Genus Cladonia Research Articles

Vertical concentration gradients of heavy metals in Cladonia lichens across different parts of thalli

Certain lichens of the genus Cladonia are effective heavy-metal-tolerant colonisers of strongly contaminated and disturbed sites. Among them, Cladonia cariosa, Cladonia pyxidata and Cladonia rei are the major components of specific cryptogamic assemblages proven to be bioindicators of soil pollution. This study examines the bioaccumulation capacity and heavy metal accumulation pattern of these species in the context of element concentration levels in various parts of their thalli at various vertical distances from the ground. The content of Zn, Pb, Cd, As and Cu in primary squamules, lower and upper parts of secondary thalli (podetia), and fruiting bodies (apothecia), as well as the corresponding substrate, was analysed using the AAS method. The substrate turned out to be the main source of heavy metals in the examined Cladonia lichens. Element accumulation in particular parts of thalli greatly depends on metal enrichment in the immediate vicinity while Cu/Zn ratios for both substrate and lichen samples were very low and comparable within the species. Concentration levels in thalli usually decrease significantly with distance from the substrate. The exception is copper, which content was frequently higher in apothecia than in the upper parts of podetia. Low bioaccumulation factors calculated for the examined Cladonia specimens classified these lichens as weak accumulators of heavy metals. Even given an extremely high level of contaminants in the substrate, the upper parts of thalli are not greatly affected. Consequently, fruticose and erect growth form, in combination with low accumulation capacity and a remarkable decrease in metal content along a vertical gradient, may be an important attribute of Cladonia lichens in the colonisation of a highly contaminated substrate. The content of elements differs significantly between particular parts of Cladonia thalli; this should be taken into account whenever burdens of heavy metals are used as indicators in biomonitoring studies.

DNA Barcodes for the Distinction of Reindeer Lichens: A Case Study UsingCladonia rangiferinaandC. stygia

Abstract: Kanz, B., Brackel, W. v., Cezanne, R., Eichler, M., Hohmann, M.-L., Teuber , D. & Printzen, C. 2015. DNA barcodes for the distinction of reindeer lichens: a case study using Cladonia rangiferina and C. stygia. — Herzogia 28: 445–464. Many groups of lichens are difficult to identify due to their phenotypical plasticity. Molecular methods are therefore increasingly used not only to elucidate phylogenetic relationships among taxa, but also to assign individuals to species. Most members of the genus Cladonia are conspicuous and frequently collected, but notoriously variable. At the same time the genus comprises species that have to be regularly monitored under EU legislation. A „DNA barcoding“ approach could thus be very helpful to correctly identify taxa. The internal transcribed spacer region of the ribosomal RNA repeat has been proposed as a universal barcoding marker for fungi. However, it has also been shown that data from this locus alone might be insufficient to reliably distinguish members o...

Materials for biota of lichens and lichenicolous fungi in the military area near Toruń, Poland

The paper presents the first species list of lichens and lichenicolous fungi found and identified in the artillery training area located near the city of Torun. Lichenological studies were conducted in that area in 1998–2001 and 2009. A total of 100 taxa were identified, including 4 species of lichenicolous fungi, with particular emphasis on epigeic lichens – mainly from the genus Cladonia and Cetraria associated with heaths and arenaceous grasslands. The identified lichens include some threatened and protected taxa, i.a. Bryoria subcana , Cetraria islandica , Physconia distorta , Peltigera canina , P. polydactylon , P. praetextata , Ramalina fraxinea , Stereocaulon condensatum.

Contribution to the knowledge of the genus "Cladonia" in Macaronesia

Sixteen species of the genus Cladonia are reported from Macaronesia from the Canary Islands. Three species are new to canarian flora and, two of them new to Macaronesia. The chemical variation of the taxa is reported. A phytogeographic distribution of the taxa with data on their habitat and ecology are presented.

Distribution of mycotoxins and usnic acid in the thalli of epigeous lichens

The distribution of mycotoxins anc unic acid in the thallus of epigenous fruticose lichens Alectoria ochroleuca; several species of the genera Cladonia, Cotraria, and Flavocetraria; foliose lichens Nephroma arcticum; and six species of the genus Peltigera were studied by mnzyme-linked immunosorbent assay. The mycotoxin content was lower in the top or apical parts of the thalli than in the basal parts and in dead areas. Differences in the distribution of usnic acid were insignificant in the majority of species, but in Cladonia stellaris and N. arcticum lichens the content of this metabolite in the upper and pc ripheral areas was higher than in the basal parts.

