Cladonia Chlorophaea Research Articles

Identification and determination of usnic acid and fatty acid from various lichen species in arequipa, Peru, as well as antibacterial and antioxidant capacity

Seven species of lichens such us Umbilicaria aff. calvescens, Hypotrachyna enderythraea, Punctelia graminicola, Cladonia chlorophaea, Xanthoparmelia farinose, Psiloparmelia distincta (rock and tree substrates; rs and ts), and Usnea durietzii were collected from the province of Arequipa - Peru. Their usnic acid (UA) and fatty acid (FA) amounts in methanol-acetone were determined by High-performance liquid chromatography with diode-array detection (HPLC-DAD) and Gas chromatography-flame ionization detector (GC-FID). The antimicrobial activities of these extracts were evaluated against Staphylococcus aureus ATCC strains (43300, 29213, 25923 and 700699), Escherichia coli strains (O157:H7 and ATCC 10536), Salmonella enterica sv typhimurium ATCC 14028, Candida albicans ATCC 90028 and Candida. tropicalis ATCC 750T. In addition, antioxidant capacity was also studied by Total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical, 2,2-Azino-bis-3-ethylbenzothiazoline-6-sulfonic Acid (ABTS) radical cation and Ferric Reducing Antioxidant Power (FRAP) assays. The UA contents in all the studied lichen species varied between 0.017 and 0.304 %. Among all the tested extracts, Usnea durietzii had the highest total FA concentration (5.81 mg/g). P. distincta (rs) was active against S. aureus strains (MIC = 0.02–0.2 mg/mL) as well as C. albicans (MIC = 4 mg/mL) and C. tropicalis (MIC = 3 mg/mL). P. distincta (ts) displayed moderate total phenol content (TPC = 29.1 ± 1.6 mg GAE/g) and antioxidant capacity evidenced by scavenging DPPH (IC50 = 1.45 ± 0.03 mg/mL) and ABTS (18.2 ± 0.2 mg TE/g) radicals, but it showed high values of FRAP (1953 ± 87 μM Fe2+/g). Our findings indicate that P. distincta (rs) could be considered as a potential antimicrobial metabolites source whereas P. distincta (ts) and Puntelia graminicole for polyphenols with antioxidant compound.

Ca oxalates in lichen genus Cladonia on the dumps of the Sugur copper deposit (South Urals)

Ca oxalate crystals (mainly, weddellite (Ca(C2O4) • (2.5-x)H2O) are found in the lichen Cladonia chlorophaea on dumps of the Sugur copper deposit underwent to aerosol emissions from the Karabash copper smelting (KCS), South Urals. The samples of lichens and substrate are studied by scanning electron microscopy, energy-dispersion microanalysis, X-ray phase analysis and inductively coupled plasma mass spectrometry. The contents of toxic elements (Cu, Zn, Pb, Cd, Sn, Sb, Se, As, Te) of substrate and lichen thalli exceed their average contents in the Earth’s crust, whereas their contents in thalli are lower than in substrate (except for Cd). The high Pb content is associated with Ca oxalate. The lichen thalli contains inclusions of minerals, which compose the dump, and spherical particles, which are associated with the activity of the KCS and which contain signifcant concentrations of heavy metals. Keywords: calcium oxalates, weddellite, biomineralization, Cladonia, Karabash copper smelter, Sugur copper deposit,

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.

Cytogenetic and oxidative effects of three lichen extracts on human peripheral lymphocytes.

In the present study, we investigated cytogenetic and oxidative [total antioxidant capacity (TAC), totaloxidant status (TOS)] effects of methanol and water extracts of Cladonia chlorophaea (Flörke ex Sommerf.) Sprengel, Dermatocarpon miniatum (L.) W.Mann and Parmelia saxatilis (L.) Ach. on cultured human lymphocytes. In addition, different phenolic compounds in the extracts were quantified by high performance liquid chromatography (HPLC) analysis. As a result of HPLC analysis, methanol extracts of all lichen species tested had higher phenolic compounds. Likewise, methanol extracts of eachlichen increased TAC levels in lymphocytes more thanwater extracts. The TOS levels of the cells treated with different concentrations (1-100mg/L) of the extracts decreased due to the increasing concentration of the extracts. Genotoxicity experiments revealed that the tested lichen extracts did not significantly increase (p>0.05) thelevel of genotoxicity on human peripheral lymphocyte culture compared to the negative control group. The results showed that C.chlorophaea, D.miniatum and P.saxatilis lichens, which were found to be a rich source of phenolic compounds, might be of interest in the pharmaceutical and food industries.

