Photosynthetic Partners Research Articles

The sound of lichens: ultrasonic acoustic emissions during desiccation question cavitation events in the hyphae.

Lichens are a mutualistic symbiosis between a fungus and one or more photosynthetic partners. They are photosynthetically active during desiccation until relative water contents (RWC) as low as 30% (on dry mass). Experimental evidence suggests that during desiccation, the photobionts have a higher hydration level than the surrounding fungal pseudo-tissues. Explosive cavitation events in the hyphae might cause water movements towards the photobionts. This hypothesis was tested in two foliose lichens by measurements of ultrasonic acoustic emissions (UAE), a method commonly used in vascular plants but never in lichens, and by measurements of photosystem II efficiency, water potential and RWC. Thallus structural changes were characterised by low-temperature scanning electron microscopy. The thalli were silent between 380% and 30% RWCs, i.e. when explosive cavitation events should cause movements of liquid water. Nevertheless, the thalli emitted UAE at approximately 5% RWC. Accordingly, the medullary hyphae were partially shrunk at about 15% RWC, whereas they were completely shrunk below 5% RWC. These results do not support the hypothesis of hyphal cavitation and suggest that the UAE originate from structural changes at hyphal level. The shrinking of hyphae is proposed as an adaptation to avoid cell damage at very low RWCs.

Bioactive Components Analysis and Pharmacological Properties of Extracts and Metabolites of Lichen Umbilicaria crustulosa

Lichens, a diverse group of organisms, have a unique structure consisting of fungal filaments and photosynthetic partner cells. This research conducted a comprehensive chemical analysis and evaluation of the anti-inflammatory and antioxidant properties of methanolic and acetone extracts from Umbilicaria crustulosa lichen, along with its isolated metabolites. The process involved separating atranorin and chloratranorin fractions, physodic acid, and gyrophoric acid. Secondary metabolites were identified using chromatographic and spectroscopic data. The total polyphenols content was determined spectrophotometrically. This study examined the antioxidant activity of extracts of the lichen U. crustulosa and the isolated fractions using three methods: DPPH scavenging activity, ABTS scavenging activity, and reducing power. This study also evaluated the acute oral toxicity and the anti-inflammatory activity of the extracts in Wistar albino rats. A higher content of the total phenolic compounds was found in the acetone extract, but antioxidant and anti-inflammatory activities were more prominent in the methanolic extract. The isolated atranorin and chloratranorin fractions and compound physodic acid showed the highest antioxidant activity. No toxic effects were noted in the acute oral toxicity study. This study highlights the potential of the investigated lichen as a valuable source of novel biological agents.

The underestimated fraction: diversity, challenges and novel insights into unicellular cyanobionts of lichens.

Lichens are remarkable and classic examples of symbiotic organisms that have fascinated scientists for centuries. Yet, it has only been for a couple of decades that significant advances have focused on the diversity of their green algal and/or cyanobacterial photobionts. Cyanolichens, which contain cyanobacteria as their photosynthetic partner, include up to 10% of all known lichens and, as such, studies on their cyanobionts are much rarer compared to their green algal counterparts. For the unicellular cyanobionts, i.e. cyanobacteria that do not form filaments, these studies are even scarcer. Nonetheless, these currently include at least 10 different genera in the cosmopolitan lichen order Lichinales. An international consortium (International Network of CyanoBionts; INCb) will tackle this lack of knowledge. In this article, we discuss the status of current unicellular cyanobiont research, compare the taxonomic resolution of photobionts from cyanolichens with those of green algal lichens (chlorolichens), and give a roadmap of research on how to recondition the underestimated fraction of symbiotic unicellular cyanobacteria in lichens.

