DNA barcoding and LC-MS metabolite profiling of the lichen-forming genus Melanelia: Specimen identification and discrimination focusing on Icelandic taxa

Taxa in the genus Melanelia (Parmeliaceae, Ascomycota) belong to a group of saxicolous lichens with brown to black foliose thalli, which have recently undergone extensive changes in circumscription. Taxa belonging to Parmeliaceae are prolific producers of bioactive compounds, which have also been traditionally used for chemotaxonomic purposes. However, the chemical diversity of the genus Melanelia and the use of chemical data for species discrimination in this genus are largely unexplored. In addition, identification based on morphological characters is challenging due to few taxonomically informative characters. Molecular identification methods, such as DNA barcoding, have rarely been applied to this genus. This study aimed to identify the Melanelia species from Iceland using DNA barcoding approach, and to explore their chemical diversity using chemical profiling. Chemometric tools were used to see if lichen metabolite profiles determined by LC-MS could be used for the identification of Icelandic Melanelia species. Barcoding using the fungal nuclear ribosomal internal transcribed spacer region (nrITS) successfully identified three Melalenlia species occurring in Iceland, together with Montanelia disjuncta (Basionym: Melanelia disjuncta). All species formed monophyletic clades in the neighbor-joining nrITS gene tree. However, high intraspecific genetic distance of M. stygia suggests the potential of unrecognized species lineages. Principal component analysis (PCA) of metabolite data gave a holistic overview showing that M. hepatizon and M. disjuncta were distinct from the rest, without the power to separate M. agnata and M. stygia due to their chemical similarity. Orthogonal partial least–squares to latent structures–discriminate analysis (OPLS-DA), however, successfully distinguished M. agnata and M. stygia by identifying statistically significant metabolites, which lead to class differentiation. This work has demonstrated the potential of DNA barcoding, chemical profiling and chemometrics in identification of Melanelia species.The study was financially supported by the People Programme (Marie Curie Actions) of the European Union’s Seventh Framework Programme FP7/2007-2013/ under REA grant agreement No. 606895 (http://cordis.europa.eu/project/rcn/109122_en.html) as well as a minor contribution from Bergthora and Thorsteinn Scheving Thorsteinsson Fund. The funders provided support in the form of salaries for author [MX], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of this author is articulated in the ‘author contributions’ section. The commercial affiliation Arctic Mass has no financial or competing interests in this study but two of the authors MT and FFE are partially affiliated there. Arctic Mass had the role of providing access to instruments (mass spectrometers) used in the study.Peer Reviewe

Phylogenetic diversity of the lichenized algal genus Trebouxia (Trebouxiophyceae, Chlorophyta): a new lineage and novel insights from fungal-algal association patterns of Icelandic cetrarioid lichens (Parmeliaceae, Ascomycota)

Publisher's version (útgefin grein)Lichens have high tolerance to harsh environmental conditions, where lichen symbiont interactions (e.g. myco- and photobionts) may play a crucial role. The characterization of fungal-algal association patterns is essential to understand their symbiotic interactions. This study investigated fungal-algal association patterns in Icelandic cetrarioid lichens using a multi-locus phylogenetic framework, including fungal nrITS, MCM7, mtSSU, RPB1 and RPB2 and algal nrITS, nrLSU, rbcL and mtCOXII data. Most Icelandic cetrarioid lichenized fungi were found to be specifically associated to the known Trebouxia clade “S” (Trebouxia simplex/suecica group), whereas the lichen-forming fungus Cetrariella delisei forms a symbiosis with a previously unrecognized lineage of Trebouxia, provisionally named as the “D” clade. This new Trebouxia lineage is supported by maximum likelihood and Bayesian phylogenetic analyses using all four included algal loci.This project was supported by the European Union’s Seventh Framework Programme for research, technological development and demonstration (grant agreement number 606895) to FP7-MCA-ITN MedPlant, “Phylogenetic Exploration of Medicinal Plant Diversity”. The Icelandic Research Fund (grant number 185442051) and the Bergthora and Thorsteinn Scheving Thorsteinsson Fund are also acknowledged for financial support.Peer Reviewe

What are the effects of herbivore diversity on tundra ecosystems? : A systematic review protocol