Anti-mutagenic and Anti-oxidant Potencies of Cetraria Aculeata(Schreb.) Fr.,Cladonia Chlorophaea(Flörke ex Sommerf.) Spreng. andCetrelia olivetorum(Nyl.) W.L. Culb. & C.F. Culb.).

In this study, the mutagenic and anti-mutagenic effects of methanol extract of three lichen species (Cetraria aculeata, Cladonia chlorophaea and Cetrelia olivetorum) were investigated by using E. coli-WP2, Ames-Salmonella (TA1535 and TA1537) and sister chromatid exchange (SCE) test systems. The results obtained from bacterial test systems demonstrated that methanol extracts of three lichen species have strong anti-mutagenic potencies on TA1535, TA1537 strains and to a lesser extent on E. coli-WP2 strain. The anti-oxidant level of human lymphocytes cells was determined in order to clarify the mechanism underlying the anti-mutagenic effects of these lichen species. Co-treatments of 5, 10 and 20 µg/mL concentrations of these three lichen species with AFB decreased the frequencies of SCE and the level of MDA and increased the amount of SOD, GSH and GPx which decreased by aflatoxin. The findings of this work have clearly demonstrated that Cetraria aculeata, Cladonia chlorophaea and Cetrelia olivetorum have significant anti-mutagenic effects which are thought to be partly due to the anti-oxidant activities and the interaction capability of lichen extracts with mutagen agents (Sodium azide, acridin, N-methyl-N'-nitro-N-nitrosoguanidine and aflatoxin B1).

Evolution of ketosynthase domains of polyketide synthase genes in the Cladonia chlorophaea species complex (Cladoniaceae)

Lichen-forming fungi synthesize a diversity of polyketides, but only a few non-reducing polyketide synthase (PKS) genes from a lichen-forming fungus have been linked with a specific polyketide. While it is a challenge to link the large number of PKS paralogs in fungi with specific products, it might be expected that the PKS paralogs from closely related species would be similar because of recent evolutionary divergence. The objectives of this study were to reconstruct a PKS gene phylogeny of the Cladonia chlorophaea species complex based on the ketosynthase domain, a species phylogeny of the complex, and to explore the presence of PKS gene paralogs among members of the species complex. DNA was isolated from 51 individuals of C. chlorophaea and allies to screen for the presence of 13 PKS paralogs. A 128 sequence PKS gene phylogeny using deduced amino acid sequences estimated from the 13 PKS paralogs and sequences subjected to BLASTx comparisons showed losses of each of two PKS domains (reducing and methylation). This research provided insight into the evolution of PKS genes in the C. chlorophaea group, species evolution in the group, and it identified potential directions for further investigation of polyketide synthesis in the C. chlorophaea species complex.

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.

Distribution of forest floor lichen communities in the mountainous area of southern Urumqi, Xinjiang, China

Forest floor lichens are sensitive to variations in habitat conditions. To understand the distribu- tional patterns of forest floor lichen communities and how habitat factors relate to dominant lichen species composition and diversity, we sampled 240 plots (50 cm×50 cm) in the mountainous area of southern Urumqi, Xinjiang, China. We detected 17 species in total, belonging to 10 genera and 5 families. Cluster analysis and detrended correspondence analysis (DCA) showed that plots clustered into four groups: (1) a Cladonia pyxidata + Cladonia cariosa + Cladonia chlorophaea community, including seven species with a total coverage of 19.7%; (2) a Solorina bispora + Cladonia cariosa + Cladonia scabriuscula community, in- cluding 11 species with a total coverage of 21.1%; (3) a Cetraria nivalis + Cladonia pyxidata community, in- cluding three species with a total coverage of 8.1%; and (4) a Peltigera rufescens + P. polydactyla commu- nity, including seven species with the total coverage of 23.83%. Canonical correspondence analysis (CCA) indicated that the species composition and diversity of floor lichens were most strongly influenced by altitude, then forest canopy density and humidity; the distribution patterns of lichen communities were apparently influenced by human disturbance. Variation in lichen communities among different elevations