Genomic analysis of Coccomyxa viridis, a common low-abundance alga associated with lichen symbioses

Lichen symbiosis is centered around a relationship between a fungus and a photosynthetic microbe, usually a green alga. In addition to their main photosynthetic partner (the photobiont), lichen symbioses can contain additional algae present in low abundance. The biology of these algae and the way they interact with the rest of lichen symbionts remains largely unknown. Here we present the first genome sequence of a non-photobiont lichen-associated alga. Coccomyxa viridis was unexpectedly found in 12% of publicly available lichen metagenomes. With few exceptions, members of the Coccomyxa viridis clade occur in lichens as non-photobionts, potentially growing in thalli endophytically. The 45.7 Mbp genome of C. viridis was assembled into 18 near chromosome-level contigs, making it one of the most contiguous genomic assemblies for any lichen-associated algae. Comparing the C. viridis genome to its close relatives revealed the presence of traits associated with the lichen lifestyle. The genome of C. viridis provides a new resource for exploring the evolution of the lichen symbiosis, and how symbiotic lifestyles shaped evolution in green algae.

The combined effects of habitat fragmentation and life history traits on specialisation in lichen symbioses

Abstract Interactions between organisms are determined by life‐history traits. Ecological strategies regarding species specialisation range from generalist to highly specialised relationships. Although it is expected that habitat fragmentation's effect on species abundance and survival depends on their degree of specialisation and life‐history traits, few studies have delved into the interplay between interaction specialisation, life‐history traits and habitat fragmentation. Here, we investigate the combined effect of habitat fragmentation, forest structure and life‐history traits (growth form and reproductive mode) on the specialisation of lichen‐forming fungi (mycobionts) toward their photosynthetic partners (photobionts) in lichen symbioses. We studied mycobiont specialisation in epiphytic lichen communities present in 10 fragments of Quercus rotundifolia forest embedded in an agricultural matrix. Both mycobionts and photobionts were identified DNA barcoding and mycobiont specialisation was measured through interaction parameters calculating the relative number of interactions (normalised degree; ND) and the specialisation of each species based on its discrimination from a random selection of partners (d'). Phylogenetic generalised linear mixed models were used to analyse the effect of patch size as well as the life history traits growth form (crustose, foliose, fruticose) and reproduction mode (sexual vs. asexual) on mycobiont specialisation. Both mycobiont and photobiont richness along the patch size gradient followed a hump‐back pattern, which was more pronounced in photobionts. Mycobionts forming crustose thalli established the largest number of interactions. Mycobiont specialisation (d') was larger for fruticose and foliose forms and species with vegetative reproduction. Along the gradient of fragment size, the relative number of interactions decreased and the specialisation of mycobionts with vegetative reproduction increased. Synthesis. The study of mycobiont specialisation towards their photobionts in epiphytic lichen communities in a fragmented Mediterranean forest revealed a complex interaction between species' life history traits and habitat fragmentation. In particular, this interplay had a significant impact on the specialisation of mycobionts. The results show the ability of some species to modulate their specialisation according to habitat conditions, suggesting that some species may be more resilient to abiotic changes than expected.

Morphological and molecular characterization of Geraldius galapagoensis (Nematoda: Chambersiellidae) associated with lichens in Argentina

Lichens are symbiotic organisms, usually composed of a fungal partner, the mycobiont, and one or more photosynthetic partners, the photobiont, which is most often either a green alga or a cyanobacterium, that harbor a diverse community of invertebrates such as rotifers, tardigrades, mites, springtails, crustaceans, and nematodes. In this work, we isolated the nematode Geraldius galapagoensis (Chambersiellidae) associated with the lichen Hyperphyscia syncolla (Physciaceae) in a region of Buenos Aires Province, Argentina. This species was discovered in a tropical forest of Ecuador and is characterized mainly by a head offset by a constriction from the rest of the body, a esophagus with a cylindrical pharyngeal corpus without a median bulb, a narrow isthmus and an oval basal pharyngeal bulb with strong transverse/butterfly valve apparatus, a tail curved ventrally, ending in dorsally hooked end; the male with seven pairs of latero-ventral pre-anal papillae and three pairs of post-anal in the following positions: one pair latero-ventral and two pairs dorso-lateral and two slightly curved spicules with asymmetric manubrium with an anterior extension. The comparison of the morphometrics of our G. galapagoensis male with that of the original description shows that the body length is shorter, as are the distance of the excretory pore to the anterior end and the tail. On the other hand, the distance from the anterior end to the nerve ring and the esophagus length are greater. The head width, body diameter and spicule length are quite similar. We provide a morphological and morphometrical characterization of a G. galapagoensis second isolate and the first world report of molecular sequences belonging to this species.

