A spectrophotometric analysis of extracted water-soluble phenolic metabolites of lichens.

MAIN CONCLUSION: Rainwater most probably constitutes a relatively effective solvent for lichen substances in nature which have the potential to provide for human and environmental needs in the future. The aims were (i) to test the hypothesis on the potential solubility of lichen phenolic compounds using rainwater under conditions that partly reflect the natural environment and (ii) to propose new and effective methods for the water extraction of lichen substances. The results of spectrophotometric analyses of total phenolic metabolites in rainwater-based extracts from epigeic and epiphytic lichens, employing the Folin-Ciocalteu (F.-C.) method, are presented. The water solvent was tested at three pH levels: natural, 3, and 9. Extraction methods were undertaken from two perspectives: the partial imitation of natural environmental conditions and the potential use of extraction for economic purposes. From an ecological perspective, room-temperature water extraction ('cold' method) was used for 10-, 60-, and 120-min extraction periods. A variant of water extraction at analogous time intervals was an 'insolation' with a 100W light bulb to simulate the heat energy of the sun. For economic purposes, the water extraction method used the Soxhlet apparatus and its modified version, the 'tea-extraction' method ('hot' ones). The results showed that those extractions without an external heat source were almost ineffective, but insolation over 60- and 120-min periods proved to be more effective. Both tested 'hot' methods also proved to be effective, especially the 'tea-extraction' one. Generally, an increase in the concentration of phenolic compounds in water extracts resulted from an increasing solvent pH. The results show the probable involvement of lichen substances in biogeochemical processes in nature and their promising use for a variety of human necessities.

A review of the potential of lichen substances as antifungal agents: the effects of extracts and lichen secondary metabolites on Fusarium fungi.

The present meta-analysis provides literature data on the effect of lichen extracts and single secondary metabolites used against Fusarium spp. moulds. Lichen extracts were obtained from 51 corticolous, 17 terricolous and 18 saxicolous lichen species and 37 secondary compounds were tested against eight fungal species, i.e., Fusarium acuminatum, F. avenaceum, F. culmorum, F. fujikuroi, F. oxysporum, F. roseum, F. solani and F. udum. The researchers used several test methods, mostly to determine MIC and IZ. Extracts were obtained using several solvents, mainly organic ones with use of the Soxhlet apparatus. The most frequently tested species was F. oxysporum, against which lichen substances from Alectoria sarmentosa, Cladonia mitis, C. rangiferina, Flavoparmelia caperata, Hypotrachyna cirrhata, Leucodermia leucomelos, Parmotrema austrosinense, P. reticulatum, Physcia aipolia, Pseudevernia furfuracea, Roccella montagnei and Umbilicaria nylanderiana and secondary metabolites such as 2-hydroxy-4-methoxy-3,6-dimethylbenzoic acid, atranorin, lecanoric and (+)-usnic acids showed the highest antifungal potential. These agencies could compete with the potential of fungicides, such as flucytosine and fluconazole. Other species have been poorly investigated. Statistical analysis of literature data showed that the fungistatic potential of lichen extracts is significantly different from individual secondary metabolites. Similarly, the potential of secondary metabolites often differs significantly from that of non-lichen substances. This meta-analysis indicates the potential of lichen substances as future anti-fusarial agents.

Does the lichenicolous fungus Heterocephalacria bachmannii affect the antimicrobial potential of its host Cladonia foliacea?

The effects of the lichenicolous fungus Heterocephalacria bachmannii on the antimicrobial potential of the lichen Cladonia foliacea demonstrated that the extracts investigated have antimicrobial potential against gram-positive and negative bacteria, and yeast, and inhibit the germination of fungal spores. Inhibition activity varied considerably depending on the extract, the bacterial species, and the absence or presence of H. bachmannii; unparasitised C. foliacea has a higher antimicrobial activity. Methanol and acetone extracts of C. foliacea alone have higher inhibition diameters than C. foliacea with H. bachmannii against Enterobacter cloacae; the methanol extract of C. foliacea showed the best inhibition (250 µg/ml). C. foliacea also has a high lysozyme potential against Streptococcus agalactiae and Staphylococcus aureus. Fungal hyphae of Alternaria alternata were more affected by the methanol extract from C. foliacea.

