Raquel Pino-Bodas

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

Phenotypic 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.

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.

Phenotypical plasticity and homoplasy complicate species delimitation in the Cladonia gracilis group (Cladoniaceae, Ascomycota)

Species delimitation in the Cladonia gracilis group has long been known to be difficult due to morphological variability of taxa. The present study addresses the circumscription of species within this group, examining a number of specimens of the currently accepted taxa Cladonia coniocraea, C. cornuta subsp. cornuta, C. cornuta subsp. groenlandica, C. ecmocyna subsp. ecmocyna, C. ecmocyna subsp. intermedia, C. gracilis subsp. gracilis, C. gracilis subsp. elongata, C. gracilis subsp. tenerrima, C. gracilis subsp. turbinata, C. gracilis subsp. vulnerata, C. macroceras, C. maxima, and C. ochrochlora using genealogical concordance phylogenetic species recognition. We employed maximum parsimony, maximum likelihood and Bayesian methods of phylogenetic reconstructions based on DNA sequences of ITS rDNA, IGS rDNA, RPB2 and partially EF1-α regions. Our results indicate that the C. gracilis group is monophyletic but that most currently accepted taxa do not form monophyletic groups, with the exception of C. ecmocyna and C. cornuta subsp. cornuta. Different tests suggest that incomplete lineage sorting and sporadic recombination events are responsible for a phylogeny that largely lacks support. Our data also strongly suggest that C. coniocraea, C. cornuta subsp. groenlandica, and C. ochrochlora are conspecific, with C. coniocraea being the oldest available name. The morphological characters in the group are shown to be highly homoplasious, causing, in tandem with phenotypical plasticity of the taxa, the difficulties in delimiting species in the C. gracilis group.

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.

Multilocus approach to species recognition in the Cladonia humilis complex (Cladoniaceae, Ascomycota)

PREMISE OF THE STUDY The members of the Cladonia humilis complex are characterized by a well-developed primary thallus and broadly scyphose podetia. In the present study, this complex was phylogenetically analyzed to test the boundaries between the species and to determine the usefulness of the phenotypic characters to distinguish them. The species C. conista, C. cyathomorpha, C. hammeri, C. humilis, C. kurokawae, C. nashii, C. pulvinella, and C. subconistea were examined. METHODS Four DNA loci were sequenced and analyzed to test the monophyly of the species. For the phylogenetic reconstructions, maximum parsimony, maximum likelihood, and Bayesian methods were employed. The genealogical sorting index was used to quantify the exclusive ancestry of the nonmonophyletic species on the tree. KEY RESULTS The performed phylogenetic analyses showed that the C. humilis complex is not monophyletic. Cladonia nashii is not closely related to the remaining taxa within the complex. Seven monophyletic lineages were identified, most of which comprise specimens belonging to more than one chemotype. Cladonia hammeri and C. pulvinella are conspecific, and this taxon is not present in Europe. CONCLUSIONS This study suggests that morphological characters and secondary metabolites have less taxonomical value than thought in the Cladonia humilis complex. Use of multilocus phylogeny is recommended to delimit species.

Cladonia subturgida and C. iberica (Cladoniaceae) form a single, morphologically and chemically polymorphic species

Cladonia subturgida and C. iberica constitute the whole of a Mediterranean problematic species, which shows great morphological polymorphism. A study was carried out in order to delimit the extant taxa within this group. The variability of the group was studied morphologically, chemically and phylogenetically. The phylogeny was based on three loci (ITS rDNA, rpb2 and mtLSU), using Maximum Parsimony and Bayesian analyses. Six chemotypes were identified, the most common one containing atranorin and protolichesterinic acid. Our results prove that C. subturgida and C. iberica constitute a single, morphologically and chemically polymorphic species. The taxonomic rank of C. turgida var. corsicana was also studied based on analyses of morphological, chemical and ITS rDNA data. The new nomenclatural combination, C. corsicana, is proposed.

The phenotypic features used for distinguishing species within the Cladonia furcata complex are highly homoplasious

The phenotypic features used for distinguishing species within the Cladonia furcata complex are highly homoplasious

Phylogenetic position of the crustose Stereocaulon species

Phylogenetic relationships of Stereocaulon with emphasis on the crustose taxa were studied based on nuclear ribosomal ITS1-5.8S-ITS2 and partial beta-tubulin sequences. The placement of four of the six crustose species currently included in the genus has previously been confirmed based on molecular data. It has, however, remained unresolved whether the crustose growth form is a plesiomorphic or apomorphic feature within Stereocaulon, due to contradictory placements of the crustose species in earlier studies. The aim of this study was to clarify the position of the crustose species by including additional data, especially of S. nivale and S. plicatile, which have not been included in previous analyses. The inclusion of S. plicatile in the genus is of particular interest as it is the only species in the genus with submurifrom to muriform ascospores. Altogether 37 specimens representing 31 species of the ingroup, including all the crustose Stereocaulon species, were incorpo- rated in the analyses. Conventional, as well as direct optimization parsimony, maximum likelihood and Bayesian analyses were performed. The results show that the crustose species do not form a monophyletic entity and that the crustose growth form is a plesiomorphic feature within Stereocaulon. The crustose S. nivale and S. plicatile are nested within the genus and their inclusion in Stereocaulon is thereby confirmed. The nested position of S. plicatile indicates that the submuriform to muriform spore type has been gained independently within the genus. Here, S. plicatile is also reported for the first time from Scandinavia.

Cladonia corymbescens consists of two species

e lichen Cladonia corymbescens (Cladoniaceae, Ascomycota) has been reported to have two main ranges, one in Melanesia and Australasia and another in the Himalayas and surrounding areas. When specimens from New Caledonia, Bhutan, and �怀ailand were subjected to molecular sequencing (ITS rDNA and RPB2), two distinct clades were detected. �怀e clades were also supported by morphological and geographical di�耀erences supporting recognition of two distinct species. Cladonia corymbescens s. str. is present in the Philippines, Melanesia, and Australasia, while the South East Asian mainland material is referred to C. pseudo�耀ssa, a new combination at species level.

New distributional records for Chilean bryophyte and lichen flora

Se reportan 10 nuevos registros de distribucion geografica para la flora brio-liquenica de Chile. Tres especies y una subespecie son nuevas para la provincia de Chiloe, cinco especies son nuevas para la Region de Los Lagos y una es nueva para Chile continental. Los especimenes fueron recolectados en turberas esfagnosas y bosques anegados de Tepualia stipularis de la Isla Grande de Chiloe (Chile) (41°- 43° S 74°-73° W). Se proveen caracteres diagnosticos de las especies, quimica (solo para liquenes), preferencias de habitat y distribucion. Se extiende el rango de distribucion de dos especies.