Exploring the influence of climatic variables on mycobiome composition and community diversity in lichens: insights from structural equation modeling analysis

BackgroundLichens are symbiotic organisms composed of a fungus and a photosynthetic partner, which are key ecological bioindicators due to their sensitivity to environmental changes. The endolichenic fungi (ELF) living inside lichen thalli, are an important but understudied component of playing crucial ecological roles such as nutrient cycling and protection against environmental stressors. Therefore ELF community investigation is vital for fostering sustainable ecosystems and leveraging their ecological benefits. Deciphering the intricate relationships between ELF and their lichen hosts, alongside the influence of environmental factors on these communities, presents a significant challenge in pinpointing the underlying drivers of community structure and diversity.ResultsOur research demonstrated that locational factors were the main drivers of the ELF community structure, rather than host haplotype. Several climatic factors affected the diversity of the ELF community and contributed to the prevalence of different types of fungal residents within the ELF community. A decrease in isothermality was associated with a greater prevalence of pathotrophic and saprotrophic fungi within the ELF community, resulting in an overall increase in community diversity. By conducting a structural equation modeling analysis, we identified a robust link between climatic variables, fungal trophic mode abundance, and the species diversity of the ELF community.ConclusionThis study’s discoveries emphasize the significance of examining climate-related factors when investigating ELF’s structure and function. The connection between fungi and climate is intricate and complex, and can be influenced by various other factors. Investigating the potential for ELF to adapt to changing climatic conditions, as well as the potential effects of changes in ELF communities on lichen function, would be valuable research areas. We anticipate that our research results will establish a basis for numerous future ELF research projects and have a significant impact on the field.

Do urban air pollutants induce changes in the thallus anatomy and affect the photosynthetic efficiency of the nitrophilous lichen Physcia adscendens?

Lichens are symbiotic organisms that are generally sensitive to air pollution due to their specific biological and physiological features. Physcia adscendens is a nitrophilous lichen well-known for being resistant to air pollution associated with progressive anthropopressure. The aim of this study was to investigate the effect of nitrogen oxides and suspended particulate matter (PM10 and PM2.5) on anatomical structure of the thallus and photobiont’s photosynthetic efficiency in P. adscendens inhabiting sites that differ in terms of air pollution level and thereby to determine the relevance of these pollutants for shaping the structure of the thallus and the physiological condition of the photosynthetic partner. We found that P. adscendens from polluted sites had increased thickness of the algal layer and the larger size of the algae cells, but a much lower ratio of the algal layer to the whole thallus. Lichens from highly polluted sites had also higher photosynthetic efficiency, which indicates a relatively good physiological condition of the photobiont. This indicates that the photobiont of P. adscendens is well-adapted to function under air pollution stress which may contribute to its success in colonizing polluted sites. Both changes in the anatomy of the lichen thallus and the efficiency of photosynthesis may be related to the enrichment of the environment with nitrogen. The increased photosynthetic efficiency as well as investment in the size of photobiont cells and growth mycobiont hyphae confirms that P. adscendens is well-adapted to urban conditions; however, the mechanism behind those adaptations needs more focus in the context of global environmental changes.

Fungal community inside lichen: a curious case of sparse diversity and high modularity