The effect of lichen secondary metabolites on Aspergillus fungi.

A systematic review of literature data on the antifungal potential of extracted lichen compounds and individual secondary metabolites against mold species of the genus Aspergillus is provided. Crude extracts from 49 epiphytic, 16 epigeic and 22 epilithic species of lichens and 44 secondary metabolites against 10 species, Aspergillus candidus, A. flavus, A. fumigatus, A. nidulans, A. niger, A. ochraceus, A. parasiticus, A. restrictus, A. stellatus and A. ustus, were analysed. Several measuring techniques were employed for such analyses. Lichen substances were extracted with alcoholic and other organic solvents mainly using the Soxhlet apparatus. Among the three most-studied mold species, the results showed that the crude extracts from the thalli of the lichens Cladonia foliacea, Hypotrachyna cirrhata, Leucodermia leucomelos, Platismatia glauca and Pseudevernia furfuracea against Aspergillus flavus, from C. foliacea, Nephroma arcticum and Parmelia sulcata against A. fumigatus and from Evernia prunastri, Hypogymnia physodes, Umbilicaria cylindrica and Variospora dolomiticola against A. niger have the greatest antifungal potential. The lichen secondary metabolites showed a higher inhibitory potential, e.g. protolichesterinic acid against A. flavus, lecanoric acid against A. fumigatus and orsellinic acid against A. niger; the other seven species of Aspergillus have been poorly studied and require further investigation. A comparison of the inhibitory potential of the tested mixtures of lichen substances and their secondary metabolites shows that they can compete with commonly used antifungal substances, such as ketoconazole and clotrimazole against A. flavus, A. nidulans, A. niger and A. parasiticus and fluconazole in the case of A. fumigatus.

Lichenochemical Screening and Antioxidant Capacity of Four Tunisian Lichen Species.

The phenolic composition and antioxidant capacity of four Tunisian lichen species, Cladonia rangiformis, Flavoparmelia caperata, Squamarina cartilaginea and Xanthoria parietina, were determined in order to provide a better understanding of their lichenochemical composition. Powdered material of F. caperata was the richest in total phenolic content (956.68 µg GAE g-1 DW) and S. cartilaginea in proanthocyanidin content (77.31 µg CE g-1 DW), while the acetone extract of X. parietina showed the highest flavonoid content (9.56 µg CE g-1 DW). The antioxidant capacity of all lichen extracts and crude material was evaluated by DPPH. scavenging, iron-chelating, and iron-reducing powers. Results showed that methanol extracts of S. cartilaginea had the highest DPPH. antioxidant capacity (IC50 =0.9 µg mL-1 ) and the highest iron-reducing power was attributed to the acetone extract of this species. All extracts of all species were further screened by Fourier-transform infrared spectroscopy (FT-IR) and nuclear resonance spectroscopy (NMR); results showed an abundance of phenols, aromatic compounds, and fatty acids. Overall, our results showed that the investigated species are a rich source of potentially bioactive compounds with valuable properties.

Secondary Metabolites and Antioxidant Capacity of the Tunisian Lichen Diploschistes ocellatus (Ascomycota).

This study, the first to assess the total phenolic, flavonoid, tannins, and proanthocyanidin content of the Tunisian lichen Diploschistes ocellatus, determined the antioxidant capacity in scavenging 2,2 diphenyl-1-picrylhydrazyl (DPPH), as well as the ferric-reducing and iron-chelating powers. The phenolic compound content of D. ocellatus was shown to be related to antioxidant activity. The highest phenolic and flavonoid contents of extracts were obtained with acetone (286.3 µg GAE/g DW and 3.24 µg CE/g DW, respectively), while the highest tannin and proanthocyanidin contents were obtained with methanol (5.5 µg TAE/g DW and 35.12 µg CE/g DW, respectively). The highest DPPH' scavenging capacity and iron-chelating power of extracts were obtained with methanol (concentration providing 50% inhibition [IC50] = 0.029 mg/mL and IC50 = 0.425 mg/mL, respectively), while acetone extracts showed a higher reducing power (IC50 = 0.118 mg/mL).

ETS and plastid sequence data indicate a spontaneous origin of Scandinavian betony, Betonica officinalis L.