Molecular phylogeny and ultrastructure of the lichen microalga Asterochloris mediterranea sp. nov. from Mediterranean and Canary Islands ecosystems.

The microalgae of the genus Asterochloris are the preferential phycobionts in Cladonia, Lepraria and Stereocaulon lichens. Recent studies have highlighted the hidden diversity of the genus, even though phycobionts hosting species of the genus Cladonia in Mediterranean and Canarian ecosystems have been poorly explored. Phylogenetic analyses were made by concatenation of the sequences obtained with a plastid - LSU rDNA - and two nuclear - internal transcribed spacer (ITS) rDNA and actin - molecular markers of the phycobionts living in several populations of the Cladonia convoluta-Cladonia foliacea complex, Cladonia rangiformis and Cladonia cervicornis s. str. widely distributed in these areas in a great variety of substrata and habitats. A new strongly supported clade was obtained in relation to the previously published Asterochloris phylogenies. Minimum genetic variation was detected between our haplotypes and other sequences available in the GenBank database. The correct identification of the fungal partners was corroborated by the ITS rDNA barcode. In this study we provide a detailed characterization comprising chloroplast morphology, and ultrastructural and phylogenetic analyses of a novel phycobiont species, here described as Asterochloris mediterranea sp. nov. Barreno, Chiva, Moya et Škaloud. A cryopreserved holotype specimen has been deposited in the Culture Collection of Algae of Charles University in Prague, Czech Republic (CAUP) as CAUP H 1015. We suggest the use of a combination of several nuclear and plastid molecular markers, as well as ultrastructural (transmission electron and confocal microscopy) techniques, both in culture and in the symbiotic state, to improve novel species delimitation of phycobionts in lichens.

A REMARKABLE CLADONIACEAE FLORA AT SUBANDEAN REGION IN CHÁMEZA (CASANARE, COLOMBIA)

A remarkable Cladoniaceae flora was discovered in the subandean region of Chameza, ( Casanare, Colombia ) at 1200 meters above sea level. Four species of the genus Cladonia (Cladoniaceae, Lichenized Fungi) are new records for Colombia : Cladonia macilentoides, C. scabriuscula, C. sipmanii and C. subdelicatula. This shows the importance of increasing diversity studies at mid elevations where suitable habitats for species of Cladonia occur.

Mycotoxins and usnic acid and their distribution in lichens belonging to the genera Cetraria, Flavocetraria, and Cladonia

The content and the pattern of distribution of mycotoxins and usnic acid in thalli of lichens belonging to the genera Cladonia (four species), Cetraria (two species), and Flavocetraria (two species) have been studied using the method of immunoenzymatic analysis. Statistically significant differences have been estimated between the upper and the lower parts of the thalli. The levels of the mycotoxins in the lower, older parts of the thalli have been higher than in the upper, younger ones. The comparison of lichen specimens belonging to the same species and collected in the same location at different years did not reveal statistically significant differences in the content of mycotoxins both in the upper and the lower parts of the thalli. Statistically significant differences in the usnic acid content have been revealed only in the case of C. stellaris. In the upper part of the podetia of this species, the usnic acid content has been higher than in the lower one.

Expectorant and antioxidant activities of purified fumarprotocetraric acid from Cladonia verticillaris lichen in mice

