NKFI K_124341 "Global patterns to molecular problems in lichenology"

(NKFI K_124341 , 2017 - 2023, running project)
Coordinator or leader of the project: 
Dr Edit Farkas PI
Coordinator or leader institution of the project: 
MTA Centre for Ecological Research, Institute of Ecology and Botany
Responsible person at ÖK: 
Short description: 

Lichenized associations are important for solving environmental problems. There are lichens applied for bioindicaton. Some are becoming extinct, others becoming more abundant as an answer for environmental changes. Lichens are thalline organisms of variable colour and compositions. They consist of at least two partners of different taxonomic position: photosynthetizing algae or cyanobacteria are surrounded by fungal hyphae. To this biological diversity c. 1000 special unique organic lichen compounds are added. Their occurrence helps identification of lichens. Sometimes previously unknown chemical varieties are discovered. Molecular genetic investigations are carried out to justify taxonomic position and infraspecific variability of some species described form Hungary. These have also importance for nature conservancy. Special microscopic diagnostic method is introduced. UV-tolerant test species are selected for the study of the effects of global environmental changes.

Apart from the high rank journal publications, as a distant aim, (partly from supplementary sources) publication of a richly illustrated identification book and distribution atlas is planned to support further research and applied fields (ecology, monitoring, bioindication, conservation), as well as education: an excellent starting point for the younger generation, students interested in lichenology or biodiversity and chemical diversity in general.

Aims: 

Revisional studies are planned to establish the current and changed distribution of lichen taxa related to environmental changes. Several taxonomic groups in Central European/Carpathian regions are studied where lichen substances have a key role in identification. An improved knowledge on foliose and fruticose macrolichens is crucial for the study of global environmental changes and monitoring work whereas a more precise identification of tiny crustose lichens is important for bioindication studies. Developing methods make detection of quality and quantity of various lichen substances possible by chromatography (HPTLC, HPLC). Endemic taxa not studied elsewhere are in the focus of our studies, molecular genetic studies are planned to justify taxonomic position and infraspecific variety. Identification key published in 1994 is revised. Digital illustrations are prepared, distributions of species are mapped before and after 1975. Our aim is to predict biodiversity of various areas due to environmental changes. Satellite research is carried out for possibilities of visualization of secondary lichen metabolites and their localization by fluorescence microscope. UV-filter cortical pigment containing test lichen is used for the study of radiation and chemical diversity.
High rank journal publications are planned on new chemical varieties, on lichen-forming and lichenicolous fungi of European and world importance, on various distribution types and the possible reasons of these. The synthesis of our results is a manuscript of an identification book and distribution atlas to be published with the involvement of supplementary sources. Online publication ways are also considered.

Results: 