BackgroundLichens represent not only the mutualism of fungal and photosynthetic partners but also are composed of microbial consortium harboring diverse fungi known as endolichenic fungi. While endolichenic fungi are known to exert a remarkable influence on lichen ecology through their crucial roles in nutrient cycling, bioprospecting and biodiversity, the enigmatic community structures of these fungal inhabitants remain shrouded in mystery, awaiting further exploration and discovery. To address knowledge gap, we conducted metabarcoding on two lichens using 18S gene amplification, Dirinara applanta and Parmotrema tinctorum, and compared their microbial communities to those found in the pine bark to which the lichens were attached. Our hypothesis was that the endolichenic communities would exhibit distinct diversity patterns, community structures, network structures, and specialist composition compared to the surrounding epiphytic community.ResultsOur investigation has shed light on the clear demarcation between the endolichenic and epiphytic fungal communities, as they exhibit markedly different characteristics that set them apart from each other. This research demonstrated that the endolichenic communities are less diverse as compared to the epiphytic communities. Through community similarity analysis, we observed that two endolichenic communities are more similar to each other in terms of community composition than with the adjacent epiphytic communities. Moreover, we unveiled a striking contrast in the network structures between the endolichenic and epiphytic communities, as the former displayed a more modular and less nested features that is evocative of a potent host-filtration mechanism.ConclusionsThrough our investigation, we have discovered that lichens harbor less intricate and interconnected fungal communities compared to the neighboring epiphytic environment. These observations provide valuable insights into the metagenomic architecture of lichens and offer a tantalizing glimpse into the unique mycobiome.

Invariant properties of mycobiont‐photobiont networks in Antarctic lichens

AbstractAimLichens are often regarded as paradigms of mutualistic relationships. However, it is still poorly known how lichen‐forming fungi and their photosynthetic partners interact at a community scale. We explored the structure of fungus‐alga networks of interactions in lichen communities along a latitudinal transect in continental Antarctica. We expect these interactions to be highly specialized and, consequently, networks with low nestedness degree and high modularity.LocationTransantarctic Mountains from 76° S to 85° S (continental Antarctica).Time PeriodPresent.Major Taxa StudiedSeventy‐seven species of lichen‐forming fungi and their photobionts.MethodsDNA barcoding of photobionts using nrITS data was conducted in 756 lichen specimens from five regions along the Transantarctic Mountains. We built interaction networks for each of the five studied regions and a metaweb for the whole area. We explored the specialization of both partners using the number of partners a species interacts with and the specialization parameter d'. Network architecture parameters such as nestedness, modularity and network specialization parameter H2' were studied in all networks and contrasted through null models. Finally, we measured interaction turnover along the latitudinal transect.ResultsWe recovered a total of 842 interactions. Differences in specialization between partners were not statistically significant. Fungus‐alga interaction networks showed high specialization and modularity, as well as low connectance and nestedness. Despite the large turnover in interactions occurring among regions, network parameters were not correlated with latitude.Main ConclusionsThe interaction networks established between fungi and algae in saxicolous lichen communities in continental Antarctica showed invariant properties along the latitudinal transect. Rewiring is an important driver of interaction turnover along the transect studied. Future work should answer whether the patterns observed in our study are prevalent in other regions with milder climates and in lichen communities on different substrates.

Carbon-concentrating mechanisms are a key trait in lichen ecology and distribution.

Carbon-concentrating mechanisms (CCMs) are a widespread phenomenon in photosynthetic organisms. In vascular plants, the evolution of CCMs ([C44-carbon compound] and crassulacean acid metabolism [CAM]) is associated with significant shifts, most often to hot, dry and bright, or aquatic environments. If and how CCMs drive distributions of other terrestrial photosynthetic organisms, remains little studied. Lichens are ecologically important obligate symbioses between fungi and photosynthetic organisms. The primary photosynthetic partner in these symbioses can include CCM-presenting cyanobacteria (as carboxysomes), CCM-presenting green algae (as pyrenoids) or green algae lacking any CCM. We use an extensive dataset of lichen communities from eastern North America, spanning a wide climatic range, to test the importance of CCMs as predictors of lichen ecology and distribution. We show that the presence or absence of CCMs leads to opposite responses to temperature and precipitation in green algal lichens, and different responses in cyanobacterial lichens. These responses contrast with our understanding of lichen physiology, whereby CCMs mitigate carbon limitation by water saturation at the cost of efficient use of vapor hydration. This study demonstrates that CCM status is a key functional trait in obligate lichen symbioses, equivalent in importance to its role in vascular plants, and central for studying present and future climate responses.

How to build a lichen: from metabolite release to symbiotic interplay.

