface sediments from across the treeline on the Kola Peninsula, Russia

We sampled and analyzed surface sediments from 31 lakes along a latitudinal transect crossing the coniferous treeline on the Kola Peninsula, Russia. The major vegetation zones along the transect were tundra, birch-forest tundra, pine-forest tundra, and forest. The results indicate that the major vegetation types in our study area have distinct pollen spectra. Sum-of-squares cluster analysis and principal components analysis (PCA) groupings of pollen sites correspond to the major vegetation zones. PCA ordination of taxa indicates that the first axis separates taxa typical of the forest zone (Pinus, Picea) from taxa typical of tundra and forest-tundra zones (Polypodiaceae, Ericaceae, and Betula). The current position of the coniferous treeline, defined in our region by Pinus sylvestris, occurs roughly where Pinus pollen values reach 35% or greater. Arboreal pollen (AP)/non-arboreal pollen (NAP) ratios were calculated for each site and plotted against geographic distance along the transect. AP/NAP ratios of 7 or greater are found within pine-forest tundra and forest vegetation zones. Pinus stomates (dispersed stomatal guard cells) are absent from sites north of the coniferous treeline and all but two samples from the forested sites contain stomates. Stomate concentrations among the samples are highly variable and range from 10 to 458 per ml and positively correlate with the changing Pinus pollen values.