Project report - NKFI K 124341
First year [01.09.2017_31.08.2018]
The part time contracts with assitant researcher fellows (Katalin Veres, Nóra Varga) and research assistants (Krisztina Szabó, Dóra Smahajcsik) were established.
The works on the project have been started according to the main aims.
Revisions
were started in various taxonomic groups (Cladonia – furcata/rangiformis, subulata/rei, pyxidata aggr., Lepraria – 12 species, Xanthoparmelia – usnic acid containing species, pokornyi/ryssolea and some other taxa) on the basis of morphology (LM) and lichen secondary chemistry. During the analysis of c. 800 specimens (from herbaria BP, DE, EGR, JPU, SZE, SZO, VBI) several (c. 50) lichen secondary substances known from earlier studies and 13 metabolites (LSMs) not extracted earlier from Hungarian lichen specimens were identified by HPTLC.
Posters has been presented on Lepraria [Farkas et al. 2017, 2018]. Papers on revisions of cetrelioid and leprarioid species are under preparation.
For a more detailed morphological investigation and analysis of localization of fluorescent lichen substances a modern SMZ18 NIKON stereo microscope was purchased. The digital camera unit is planned to be ordered in the second year of the project.
Rare and protected species
were investigated and their distribution studied. The results were presented as posters on protected species and Cladina species in region Balaton-felvidék and as a journal publication on Solorina saccata [Farkas et al.2017, Sinigla et al.2018a, b].
Further lichenicolous species were identified form Hungary and the Carpathian Basin. The manuscripts of journal publications are under preparation.
Results were presented
in Hungarian and international scientific meetings as posters.
Although it was planned, at last we had to make the decision not to participate in IMC11 congress because of its high costs, however our participation in the 2nd Lichen Genomics Workshop (Graz, Austria:2-5.11.2017) was considered very important and useful.
Molecular genetic studies were initiated.
Taxa and methodology for additional studies by molecular genetic methods were discussed. Analysis of Cladonia magyarica was selected as first priority. Since this species is legally protected in Hungary, permission for collection and scientific investigation were requested and received.
The background and plans of molecular genetic research was presented as oral presentation during 2nd Lichen Genomics Workshop [Farkas 2017]. First DNA extractions have been carried out in the Molecular Taxonomic Laboratory of the Hungarian Natural History Museum, Budapest. Specimens from lowland steppe and rocky grassland habitats were collected, samples of apothecial and sterile parts were selected for isolation.
Several DNA extraction methods and protocols were tested and compared. PCR reaction for ITS region was optimized and products were sequenced using the Sanger sequencing method. Further samples, genes and primers are going to be tested according to the most recent literature data to combine our previous results for a multigene analysis. The methods, applied parameters were consulted with colleagues R. Pino-Bodas and S. Stenroos in Finnish Museum of Natural History, Helsinki.
A field experiment was established
to justify the production of UV-protecting LSMs after acetone rinsing in the experimental field of the National Botanical Garden, Vácrátót (6 repeated investigations from 2018 to 2020 in each spring and autumn). The preliminary studies for this experiment have already been started and supported also by OTKA K81232. It was essential to know if the selected species Cladonia foliacea survives the acetone treatment [Farkas et al. 2017a, b, Veres et al. 2018]. Its vitality was analysed by chlorophyll fluorescence measurements (Fv/Fm values).
The first data show differences between control and acetone rinsed samples, as well as between samples collected from the lowland and the mountain site, however a further more detailed evaluation is necessary.
Currently a manuscript is under preparation on the results of chlorophyll fluorescence analysis (Fv/Fm measurements) of the preliminary studies.
We had 6 posters and 4 oral presentations in the following conferences:
XXth Symposium of Baltic Mycologists and Lichenologists, Gdansk, Poland, 25–29.09.2017
4th Conference on Cryptogams, Eger, Hungary, 1.11–30.12.2017
12th International Conference Advances in research on the flora and vegetation of the Carpato ‐ Pannonian region, Debrecen, Hungary, 23–25.02.2018
11. Magyar Ökológus Kongresszus [11th Congress of Hungarian Ecologists] Nyíregyháza, Hungary, 28–30.08.2018
2 papers have been published (Lőkös et al. 2018, Sinigla et al. 2018) and further 5 manuscripts are near to submission.