The lichen Cladonia verticillaris produces bioactive secondary metabolites, such as fumarprotocetraric (FUM) and protocetraric acids. Species of the genus Cladonia demonstrate anti-tumor, anti-inflammatory and antipyretic activities and have been used in folk medicine to treat respiratory diseases (throat irritation, cough, asthma and tuberculosis). The aim of the present study was to evaluate the expectorant and mucolytic activities of fumarprotocetraric acid in albino Swiss mice. FUM was extracted and purified from an acetone extract of C. verticillaris. The phenol red quantification method was used on the bronchoalveolar lavage fluid following the administration of FUM (25, 50 or 100 mg/kg orally or intraduodenally and 12.5, 25 or 50 mg/kg, intraperitoneally) for the evaluation of expectorant activity. Control groups received either saline solution (7.5 mL/kg) or ambroxol (1 mg/kg) through the same administration routes. Antioxidant activity was evaluated using the thiobarbituric acid reactive species assay in mouse lung tissue treated with the FUM at 25, 50 or 100 mg/kg orally, followed by a lipopolysaccharide solution at 1 mg/kg intrapleurally. The same protocol was used for the control groups using either saline solution (7.5 mL/kg, orally) or N-acetylcysteine (20 mg/kg, orally). Results: Orally administered FUM at doses of 25 and 50 mg/kg promoted significantly greater dose-dependent phenol red activity in the bronchoalveolar lavage and expectorant activity in comparison to the controls (p < 0.05). Lipid peroxidation (malondialdehyde equivalent) was reduced by 50% in the lung tissue. Conclusion: The results confirm the expectorant and antioxidant properties of fumarprotocetraric acid produced by the lichen C. verticillaris.

Water-soluble phenolic compounds in lichens

The quantity and the qualitative composition (for some species) of phenolic compounds (PC) washed out of the intact thalli of lichens of the orders Peltigerales (the genera Peltigera, Solorina, and Nephroma) and Lecanorales (the genera Cladonia, Alectoria, and Cetraria) were studied. It was shown that the quantity of leachable PCs in Peltigerales was on average 2–3 times higher than in Lecanorales. At the same time, the extractability of PC from intact thalli by water was higher in Lecanorales than in Peltigerales: 48–88% and 34–70%, respectively, of the PC content in ethanol extracts from crushed thalli (i.e., of the total content of soluble PC). Water-soluble PC in the lichens Peltigera aphthosa, Solorina crocea, Cetraria islandica, Flavocetraria nivalis, Cladonia uncialis, and Cladonia arbuscula were represented by 7–12 phenolic compounds with similar qualitative composition in the species of the same order. The most part of water soluble PC were phenylpropanoids. All of the studied species showed the presence of p-hydroxybenzoic acid derivatives; vanillic and protocatechuic acid derivatives were found in Cetraria and Cladonia species, respectively.

A Morphometric Evaluation of theCladonia chlorophaeaGroup and Allied Taxa (Cladoniaceae, Ascomycota)

OSYCZKA, P. 2013. A morphometric evaluation of the Cladonia chlorophaea group and allied taxa (Cladoniaceae, Ascomycota). — Herzogia 26: 49– 64.The Cladonia chlorophaea group represents a complex of morphologically and chemically similar taxa that have long perplexed taxonomists. Until now, few studies have systematically evaluated morphological characters in the group. A morphometric analysis of samples belonging to the taxa of the Cladonia chlorophaea and C. humilis groups as well as C. fimbriata, was performed using various fundamental statistical methods. It revealed four unambiguously distinct morphological aggregates of samples, corresponding to four well known, cosmopolitan species, namely C. asahinae, C. chlorophaea, C. fimbriata and C. humilis. The analysis highlights the most reliable diagnostic characters distinguishing the obtained groups which can be applied for species determination. A chemical race of C. asahinae with fumarprotocetraric acid chemosyndrome only is reported from Europe and a chemotype of C. fimbriata with fumarprotocetraric and bourgeanic acids was revealed during the study. Morphometric analyses may be a very useful tool for evaluation of features within the genus Cladonia and other lichen genera containing cryptic species.

Species delimitation in Cladonia (Ascomycota): a challenge to the DNA barcoding philosophy

The lichen-forming fungal genus Cladonia is species-rich with approximately 500 described species. The accepted barcode for fungi (ITS rDNA) often fails in identifying Cladonia spp. In order to find other markers that, in combination with the ITS rDNA region can be used for species identification in Cladonia, we studied the loci IGS rDNA, ef1α, rpb2 and cox1. A total of 782 sequences from 36 species have been analyzed. PCR amplification success rate, intraspecific and interspecific genetic distance variation, calculated using the K2P model, and the correct identification percentage (PCI) were taken into account to assess possible barcode regions. The marker showing the least intraspecific genetic distance range was cox1, followed by ITS rDNA and ef1α. Of the five studied markers only cox1 showed a barcoding gap. The rpb2 locus showed the highest PCI values, but it was the most difficult to amplify. The highest correct identification rates using blast method were obtained with rpb2.

