Silke Werth

QUANTIFYING DISPERSAL AND ESTABLISHMENT LIMITATION IN A POPULATION OF AN EPIPHYTIC LICHEN

Dispersal is a process critical for the dynamics and persistence of metapopulations, but it is difficult to quantify. It has been suggested that the old-forest lichen Lobaria pulmonaria is limited by insufficient dispersal ability. We analyzed 240 DNA extracts derived from snow samples by a L. pulmonaria-specific real-time PCR (polymerase chain reaction) assay of the ITS (internal transcribed spacer) region allowing for the discrimination among propagules originating from a single, isolated source tree or propagules originating from other locations. Samples that were detected as positives by real-time PCR were additionally genotyped for five L. pulmonaria microsatellite loci. Both molecular approaches demonstrated substantial dispersal from other than local sources. In a landscape approach, we additionally analyzed 240 snow samples with real-time PCR of ITS and detected propagules not only in forests where L. pulmonaria was present, but also in large unforested pasture areas and in forest patches where L. pulmonaria was not found. Monitoring of soredia of L. pulmonaria transplanted to maple bark after two vegetation periods showed high variance in growth among forest stands, but no significant differences among different transplantation treatments. Hence, it is probably not dispersal limitation that hinders colonization in the old-forest lichen L. pulmonaria, but ecological constraints at the stand level that can result in establishment limitation. Our study exemplifies that care has to be taken to adequately separate the effects of dispersal limitation from a limitation of establishment.

Effect of disturbances on the genetic diversity of an old-forest associated lichen

Lichens associated with old forest are commonly assumed to be negatively affected by tree logging or natural disturbances. However, in this study performed in a spruce‐dominated sylvopastoral landscape in the Swiss Jura Mountains, we found that genetic diversity of the epiphytic old‐forest lichen Lobaria pulmonaria depends on the type of disturbance. We collected 923 thalli from 41 sampling plots of 1 ha corresponding to the categories stand‐replacing disturbance (burnt), intensive logging (logged) and uneven‐aged forestry (uneven‐aged), and analysed the thalli at six mycobiont‐specific microsatellite loci. We found evidence for multiple independent immigrations into demes located in burnt and logged areas. Using spatial autocorrelation methods, the spatial scale of the genetic structure caused by the clonal and recombinant component of genetic variation was determined. Spatial autocorrelation of genotype diversity was strong at short distances up to 50 m in logged demes, up to 100 m in uneven‐aged demes, with the strongest autocorrelation up to 150 m for burnt demes. The spatial autocorrelation was predominantly attributed to clonal dispersal of vegetative propagules. After accounting for the clonal component, we did not find significant spatial autocorrelation in gene diversity. This pattern may indicate low dispersal ranges of clonal propagules, but random dispersal of sexual ascospores. Genetic diversity was highest in logged demes, and lowest in burnt demes. Our results suggest that genetic diversity of epiphytic lichen demes may not necessarily be impacted by stand‐level disturbances for extended time periods.

Landscape‐level gene flow in Lobaria pulmonaria, an epiphytic lichen

Epiphytes are strongly affected by the population dynamics of their host trees. Owing to the spatio‐temporal dynamics of host tree populations, substantial dispersal rates — corresponding to high levels of gene flow — are needed for populations to persist in a landscape. However, several epiphytic lichens have been suggested to be dispersal‐limited, which leads to the expectation of low gene flow at the landscape scale. Here, we study landscape‐level genetic structure and gene flow of a putatively dispersal‐limited epiphytic lichen, Lobaria pulmonaria. The genetic structure of L. pulmonaria was quantified at three hierarchical levels, based on 923 thalli collected from 41 plots situated within a pasture–woodland landscape and genotyped at six fungal microsatellite loci. We found significant isolation by distance, and significant genetic differentiation both among sampling plots and among trees. Landscape configuration, i.e. the effect of a large open area separating two forested regions, did not leave a traceable pattern in genetic structure, as assessed with partial Mantel tests and analysis of molecular variance. Gene pools were spatially intermingled in the pasture–woodland landscape, as determined by Bayesian analysis of population structure. Evidence for local gene flow was found in a disturbed area that was mainly colonized from nearby sources. Our analyses indicated high rates of gene flow of L. pulmonaria among forest patches, which may reflect the historical connectedness of the landscape through gene movement. These results support the conclusion that dispersal in L. pulmonaria is rather effective, but not spatially unrestricted.