Project report - NKFI K 124341
Second year [01.09.2018_31.08.2019]
The works on the project has been continued according to the main aims.
Revisions based on HPTLC analysis of lichen secondary metabolites (LSMs) were continued and carried out in further groups (Xanthoparmelia – brown species not containing usnic acid; Cladonia chlorophaea group; various Cladonia species; Cladonia magyarica forms sensu Gallé). Manuscripts were prepared on earlier studied groups (Cetrelia, Lepraria). Additional specimens and fields were analysed: α-alectoronic and β-alectoronic, furthermore α- and β-collatolic acids were identified in Cetrelia chicitae applying anysaldehide for visualization of the TLC plate; comparative morphometric studies on the soredia of the 4 Cetrelia species were carried out, IUCN red list categories of the species were established. A form described by Gyelnik is lectotypified (Farkas et al. 2019, Cryptogamie, Mycologie – submitted, revised – minor revision).
Identification key for species of Lepraria s.l. was compiled.
Various Cladonia species (c. 180 specimens) collected in Hungary during the last decades were identified using HPTLC. Fresh collections from 34 coenological sampling sites (Mt. Bakony) containing Cladina species and urban collections (on roof of high buildings in Debrecen) of Cladonia and Xanthoparmelia specimens were analysed (Matus et al. 2018, 2019a, 2019b).
Albanian Cladonia species (152 specimens of 15 species) were identified as a contribution to a joint publication with Spanish colleagues. Short characterization of species was published with distribution maps for each species, chemical variability for many of the species was discussed (Burgaz et al. 2019).
Chemical diversity within Cladonia chlorophaea species group was unknown in Hungarian specimens– 5 species were differentiated: morphometric studies are carried out – based on parameters of primary thalli, podetia, soredia – to compare chemically different species.
Lichenicolous fungi living on Cladonia species were studied in details and several of them were identified for the first time from Hungary. Most of the lichenicolous fungi on frequent Cladonia species (e.g., C. pyxidata, C. foliacea) appear as black dots, macroscopically impossible to differentiate them though these species are taxonomically very diverse belonging to ascomycetes, basidiomycetes or hyphomycetes. Seven of the most frequent taxa (like Didymocyrtis sp., Lichenoconium sp and others) found on podetia or primary thallus were presented on a poster illustrating their characteristic propagules.
Brown Xanthoparmelia species contains stenosporic, divaricatic and gyrophoric acids and terpenoids in different combinations. The preliminary results of HPTLC studies suggest that the morphological characters might have priority in identifications over chemical differences during identification process.
Two posters were prepared on the above topics for the 18th Congress of European Mycologists by the titles "Morphologically similar, cryptic taxa of macrolichens of conservation importance in the Carpathian Basin" and "Cladoniicolous fungi in the Carpathian Basin". (Farkas et al. 2019, Varga et al. 2019).
New records of Solenopsora candicans, S. cesatii, S. grisea, S. marina, S. olivacea subsp. olbiensis and S. olivacea subsp. olivacea from calcareous rocks, and S. liparina from ultramafic rocks in the Balkan Peninsula and adjacent islands were recorded. Their distribution, ecology and key morphological features are discussed in a paper published together with Slovakian colleagues (Fackovcova et al. 2019). The taxa S. candicans, S. cesatii, S. liparina, S. marina and S. olivacea subsp. olbiensis are reported for the first time from Albania, and S. cesatii from Bulgaria due to our contribution.
Cyphelium, Brodoa collected in Transsylvanian alps „Kelemen havasok” ( Eastern Carpathians) were identified after HPTLC studies.
35 lichen species and 5 lichenicolous fungi were presented from a microclimatically special habitat – considered as a refugium for cryptogams - in Jablanica Mts. (North Macedonia) on a poster for 13th Symposium on the Flora of Southeastern Serbia and Neighboring Regions (Farkas et al. 2019).
Despite of the intensive floristical investigations of the Balkan Peninsula in the last decades, there are many undiscovered or unpublished taxa, especially among microscopical fungi invading lichen thalli. In higher altitudes in the alpine regions cryptogamic organisms like lichens are conspicuous both in diversity and biomass. Our results show that several lichenicolous taxa seem to be new for the Midžor region (Serbia) as it was presented on a poster prepared for the 7th Balkan Botanical Congress (Varga et al. 2018).
