Search

PREMISE OF THE STUDY High-quality information about threatened species is required to prevent current global biodiversity losses. Lichens are important components of forest biodiversity and help to maintain ecosystem functioning. The epiphytic cyanolichen Lobarina scrobiculata is red-listed in Europe and North America, but knowledge of its ecology and distribution in Southern Europe is scarce. METHODS We used a multispatial scale design to investigate the effects of macroclimate, forest structure, tree features, and microhabitat on the occurrence, abundance, spatial distribution, and performance of Lobarina scrobiculata in the Iberian Peninsula at plot and tree scales. Generalized linear models and mixed models were used for analysis. KEY RESULTS We recorded ca 14000 individuals of the threatened species Lobarina scrobiculata from 22 populations in the Iberian Peninsula. Our results suggest that L. scrobiculata thrives mainly in oak forests with highly variable annual precipitation levels. At the plot scale, the L. scrobiculata abundance increased with annual precipitation and tree density (habitat quantity). At the tree scale, our models highlighted the importance of tree size and bark roughness (habitat quality) as the main drivers of species occurrence and abundance. We detected a marked spatial pattern on tree trunks, i.e., L. scrobiculata occurred preferentially on north-facing surfaces and close to the ground where humidity is higher. CONCLUSIONS By integrating multiscale modeling, we analyzed a unique large dataset and these results are essential for understanding the ecology of this threatened cyanolichen. There is an urgent need to preserve the forests that this species currently inhabits as well as potential colonization sites.

Molybdenum (Mo) nitrogenase has long been considered the predominant isoenzyme responsible for dinitrogen fixation worldwide. Recent findings have challenged the paradigm of Mo hegemony, and highlighted the role of alternative nitrogenases, such as the vanadium-nitrogenase. Here, we first characterized homeostasis of vanadium (V) along with other metals in situ in the dinitrogen fixing cyanolichen Peltigera aphthosa. These lichens were sampled in natural sites exposed to various levels of atmospheric metal deposition. These results were compared with laboratory experiments where Anabaena variabilis, which is also hosting the V-nitrogenase, and a relatively close relative of the lichen cyanobiont Nostoc, was subjected to various levels of V. We report here that V is preferentially allocated to cephalodia, specialized structures where dinitrogen fixation occurs in tri-membered lichens. This specific allocation is biologically controlled and tightly regulated. Vanadium homeostasis in lichen cephalodia exposed to various V concentrations is comparable to the one observed in Anabaena variabilis and other dinitrogen fixing organisms using V-nitrogenase. Overall, our findings support current hypotheses that V could be a more important factor in mediating nitrogen input in high latitude ecosystems than previously recognized. They invite the reassessment of current theoretical models linking metal dynamics and dinitrogen fixation in boreal and subarctic ecosystems.

Foliose lichens with cyanobacterial bionts (bipartite and tripartite) form a distinct assemblage of epiphytes strongly associated with humid microclimatic conditions in inland British Columbia. Previous research showed that these cyano- and cephalolichen communities are disproportionately abundant and species-rich on conifer saplings beneath Populus compared to beneath other tree species. More revealing, lichens with cyanobacterial bionts were observed beneath Populus even in stands that did not otherwise support them. We experimentally test the hypothesis that this association is due to the interception of glucose-rich nectar that is exuded from Populus extra-floral nectaries (EFN). Using CO2 flux measurements and phospholipid fatty acid (PLFA) analysis with experimental applications of 13C6-labeled glucose, we demonstrate that cyano- and cephalolichens have a strong respiratory response to glucose. Lichens treated with glucose had lower net photosynthesis and higher establishment rates than control thalli. Furthermore, lichens with cyanobacterial bionts rapidly incorporate exogenous 13C into lichen fatty acid tissues. A large proportion of the 13C taken up by the lichens was incorporated into fungal biomarkers, suggesting that the mycobiont absorbed and assimilated the majority of applied 13C6 glucose. Our observations suggest that both cyanolichens and cephalolichens may utilize an exogenous source of glucose, made available by poplar EFNs. The exogenous C may enable these lichens to become established by providing a source of C for fungal respiration despite drought-induced inactivity of the cyanobacterial partner. As such, the mycobiont may adopt an alternative nutritional strategy, using available exogenous carbon to extend its realized niche.