Radiological investigations at the “Taiga” nuclear explosion site, part II: man-made γ-ray emitting radionuclides in the ground and the resultant kerma rate in air

Samples of soil and epigeic lichens were collected from the “Taiga” peaceful nuclear explosion site (61.30°N 56.60°E, the Perm region, Russia) in 2009 and analyzed using high resolution γ-ray spectrometry. For soil samples obtained at six different plots, two products of fission (137Cs and 155Eu), five products of neutron activation (60Co, 94Nb, 152Eu, 154Eu, 207Bi) and 241Am have been identified and quantified. The maximal activity concentrations of 60Co, 137Cs, and 241Am for the soils samples were measured as 1650, 7100, and 6800 Bq kg−1 (d.w.), respectively. The deposit of 137Cs for the top 20 cm of soil on the tested plots at the “Taiga” site ranged from 30 to 1020 kBq m−2; the maximal value greatly (by almost 3 orders of magnitude) exceeded the regional background (from global fallout) level of 1.4 kBq m−2. 137Cs contributes approximately 57% of the total ground inventory of the man-made γ-ray emitters for the six plots tested at the “Taiga” site. The other major radionuclides –241Am and 60Co, constitute around 40%. Such radionuclides as 60Co, 137Cs, 241Am, and 207Bi have also been determined for the epigeic lichens (genera Cladonia) that colonized certain areas at the ground lip produced by the “Taiga” explosion. Maximal activity concentrations (up to 80 Bq kg−1 for 60Co, 580 Bq kg−1 for 137Cs, 200 Bq kg−1 for 241Am, and 5 Bq kg−1 for 207Bi; all are given in terms of d.w.) have been detected for the lower dead section of the organisms. The air kerma rates associated with the anthropogenic sources of gamma radiation have been calculated using the data obtained from the laboratory analysis. For the six plots tested, the kerma rates ranged from 50 to 1200 nGy h−1; on average, 51% of the dose can be attributed to 137Cs and 45% to 60Co. These estimates agree reasonably well with the results of the in situ measurements made during our field survey of the “Taiga” site in August 2009.

Enzyme immunoassay of the secondary metabolites of micromycetes as components of lichen substances

The composition of low-molecular biologically active metabolites typical of microscopic fungi has been studied in blastemas of fruticose lichens of the genera Cladonia, Cetraria, Evernia, Bryoria, and Usnes. The enzyme immunoassay method showed the presence of sterigmatocystin, emodin, mycophenolic acid, citrinin, alternariol, and diacetoxyscirpenol, which occurred regularly and, in most cases, at a frequency of 55 to 100%. The highest levels of accumulation were 0.001-0.003% for emodin, 0.0002% for alternariol and citrinin, 0.0001% for sterigmatocystin and mycophenolic acid, and 0.00005% of the weight of air-dry material for diacetoxyscirpenol. Other metabolites (cyclopiazonic acid, ergot alkaloids, ochratoxin A, PR toxin, deoxynivalenol, zearalenone, and fumonisins) were detected in these lichens less frequently (sometimes only upon the expansion of the territory of sampling), and their content was no more than 0.00005%. The peculiarities of the component composition and the levels of accumulation of fungal metabolites in lichens of different taxonomic affiliation were discussed.

Species delimitations in the Cladonia cariosa group (Cladoniaceae, Ascomycota)

AbstractPhenotypic characters, either morphological or chemical, have shown to be insufficient to delimit species boundaries in the genus Cladonia. The present study addresses the circumscription of species within the Cladonia cariosa group, examining a number of specimens of the currently accepted taxa Cladonia cariosa, C. symphycarpa, C. acuminata, C. subcariosa and C. latiloba. We employed maximum likelihood and Bayesian methods of phylogenetic reconstructions based on DNA sequences of ITS, rpb2 and ef1α regions. Our results show that the C. cariosa group consists of at least four phylogenetic lineages. It is also shown that each of these lineages is chemically variable, which restricts the taxonomic value of the chemical differences within the group. However, anatomical differences, such as squamule surface and cortex structure, were found to correlate with the distinct lineages found in the phylogenetic analysis. This result confirms the taxonomic value of the cortical surface under SEM, as was found in other lichen groups.