We decided on multigene approach for molecular studies of the planned taxa. Most probably next generation sequencing will not be available for our studies – there was insufficient information available about this method during the planning of this project. The role and possibility for application of PKS genes is also under consideration since it is not leading to equally useful results in all groups of lichens. However markers (nuc ITS, IGS, EF1a, Rpb2 partial) – including those for photobionts (originally not planned) ¬ were selected on the basis of relevant literature, primers were bought accordingly. PCR reactions of DNA isolated from Cladonia magyarica specimens of Hungarian lowland and mountain sites have been processed. DNA was isolated from Vezdaea species and Xanthoparmelia mougeotii, samples are ready for further investigations .
From the combined phylogenetic analysis of multi-locus sequence data of the Lecanoraceae including two nuclear protein-coding markers (RPB2 and RPB1), the internal transcribed spacer and a fragment of the mitochondrial small subunit, it was found that the originally monotypic eastern Asian genus Verseghya is positioned within the Verseghya-Lecidella-Pyrrhospora clade of the Lecanoraceae and includes one more taxon Verseghya thysanophora widely distributed in Northern Hemisphere. Validation of one name as Polyozosia perpruinosa Fröberg ex S. Y. Kondr., L. Lőkös et Farkas is also proposed (Kondratyuk et al. 2019).
For testing diagnostic possibilities for visualisation of LSMs, NIKON SMZ18 research stereo microscope with photographic unit has been bought and ready to work with.
LSM localization investigations have been started on common foliose and fruticose lichens. Microscopic studies were carried out on usnic acid and atranorin containing cortical layers of various Usnea and Cladonia species. Specimens with lichenicolous fungi were also included in the study.
A cooperation has been developed with Dr. László Márk (University of Pécs, Medical School) for further examinations by a more sensitive method MALDI-MS imaging. The work covering a great number of taxa will hopefully increase our chances for valuable publications.
The preliminary studies for the ongoing acetone rinsing experiment justified that the selected species Cladonia foliacea survives the acetone treatment. Its vitality was analysed by chlorophyll fluorescence measurements and found that the long-term tolerance of this species is higher than any other species checked earlier under more humid conditions (Farkas et al. 2019, submitted).
Significant seasonal differences were found in lichen secondary metabolites (usnic acid, fumarprotocetraric acid) analysed by HPTLC and HPLC-PDA between summer and winter collected thalli (Farkas et al. 2019, submitted).
Data of the first year basically show sligthly decreasing differences between control and acetone rinsed samples in the re-production of LSMs, while Fv/Fm values indicate good condition of the thalli, though results of fumarprotocetraric acid are less clear and further masurements (from samples of spring and autumn 2019) are necessary for the interpretations. First results are summarised in a short publication (manuscript under preparation).
The effect of seasonal variation and different micro-environmental conditions on the metabolic activity of five terricolous lichen species, representing different growth forms were explored in temperate semiarid grasslands. The results of two years chlorophyll fluorescence analysis and background parameters (air temperature, photosynthetically active radiation, soil temperature and vapour pressure deficit) were analysed and summarised in a detailed paper submitted recently (Veres et al. 2019, submitted). The results direct attention to the importance of investigating more than one species, representing different growth forms, to find the response of lichens to changing environment in space and time.
Relation of distribution and climatic parameters is investigated on selected species. Concerning to prediction of lichen distribution due to global climatic change a cooperation has been built within the institute – species were selected for further studies.
Submitted papers mentioned in the report and listed among publications are indicated with their status, pdf will be deposited in the repository "mtmt" upon acceptance.