Summary Development of the symbiotic association in the bipartite lichen Pseudocyphellaria crocata was investigated by characterizing two regions of the thallus. Thallus organization was examined using microscopy. A HIP1-based differential display technique was modified for use on Nostoc strains, including lichenized strains. Northern hybridization and quantitative real-time polymerase chain reaction were used to confirm differential display results, and determine expression levels of key cyanobacterial genes. Photosystem II yield across the thallus was measured using pulse-amplitude modulated fluorescence. Microscopy revealed structural differences in the thallus margins compared with the centre and identified putative heterocysts in both regions. Differential display identified altered transcript levels in both Nostoc punctiforme and a lichenized Nostoc strain. Transcript abundance of cox2, atpA, and ribA was increased in the thallus margin compared with the centre. Expression of cox2 is heterocyst specific and expression of other heterocyst-specific genes (hetR and nifK) was elevated in the margin, whereas, expression of psbB and PSII yield were not. Structural organization of the thallus margin differed from the centre. Both regions contained putative heterocysts but gene expression data indicated increased heterocyst differentiation in the margins where photosystem II yield was decreased. This is consistent with a zone of heterocyst differentiation within the thallus margin.

Growth in two old forest lichens was studied to evaluate how temporal (seasonal) and spatial (aspect-wise) partitioning of biomass and area growth respond to seasonal changes in light and climate. We monitored relative growth rates during annual courses in the cephalolichen Lobaria pulmonaria and the cyanolichen Lobaria scrobiculata transplanted in boreal clear-cut to five fixed aspects in winter, spring, summer, and autumn. For each annual set, growth was quantified in January-March, April-June, July-September and October-December. Mean biomass and area increased in all seasons, but growth was highest in July-September. Mass growth did not follow area increment during a year. As a result, mass per area (specific thallus mass (STM)) declined (L. scrobiculata) or stayed constant (L. pulmonaria) in the dark, humid October-December season, whereas it strongly increased in the dry, sunny April-June season. Aspect influenced growth in species-specific ways. Seasonality in biomass growth mainly followed light availability, whereas area growth was strongest during humid seasons. The substantial STM changes across seasons, species, and aspects can be explained as passive responses to seasonal climate. However, as STM, according to the literature, is a driver of water storage, recorded changes probably improve fitness by prolonging hydration in places or during times with high evaporative demands.

Question: What are the responses of epiphytic lichens to the intensity of management along a large environmental gradient in Mediterranean Quercus forests? Location: Central Spain. Methods: This study was carried out on 4590 trees located in 306 forest stands dominated by Quercus faginea or Quercus ilex ssp. ballota. The effect of forest management and other predictor variables on several species diversity indicators were studied. Variables modelled were total species richness, cyanolichen richness and community composition. A large number of predictor variables were included: forest fragmentation (patch size, stand variability), climate and topographic (altitude, slope, sun radiation, annual rainfall and mean annual temperature) and intensity of management. General linear models and constrained ordination techniques were used to model community traits and species composition, respectively. Results: Total richness and especially cyanolichens richness were significantly and negatively affected by the intensity of management. Lichen composition was influenced by management intensity, climatic and topographic variables and stand variability. Conclusions: In Mediterranean forests, human activities related to forestry, agricultural and livestock use cause impoverishment of lichen communities, including the local disappearance of the most demanding species. The conservation of unmanaged forests with a dense canopy is crucial for lichen diversity.