Funding Information: The project was funded by the Icelandic Research Fund (Grant Nr. 217754) and the European Union’s Horizon 2020 programme (CHARTER project, Grant Agreement Nr. 869471). Funding for open access publication was provided by the Agricultural University of Iceland. The funding bodies had no influence in the design of the study and collection, analysis and interpretation of data. Funding Information: This study is a contribution of the Herbivory Network (http://herbivory.lbhi.is), a UArctic Thematic Network. Publisher Copyright: © 2022, The Author(s).Background: Changes in the diversity of herbivore communities can strongly influence the functioning of northern ecosystems. Different herbivores have different impacts on ecosystems because of differences in their diets, behaviour and energy requirements. The combined effects of different herbivores can in some cases compensate each other but lead to stronger directional changes elsewhere. However, the diversity of herbivore assemblages has until recently been a largely overlooked dimension of plant–herbivore interactions. Given the ongoing environmental changes in tundra ecosystems, with increased influx of boreal species and changes in the distribution and abundance of arctic herbivores, a better understanding of the consequences of changes in the diversity of herbivore assemblages is needed. This protocol presents the methodology that will be used in a systematic review on the effects of herbivore diversity on different processes, functions and properties of tundra ecosystems. Methods: This systematic review builds on an earlier systematic map on herbivory studies in the Arctic that identified a relatively large number of studies assessing the effects of multiple herbivores. The systematic review will include primary field studies retrieved from databases, search engines and specialist websites, that compare responses of tundra ecosystems to different levels of herbivore diversity, including both vertebrate and invertebrate herbivores. We will use species richness of herbivores or the richness of functional groups of herbivores as a measure of the diversity of the herbivore assemblages. Studies will be screened in three stages: title, abstract and full text, and inclusion will follow clearly identified eligibility criteria, based on their target population, exposure, comparator and study design. The review will cover terrestrial Arctic ecosystems including the forest-tundra ecotone. Potential outcomes will include multiple processes, functions and properties of tundra ecosystems related to primary productivity, nutrient cycling, accumulation and dynamics of nutrient pools, as well as the impacts of herbivores on other organisms. Studies will be critically appraised for validity, and where studies report similar outcomes, meta-analysis will be performed.Peer reviewe

Alkaloid fingerprinting resolves Huperzia selago genotypes in Iceland

Pre-print (óritrýnt handrit)The club moss family Lycopodiaceae produces a diverse array of bioactive lycopodium alkaloids (LAs). In particular, the alkaloid huperzine A (hupA) has grasped attention since it is a potent acetylcholinesterase inhibitor of medical interest in Alzheimer's disease. Although the structural diversity and bioactivities of LAs have been studied to some extent, their chemotaxonomic value is mostly unexplored, especially to a lower taxonomic unit (e.g. subspecies or genotypes). This study focused on previously reported Icelandic Huperzia selago genotypes, and aimed to evaluate the chemotaxonomic value of LAs in resolving them. Using liquid chromatography-mass spectrometry (LC-MS), alkaloid fingerprints of H. selago taxa were subjected to principal component analysis (PCA). Our results revealed that each genotype tends to have its own alkaloid profile. Genotype 1 and 3 form distinct groups in a PCA plot, where genotype 2 is an intermediate between the other two genotypes. HupA and its derivative, huperzine B, both contribute to the differentiation of genotype 3 from the others. Therefore, our study demonstrated the potential of alkaloid fingerprints in resolving deep taxonomic groups and selecting plant taxa of medicinal importance.This work was supported by The People Program (Marie Curie Actions) of the European Union’s Seventh Framework Program FP7/2007-2013 (grant number 606895); Icelandic Research Fund (grant number 152604051); and the Bergthora and Thorsteinn Scheving Thorsteinsson Fund

Infraspecific Variation of Huperzine A and B in Icelandic Huperzia selago Complex