Project report - NKFI K 124341
Third year [01.09.2019_31.08.2020]
The works on the project have been continued according to the main aims.
Laboratory work (HPTLC analysis of lichen secondary metabolites (LSMs), microscopical studies) were possible in limited time due to COVID-19 situation. Additional specimens and fields were analysed to improve former manuscripts.
Field studies, morphological and chemical investigations of brown Xanthoparmelia species were carried out to clarify our concept about the priority of morphological characters over differences in LSM production.
Morphometric studies are carried out to compare chemically different species within Cladonia chlorophaea species group.
New localities of Parmelia submontana in Hungary were discovered (abstract submitted for MMK7, 2020 – cancelled meeting), journal publication manuscript was prepared. Parmeliella triptophylla was revealed as new species to Hungary, new localities of further species (Cladonia mitis for Hungary, Multiclavula mucida and Xanthoparmelia protomatrae for Romania) were discovered (manuscript submitted/published – Papp et al. 2020).
20 numbers (61–80) of Lichenes Delicati Exsiccati Editae (LDEE5) containing collections from the Carpathians and various exotic sites were prepared and published in honour and memory of Dr Antonín Vězda born 100 years ago (Farkas 2020). A further 20 numbers (81–100) including specimens from natural and urban localities are compiled according to former plans, publication of LDEE6 is under preparation for Acta Bot. Hung. 63(1–2), 2021.
ITS sequences have been gained from DNA isolated from Vezdaea species, samples are studied for further sequences. A manuscript is under preparation.
LSM localization investigations have been carried out on common foliose and fruticose lichens. Microscopical studies were continued on usnic acid and atranorin containing cortical layers of various Usnea and Cladonia species. For detection of zeorin, Leucodermia leucomelos was compared to Physcia and Phaeophyscia species by various methods and chemical reagents.
In a cooperation with Dr László Márk (University of Pécs, Medical School) terpenoid containing taxa (Peltigera spp., Pseudocyphellaria spp.) were analysed by MALDI-MS (manuscript under preparation).
The preliminary studies for the ongoing acetone rinsing experiment justified that the selected species Cladonia foliacea survives the acetone rinsing treatment (Farkas et al. 2020).
Significant seasonal differences were found in lichen secondary metabolites (usnic acid, fumarprotocetraric acid) analysed by HPTLC and HPLC-PDA between summer and winter collected thalli (Farkas et al. 2020).
According to the first results of the long-term light protection experiment, the LSMs might decrease the damaging effects of the radiation in the photosynthetic system of the symbiotic algae. It seems so, that the amount of usnic acid is increasing towards a saturation value at a supposedly lower level than the original control value. The re-production of fumarprotocetraric acid is uncertain under current experimental arrangements (abstract submitted for MMK7, 2020 – cancelled meeting; manuscript is under preparation). Samples were recollected from the experimental field after 2.5 and 3 years and prepared for HPLC analysis.
A paper on the results of two years chlorophyll fluorescence analysis and background parameters (air temperature, photosynthetically active radiation, soil temperature and vapour pressure deficit) was published (Veres et al. 2020).
Results on investigation of environmental variables affecting terricolous lichen communities on different scales in a duneland ecosystem were summarised (abstract submitted for MMK7, 2020 – cancelled meeting; a manuscripts is submitted (Lichenologist, major revision) and
another manuscript is under preparation – both by Veres et al.).
The distribution and habitat preference of three protected Cladonia species were compared in the Balaton Uplands region. It was established that while C. mitis is the most frequent, C. rangiferina has the narrowest requirements in a series of environmental variables (Sinigla et al. – Lichenologist, major revision).
The cryptogamic communities and the environmental conditions on flatroofs in the city of Debrecen (East Hungary) were studied (a publication is under preparation - major revision for Biologia Futura by Aszalósné Balogh et al. 2020).
Concerning to prediction of lichen distribution due to global climatic changes selected species and climatic and other environmental parameters are under study in a cooperation within the institute.
The current PI was accepted as a symposium organiser on “Progress in classical and genomic aspects of lichen secondary chemistry” for IAL9 international symposium, the major scientific meeting of the special field of lichenology (https://doity.com.br/ial9/blog/symposia).
Registration fee was transferred and accepted for the postponed meeting for 1–6 August 2021. (Fortunately air ticket and accommodation were not arranged yet for the 2020 meeting, therefore unnecessary costs were not spent.) A lecture was in preparation for IAL9, 2020 (cancelled meeting) on the development of analytical methods in the study of LSMs, abstracts of further 3 posters were also submitted for the meeting: 1) on the knowledge on extreme tolerance of Cladonia foliacea, a temperate continental grassland lichen species, 2) various and changing distribution patterns of lichens in the Carpathian Basin, 3) on the checklist of lichenicolous fungi in Hungary in preparation.
7 abstracts (1 lecture, 6 posters) were submitted for IAL9, 2020 (1+3) and MMK7, 2020 (3). Further planned participations in AFVK13, 2020 (currently planned for 2021), 2020 and 5th Conference on Cryptogams, 2020 (currently planned for 25–26 August 2021) were
impossible either, because also these meetings were postponed.
Five of six earlier submitted papers have been published, one has been expected acceptance after minor revision since 18.05.2020, and seven further journal publications are under preparation.
Submitted papers mentioned in the report and listed among publications are indicated with their status, pdf will be deposited in the repository “mtmt” upon acceptance.