Nitrogen-fixing epiphytes (especially lichens with a cyanobacterial symbiont-cyanolichens) have the potential to contribute significant amounts of nitrogen (N) to montane tropical forests, which are typically low in N-but the factors controlling the abundance and distribution of epiphytic cyanolichens are poorly understood. In long-term fertilization experiments in montane forests on a 4.1 million-yr-old Oxisol on the island of Kaua' i and on a 152-yr-old lava flow on the island ofHawai'i, the epiphytic cyanolichen Pseudocyphellaria crocata increased significantly in abundance in canopies of host trees fertilized with phosphorus (P). There was no response to fertilization with N or other essential elements. Nitrogen-fixation rates were also elevated in lichens in P-fertilized plots at both sites. Phosphorus supply to host trees may be an important factor controlling N inputs to montane tropical forests by N-fixing epiphytes.

Dinitrogen (N2)-fixing lichens (cyanolichens) have long been recognized as a source of exogenous N in various ecosystems, yet the effects of nitrogen inputs from these lichens have been little studied. Cyanolichens of the genus Peltigera Willd. are often the dominant members of the earth-dwelling (terricolous) cyanolichen community. They occur in diverse habitats around the globe, but no studies of their influence on soil N have been reported. We conducted field and laboratory investigations with the primary objective of determining whether soil N availability is increased near healthy thalli of terricolous dinitrogen-fixing lichens. We measured available soil N in situ with ion-exchange resin bags, potentially mineralizable N with laboratory incubations, total soil N, and soil temperature. Measurements were taken along transects that extended away from thalli of Peltigera, perpendicular to and parallel to topographic contours. Studies were conducted in ten types of forest across northern Minnesota, including two contrasting sites that represent extremes of habitat for Peltigera. Soil N availability, potentially mineralizable N, and soil %N increased significantly with proximity to healthy thalli of Peltigera (P-values

Question/Location: We modelled lichen epiphyte biomass in relation to topography, stand structure, and lichen community composition in the central Cascades of western Oregon. Methods: Sampling was stratified by stand structure. Epiphyte biomass was estimated based on lichen litter for three functional groups: cyanolichens, forage lichens, and matrix lichens. Regression models for estimating lichen biomass (log10 kg.ha−1) were developed based on three pools of predictors, each pool demanding different levels of effort to obtain the data. First, we created models from topographic predictors that are easily extracted from GIS data. We then developed models based on both topographic and stand-structure variables. Finally, we developed models using topography, stand structure, and lichen community data. Results/Conclusions: Lichen biomass changed with elevation, cyanolichen biomass highest at low elevations (470–950 m) and forage lichen biomass highest at higher elevations (950–1470 m). Lichen biomass...

Summary • Cyanobacteria were isolated from bipartite cyanolichen species of Pseudocyphellaria and the identity of the major photobionts established. The specificity of the cyanobacterial–fungal association was also examined. • Comparison of 16S rRNA gene sequences distinguished cyanobacterial and green algal isolates, and both 16S rRNA gene and tRNALeu (UAA) intron sequences of isolates and lichen thalli identified candidate photobionts. In addition, the genetic diversity of both the cyanobiont and mycobiont was investigated using the comparison of tRNALeu (UAA) intron sequences and ITS sequences, respectively. • The 16S rRNA gene sequences identified two species-specific photobionts with similar sequences; however, the tRNALeu (UAA) intron sequences unambiguously discriminated between the two symbiotic cyanobacterial strains. Moreover, the fungal ITS sequences of the two corresponding lichens, Pseudocyphellaria crocata and Pseudocyphellaria neglecta, showed little variation. • The cyanobacterial–fungal associations of P. crocata and P. neglecta were specific for all samples. However, the similarity of the ITS sequences raised the possibility that they represent the same species and that their different morphology is influenced by the cyanobacterial symbiont.