Publisher's version (útgefin grein)The alkaloids huperzine A and huperzine B were originally isolated from the Chinese club moss Huperzia serrata. They are known inhibitors of acetylcholinesterase, and especially huperzine A shows pharmaceutical potential for the treatment of Alzheimerʼs disease. Its supply heavily relies on natural plant sources belonging to the genus Huperzia, which shows considerable interspecific huperzine A variations. Furthermore, taxonomic controversy remains in this genus, particularly in the Huperzia selago group. With focus on Icelandic H. selago taxa, we aimed to explore the relatedness of Huperzia species using multi-locus phylogenetic analysis, and to investigate correlations between huperzine A contents, morphotypes, and genotypes. Phylogenetic analysis was performed with five chloroplastic loci (the intergenic spacer between the photosystem II protein D1 gene and the tRNA-His gene, maturase K, ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit, tRNA-Leu, and the intergenic spacer region between tRNA-Leu and tRNA-Phe). Huperzine A and huperzine B contents were determined using an HPLC-UV method. The phylogenetic analysis suggests that previously proposed Huperzia appressa and Huperzia arctica should not be considered species, but rather subspecies of H. selago. Three genotypes of Icelandic H. selago were identified and presented in a haplotype networking diagram. A significantly (p < 0.05) higher amount of huperzine A was found in H. selago genotype 3 (264 – 679 µg/g) than genotype 1 (20 – 180 µg/g), where the former shows a typical green and reflexed “selago” morphotype. The huperzine A content in genotype 3 is comparable to Chinese H. serrata and a good alternative huperzine A source. Genotype 2 contains multiple morphotypes with a broad huperzine A content (113 – 599 µg/g). The content of huperzine B in Icelandic taxa (6 – 13 µg/g) is much lower than that in Chinese H. serrata (79 – 207 µg/g).The People Programme (Marie Curie Actions) of the European Unionʼs Seventh Framework Programme FP7/2007–2013 (grant number 606895) and the Icelandic Research Fund (grant number 152604051) are acknowledged for financial support. This study was also funded from the Bergthora and Thorsteinn Scheving Thorsteinsson Fund.Peer Reviewe

Microbial sucession dynamics in the forefield of Breiðamerkurjokull Glacier (Iceland)

FEMS 2017 (7th. 2017. Valencia)Backgrounds One key consequence of glacier recession, as effect of climatic change, is the creation of new habitats for colonization. In glacier forefields, primary succession occurs simultaneously in soils and rocks recently discovered offering a type of natural experiment in which temporal colonization dynamics can be analyzed. Objectives A chronosequence established at Breiðamerkurjökull Glacier forefield, was used as a framework to analyze primary microbial succession processes in subarctic regions. This outlet glacier stretches to southeast from Vatnajökull Glacier and has been dramatically retreating during the 20th century. Methods Soil samples from different succession stages were collected. Microbial community structure was analyzed by high-throughput amplicon sequencing. Potential microbial activity (microbial respiration, N mineralization) as well as different soil attributes were also measured in these samples

Are recently deglaciated areas at both poles colonised by the same bacteria?

Polar glacier forefields offer an unprecedented framework for studying community assembly processes in regions that are geographically and climatically isolated. Through amplicon sequence variant (ASV) inference, we compared the composition and structure of soil bacterial communities from glacier forefields in Iceland and Antarctica to assess overlap between communities and the impact of established cryptogamic covers on the uniqueness of their taxa. These pioneer microbial communities were found to share only 8% of ASVs and each taxonomic group's contribution to the shared ASV data subset was heterogeneous and independent of their relative abundance. Although the presence of ASVs specific to one glacier forefield and/or different cryptogam cover values confirms the existence of habitat specialist bacteria, our data show that the influence of cryptogams on the edaphic bacterial community structure also varied also depending on the taxonomic group. Hence, the establishment of distinct cryptogamic covers is probably not the only factor driving the uniqueness of bacterial communities at both poles. The structure of bacterial communities colonising deglaciated areas seems also conditioned by lineage-specific limitations in their dispersal capacity and/or their establishment and persistence in these isolated and hostile regions

Status and trends in Arctic vegetation: Evidence from experimental warming and long-term monitoring

Changes in Arctic vegetation can have important implications for trophic interactions and ecosystem functioning leading to climate feedbacks. Plot-based vegetation surveys provide detailed insight into vegetation changes at sites around the Arctic and improve our ability to predict the impacts of environmental change on tundra ecosystems. Here, we review studies of changes in plant community composition and phenology from both long-term monitoring and warming experiments in Arctic environments. We find that Arctic plant communities and species are generally sensitive to warming, but trends over a period of time are heterogeneous and complex and do not always mirror expectations based on responses to experimental manipulations. Our findings highlight the need for more geographically widespread, integrated, and comprehensive monitoring efforts that can better resolve the interacting effects of warming and other local and regional ecological factors