Project report - NKFI K 124341
Fourth year [01.09.2020_31.08.2021]
The works on the project have been continued according to the main aims, though COVID19 pandemia delayed our work. The older generation had to stay in home-office, they were not allowed to visit institutions in person only from the middle of June 2021. Younger researchers were applied only in part-time due to various reasons. Changes of the staff in the Molecular Taxonomic Laboratory of the Hungarian Natural History Museum, Budapest also caused unforeseen difficulties. Therefore the closing date of the project was extended.
Accordingly the laboratory work (molecular genetic work, HPTLC analysis of lichen secondary metabolites (LSMs), microscopical studies) were possible in limited time only. Additional specimens and fields were analysed to improve former manuscripts.
The European occurrence of Cetrelia chicitae – described from America – is confirmed on the basis of identical secondary metabolite composition by 7 lichen substances. Four species of the genus were revealed and mapped in Hungary. C. chicitae and C. monachorum are new for the Hungarian lichen flora. From the originally known two taxa C. cetrarioides proved to be rare, C. olivetorum is less frequent than regarded earlier. Currently C. monachorum is the most frequent Cetrelia species in Hungary. C. chicitae and C. cetrarioides may need protection measures. Parmelia cetrarioides f. pseudofallax (Gyeln.) Gyeln. has been lectotypified (Farkas et al. 2021).
Further field studies, morphological and chemical investigations of brown Xanthoparmelia species confirmed our concept about the priority of morphological characters over differences in LSM production –a poster was presented earlier, journal publications on usnic acid containing and brown species of this genus with different secondary chemistry is planned separately.
Morphometric studies are carried out to compare chemically different species within Cladonia chlorophaea species group – a poster presentation and a manuscript is under preparation.
20 numbers (81–100) of Lichenes Delicati Exsiccati Editae (LDEE6) containing collections from natural and urban localities were prepared and published in honour and memory of Dr Antonín Vězda born 100 years ago (Farkas 2021). The alphabetical list of species – with updated nomenclature – distributed in the 6 fascicles with indication of the country of origin was presented. Three new species were described in the 6 publications attached to the exsiccate and 15 species were found to be new to the given countries.
Further sequences have been gained from DNA isolated from Vezdaea species, samples were also collected for the study of further sequences. A manuscript is under preparation. Molecular genetic analysis of Cladonia magyarica samples was also continued.
The checklist of Hungarian lichenized and lichenicolous fungi is continuously updated with new occurrences and revised nomenclaturally. Currently it contains 921 taxa. A manuscript on annotated checklist of lichenicolous fungi – with nearly 100 species– is going to be submitted in September 2021 by Varga et al.
LSM localization investigations have been carried out on common foliose and fruticose lichens. In a cooperation with Dr László Márk (University of Pécs, Medical School) Cetrelia, Cladonia and Xanthoparmelia species were analysed by MALDI-MS, results on spectra were presented in IAL9 symposium oral presentation. These are planned to be analysed further and images are also under preparation.
All specimens of acetone rinsing experiment were collected from experimental field after two and half and three years, HPLC and chlorophyll a fluorescence studies were carried out. The long term results – presented in Hungarian and international symposia – confirmed that the selected species Cladonia foliacea tolerated the acetone rinsing treatment (collected either from lowland or mountain sites) and re-produced the secondary metabolites, though the three years were not enough to reach the original concentrations (Farkas et al. 2021a,b). The amount of usnic acid is increasing towards a saturation value at a lower level than the original control value. Manuscripts are under preparation.
Further studies revealed the relation of lichen populations to a series of environmental conditions and confirmed the bioindicator nature of lichens at various scales by combined methods (of ecophysiology, coenology, multivariate statistical analysis, etc.).
Terricolous lichens are relevant associates of biological soil crusts in arid and semiarid environments. Dunes are ecosystems of high conservation interest, because of their unique, vulnerable and threatened features. The function of lichens is affected by the changing seasons and different microhabitat conditions. We explored the effect of seasonal variation and different micro-environmental conditions (aspect) on the metabolic activity of five terricolous lichen species (Cladonia foliacea, C. furcata, C. pyxidata group, Diploschistes muscorum and Thalloidima physaroides), representing various growth forms, in temperate semiarid grasslands. As a result, the air temperature, photosynthetically active radiation, soil temperature and vapour pressure deficit were significantly higher on south-west than on north-east facing microsites, where relative humidity and water content of soil proved to be considerably higher. Higher photosynthetic activity, as well as a higher level of photoprotection, was detected in lichens from north-east-oriented microsites compared with south-west populations. In addition, the difference between sun and shade populations varied seasonally. Since a species-specific response to both aspect and season was detected, we proposed to investigate more than one species of different growth forms, to reveal the response of lichens to the changing environment in space and time. (Veres et al. 2020).