The GenusCladonia, GroupCocciferae, in Poland

Osyczka, P. 2011. The genus Cladonia, group Cocciferae, in Poland. — Herzogia 24: 231–249.A revision of the genus Cladonia, group Cocciferae, is presented for Poland. The morphology, habitat and distribution of the taxa are treated, and the chemical races of each species and their frequency in the country have been indicated. The study is based on the examination of approximately 3300 herbarium specimens originating from Poland. Seventeen species have been accepted; the occurrence of the two previously reported species Cladonia bacilliformis and C. cyanipes has not been confirmed. Keys for all species known in Poland are provided.

A note about new and rarely recorded taxa of the genus Cladonia from the Arctic island Spitsbergen (Svalbard)

The paper is based on field studies dealing with the lichen genus &lt;em&gt;Cladonia&lt;/em&gt; Hill ex &lt;em&gt;P. Browne&lt;/em&gt; (&lt;em&gt;Cladoniaceae&lt;/em&gt;) carried out by the author in the region of Spitsbergen in 2002 as well as taxonomical work and revision of herbarium material, collected during several earlier Polish polar expeditions to this part of the Arctic. As the result not reported so far from the island species &lt;em&gt;Cladonia stygia&lt;/em&gt; (Fr.) Ruoss, &lt;em&gt;C. trassii&lt;/em&gt; Ahti, &lt;em&gt;C. verticillata&lt;/em&gt; (Hoffm.) Schaer. have been determined. Besides, the presence of very rarely recorded taxa &lt;em&gt;C. crispata&lt;/em&gt; var. &lt;em&gt;cetrariiformis&lt;/em&gt; (Delise) Vain., &lt;em&gt;C. merochlorophaea&lt;/em&gt; Asahina and &lt;em&gt;Cladonia symphycarpia&lt;/em&gt; (Flörke) Fr. is confirmed and their new localities are given. Furthermore, the presence of &lt;em&gt;C. coccifera&lt;/em&gt; s.str. is also substantiated.

Insight into the Cladonia convoluta-C. foliacea (Cladoniaceae, Ascomycota) complex and related species, revealed through morphological, biochemical and phylogenetic analyses

Species boundaries are often not easy to establish in lichens of the genus Cladonia, where each species displays a great morphological variability. Cladonia convoluta and C. foliacea are two commonly accepted, morphologically similar lichen taxa. Molecular (nuclear ITS rDNA, rpb2 partial gene, cox1 partial gene), chemical and morphological characters were used to elucidate the taxonomic rank of these taxa. Our analyses were carried out on material from several European countries. Other related species, viz. C. firma, C. cervicornis and C. pulvinata were also studied. Maximum Parsimony, Maximum Likelihood and Bayesian analyses were based on 71 ITS, 39 cox1 and 33 rpb2 sequences. Neither morphological characters nor phylogenetic analyses gave evidence to delimit two taxa in the C. convoluta/C. foliacea complex; however C. firma, C. cervicornis and C. pulvinata formed three distinct monophyletic groups.

Biota of lichens on the Zadroże Dune and its immediate surroundings

The present paper aims at describing the biota of lichens of the Zadroże Dune and its adjacent area in Toruń. The research on lichens was conducted during the years 2005-2006. The data on lichens come from my own research and literature. Altogether, the occurrence of 82 lichen taxa was recorded in the investigated area, including 74 species recorded during my own research. Due to the availability of habitats and substrata, terricolous species dominate here, mainly from the genus Cladonia and Cetraria. Particularly noteworthy are rare, endangered and protected species from this area e.g. Peltigera canina, Polyblastia agraria, Rinodina conradii and Stereocaulon condensatum

Biota of lichens on the Zadroże Dune and its immediate surroundings

The present paper aims at describing the biota of lichens of the Zadroże Dune and its adjacent area in Toruń. The research on lichens was conducted during the years 2005-2006. The data on lichens come from my own research and literature. Altogether, the occurrence of 82 lichen taxa was recorded in the investigated area, including 74 species recorded during my own research. Due to the availability of habitats and substrata, terricolous species dominate here, mainly from the genus Cladonia and Cetraria. Particularly noteworthy are rare, endangered and protected species from this area e.g. Peltigera canina, Polyblastia agraria, Rinodina conradii and Stereocaulon condensatum