Identity and genetic structure of the photobiont of the epiphytic lichen Ramalina menziesii on three oak species in southern California.

Lichens, a classic example of an obligate symbiosis between fungi and photobionts (which could be algae or cyanobacteria), are abundant in many terrestrial ecosystems. The genetic structure of the photobiont population found in association with a lichen-forming fungal species could be affected by fungal reproductive mode and by the spatial extent of gene flow in the photobiont. Using DNA sequences from one nuclear ribosomal and two chloroplast loci, we analyzed the genetic structure of the photobiont associated with the fungus Ramalina menziesii at an oak woodland study site in southern California. We had previously shown that the fungus exhibited no genetic structure among four local sites or three phorophyte species. Our goals were to identify the photobiont species and assess its genetic structure. We found that R. menziesii was highly specific in its photobiont choice and associated with one alga, Trebouxia decolorans. In contrast to the fungal population, we found significant differentiation among the algae sampled on three oak species and little genetic structure among the sites for two of the three algal loci. We hypothesize that R. menziesii is locally adapted to the phorophyte species through habitat specialization in the algal partner of the symbiosis.

Modelling forest recolonization by an epiphytic lichen using a landscape genetic approach

The process of recolonization after disturbance is crucial for the persistence and dynamics of patch-tracking metapopulations. We developed a model to compare the spatial distribution and spatial genetic structure of the epiphytic lichen Lobaria pulmonaria within the perimeter of two reconstructed 19th century disturbances with a nearby reference area without stand-level disturbance. Population genetic data suggested that after stand-replacing disturbance, each plot was colonized by one or a few genotypes only, which subsequently spread clonally within a local neighborhood. The model (cellular automaton) aimed at testing the validity of this interpretation and at assessing the relative importance of local dispersal of clonal propagules vs. long-distance dispersal of clonal and/or sexual diaspores. A reasonable model fit was reached for the empirical data on host tree distribution, lichen distribution, and tree- and plot-level genotype diversity of the lichen in the reference area. Although model calibration suggested a predominance of local dispersal of clonal propagules, a substantial contribution of immigration of non-local genotypes by long-distance dispersal was needed to reach the observed levels of genotype diversity. The model could not fully explain the high degree of clonality after stand-replacing disturbance, suggesting that the dispersal process itself may not be stationary but depend on the ecological conditions related to disturbance.

Population genetics of lichen-forming fungi - a review

Population genetics investigates the distribution of genetic variation in natural populations and the genetic differentiation among populations. Lichen-forming fungi are exciting subjects for population genetic studies due to their obligate symbiosis with a green-algal and/or cyanobacterial photobiont, and because their different reproductive strategies could influence fungal genetic struc- tures in various ways. In this review, first, I briefly summarize the results from studies of chemotype variation in populations of lichen-forming fungi. Second, I compare and evaluate the DNA-based molecular tools available for population genetics of lichen-forming fungi. Third, I review the literature available on the genetic structure of lichen fungi to show general trends. I discuss some fascinating examples, and point out directions for future research.

Congruent Genetic Structure in the Lichen-Forming Fungus Lobaria pulmonaria and Its Green-Algal Photobiont

The extent of codispersal of symbionts is one of the key factors shaping genetic structures of symbiotic organisms. Concordant patterns of genetic structure are expected in vertically transmitted symbioses, whereas horizontal transmission generally uncouples genetic structures unless the partners are coadapted. Here, we compared the genetic structures of mutualists, the lichen-forming fungus Lobaria pulmonaria and its primary green-algal photobiont, Dictyochloropsis reticulata. We performed analysis of molecular variance and variogram analysis to compare genetic structures between symbiosis partners. We simulated the expected number of multilocus-genotype recurrences to reveal whether the distribution of multilocus genotypes of either species was concordant with panmixia. Simulations and tests of linkage disequilibrium provided compelling evidence for the codispersal of mutualists. To test whether genotype associations between symbionts were consistent with randomness, as expected under horizontal transmission, we simulated the recurrence of fungal-algal multilocus genotype associations expected by chance. Our data showed nonrandom associations of fungal and algal genotypes. Either vertical transmission or horizontal transmission coupled with coadaptation between symbiont genotypes may have created these nonrandom associations. This study is among the first to show codispersal and highly congruent genetic structures in the partners of a lichen mutualism.

