The evolution and biogeographic history of epiphytic thalloid liverworts.

Among liverworts, the epiphytic lifestyle is not only present in leafy forms but also in thalloid liverworts, which so far has received little attention in evolutionary and biogeographical studies. Metzgeria, with about 107 species worldwide, is the only genus of thalloid liverworts that comprises true epiphytes. In the present study, we provide the first comprehensive molecular phylogeny, including estimated divergence times and ancestral ranges of this genus. Analyses are based on a plastid marker dataset representing about half of the Metzgeria species diversity. We show for the first time with molecular data that Austrometzgeria is indeed a member of Metzgeria and that two morpho-species M. furcata and M. leptoneura are not monophyletic, but rather represent geographically well-defined clades. Our analyses indicate that Metzgeria started to diversify in the Cretaceous in an area encompassing today's South America and Australasia. Thus, Metzgeria is one of the few known epiphytic liverwort genera whose biogeographic history was directly shaped by Gondwana vicariance. Subsequent dispersal events in the Cenozoic resulted in the colonization of Asia, Africa, North America, and Europe and led to today's worldwide distribution of its species. We also provide the first reliable stem age estimate for Metzgeria due to the inclusion of its sister taxon Vandiemenia in our dating analyses. Additionally, this stem age estimate of about 240 million years most likely marks the starting point of a transition from a terrestrial to an epiphytic lifestyle in thalloid liverworts of the Metzgeriales. We assume that the Cretaceous Terrestrial Revolution played a key role in the evolution of epiphytic thalloid liverworts similar to that known for leafy liverworts.

Quantitative analysis of fine dust particles on moss surfaces under laboratory conditions using the example of Brachythecium rutabulum.

The identification of a model organism for investigations of fine dust deposits on moss leaflets was presented. An optical method with SEM enabled the quantitative detection of fine dust particles in two orders of magnitude. Selection criteria were developed with which further moss species can be identified in order to quantify the number of fine dust particles on moss surfaces using the presented method. Among the five moss species examined, B. rutabulum had proven to be the most suitable model organism for the method presented here. The number of fine dust particles on the moss surface of B. rutabulum was documented during 4 weeks of cultivation in the laboratory using SEM images and a counting method. The fine dust particles were recorded in the order of 10 µm-0.3 µm, divided into two size classes and counted. Under laboratory conditions, the number of particles of the fine fraction 2.4 µm-0.3 µm decreased significantly.

Different Predictors Shape the Diversity Patterns of Epiphytic and Non-epiphytic Liverworts in Montane Forests of Uganda.

We studied the influence of regional and local variables on the liverwort diversity within natural forest vegetation of Uganda to contribute to our understanding of the mechanisms and processes determining species richness. To this end, we compared the species richness distribution patterns of epiphytic and non-epiphytic liverworts (Marchantiophytina) in 24 plots in the forests of four Ugandan national parks. We recorded a total of 119 species and subspecies from 18 families, including 16 new species records for the country. We used generalized linear models (GLMs) and the relative variable importance of regional and local climatic and environmental variables to assess their respective impact on the species diversity. We found that the richness patterns of total and epiphytic richness were largely driven by regional climatic factors related to temperature and water-availability. In contrast, species diversity of non-epiphytic and rare species was additionally strongly determined by local-scale microhabitat factors such as height of forest canopy and slope inclination, reflecting the availability of suitable microhabitats. We conclude that macroclimatic variables perform well in predicting epiphytic liverwort richness, whereas the adequate prediction of non-epiphytic richness requires site-specific variables. Also, we propose that richness of epiphytic liverworts will be impacted more directly by climate change than richness of non-epiphytic and rare species.

World checklist of hornworts and liverworts.

A working checklist of accepted taxa worldwide is vital in achieving the goal of developing an online flora of all known plants by 2020 as part of the Global Strategy for Plant Conservation. We here present the first-ever worldwide checklist for liverworts (Marchantiophyta) and hornworts (Anthocerotophyta) that includes 7486 species in 398 genera representing 92 families from the two phyla. The checklist has far reaching implications and applications, including providing a valuable tool for taxonomists and systematists, analyzing phytogeographic and diversity patterns, aiding in the assessment of floristic and taxonomic knowledge, and identifying geographical gaps in our understanding of the global liverwort and hornwort flora. The checklist is derived from a working data set centralizing nomenclature, taxonomy and geography on a global scale. Prior to this effort a lack of centralization has been a major impediment for the study and analysis of species richness, conservation and systematic research at both regional and global scales. The success of this checklist, initiated in 2008, has been underpinned by its community approach involving taxonomic specialists working towards a consensus on taxonomy, nomenclature and distribution.

