Global distribution of Trebouxiophyceae diversity explored by high-throughput sequencing and phylogenetic approaches.

Trebouxiophyceae are a ubiquitous class of Chlorophyta encountered in aquatic and terrestrial environments. Most taxa are photosynthetic, and many acts as photobionts in symbiotic relationships, while others are free-living. Trebouxiophyceae have also been widely investigated for their use for biotechnological applications. In this work, we aimed at obtaining a comprehensive image of their diversity by compiling the information of 435 freshwater, soil and marine environmental DNA samples surveyed with Illumina sequencing technology in order to search for the most relevant environments for bioprospecting. Freshwater and soil were most diverse and shared more than half of all operational taxonomic units (OTUs), however, their communities were significantly distinct. Oceans hosted the highest genetic novelty, and did not share any OTUs with the other environments; also, marine samples host more diversity in warm waters. Symbiotic genera usually found in lichens such as Trebouxia, Myrmecia and Symbiochloris were also abundantly detected in the ocean, suggesting either free-living lifestyles or unknown symbiotic relationships with marine planktonic organisms. Altogether, our study opens the way to new prospection for trebouxiophycean strains, especially in understudied environments like the ocean.

From one to six: unrecognized species diversity in the genus Acantholichen (lichenized Basidiomycota: Hygrophoraceae).

We present a taxonomic revision of the lichenized basidiomycete genus Acantholichen, species of which produce a characteristic blue-gray, microsquamulose thallus with spiny apical hyphal cells known as acanthohyphidia. Since its discovery, the genus was thought to be monospecific, only including the generic type, A. pannarioides. However, a detailed morphological and anatomical study of recently collected specimens from the Galápagos, Costa Rica, Brazil and Colombia, combined with a molecular phylogenetic analysis of the internal transcribed spacer (ITS1-5.8S-ITS2) region and 28S of the nuc rDNA and RPB2 sequences, revealed a much more diverse and widespread species assemblage. Based on the results of these analyses, we describe five new species in the genus: A. albomarginatus, A. campestris, A. galapagoensis, A. sorediatus and A. variabilis. We also provide an identification key to all species, anatomical and morphological descriptions, photographs and a table comparing main characters of each species.

A new species of Phyllopsora (Lecanorales, lichen-forming Ascomycota) from Dominican amber, with remarks on the fossil history of lichens.

Phyllopsora dominicanus sp. nov. (Bacidiaceae, Lecanorales, lichen-forming Ascomycota) is described and illustrated from Dominican amber. The diagnostic features of the lichen include a minute subfolious thallus of lacinulate, ascending squamules, a well-developed upper cortex, and a net-like pseudocortex on the lower surface. The algal symbionts are unicellular green algae, forming a distinct layer immediately below the upper cortex. The fossil demonstrates that distinguishing features of Phyllopsora have remained unchanged for tens of millions of years. The fossil also provides the first detailed views of mycobiont-photobiont contacts in Tertiary green algal lichens. The mycobiont hyphae formed apical and intercalary appressoria by pressing closely against the photobiont cells. This indicates that a conserved maintenance of structure is also seen in the fine details of the fungal-algal interface.

Life in extreme environments: survival strategy of the endolithic desert lichen Verrucaria rubrocincta.

Verrucaria rubrocincta Breuss is an endolithic lichen that inhabits caliche plates exposed on the surface of the Sonoran Desert. Caliche surface temperatures are regularly in excess of 60 degrees C during the summer and approach 0 degrees C in the winter. Incident light intensities are high, with photosynthetically active radiation levels typically to 2,600 micromol/m(2) s(-1) during the summer. A cross-section of rock inhabited by V. rubrocincta shows an anatomical zonation comprising an upper micrite layer, a photobiont layer containing clusters of algal cells, and a pseudomedulla embedded in the caliche. Hyphae of the pseudomedulla become less numerous with depth below the rock surface. Stable carbon and oxygen isotopic data for the caliche and micrite fall into two sloping, well-separated arrays on a delta(13)C-delta(18)O plot. The delta(13)C(PDB) of the micrite ranges from 2.1 to 8.1 and delta(18)O(SMOW) from 25.4 to 28.9, whereas delta(13)C(PDB) of the caliche ranges from -4.7 to 0.7 and delta(18)O(SMOW) from 23.7 to 29.2. The isotopic data of the micrite can be explained by preferential fixing of (12)C into the alga, leaving local (13)C enrichment and evaporative enrichment of (18)O in the water. The (14)C dates of the micrite range from recent to 884 years b.p., indicating that "dead" carbon from the caliche is not a significant source for the lichen-precipitated micrite. The endolithic growth is an adaptation to the environmental extremes of exposed rock surfaces in the hot desert. The micrite layer is highly reflective and reduces light intensity to the algae below and acts as an efficient sunscreen that blocks harmful UV radiation. The micrite also acts as a cap to the lichen and helps trap moisture. The lichen survives by the combined effects of biodeterioration and biomineralization. Biodeterioration of the caliche concomitant with biomineralization of a protective surface coating of micrite results in the distinctive anatomy of V. rubrocincta.