BACKGROUND AND AIMS: Betony (Betonica officinalis L.) is one of the rarest and most spectacular plants in the Scandinavian flora. A long-term question has been whether it is spontaneous or introduced, or whether it comprises both spontaneous and introduced populations. This study aimed to answer this question by analyzing sequence data from the nuclear external transcribed spacer (ETS) region and three regions of the plastid genome, the trnT-trnL intergenic spacer (IGS) region, tRNA-Leu (trnL) intron, and the trnS-trnG IGS. MATERIALS AND METHODS: Altogether 41 samples from 11 European countries were analyzed. A unique duplication in the trnT-trnL IGS was detected in material from Skåne (southern Sweden), the "Skåne-duplication." Populations with this duplication are united on a moderately supported branch in the phylogeny based on plastid sequences. A distinct heath genotype from Yorkshire was discovered in the phylogeny based on plastid sequences and in a comparative cultivation. RESULTS: Phylogeny based on ETS sequences does not support any Scandinavian group, whereas a principal coordinates analysis ordination based on variable ETS positions indicated a spontaneous origin for all Scandinavian populations, which comprise a genetically well-defined subgroup of the species, most closely related to other spontaneous populations from adjacent parts of continental parts of northern Europe. DISCUSSION: Seven possible naturally occurring localities remain in Scandinavia, five in central Skåne, southernmost Sweden, and two on the southwestern part of the Danish island of Lolland.

Raman spectroscopic analysis of the effect of the lichenicolous fungus Xanthoriicola physciae on its lichen host.

Lichenicolous (lichen-dwelling) fungi have been extensively researched taxonomically over many years, and phylogenetically in recent years, but the biology of the relationship between the invading fungus and the lichen host has received limited attention, as has the effects on the chemistry of the host, being difficult to examine in situ. Raman spectroscopy is an established method for the characterization of chemicals in situ, and this technique is applied to a lichenicolous fungus here for the first time. Xanthoriicola physciae occurs in the apothecia of Xanthoria parietina, producing conidia at the hymenium surface. Raman spectroscopy of apothecial sections revealed that parietin and carotenoids were destroyed in infected apothecia. Those compounds protect healthy tissues of the lichen from extreme insolation and their removal may contribute to the deterioration of the apothecia. Scytonemin was also detected, but was most probably derived from associated cyanobacteria. This work shows that Raman spectroscopy has potential for investigating changes in the chemistry of a lichen by an invading lichenicolous fungus.

¹³7Cs concentrations in foliose lichens within Tsukuba-city as a reflection of radioactive fallout from the Fukushima Dai-ichi Nuclear Power Plant accident.

(137)Cs concentrations in ten species of foliose lichens collected within Tsukuba-city in August 2013 ranged from 1.7 to 35 kBq/kg. The relationships between (137)Cs in two dominant species, Dirinaria applanata and Physcia orientalis, and the air dose rate (µSv/h) at the sampling sites were investigated. (137)Cs in P. orientalis measured about 1 year after the Fukushima nuclear accident was correlated (r(2) = 0.80) more closely with the air dose rate than those measured after about 2 years (r(2) = 0.65), possibly demonstrating its continued value as a biomonitor to reflect ambient fall-out levels. In contrast, those of Dirinaria applanata were not correlated with the air dose rate in either year.

Raman spectroscopy of hot desert, high altitude epilithic lichens.

Twenty-three highly-coloured lichen specimens belonging to the genera Candelariella, Aspicilia and Xanthoria from high altitude sites in the Atacama Desert, Chile, 2300-4500 metres, have been analysed non-destructively by Raman spectroscopy. The vibrational band assignments in the spectra of the specimens, which were still attached to their limestone or sandstone substrata, were accomplished by comparison with the chemical compositions obtained from wet chemical extraction methods. Carotenoids and chlorophyll were found in all specimens as major components and the characteristic spectral signatures of calcium oxalate monohydrate (whewellite) and dihydrate (weddellite) could be identified; chemical signatures were found for these materials even in lichen thalli growing on the non-calcareous substrata, indicating probably that the calcium was provided here from wind- or rain-borne sources. The Raman spectral biomarkers for a variety of protective biomolecules and accessory pigments such as usnic acid, calycin, pulvinic acid dilactone and rhizocarpic acid have been identified in the lichens, in broad agreement with the chemical extraction profiles. The present study indicates that some form of non-destructive taxonomic identification based on Raman spectroscopy was also possible.