A study of the epiphytic communities of Atlantic oak woods along an atmospheric nitrogen deposition gradient

1 Atlantic oak woods are of high conservation value and contain many rare lichens and bryophytes. The effects of nitrogen pollution on these epiphytic communities have not been previously studied. We investigated the composition of Atlantic oak wood epiphytic communities in relation to atmospheric N deposition in order to identify N indicator species and propose a critical load for such communities. 2 The epiphytic communities of seven Atlantic oak woods receiving estimated total nitrogen deposition in the range 10–53 kg N ha−1 year−1 were surveyed. Exposure of epiphytes to atmospheric N over 7 months was measured in terms of stemflow flux and concentration and airborne NH3. 3 Redundancy analysis (RDA) related the species to stemflow chemistry and bark pH. Different groups of species were found at (i) a coastal low N deposition site (Graphina ruiziana, Lecania cyrtella, Lobaria pulmonaria, Opegrapha atra, Orthotrichum affine, Melanelia fuliginosa ssp. glabratula and Pertusaria hymenea), (ii) inland low N deposition sites (I. myosuroides, F. tamarisci, Plagiochila atlantica, Cladonia chlorophaea, C. squamosa, Hypotrachyna laevigata and Thelotrema lepadinum) and (iii) high N deposition sites (Hypnun andoi, Hypnum cupressiforme, Calicium viride, Chrysothrix candelaris, C. coniocraea, Hypogymnia physodes, Parmelia saxatilis, Phyllospora rosei and Usnea subfloridana). 4 RDA analysis showed that bark pH and concentration in the stemflow explained the greatest amount of variation in the species composition among the sites. The sites separated into two groups (low and high concentration) leading to an estimate for the critical load for N deposition for epiphytes in Atlantic oak woods of 11–18 kg N ha−1 year−1. 5 Analysis of the occurrence of species against N levels suggested that I. myosuroides and F. tamarisci are indicators of small N inputs, whereas H. andoi, C. coniocraea, P. saxatilis and H. physodes are tolerant of increased N loads. 6 Different forms of N (, , NH3) were found to significantly affect the occurrence of different species. Some species were sensitive to total flux while others were sensitive to concentration.

Genetic Variation Within and Among Mats of the Reindeer Lichen, Cladina Subtenuis

Cladina subtenuis is the most commonly found ‘ reindeer lichen ’ in the southeastern United States. In the present study C. subtenuis was examined for polymorphism in the small subunit (SSU) ribosomal DNA (rDNA) within and among small clumps of podetia, here referred to as mats, from five geographical locations. Polymorphism in the SSU rDNA was not detected among podetia within mats, but polymorphism was detected among mats from a single location. Therefore, mats of C. subtenuis may represent a single genetic individual, although genetically distinct mats may occur in close proximity. Their proximity couldencourage sexual reproduction between genetically distinct individuals, which would maintain variation in these populations. The SSU rDNA types represented three different lengths and restriction-site patterns, reflecting the presence of sequence insertions of at least 200 nucleotides in the SSU rDNA coding region. Thei nsertions, differing in position, number and sequence, are group I introns similar to those previously reported in the Cladonia chlorophaea complex. The intron variation among the SSU rDNA types indicates that molecular evolution has occurred within this species, perhaps since its divergence from related taxa and dispersal across the southeastern United States.