Nordic LifeWatch cooperation, final report: A joint initiative from Denmark, Iceland, Finland, Norway and Sweden

The main goal of the present report is to outline the possibilities for an enhanced cooperation between the Nordic countries within eScience and biodiversity. LifeWatch is one of several ESFRI projects which aim to establish eInfrastructures and databases in the field of biodiversity and ecosystem research. Similarities between Nordic countries are extensive in relation to a number of biodiversity related issues. Most species in Nordic countries are common, and frequently the same challenges concerning biodiversity and ecosystem services are addressed in the different countries. The present report has been developed by establishing a Nordic LifeWatch network with delegates from each of the Nordic countries. The report has been written jointly by the delegates, and the work was organized by establishing working groups with the following themes: strategic issues, technical development, legal framework and communication. Written during two workshops, Skype meetings and emailing, the following main issues are discussed in the present report: * Scientific needs for improved access to biodiversity data and advanced eScience research infrastructure in the Nordic countries. * Future challenges and priorities facing the international biodiversity research community. * Scientific potential of openly accessible biodiversity and environmental data for individual researchers and institutions. * Spin-off effects of open access for the general public. * Internationally standardized Nordic metadata inventory. * Legal framework and challenges associated with environmental-, climate-, and biodiversity data sharing, communication, training and scientific needs. * Finally, some strategic steps towards realizing a Nordic LifeWatch construction and operational phase are discussed. Easy access to open data on biodiversity and the environment is crucial for many researchers and research institutions, as well as environmental administration. Easy access to data from different fields of science creates an environment for new scientific ideas to emerge. This potential of generating new, interdisciplinary approaches to pre-existing problems is one of the key features of open-access data platforms that unify diverse data sources. Interdisciplinary elements, access to data over larger gradients, compatible eSystems and eTools to handle large amounts of data are extremely important and, if further developed, represent significant steps towards analysis of biological effects of climate change, human impact and development of operational ecosystem service assessment techniques. It is concluded that significant benefits regarding both scientific potential, technical developments and financial investments can be obtained by constructing a common Nordic LifeWatch eInfrastructure. Several steps concerning organizing and funding of a future Nordic LifeWatch are discussed, and an action plan towards 2020 is suggested. To analyze the potential for future Nordic LifeWatch in detail, our main conclusion is to arrange a Nordic LifeWatch conference as soon as possible. This conference should involve Nordic research councils, scientists and relevant stakeholders. The national delegates from the participating countries in the Nordic LifeWatch project are prepared to present details from the report and developments so far as a basis for further development of Nordic LifeWatch. The present work is financed by NordForsk and in-kind contributions from participating institutions

Moss and Liverwort Covers Structure Soil Bacterial and Fungal Communities Differently in the Icelandic Highlands

Cryptogamic covers extend over vast polar tundra regions and their main components, e.g., bryophytes and lichens, are frequently the first visible colonizers of deglaciated areas. To understand their role in polar soil development, we analyzed how cryptogamic covers dominated by different bryophyte lineages (mosses and liverworts) influence the diversity and composition of edaphic bacterial and fungal communities as well as the abiotic attributes of underlying soils in the southern part of the Highlands of Iceland. For comparison, the same traits were examined in soils devoid of bryophyte covers. We measured an increase in soil C, N, and organic matter contents coupled with a lower pH in association with bryophyte cover establishment. However, liverwort covers showed noticeably higher C and N contents than moss covers. Significant changes in diversity and composition of bacterial and fungal communities were revealed between (a) bare and bryophyte-covered soils, (b) bryophyte covers and the underlying soils, and (c) moss and liverworts covers. These differences were more obvious for fungi than bacteria, and involved different lineages of saprotrophic and symbiotic fungi, which suggests a certain specificity of microbial taxa to particular bryophyte groups. In addition, differences observed in the spatial structure of the two bryophyte covers may be also responsible for the detected differences in microbial community diversity and composition. Altogether, our findings indicate that soil microbial communities and abiotic attributes are ultimately affected by the composition of the most conspicuous elements of cryptogamic covers in polar regions, which is of great value to predict the biotic responses of these ecosystems to future climate change.Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. This work was supported by the grant PID2019-105469RB-C22 (AEI, MICINN).Peer reviewe