Local genetic structure in a North American epiphytic lichen, Ramalina menziesii (Ramalinaceae).

Epiphytic lichens possess unique life history traits that can have conflicting effects on genetic structure: symbiotic mutualism between a fungus with its algal or cyanobacterial photobiont, association with a host plant, and ability to reproduce sexually and asexually. Our study species, Ramalina menziesii, has small ascospores that can facilitate long-distance gene movement, and it is capable of clonal reproduction. The goals of this study are to test whether different haplotypes were differentially distributed across host plant species, to look for evidence of asexual vs. sexual reproduction, and to assess the local genetic structure of the population. We sampled individuals from multiple trees of three oak species in four lichen subpopulations within a savanna ecosystem. Using DNA sequence data from four fungal nuclear loci, we found no tendency for host specialization. Alleles were randomly distributed across subpopulations. The frequency of multilocus genotypes was consistent with a randomly mating population. Sexual reproduction involving relichenization appeared to be the predominant mode of reproduction of R. menziesii at this study site. We found no significant local genetic structure suggesting widespread gene flow at the local scale. The genetic structure of this lichen is comparable to that of widely distributed epiphytic plants.

Hitchhiking with forests: population genetics of the epiphytic lichen Lobaria pulmonaria in primeval and managed forests in southeastern Europe

Availability of suitable trees is a primary determinant of range contractions and expansions of epiphytic species. However, switches between carrier tree species may blur co-phylogeographic patterns. We identified glacial refugia in southeastern Europe for the tree-colonizing lichen Lobaria pulmonaria, studied the importance of primeval forest reserves for the conservation of genetically diverse populations and analyzed differences in spatial genetic structure between primeval and managed forests with fungus-specific microsatellite markers. Populations belonged to either of two genepools or were admixed. Gene diversity was higher in primeval than in managed forests. At small distances up to 170 m, genotype diversity was lower in managed compared with primeval forests. We found significant associations between groups of tree species and two L. pulmonaria genepools, which may indicate “hitchhiking” of L. pulmonaria on forest communities during postglacial migration. Genepool B of L. pulmonaria was associated with European Beech (Fagus sylvatica) and we can hypothesize that genepool B survived the last glaciation associated within the refuge of European Beech on the Coastal and Central Dinarides. The allelic richness of genepool A was highest in the Alps, which is the evidence for a northern refuge of L. pulmonaria. Vicariant altitudinal distributions of the two genepools suggest intraspecific ecological differentiation.

Phylogeography of Ramalina menziesii, a widely distributed lichen-forming fungus in western North America

The complex topography and climate history of western North America offer a setting where lineage formation, accumulation and migration have led to elevated inter‐ and intraspecific biodiversity in many taxa. Here, we study Ramalina menziesii, an epiphytic lichenized fungus with a range encompassing major ecosystems from Baja California to Alaska to explore the predictions of two hypotheses: (i) that the widespread distribution of R. menziesii is due to a single migration episode from a single lineage and (ii) that the widespread distribution is due to the formation and persistence of multiple lineages structured throughout the species range. To obtain evidence for these predictions, we first construct a phylogenetic tree and identify multiple lineages structured throughout the species range – some ancient ones that are localized and other more recent lineages that are widely distributed. Second, we use an isolation with migration model to show that sets of ecoregion populations diverged from each other at different times, demonstrating the importance of historical and current barriers to gene flow. Third, we estimated migration rates among ecoregions and find that Baja California populations are relatively isolated, that inland California ecoregion populations do not send out emigrants and that migration out of California coastal and Pacific Northwest populations into inland California ecoregions is high. Such intraspecific geographical patterns of population persistence and dispersal both contribute to the wide range of this genetically diverse lichen fungus and provide insight into the evolutionary processes that enhance species diversity of the California Floristic Province.