Lichen biodeterioration of ecclesiastical monuments in northern Spain.

Seven highly-coloured lichen species belonging to the genera Caloplaca, Candelariella, Aspicilia and Xanthoria from ecclesiastical buildings in northern Spain have been analysed non-destructively by FT-Raman spectroscopy. The vibrational band assignments in the spectra of the specimens, which were still attached to their limestone or sandstone substrata, were accomplished with the assistance of the chemical compositions obtained from wet chemical extraction methods. beta-Carotene was found in all specimens as the major pigment, and the characteristic spectral signatures of calcium oxalate monohydrate (whewellite) and dihydrate (weddelite) could be identified; chemical signatures were found for these materials even in lichen thalli growing the non-calcareous substrata, indicating, probably, that the calcium was provided here from wind-or-rain-borne sources. The Raman spectral biomarkers found in the lichens broadly agreed with the chemical extraction profiles as expected, but the present study indicates that some form of non-destructive taxonomic identification based on Raman spectroscopy was possible.

FT-Raman spectroscopy of lichens on dolomitic rocks: an assessment of metal oxalate formation.

The FT-Raman spectra of five epilithic lichen taxa growing on dolomite and magnesium-rich carbonate rocks have been analysed and interpreted for the key molecular marker bands associated with calcium oxalate monohydrate (whewellite), calcium oxalate dihydrate (weddelite) and magnesium oxalate dihydrate. From the results, it can be concluded that the biomineral product of lichen biodeterioration involves the calcareous part of the substratum only; no trace of magnesium oxalate has been found in the Raman spectra. Two of the species, Lecanora sulfurea and Aspicilia calcarea, produce calcium oxalate monohydrate exclusively, but Dirina massiliensis f. sorediata, D. massiliensis f. massiliensis and Tephromela atra produce significant quantities of the dihydrate. An explanation is advanced for the exclusive accumulation of calcium oxalate into the lichen thallus despite the significant presence of magnesium ions.

Quantitative EPR study on free radicals in the natural polyphenols interacting with metal ions and other environmental pollutants.

Quantitative electron paramagnetic resonance (EPR) method was applied to characterise radicals stabilised in polyphenolic matrices of various biogenic materials: lichens, mosses, composts, soils, peats, brown coals and sewage sludge sediments. The investigations were carried out on raw materials and extracted fractions of humic acids (HA). General trends of g value and spin concentration changes were found. These parameters in lichens strongly depend on lichen species and air pollution. Determination of the g value and semiquinone spin concentration allows to assess degree of transformation of organic matter in compost, soil, peat and lignite. Application of gaseous ammonia as a base penetrating the organic matrices extends the possibilities and usefulness of the method. Interaction of metal ions with humic materials is illustrated by interaction of VO2+ ion with peat and lignite HA as well as demineralised (raw and carbonised) brown coal. Our investigations demonstrate that quantitative EPR is a rapid and effective monitoring method to study the influence of various environmental factors on substances containing polyphenolic matrices.

Richard Spruce, botânico e desbravador da América do Sul / Richard Spruce, botanist-South America's explorer

No período entre 1849 e 1864, o botânico e explorador inglês Richard Spruce promoveu minucioso estudo da flora amazônica e dos costumes dos povos que habitavam essa regiäo. Ainda hoje, grande parte do conhecimento sobre várias famílias botânicas daquela regiäo advém do esforço desenvolvido por esse cientista. A amplitude de seus interesses, a meticulosidade e a exatidäo de suas descriçöes foram fenomenais: nada parece ter escapado à sua atençäo e capacidade de documentaçäo. Spruce era näo apenas notável botânico, mas também admirável antropólogo, lingüista(sabia francês, espanhol e português), geólogo e geógrafo, bem como arguto observador sociológico dos sistemas políticos e dos hábitos das tribos amazônicas e andinas entre as quais esteve, trazendo considerável contribuiçäo para o entendimento das crenças e práticas nativas e para o conhecimento das propriedades e usos das plantas, no contexto amazônico. Sua participaçäo na exploraçäo econômica de espécies locais também foi importante, particularmente em relaçäo aos gêneros "Hevea e Cinchona"