We explored also the composition of these communities and the environmental factors affecting them. We found that the site had a significant effect on species richness that might reflect the different types and severity of previous disturbance events at the studied sites. On a smaller, ‘dune’ scale, in general lower herb cover and height and a higher moss cover were characteristic of arid dune sides. Most of the frequent species were negatively affected by higher moss cover. Some lichen species were more abundant (e.g. Cladonia furcata) or found only (e.g. Xanthoparmelia subdiffluens, Gyalolechia fulgens) on arid dune sides, while others preferred (e.g. C. pyxidata) or occurred only on (e.g. Peltigera species, C. rei) humid sides. The diverse microhabitat types, microclimate and landscape structure, results in species-rich and valuable terricolous lichen communities forming in inland dune ecosystems. (Veres et al. 2021a)
In lichens, the protection of the photosynthetic apparatus is essential for the survival of its thallus, especially under harsh environmental conditions. This was investigated to reveal the various photo-adaptation and -protection mechanisms occurring in the heterogeneous/polymorphic thallus structure during four seasons in two (exposed and shaded) microhabitat types. The changes in the concentrations of atranorin, fumarprotocetraric acid and usnic acid (UV protectant metabolites) of Cladonia foliacea, C. furcata and C. magyarica, as well as the plastid pigment concentration in these and in three other species, were determined. Both the UV protectant metabolite and plastid pigment concentrations showed seasonal fluctuations according to changing light and humidity conditions. Exposure of the microhabitat had no significant influence on the concentrations. The interspecific difference of the adaptation were more pronounced than that of the intraspecific one among seasons and microhabitat types. The complementary yield of photochemical quenching, both regulated and non-regulated non-photochemical quenching also showed a seasonal and species-specific picture in their proportions. (Veres et al. 2021b)
Habitat preferences of three protected Cladonia species (C. arbuscula, C. mitis and C. rangiferina) growing on underlying rocks of red sandstone, basalt, Pannonian sandstone and gravel were investigated by detailed sampling in the Bakony Mts including the Balaton Uplands region. Sporadic populations of these species mostly exist at the top of hills and mountains in open acidofrequent oak forests, but they may occur in other habitats, such as closed acidofrequent oak forests, slope steppes on stony soils, siliceous open rocky grasslands, open sand steppes, wet and mesic pioneer scrub and dry Calluna heaths. Cladonia rangiferina was found to grow beneath higher canopy cover than either C. arbuscula or C. mitis. Furthermore, there were significant differences in canopy cover between occupied and unoccupied quadrats in the case of all three species. It was established that Cladonia rangiferina is a good indicator species of natural habitats in Hungary due to its restricted distribution and low ecological tolerance. These results may lead to the adoption of effective conservation methods (e.g. game exclusion, artificial dispersal) in the future. (Sinigla et al. 2021)
The current PI was a symposium organiser in “The expanding and diversifying field of lichen secondary chemistry” for IAL9 international symposium, the major scientific meeting of the special field of lichenology (https://doity.com.br/ial9/blog/symposia). One lecture and three posters were presented during IAL9 Symposium of International Association for Lichenology, 1–6 August 2021. A lecture (Farkas et al. 2021c)was presented on the development of analytical methods in the study of LSMs, posters were shown
1) on the knowledge on extreme tolerance of Cladonia foliacea, a temperate continental grassland lichen species – the photosynthetic activity (Fv/Fm) remained relatively high, indicating a well-functioning photosynthetic system in both lowland and mountain samples for years after acetone treatment (Veres et al. 2021c),
2) on various and changing distribution patterns of lichens in the Carpathian Basin – the various trends are known for some species at a global scale or European level, other examples are rather characteristic for Central Europe or Hungary. As the distribution is strongly correlated to background environmental conditions (e.g. air pollution, global warming, etc.), the analyses of distribution patterns have a great value for bioindication (Farkas et al. 2021),
3) on the checklist of lichenicolous fungi in Hungary in preparation – data of c. 100 species originate from sandy grasslands, areas with more anthropogenic influence (tree plantations around townships) and from our mountain regions representing less studied vegetation types (Varga et al. 2021).
Four posters (Farkas et al. 2021, Muhoro and Farkas 2021a, Veres et al. 2021d, e) were also presented in MÖK12 Hungarian Ecological Congress, 24–26 August 2021, partly in the above mentioned topics. Manuscripts are under preparation.
Additionally in a review paper we focused on lichens with potential insecticidal and antiprotozoal activity due to their LSMs. Literature sources were analyzed according to the taxonomic group of the insect, the protozoal disease and the lichen, the type of bioactive compounds (including method of application and mount applied), and the potential bioactivity based on mortalities caused after 24 h of exposure on insects and on parasitic protozoa. Insecticidal and antiprotozoal effects of crude extracts and seven lichen secondary metabolites (mostly usnic acid) of 32 lichen species were determined. The results are based on laboratory experiments; however, the efficacy of metabolites should be confirmed in the field. (Muhoro and Farkas 2021b). Based on these results, further comparative studies are planned on lichen species – occurring both in temperate and tropical region – containing usnic acid and other LSMs indicating effects of major macroclimatic differences by their quantitative analysis.
Five papers have been published, one has been accepted and one is under major revision, further journal publications are under preparation.
Submitted papers mentioned in the report and listed among publications are indicated with their status, pdf will be deposited in the repository “mtmt” upon acceptance.