Exposing their vegetative bodies to the light, lichens are outstanding amongst other fungal symbioses. Not requiring a pre-established host, 'lichenized fungi' build an entirely new structure together with microbial photosynthetic partners that neither can form alone. The signals involved in the transition of a fungus and a compatible photosynthetic partner from a free-living to a symbiotic state culminating in thallus formation, termed 'lichenization', and in the maintenance of the symbiosis, are poorly understood. Here, we synthesise the puzzle pieces of the scarce knowledge available into an updated concept of signalling involved in lichenization, comprising five main stages: (1) the 'pre-contact stage', (2) the 'contact stage', (3) 'envelopment' of algal cells by the fungus, (4) their 'incorporation' into a pre-thallus and (5) 'differentiation' into a complex thallus. Considering the involvement of extracellularly released metabolites in each phase, we propose that compounds such as fungal lectins and algal cyclic peptides elicit early contact between the symbionts-to-be, whereas phytohormone signalling, antioxidant protection and carbon exchange through sugars and sugar alcohols are of continued importance throughout all stages. In the fully formed lichen thallus, secondary lichen metabolites and mineral nutrition are suggested to stabilize the functionalities of the thallus, including the associated microbiota.

Symbiont-specific responses to environmental cues in a threesome lichen symbiosis.

Photosymbiodemes are a special case of lichen symbiosis where one lichenized fungus engages in symbiosis with two different photosynthetic partners, a cyanobacterium and a green alga, to develop two distinctly looking photomorphs. We compared gene expression of thallus sectors of the photosymbiodeme-forming lichen Peltigera britannica containing cyanobacterial photobionts with thallus sectors with both green algal and cyanobacterial photobionts and investigated differential gene expression at different temperatures representing mild and putatively stressful conditions. First, we quantified photobiont-mediated differences in fungal gene expression. Second, because of known ecological differences between photomorphs, we investigated symbiont-specific responses in gene expression to temperature increases. Photobiont-mediated differences in fungal gene expression could be identified, with upregulation of distinct biological processes in the different morphs, showing that interaction with specific symbiosis partners profoundly impacts fungal gene expression. Furthermore, high temperatures expectedly led to an upregulation of genes involved in heat shock responses in all organisms in whole transcriptome data and to an increased expression of genes involved in photosynthesis in both photobiont types at 15 and 25°C. The fungus and the cyanobacteria exhibited thermal stress responses already at 15°C, the green algae mainly at 25°C, demonstrating symbiont-specific responses to environmental cues and symbiont-specific ecological optima.

Assessing Identification Accuracy of Research Grade iNaturalist Observations in Lichens and other Taxonomically Difficult Organisms

Community science-generated biodiversity data can provide essential information for understanding species distributions, behaviors and conservation statuses. However, their utility can be limited due to high uncertainty and variability in quality, especially for small taxonomically difficult organisms like fungi and insects. One important set of community-generated data that are increasingly used by scientists are Research Grade (RG) iNaturalist observations. These observations are aggregated into the Global Biodiversity Information Facility database. Here we assessed the accuracy of RG lichen observations in iNaturalist. Lichens are mutualistic symbioses formed between fungi and a photosynthetic partner, either algae or cyanobacteria that occur in every terrestrial ecosystem on the planet (Brodo et al. 2001). They are sensitive indicators of environmental health, especially air quality, and provide essental food and nesting material for animals, along with performing many other ecosystem services (Allen and Lendemer 2021, Brodo et al. 2001, Nimis et al. 2002). We examined hundreds of observations and determined if the identification was correct, if it was not possible to identify the observation given the data provided, or if the identification was incorrect. Identification accuracy of selected species varied widely, from zero observations with enough information for correct identification (e.g., Rhizocarpon geographicum and Cladonia chlorophaea) to 100% correct identifications (e.g., Cetradonia linearis and Physconia subpallida, McMullin and Allen 2022). Most frequently, species that require microscopic examination or chemical tests for accurate identification were unable to be verified versus those that require only macromorphology. We provide a series of suggestions for best practices to improve the quality of RG observations and thus the utility of community-generated observation data for taxonomically difficult organisms.