Juvenile development from sown vegetative diaspores of the cyanobacterial lichen Lobaria scrobiculata and the green algal lichens Platismatia glauca and P. norvegica was studied through a four-year field experiment in a boreal spruce forest. All three species developed juvenile thalli within the period of observation. The largest lobules of L. scrobiculata and Platismatia were 0.4 mm and 1.3 mm, respectively, four years after sowing the diaspores. The first very tiny lobule (0.1 mm) of L. scrobiculata was observed 29 months after sowing the soredia. This period of juvenile development is the longest period so far reported for lichens. A long phase of dormancy, after the diaspores were attached to the substrate, was characteristic for the investigated species. Apical growth of the isidia of Platismatia glauca and P. norvegica was not observed until 21 months after sowing. Low light levels, low temperatures and snow covering the diaspores during winter, causes a short growth season in boreal spruce forests. These factors might explain the long period for juvenile development. The results indicate a slow rate of colonization of a new habitat for the investigated species since they need several years to grow to visible thalli.

We examined epiphytic macrolichen communities in Pseudotsuga menziesii (Douglas-fir) forests across the western Oregon landscape for relationships to environmental gradients, stand age and structure, and commercial thinning. We used a retrospective, blocked design through the Coast and the western Cascade ranges of Oregon. Each of our 17 blocks consisted of a young, unthinned stand (age 50–110 yr); an adjacent, thinned stand of equivalent age; and an old-growth stand (age > 200 yr). We found 110 epiphytic macrolichen taxa in the stands. Forage-providing alectorioid lichens and the nitrogen-fixing cyanolichen Lobaria oregana associated strongly with old-growth stands and remnant old trees in younger stands (unthinned + thinned). Relative to unthinned stands, thinned stands had a slightly higher abundance of alectorioid lichens and a greater presence of Hypogymnia imshaugii. However, thinned stands hosted a lower landscape-level (γ) diversity, lacking many species that occurred infrequently in the unthinned stands. Patterns in the lichen community composition correlated strongly with climatic gradients; the greatest variation in composition was between the Coast and Cascade ranges. The difference in communities between mountain ranges was greatest among stands 70–110 yr old, suggesting a difference in lichen successional dynamics between the ranges.

The “New Forestry” practice of green-tree retention is becoming an important management tool for publicly owned lands, yet few data exist to demonstrate that this tool can succeed at enhancing biodiversity. We addressed this issue by using a retrospective approach to compare canopy lichen litter in adjacent, paired stands of rotation age (55–120 yr): one with and one without old-growth (>300 yr) remnant trees. We sampled three functional groups of lichens in 17 stands in western Oregon: alectorioid lichens, cyanolichens, and green-algal foliose lichens. Thirteen stands were low elevation (520–850 m) and four were mid-elevation (1220–1340 m). Biomass of cyanolichen and green-algal foliose lichen litter was greater in low-elevation sites, whereas alectorioid lichen litter biomass was greater in mid-elevation sites. Cyanolichens were absent from all mid-elevation sites. Biomass of alectorioid lichen and cyanolichen litter was greater in low-elevation sites with remnant trees than in those without remnant tre...

The 400 million-year-old Rhynie chert has provided a wealth of information about various types of fungal interactions that existed in this Early Devonian paleoecosystem. In this paper we report the first unequivocal evidence of a lichen symbiosis from the Rhynie chert. Specimens of a new genus, Winfrenatia, consist of a thallus of superimposed layers of aseptate hyphae and, on the upper surface, numerous uniform depressions. Extending into the base of each depression are hyphae that form a three-dimensional netlike structure. Enclosed within each of the net spaces is a coccoid cyanobacterium, each cell of which is surrounded by a thick sheath. These photobiont cells divide in three planes, resulting in cell clusters of up to perhaps 64 individuals. The photobiont is parasitized by the fungus in the base of each net as new cyanobacterial cells are formed distally. Reproduction is by endospores and soredia. Affinities of the mycobiont appear closest to members of the Zygomycetes, while the photobiont is most similar to coccoid cyanobacteria of the Gloeocapsa and Chroococcidiopsis types. We speculate that this cyanobacterial symbiosis was well adapted to exploit and colonize new ecological niches, especially in the periodically desiccated environment postulated for the Rhynie chert paleoecosystem. Lichens are symbiotic associations that involve a mycobiont, typically an ascomycete, and a green alga or

