Microclimatic differentiation of gene pools in the Lobaria pulmonaria symbiosis in a primeval forest landscape

Population genetics of the tree‐colonizing lichen Lobaria pulmonaria were studied in the largest primeval beech forest of Europe, covering 10 000 ha. During an intensive survey of the area, we collected 1522 thallus fragments originating from 483 trees, which were genotyped with eight mycobiont‐ and 14 photobiont‐specific microsatellite markers. The mycobiont and photobiont of L. pulmonaria were found to consist of two distinct gene pools, which are co‐existing within small areas of 3–180 ha in a homogeneous beech forest. The small‐scale distribution pattern of the symbiotic gene pools show habitat partitioning of lineages associated with either floodplains or mountain forests. Using approximate Bayesian computation (ABC), we dated the divergence of the two fungal gene pools of L. pulmonaria as the Early Pleistocene. Both fungal gene pools survived the Pleistocene glacial cycles in the Carpathians, although possibly in climatically different refugia. Fungal diversification prior to these cycles and the selection of photobionts with different altitudinal distributions explain the current sympatric, but ecologically differentiated habitat partitioning of L. pulmonaria. In addition, the habitat preferences of the mycobiont are determined by other factors and are rather independent of those of the photobiont at the landscape level. The distinct gene pools should be considered evolutionarily significant units and deserve specific conservation priorities in the future, for example gene pool A, which is a Pliocene relict.     

Cytokines induce nitric oxide-mediated mtDNA damage and apoptosis in oligodendrocytes. Protective role of targeting 8-oxoguanine glycosylase to mitochondria

Nitric oxide (NO) that is produced by inducible NO synthase (iNOS) in glial cells is thought to contribute significantly to the pathogenesis of multiple sclerosis. Oligodendrocytes can be stimulated to express iNOS by inflammatory cytokines, which are known to accumulate in the multiple sclerotic brain. The potentially pathological levels of NO produced under these circumstances can target a wide spectrum of intracellular components. We hypothesized that one of the critical targets for damage that leads to disease is mtDNA. In this study, we found that cytokines, in particular a combination of tumor necrosis factor-alpha (50 ng/ml) and IFNgamma (25 ng/ml), cause elevated NO production in primary cultures of rat oligodendrocytes. Western blot analysis revealed a strong enhancement of iNOS expression 48 h after cytokine treatment. Within the same time period, NO-mediated mtDNA damage was shown by Southern blot analysis and by ligation-mediated PCR. Targeting the DNA repair enzyme human 8-oxoguanine DNA glycosylase (hOGG1) to the mitochondria of oligodendrocytes had a protective effect against this cytokine-mediated mtDNA damage. Moreover, it was shown that mitochondrial transport sequence hOGG1-transfected oligodendrocytes had fewer apoptotic cells compared with cells containing vector only following treatment with the cytokines. Subsequent experiments revealed that targeting hOGG1 to mitochondria reduces the activation of caspase-9, showing that this recombinant protein works to reduce apoptosis that is occurring through a mitochondria-based pathway.     

Distribution and dispersal ecology of Lobaria pulmonaria in the largest primeval beech forest of Europe

Occupancy and density of the epiphytic lichen L. pulmonaria were studied in the mountains of Uholka–Shyrokyi Luh (Ukraine), which include the largest primeval beech forest in Europe. The lichen occupancy was assessed on 314 plots laid out on a systematic grid. Additional data on population density were collected from 483 trees growing both, on and between these plots. The trees harbouring L. pulmonaria were distributed very sparsely within Uholka–Shyrokyi Luh, and occupy nearly 10 % of the studied perimeter. The generalized linear models showed that area of occupancy of L. pulmonaria was significantly influenced by altitude and canopy cover, whereas the species’ density was explained by habitat types and slope exposition. Population density is higher at the timberline than in the interior forest or on lowland meadows. We found a bimodal altitudinal distribution of L. pulmonaria, with maxima below and above 900 m a.s.l., where it prefers forest stands with loose or scattered canopy. The preferred position of L. pulmonaria on host tree trunks depends on stand density and allows the species to get the necessary level of insolation also in shaded sites where it grows higher up on the trunk than in open stands. While L. pulmonaria occupied trees with various diameters, juvenile individuals are more frequent on small trees, but mature lichen individuals are predominantly found on trees of average or large sizes. Fertile individuals require specific environmental conditions, which are available at intermediate altitudes, related with sheltered light, and horizontal terraces on slopes with eastern exposition. In general, the primeval beech forest of Uholka–Shyrokyi Luh harbours a high percentage of juvenile thalli of L. pulmonaria, which lack vegetative propagules. Mature individuals have a low frequency of fruit bodies and reproduce mainly with vegetative diaspores. We interpret this as an indication of a currently growing population of L. pulmonaria within the area. We hypothesize that transboundary air pollution has decreased the lichens’ population frequency and density and has altered the ratio of developmental stages in L. pulmonaria during earlier decades.