Publications: 

2021

Farkas E., Biró B., Varga N., Sinigla M., Lőkös L. (2021): Analysis of lichen secondary chemistry doubled the number of Cetrelia W.L. Culb. & C.F. Cub. species (Parmeliaceae, lichenised Ascomycota) in Hungary CRYPTOGAMIE MYCOLOGIE 42(1): pp. 1-16.; Q1
Sinigla, M., Szurdoki, E., Lőkös, L., Bartha, D., Galambos, I., Bidló, A. & Farkas, E.: (2021): Distribution and habitat preference of protected reindeer lichen species (Cladonia arbuscula, C. mitis and C. rangiferina) in the Balaton Uplands (Hungary) Lichenologist 53(3): pp. 271-282; Q2
Varga, N.; Lőkös, L.; Farkas, E. (2021): Annotated Checklist of the Lichenicolous Fungi of Hungary. Diversity 13, 557; Q1

2020

Farkas, E., Biró, B., Csintalan, Zs., Veres, K. (2020): Acetone rinsing tolerance of the lichen species Cladonia foliacea is considerable Lichenologist 52, 325-327
Katalin Veres, Edit Farkas, Zsolt Csintalan (2020): The bright and shaded side of duneland life: the photosynthetic response of lichens to seasonal changes is species-specific. Mycol Progress 19, 629–641

2019

Burgaz, A. R., Luna-González, S., Gutiérrez-Larruga, B., Pino-Bodas, R., Lőkös, L. & Farkas, E. (2019): Diversity of Albanian Cladoniaceae. Bot. Complut.43: pp. 15–40.
Fačkovcová, Z., Lőkös, L., Farkas, E. & Guttová, A. (2019): New records of the lichen genus Solenopsora A. Massal in the Balkan Peninsula and adjacent islands. Herzogia 32(1): pp. 101–110.