Polysaccharides present in cultivated Teloschistes flavicans symbiosis: comparison with those of the thallus.

The chemical structures of polysaccharides present in aposymbiotically cultured myco- and photobionts of the lichen Teloschistes flavicans were determined, in order to compare them with those previously found in the intact thallus. The mycobiont was cultured on a solid Lilly and Barnett medium and the resulting colonies were freeze dried, defatted, and their polysaccharides were extracted successively with 2%, 10% and 30% aq. KOH, each at 100 degrees C. The extracts were neutralized (HOAc) and fractionated, giving rise to three homogeneous fractions, PFSK2 from 2% KOH, which contained a (1-->4),(1-->6)-linked alpha-glucan (1:1 ratio, pullulan), fraction PK10 from 10% KOH extraction, which was a linear (1-->3)-linked linear beta-glucan (laminaran), and fraction PK30 from 30% KOH extraction, being a branched (1-->3),(1-->6)-linked beta-glucan. The photobiont (Trebouxia sp. de Puymaly) was cultured in liquid nutrient medium, and after purification, a linear (1-->5)-linked beta-galactofuranan was characterized. The galactofuranan and the laminaran were not present in the symbiotic thallus, in contrast to the glucans, showing that the mycobiont alone produces them without participation of the photobiont.

Pseudocyphellaria crocata, P. neglecta and P. perpetua from the Northern and Southern Hemispheres are a phylogenetic species and share cyanobionts.

Pseudocyphellaria crocata, P. neglecta and P. perpetua specimens were examined to investigate links between genetic variation and morphology, geographical distribution and cyanobiont specificity. Fungal internal transcribed spacer (ITS), beta-tubulin and cyanobacterial tRNA(Leu) (UAA) intron sequences were used to investigate symbiont diversity in these lichens. Specimens were morphologically distinct but could not be distinguished by ITS sequences. Phylogenetic analyses split the P. crocata specimens into two clades, the larger of which contained P. neglecta and P. perpetua. Five cyanobionts were identified; two of these were in a number of specimens, while three were each restricted to a single lichen thallus. Fungus-specific molecular markers indicated that all specimens belonged to a single phylogenetic species. However, this may contain a cryptic species. Geography was linked to genetic diversity with Canadian specimens forming a monophyletic group, and most Southern Hemisphere specimens grouping together, although Chile represented a hot spot of genetic diversity. There was no connection between fungal genetic diversity and cyanobiont choice, consistent with the presence of a common pool of cyanobionts.

Molecular studies of photobionts of selected lichens from the coastal vegetation of Brazil.

A light microscopic and molecular analysis of photobionts in Ramalina and Cladonia from coastal habitats of Brazil is presented. A Bayesian phylogenetic analysis of ITS rDNA sequences suggests a Trebouxia lineage which is preferentially tropical in geographic distribution. This highly diverse clade also includes the morphological similar species Trebouxia higginsiae and galapagensis. Within the predominantly tropical clade of Trebouxia we distinguish several subclades, three of which are represented in our samples of Ramalina species. Since sexuality has not been recognized in coccal lichenised photobionts until recently, we cannot apply a biological species concept, but when compared with the sequence diversity between known species we conclude that several new species need to be described in this clade. The mutually exclusive presence of other Trebouxia lineages in temperate samples of Ramalina suggests an evolution towards higher selectivity in this genus. A strictly tropical lineage is not conspicuous in the photobionts of the genus Asterochloris sampled from Cladonia so far.

The foliicolous lichen flora of Mexico IV: a new, foliicolous species of Pyrenothrix (Chaetothyriales: Pyrenothrichaceae).

Pyrenothrix mexicana Herrera-Campos, Huhndorf & Lücking spec. nova is described from leaves in the upper montane rainforest of Oaxaca State, Mexico. It is the second species in the genus Pyrenothrix Riddle, established at the beginning of the twentieth century for the single species, P. nigra Riddle, a corticolous lichen restricted to southeastern United States. Both taxa have the same thallus and perithecial morphology and anatomy, but P. mexicana differs by its longer, transversally septate ascospores. The perithecial anatomy of Pyrenothrix is documented and its systematic affinities are discussed, and we conclude that the family Pyrenothrichaceae Zahlbr. should be placed in the order Chaetothyriales.

Niebla ceruchis from Laguna Figueroa: dimorphic spore morphology and secondary compounds localized in pycnidia and apothecia.

During and after the floods of 1979-80 Niebla ceruchis growing epiphytically on Lycium brevipes was one of the dominant aspects of the vegetation in the coastal dunal complex bordering the microbial mats at Laguna Figueroa, Baja California Norte, Mexico. The lichen on denuded branches of Lycium was far more extensively distributed than Lycium lacking lichen. Unusual traits of this Niebla ceruchis strain, namely localization of lichen compounds in the mycobiont reproductive structures (pycnidia and apothecia) and simultaneous presence of bilocular and quadrilocular ascospores, are reported. The abundance of this coastal lichen cover at the microbial mat site has persisted through April 1988.