Lichen cellulase may participate in the degradation of the external substrata and/or modification of the photobiont cell wall. To promote a better understanding of the roles of cellulases in lichens, a cyanolichen was chosen because of the absence of cellulose in its symbionts. Freshly-collected thalli of Peltigera canina (L.) Wild, produce β-1,4-glucanase (EC 3.2.1.4, β-1,4-D-glucanohydrolase). This enzyme's activity was detected in the soluble and cell wall fractions and it was found to be secreted to the incubation medium when thalli were floated on water or on cellobiose. Several forms of the enzyme were detected by isoelectrofocusing. In preparative isoelectrofocusing, a single peak was obtained in each fraction, characterized by pls of 5.05, 5.25 and 4.75 in the soluble, cell wall and medium fractions, respectively. These differences were in agreement with the different pattern of bands obtained in slab-isoelectrofocusing, where the most acidic band (pl of 4.45) was present only in the soluble fraction and the band with higher pl (6.17) was more intense in the cell wall fraction. Since both symbionts in a cyanolichen lack cellulose, cellulases cannot participate in the modification of their cell wall; the presence of cellulase in Peltigera canina must therefore be related to the degradation of the tissues of the moss substratum.

Nitrogen-fixing lichens (cyanolichens) are an important source of nitrogen (N) in Pacific Northwest forests, but limitation of lichen growth by elements essential for N fixation is poorly understood. To investigate how nutrient limitation may affect cyanolichen growth rates, we fertilized a tripartite cyanobacterial lichen (Lobaria pulmonaria) and a green algal non-nitrogen fixing lichen (Usnea longissima) with the micronutrients molybdenum (Mo) and vanadium (V), both known cofactors for enzymes involved in N fixation, and the macronutrient phosphorus (P). We then grew treated lichens in the field for one year in western Oregon, USA. Lichen growth was very rapid for both species and did not differ across treatments, despite a previous demonstration of P-limitation in L. pulmonaria at a nearby location. To reconcile these disparate findings, we analyzed P, Mo, and V concentrations, natural abundance δ15N isotopes, %N and change in thallus N in Lobaria pulmonaria from both growth experiments. Nitrogen levels in deposition and in lichens could not explain the large difference in growth or P limitation observed between the two studies. Instead, we provide evidence that local differences in P availability may have caused site-specific responses of Lobaria to P fertilization. In the previous experiment, Lobaria had low background levels of P, and treatment with P more than doubled growth. In contrast, Lobaria from the current experiment had much higher background P concentrations, similar to P-treated lichens in the previous experiment, consistent with the idea that ambient variation in P availability influences the degree of P limitation in cyanolichens. We conclude that insufficient P, Mo, and V did not limit the growth of either cyanolichens or chlorolichens at the site of the current experiment. Our findings point to the need to understand landscape-scale variation in P availability to cyanolichens, and its effect on spatial patterns of cyanolichen nutrient limitation and N fixation.

Free-living and lichenized cyanobacterial (cyanolichen) colonies occupy 12-29% of the surface area in the Sand Hills grasslands on the Arapaho Prairie in southwestern Nebraska. During brief periods of favorable conditions (i.e., moderate temperature and high moisture), these diazotrophs are capable of high rates of dinitrogen fixation. Maximum rates of acetylene reduction measured during 1981 and 1982 were 2,237 ,g N m-2 hr. The duration of dinitrogen fixation activity is dependent upon moisture conditions; rates of acetylene reduction were shown to decline 90% in as little as three hr. Desiccated colonies resumed acetylene reduction activity within one hr and achieved equilibrium rates within four hr following rehydration. Associative, heterotrophic diazotrophs were isolated from the rhizospheres of several grass species. These bacteria exhibited a range of culturing characteristics which may reflect differing plant-bacterial interaction potentials important to the establishment and maintenance of the plant species. None of the field assays for heterotrophic dinitrogen fixation performed over the two-yr period indicated a capacity of this group to provide ecologically significant levels of N