Small subunit rDNA variation in a population of lichen fungi due to optional group-I introns

A natural population of the lichen-forming ascomycetous fungus, Cladonia chlorophaea, contained individuals with small subunit ribosomal DNA (SSU rDNA) of at least four different size classes and nine restriction-site patterns. The source of these differences was the variable occurrence of 200–400-nucleotide insertions, previously identified as small group-I introns, at five different positions within the SSU coding region. By specific amplification of the sequences flanking these five intron positions with the polymerase chain reaction (PCR), a minimum of nine types of rDNA repeats were defined that differ in number, position, restriction pattern and size of introns. The positions of the introns were verified by sequence analysis. The variable distribution of these introns suggests that they are currently mobile — either by intron insertion, deletion or both — within this species complex.

Variation in the Cladonia chlorophaea Complex. I: Morphological and Chemical Variation in Southern Appalachian Populations

Variation in the Cladonia chlorophaea Complex. I: Morphological and Chemical Variation in Southern Appalachian Populations

Molecular Innovations in Lichen Systematics: The Use of Ribosomal and Intron Nucleotide Sequences in the Cladonia chlorophaea Complex

In the mycobiont of the Cladonia chlorophaea complex, length variation in the ribo- somal DNA is explained, in part, by the variable occurrence of multiple group I introns that may have been mobile during the evolution of this group. These potentially mobile introns can serve as a model for examining the variation detected in the ribosomal DNA from other lichens and, perhaps, as a model for examining variation in other genes, genotypes, and phenotypes. Because group I introns have been found also in the ribosomal DNA of some green algae, lichens may provide a good system to search for evidence of intron transfer between distantly related organisms. Not only can molecular systematics change the way we look at lichens, but lichens may change the way we look at molecular evolution.

Numerous group I introns with variable distributions in the ribosomal DNA of a lichen fungus

The length of the small subunit ribosomal DNA (SSU rDNA) differs significantly among individuals from natural populations of the ascomycetous lichen complex Cladonia chlorophaea. The sequence of the 3′ region of the SSU rDNA from two individuals, chosen to represent the shortest and longest sequences, revealed multiple insertions within a region that otherwise aligned with a 520-nucleotide sequence of the SSU rDNA in Saccharomyces cerevisiae. The high degree of variability in SSU rDNA size can be accounted for by different numbers of insertions; one individual had two group I introns and the second had five introns, two of which were clearly related to introns at identical positions in the other individual. Yet, introns in different positions, whether within an individual or between individuals, were not similar in sequence. The distribution of introns at three of the positions is consistent with either intron loss or acquisition, and clearly indicates the dynamic variability in this region of the nuclear genome. All seven insertions, which ranged in size from 210 to 228 nucleotides, had the conserved sequence and secondary structural elements of group I introns. The variation in distribution and sequence of group I introns within a short highly conserved region of rDNA presents a unique opportunity for examining the molecular evolution and mobility of group I introns within a systematics framework.

Cladonia chlorophaea complex at Dungeness

Cladonia chlorophaea complex at Dungeness Get access B. W. Ferry B. W. Ferry Search for other works by this author on: Oxford Academic Google Scholar Botanical Journal of the Linnean Society, Volume 101, Issue 1, September 1989, Page 174, https://doi.org/10.1111/j.1095-8339.1989.tb00147.x Published: 13 May 2008

Studies on the Cladonia Chlorophaea Complex at Dungeness, England

AbstractThe terricolous lichen flora on the Dungeness shingle, in south-east England, includes an abundance of members of the Cladonia chlorophaea complex. From a detailed examination of morphological and chemical attributes, the ‘species’ present are identified as Cladonia chlorophaea s. str., C. fimbriata, C. humilis, C. merochlorophaea, C. cryptochlorophaea and C. merochlorophaea var. novochlorophaea. Cladonia fimbriata and C. chlorophaea are dominant on the younger, more seaward ridges, whilst C. merochlorophaea becomes important on the older ridges. Cladonia humilis is confined to sandy substrata.