Aneuraceae (Metzgeriales) and tulasnelloid fungi (Basidiomycota): a model for early steps in fungal symbiosis.

A total of 35 population samples of the liverwort genera Aneura (A. pinguis) and Riccardia (R. latifrons, R. multifida, and R. palmata) were sampled from diverse habitats and geographical provenances in Germany, Austria, and Switzerland. Light and transmission electron microscopy were used to characterise the morphological features of the associations, and phylogenetic analyses based on internal transcribed spacers (ITS) and the D1/D2 regions of the fungal 28S rDNA were used to address diversity and phylogenetic relationships. By comparing the cellular structures of the plant-fungus interactions, we recognised the following states of fungal colonisation within the thalli: fungus-free, epiphytic, intercellular, and intracellular. Colonising hyphae showed dolipores with imperforate parenthesomes, slime bodies, and multilayered walls. Colonised liverwort cells had pleomorphic nuclei and elongated starch-free chloroplasts with distinctive grana. Our analyses revealed six phylogenetic groups of tulasnelloid fungi associated with liverworts, where major lineages mostly share similar host and/or ecological specialisations. The mode of colonisation of the tulasnelloid mycobionts in Aneura and Riccardia sharing identical fungal sequences is different. Consequently, the mode of colonisation may be host-dependent. Finally, our findings demonstrate that Aneuraceae are model organisms for evolutionary studies of symbiotic associations between liverworts and fungi.

Diverging diversity patterns in the Tulasnella (Basidiomycota, Tulasnellales) mycobionts of Aneura pinguis (Marchantiophyta, Metzgeriales) from Europe and Ecuador.

Aneura pinguis (Aneuraceae) is a cosmopolitan thalloid liverwort that shows a specific mycorrhiza-like interaction with basidiomycetes. To date, tropical specimens have not been studied in great depth. Samples of A. pinguis were collected from 48 individuals in one plot in South Ecuador and 54 individuals in five European countries. Light and transmission electron microscopy and molecular analyses based on nuclear rDNA coding for the ribosomal large subunit (nucLSU) and from the 5.8s-ITS2 regions were carried out to identify the associated mycobionts and to study their phylogenetic relationships. Microscopic and ultrastructural investigations of the fungal colonisation showed a high congruence between the European and the Ecuadorian sites and confirmed previous results. Tulasnellales are the only mycobionts that could be detected from ultrastructural characters with certainty. Molecular phylogenetic analysis indicated the presence of tulasnelloid fungi from at least 13 distinct clades. The composition of the communities of tulasnelloid fungi in A. pinguis differs between Ecuador and Europe. The diversity of tulasnelloid fungal partners was much higher at the Ecuadorian site.

Heterobasidiomycetes form symbiotic associations with hepatics: Jungermanniales have sebacinoid mycobionts while Aneura pinguis (Metzgeriales) is associated with a Tulasnella species.

In order to evaluate substrate dependence of the symbiotic fungal associations in leafy liverworts (Jungermanniopsida), 28 species out of 12 families were investigated by transmission electron microscopy and molecular methods. Samples were obtained from the diverse substrates: from naked soil, from the forest floor on needle litter, from between peat moss, from rotten bark of standing trees, and from stumps and rotten wood. Associations with ascomycetes were found in most of the specimens independent from the substrate. Seven species sampled from soil were found to contain basidiomycete hyphae. Ultrastructure consistently showed dolipores with imperforate parenthesomes. Molecular phylogenetic studies revealed that three specimens belonging to the Jungermanniales were associated with members of Sebacinaceae, while Aneura pinguis (Metzgeriales) was associated with a Tulasnella species. These taxa are so far the only basidiomycetes known to be symbiotically associated with leafy liverworts. The probability that the associations with Sebacinaceae are evolutionary old, but the Tulasnella associations more derived is discussed. The sebacinoid mycobionts form a similar interaction type with the jungermannialian leafy liverworts as do the associated ascomycetes. The term 'jungermannioid mycorrhiza' is proposed for this distinctive symbiotic interaction type.