Nitrate uptake by isolated bionts of the lichen <Emphasis Type="Italic">Parmelia sulcata</Emphasis>

It was found that bionts isolated from the lichen Parmelia sulcata Taylor had a marked difference with respect to nitrate assimilation. Isolated and purified photobiont, the green alga Trebouxia sp. showed no ability of nitrate absorption. Mycobiont and thallus fragments containing both bionts absorbed nitrate. Illumination had no essential influence on the rate of nitrate uptake. The respiratory inhibitor sodium azide decreased the rate of nitrate uptake by 80–100%, whereas the photosynthetic inhibitor dichlorophenyldimethylurea did not reduce it. Using mass-spectroscopic technique, it was shown that, in the intact thallus, nitrate was first absorbed by the mycobiont, and only later appeared in the photobiont. Probably such nutritional difference between bionts serves as one of the mechanisms by which the host fungi control the associated green algae and support their symbiosis.     

The marine algae of Rhodos island,Greece

The main purpose of this research is to describe the distribution of marine algae along the shores of Rhodos island. In the investigated medio- and infralittoral zones, collections revealed 131 species distributed as follows: Chlorophyceae 22·9%, Phaeophyceae 20·6% and Rhodophyceae 56·5%. The ratio Rhodophyceae/Phaeophyceae is 2·7, similar to that found by Gerloff & Geissler (1974) and lower than in other parts of the Mediterranean sea (Feldmann, 1938; Giaccone, 1968).     

Photobiont selectivity leads to ecological tolerance and evolutionary divergence in a polymorphic complex of lichenized fungi

Background and Aims

The integrity and evolution of lichen symbioses depend on a fine-tuned combination of algal and fungal genotypes. Geographically widespread species complexes of lichenized fungi can occur in habitats with slightly varying ecological conditions, and it remains unclear how this variation correlates with symbiont selectivity patterns in lichens. In an attempt to address this question, >300 samples were taken of the globally distributed and ecologically variable lichen-forming species complex Tephromela atra, together with closely allied species, in order to study genetic diversity and the selectivity patterns of their photobionts.

Methods

Lichen thalli of T. atra and of closely related species T. grumosa, T. nashii and T. atrocaesia were collected from six continents, across 24 countries and 62 localities representing a wide range of habitats. Analyses of genetic diversity and phylogenetic relationships were carried out both for photobionts amplified directly from the lichen thalli and from those isolated in axenic cultures. Morphological and anatomical traits were studied with light and transmission electron microscopy in the isolated algal strains.

Key Results

Tephromela fungal species were found to associate with 12 lineages of Trebouxia. Five new clades demonstrate the still-unrecognized genetic diversity of lichen algae. Culturable, undescribed lineages were also characterized by phenotypic traits. Strong selectivity of the mycobionts for the photobionts was observed in six monophyletic Tephromela clades. Seven Trebouxia lineages were detected in the poorly resolved lineage T. atra sensu lato, where co-occurrence of multiple photobiont lineages in single thalli was repeatedly observed.

Conclusions

Low selectivity apparently allows widespread lichen-forming fungi to establish successful symbioses with locally adapted photobionts in a broader range of habitats. This flexibility might correlate with both lower phylogenetic resolution and evolutionary divergence in species complexes of crustose lichen-forming fungi.     

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.     

Tissue storage and primer selection influence pyrosequencing‐based inferences of diversity and community composition of endolichenic and endophytic fungi

Next‐generation sequencing technologies have provided unprecedented insights into fungal diversity and ecology. However, intrinsic biases and insufficient quality control in next‐generation methods can lead to difficult‐to‐detect errors in estimating fungal community richness, distributions and composition. The aim of this study was to examine how tissue storage prior to DNA extraction, primer design and various quality‐control approaches commonly used in 454 amplicon pyrosequencing might influence ecological inferences in studies of endophytic and endolichenic fungi. We first contrast 454 data sets generated contemporaneously from subsets of the same plant and lichen tissues that were stored in CTAB buffer, dried in silica gel or freshly frozen prior to DNA extraction. We show that storage in silica gel markedly limits the recovery of sequence data and yields a small fraction of the diversity observed by the other two methods. Using lichen mycobiont sequences as internal positive controls, we next show that despite careful filtering of raw reads and utilization of current best‐practice OTU clustering methods, homopolymer errors in sequences representing rare taxa artificially increased estimates of richness c. 15‐fold in a model data set. Third, we show that inferences regarding endolichenic diversity can be improved using a novel primer that reduces amplification of the mycobiont. Together, our results provide a rationale for selecting tissue treatment regimes prior to DNA extraction, demonstrate the efficacy of reducing mycobiont amplification in studies of the fungal microbiomes of lichen thalli and highlight the difficulties in differentiating true information about fungal biodiversity from methodological artefacts.     

地衣标本的采集、制作与保存

详细论述了地衣标本的采集、制作和保存方法及注意事项,同时阐明了在采集标本的同时进行共生菌和光合共生物分离培养的重要性。     

Photobiont Selectivity and Interspecific Interactions in Lichen Communities. I. Culture Experiments with the Mycobiont Fulgensia bracteata

Abstract: Lichen communities are characterised by interspecific interactions that not only include interactions between different lichen species but also between the symbionts within a single lichen species. The community “Bunte Erdflechtengesellschaft”, growing on weathered calciferous rocks known as Gravel Alvar on Gotland (Baltic Sea, Sweden), shows a high complexity of inter‐ and intraspecific interactions, including Fulgensia bracteata, F. fulgens, Toninia sedifolia, Squamarina cartilaginea, Psora decipiens and Lecidea lurida. F. bracteata and F. fulgens are the dominant species of this community, showing a tendency to overgrow the other species involved and even parasitic behaviour. Culture experiments have been performed to investigate the selectivity of the mycobiont of F. bracteata towards a variety of potential photobionts. The results provide evidence for the selectivity of the mycobiont and varying compatibility of the respective symbionts that can be interpreted as a cascade of interdependent processes of specific and non‐specific reactions of the symbionts involved.     

Two new guaiane-sesquiterpenes from the cultured lichen mycobiont of Diorygma pruinosum

Lichens and the mycobionts derived from lichen-source are believed to be a valuable source of bioactive compounds. Diorygma pruinosum is a native lichen in Vietnam. A mycobiont of Diorygma pruinosum was separated, then cultivated. Phytochemical data on this lichen and its cultured mycobionts are scarce. The present study described the isolation and structural elucidation of two new guaiane-type sesquiterpenes, namely pruinosone (1) and hydroxypruinosone (2). Their absolute chemical structures were elucidated by extensive 1D and 2D NMR analysis, high-resolution mass spectroscopy, electronic circular dichroism (ECD), and comparisons in the literatures. Compounds 1 and 2 were evaluated for alpha-glucosidase inhibition and antimicrobial activity.