A comparative genomic analysis of lichen-forming fungi reveals new insights into fungal lifestyles

Lichen-forming fungi are mutualistic symbionts of green algae or cyanobacteria. We report the comparative analysis of six genomes of lichen-forming fungi in classes Eurotiomycetes and Lecanoromycetes to identify genomic information related to their symbiotic lifestyle. The lichen-forming fungi exhibited genome reduction via the loss of dispensable genes encoding plant-cell-wall-degrading enzymes, sugar transporters, and transcription factors. The loss of these genes reflects the symbiotic biology of lichens, such as the absence of pectin in the algal cell wall and obtaining specific sugars from photosynthetic partners. The lichens also gained many lineage- and species-specific genes, including those encoding small secreted proteins. These genes are primarily induced during the early stage of lichen symbiosis, indicating their significant roles in the establishment of lichen symbiosis.Our findings provide comprehensive genomic information for six lichen-forming fungi and novel insights into lichen biology and the evolution of symbiosis.

Diversity of Saxicolous Lichens along an Aridity Gradient in Central M閤ico

Lichens are symbiotic organisms that comprise a fungus and a photosynthetic partner wich are recognized as a good indicator of climate change. However, our understanding of how aridity affects the diversity of saxicolous lichens in drylands is still limited. To evaluate the relationship between saxicolous lichen diversity and aridity in a central México dryland, a geographical transect was established of 100 km to build an aridity gradient in the semiarid zone of the State of Querétaro, Mexico, comprising ten sampling sites with a 10 km separation. Species richness, abundance and diversity of soil lichen species were recorded using two sampling methods: the quadrat-intercept and the line-intercept method, to compare their performance in assessing soil lichen diversity in drylands. The number of species and Shannon diversity of saxicolous lichens were higher at intermediate values of the aridity index (AI = 0.10–0.34). Quadrat intercept and point intercept methods gave quite similar results, which means that the selected method does not influence the results in a significant way. This study confirms the role of saxicolous lichens as climate change indicators and reveals the importance of the sampling method selection in the evaluation of different parameters of soil lichen diversity in drylands.

Fungal biomineralization

Fungi are key organisms of the biosphere with major roles in organic-matter decomposition, element cycling, plant pathogenicity, and symbioses in aquatic and terrestrial habitats. The vast majority exhibit a filamentous, branching growth form and are aerobic chemoorganotrophs that derive carbon and energy from organic substances, and are particularly associated with soil, the plant-root zone, and rock surfaces. It is now known that some fungi are lithotrophs, deriving energy from the oxidation of inorganic materials, whereas others are photoheterotrophs, deriving additional energy from light for organic matter utilization when oxygen is limited. This means that fungi are of much wider environmental significance than previously thought and explains their ubiquity in locations previously thought to be inimical to fungal existence, such as the deep subsurface and other anaerobic locations. In addition to such free-living species, fungi associated with photosynthetic partners are also of profound biosphere importance. For example, lichens, which are composed of a symbiotic association between a fungus and a phototrophic alga and/or cyanobacterium, are pioneer colonizers and bioweathering agents of rocks and minerals. Mycorrhizas are symbiotic, plant-root-associated fungi found to colonize the majority of plant genera, where they improve plant nutrition through solubilization of essential metals and phosphate from soil minerals. Biomineralization in the soil can also immobilize toxic metals in the vicinity of plant roots, thereby benefiting plant colonization and facilitating revegetation of contaminated habitats. Wherever fungi are found, transformation of metals and minerals is a key aspect of their activity, with biomineralization an important feature. Fungal biomineralization is an important facet of geomycology - namely the roles of fungi in geochemical and geophysical processes. This article seeks to highlight the concept of biomineralization as applied to fungi, the occurrence and significance of important fungal biominerals in natural and synthetic environments, and the applied potential of fungal biomineralization in nanobiotechnology.