Gene Flow in Lichens

Experimental studies of the evolutionary biology of lichen fungi have been hampered by massive difficulties of in vitro culture and artificial crosses are still not possible. Gene flow in these organisms is demonstrated here for the first time by the analysis of secondary products in the progeny of individuals from natural populations of mixed chemotypes of the Cladonia chlorophaea complex. All of the chemotypes in this study have been interpreted as distinct sibling species. In the Appalachian Mountains of North Carolina, however, the grayi and merochlorophaea chemotypes are found to belong to a single interbreeding populations that is reproductively isolated from the cryptochlorophaea chemotype. In the Coastal Plain, the cryptochlorophaea chemotype hybridizes with the local endemic perlomera chemotype. This study has major impact for species concept in lichens because consideration of neither morphologial tendencies nor biogenetic relationships of the secondary products could have predicted its result.

GENE FLOW IN LICHENS

Experimental studies of the evolutionary biology of lichen fungi have been hampered by massive difficulties of in vitro culture and artificial crosses are still not possible. Gene flow in these organisms is demonstrated here for the first time by the analysis of secondary products in the progeny of individuals from natural populations of mixed chemotypes of the Cladonia chlorophaea complex. All of the chemotypes in this study have been interpreted as distinct sibling species. In the Appalachian Mountains of North Carolina, however, the grayi and merochlorophaea chemotypes are found to belong to a single interbreeding populations that is reproductively isolated from the cryptochlorophaea chemotype. In the Coastal Plain, the cryptochlorophaea chemotype hybridizes with the local endemic perlomera chemotype. This study has major impact for species concept in lichens because consideration of neither morphologial tendencies nor biogenetic relationships of the secondary products could have predicted its result.

Biogenetic Relationships of the Lichen Substances in the Framework of Systematics

Biogenetic relationships of secondary products in lichens can be used for cladistic analyses of evolutionary relationships among taxa. Character states are derived from biogenetic hypotheses for the origin of the steps required to generate the final products. These biogenetic characters are then used instead of the final products themselves to propose possible evolutionary histories. The Cladonia chlorophaea group provides an example of how preliminary biogenetic hypotheses can lead to a cladogram for the 14 chemotypes presently known in the complex. Secondary products of lichens are immensely valuable in constructing taxonomies. Although li- chenologists may disagree upon the interpretation of chemical variation for classification, no one today would undertake a serious systematic study of a chemically variable group without consideration of the chemical characters. In order for these data to have maximum value for systematics, we need a biogenetic theory for the substances. There have been relatively few biosynthetic studies on lichens, but evidence from pathways to similar products in nonlichen fungi and higher plants allows us to con- struct preliminary biogenetic hypotheses. These hy- potheses may shed light on the evolutionary origin of the compounds and, by extension, upon the evo- lutionary relationships among the species producing them. These ideas have indeed already influenced the classification of the lichen products themselves, and some lichen systematists have found such bio- genetically based classifications valuable in com- paring the relationships of taxa. Recent studies on secondary products of higher plants have shown that biogenetic theory can be readily incorporated into cladistic analyses. Two good examples involve flavonoids (Richardson, 1983) and sesquiterpene lactones (Seaman & Funk 1983). The present paper will briefly summarize biogenetic theory for lichen products and apply these ideas to an analysis of chemical variation in the Cladonia chlorophaea complex. Early classification of lichen products (e.g., Zopf 1907; Asahina & Shibata 1954) were constructed

Chemistry and chorology of the Cladonia chlorophaea-pyxidata complex (Lichenes, Cladoniaceae) in Italy

Summary 252 samples of lichens of the Cladonia chlorophaea—pyxidata complex collected in Italy were analyzed by thin layer chromatography. Six chemospecies occur in Italy; they are: Cladonia chlorophaea s.str. (80.9% of the samples), throughout the Peninsula, Cladonia grayi (4.0%) restricted to mountain areas on acid substrate in the North, Cladonia merochlorophaea (5.5%) and Cladonia cryptochlorophaea (2.4%), most frequent in Central Italy, at the western side of the peninsula, Cladonia conoidea (5.5%) most frequent in Central and Southern Italy, in lowlands and within mediterranean vegetation, Cladonia perlomera (1.6%), very rare throughout the country. Five of the six chemospecies are new to Italy; one (C. perlomera) is new to Europe. The higher frequency of C. chlorophaea s.str. is probably due to the fact that this is the only chemospecies within the complex that grows on limestone soils.