Antimicrobial and antioxidant activity of Evernia prunastri extracts and their isolates

Lichens are symbiotic organisms formed by a fungus and one or more photosynthetic partners which are usually alga or cyanobacterium. Their diverse and scarcely studied metabolites facilitate adaptability to extreme living conditions. We investigated Evernia prunastri (L.) Ach., a widely distributed lichen, for its antimicrobial and antioxidant potential. E. prunastri was sequentially extracted by hexane (Hex), dichloromethane (DCM) and acetonitrile (ACN) that were screened for their antioxidant and antimicrobial (against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Candida albicans) activities. The Hex extract possessed the highest antioxidant capacity (87 mg ascorbic acid/g extract) corresponding to the highest content of phenols (73 mg gallic acid/g extract). The DCM and Hex extracts were both active against S. aureus (MICs of 4 and 21 µg/ml, respectively) but were less active against Gram-negative bacteria and yeast. The ACN extract exhibited activity on both S. aureus (MIC 14 µg/ml) and C. albicans (MIC 38 µg/ml) and was therefore further fractionated by silica gel column chromatography. The active compound of the most potent fraction was subsequently characterized by 1H and 13C-NMR spectroscopy and identified as evernic acid. Structural similarity analyses were performed between compounds from E. prunastri and known antibiotics from different classes. The structural similarity was not present. Antioxidant and antimicrobial activities of E. prunastri extracts originate from multiple chemical compounds; besides usnic acid, most notably evernic acid and derivatives thereof. Evernic acid and its derivatives represent possible candidates for a new class of antibiotics.Graphic abstract

Contribution of Cyanotoxins to the Ecotoxicological Role of Lichens.

The fascinating world of lichens draws the attention of the researchers because of the numerous properties of lichens used traditionally and, in modern times, as a raw material for medicines and in the perfumery industry, for food and spices, for fodder, as dyes, and for other various purposes all over the world. However, lichens being widespread symbiotic entities between fungi and photosynthetic partners may acquire toxic features due to either the fungi, algae, or cyano-procaryotes producing toxins. By this way, several common lichens acquire toxic features. In this survey, recent data about the ecology, phytogenetics, and biology of some lichens with respect to the associated toxin-producing cyanoprokaryotes in different habitats around the world are discussed. Special attention is paid to the common toxins, called microcystin and nodularin, produced mainly by the Nostoc species. The effective application of a series of modern research methods to approach the issue of lichen toxicity as contributed by the cyanophotobiont partner is emphasized.

Establishment of Agrobacterium tumefaciens-Mediated Transformation of Cladonia macilenta, a Model Lichen-Forming Fungus.

Despite the fascinating biology of lichens, such as the symbiotic association of lichen-forming fungi (mycobiont) with their photosynthetic partners and their ability to grow in harsh habitats, lack of genetic tools manipulating mycobiont has hindered studies on genetic mechanisms underpinning lichen biology. Thus, we established an Agrobacterium tumefaciens-mediated transformation (ATMT) system for genetic transformation of a mycobiont isolated from Cladonia macilenta. A set of combinations of ATMT conditions, such as input biomass of mycobiont, co-cultivation period with Agrobacterium cells, and incubation temperature, were tested to identify an optimized ATMT condition for the C. macilenta mycobiont. As a result, more than 10 days of co-cultivation period and at least 2 mg of input biomass of the mycobiont were recommended for an efficient ATMT, owing to extremely slow growth rate of mycobionts in general. Moreover, we examined T-DNA copy number variation in a total of 180 transformants and found that 88% of the transformants had a single copy T-DNA insertion. To identify precise T-DNA insertion sites that interrupt gene function in C. macilenta, we performed TAIL-PCR analyses for selected transformants. A hypothetical gene encoding ankyrin repeats at its C-terminus was interrupted by T-DNA insertion in a transformant producing dark-brown colored pigment. Although the identification of the pigment awaits further investigation, this proof-of-concept study demonstrated the feasibility of use of ATMT in construction of a random T-DNA insertion mutant library in mycobionts for studying genetic mechanisms behind the lichen symbiosis, stress tolerance, and secondary metabolite biosynthesis.