LECANORA COMPALLENS AND L. SINUOSA,TWO NEW OVERLOOKED CORTICOLOUS LICHEN SPECIES FROM WESTERN EUROPE

Lecanora compallens and L. sinuosa, two corticolous lichens, are described as species new to science on the basis of numerous collections. Lecanora compallens is a sorediate species, known only as sterile. It is probably common in Western Europe, but overlooked because it is very similar to L. expallens. Chemically, the new species is identical with L. strobilina, of which it may represent a sorediate counterpart. Lecanora sinuosa is a new species similar to L. chlarotera and so far only known from a limited area in the Netherlands and Germany, although it may be overlooked elsewhere. Lecanora sinuosa is very close to L. hybocarpa, with which it shares thepulicaris -type epihymenium inspersed with fine crystals but differs markedly by its thick thallus and thick and sinuous apothecium margin.     

LECANORA VACILLANSH. MAGN., A REMARKABLE SPECIES AND ITS TAXONOMIC SIGNIFICANCE

The anatomy and chemistry ofLecanora vacillansH. Magn. has been studied. This saxicolous species combines key characters of both theL. subfuscaandL. subcarneagroups and indicates that the representative groups should not be segregated at generic level. More particularlyL. vacillansresembles theL. subfuscagroup in its apothecial anatomy but contains chemical substances typical of theL. subcarneagroup, whereas the morphology is reminiscent of both groups.Lecanora vacillansf.infuscatais reduced to synonymy withL. vacillans.     

LECANORA BARKMANEANAA NEW NITROPHILOUS SOREDIATE CORTICOLOUS LICHEN FROM THE NETHERLANDS

Lecanora barkmaneana, a sorediate corticolous lichen belonging to theLecanora subfuscagroup, is described as a species new to science on the basis of numerous sterile collections. It is known so far as only from the Netherlands, where it often grows on roadside trees. it seems to be spreading rapidly at nutrient-rich sites.     

A molecular phylogeny of the <Emphasis Type="Italic">Lecanora varia</Emphasis> group,including a new species from western North America

The phylogeny of the Lecanora varia group is inferred from ITS sequences using Bayesian and ML phylogenetic analysis methods. According to our data, usnic acid-containing, corticolous Lecanora species do not form a monophyletic group but occur in three major clades together with other groups of Lecanora and Protoparmeliopsis. The new combination Lecanora filamentosa (Stirt.) Elix & Palice is proposed. The new species Lecanora schizochromatica is described from northwestern North America. It produces atranorin as a major substance and is closely related to the morphologically and chemically similar L. filamentosa. The American Biatora pullula Tuck. is synonymised with Lecanora anopta Nyl., which is hereby reported for continental North America for the first time. The phylogenetic relationships between the major clades of Lecanora are still largely unresolved and require more intensive taxon and character sampling.     

Phylogenetic relationships ofSphaerophoraceae (Ascomycetes) inferred from SSU rDNA sequences

SSU rDNA was sequenced from the lichenized fungiBunodophoron scrobiculatum andLeifidium tenerum (Sphaerophoraceae), andStereocaulon ramulosum andPilophorus acicularis (Stereocaulaceae) and analysed by maximum parsimony with 44 homologous ascomycete sequences in a cladistic study. A small insertion (c. 60 nt.) was found in the sequence ofLeifidium tenerum. Sphaerophoraceae constitutes a strongly supported monophyletic group which groups together withLecanora dispersa and theStereocaulaceae. Together withPorpidia crustulata, this larger group is a sistergroup to thePeltigerineae. This analysis thus supports theLecanorales as monophyletic, includingSphaerophoraceae and thePeltigerineae, but does not provide strong support for this monophyly. The analysis also suggests that the prototunicate ascus in theSphaerophoraceae is a reversion to the plesiomorphic state. Based on morphological, anatomical and chemical reasons,Sphaerophoraceae is proposed to belong to one of the groups presently included in the paraphyletic suborderCladoniineae within theLecanorales.     

PHOTOBIONT INVENTORY OF A LICHEN COMMUNITY GROWING ON HEAVY-METAL-RICH ROCK

Abstract: The photobiont inventory of a stand of the Acarosporetum sinopicae, a lichen community comprising saxicolous, chalcophilous lichens, has been analysed. Investigated lichen species were Acarospora rugulosa, A. sinopica, Bellemerea diamartha, Lecanora polytropa, L. subaurea, Lecidea silacea, L. lapicida, Rhizocarpon geographicum, and Umbilicaria cylindrica. For all these lichen species this is the first record of the photobionts, except for L. lapicida. The photobionts were cultured axenically and investigated using light microscopical and molecular methods (ITS-sequence analyses). Every lichen species contained only one photobiont species. All photobionts belong toTrebouxia jamesii , but two different subspecies were found with the morphological differences corresponding to molecular differences. The new subspecies T. jamesii subsp. angustilobata is described, differing from the typical T. jamesii by a crenulate chloroplast but identical to the latter taxon in respect to the pyrenoid structure in the light microscope. These results are discussed with respect to the photobiont inventory of the Physcietum adscendentis, analysed in an earlier study.     

An ITS-based phylogenetic analysis of theEuryptera species group inLomatium (Apiaceae)

Lomatium, the largest genus of Apiaceae in western North America, includes many narrow endemics whose relationships are uncertain. Although no infrageneric classification exists forLomatium, several informal groups have been recognized. TheEuryptera group comprises seven narrowly endemic species distributed primarily in California. We conducted parsimony and maximum likelihood (ML) analyses using sequences of the internal transcribed spacers (ITS) of the nuclear ribosomal DNA from species of theEuryptera group and several other species ofLomatium. When considered with distribution, morphological, and cpDNA data, the ITS analyses are consistent with the monophyly of theEuryptera group and suggests that speciation in this group has occurred through geographical divergence. Inferences from ITS data also identify putative progenitors of the polyploidEuryptera species.     

The new species Lecanora bicinctoidea, its position and considerations about phenotypic evolution in the Lecanora rupicola group

A phylogenetic analysis of the Lecanora rupicola group based on combined nITS rDNA and beta-tubulin sequences and a combined dataset of ITS, beta-tubulin and partial sequences of polyketide synthase genes reveals a previously unrecognized species, which here is introduced under the name Lecanora bicinctoidea. The new species is a sister group of the L. swartzii complex (including L. swartzii and L. lojkaeana), which is characterized by eucorticate ascomata, and a morphological diversity that includes also a dwarf-fruticose lineage. The preferential occurrence on vertical to overhanging siliceous rocks corresponds more closely to L. swartzii. A detailed investigation of phenotypic characters reveals that the new species differs from the superficially similar morphospecies L. bicincta in several ways, such as a thallus of comparatively small areoles and broadly sessile ascomata and the development of an amphithecial cortex devoid of algal remnants (i.e. an eucortex). L. bicinctoidea contains methyl 3alpha-hydroxy-4-O-demethylbarbatate, a chemical compound not known from other members of the L. rupicola group. We also discuss the importance of eucortex formation as one of several factors that are involved in the evolution of substrate-detached thallus structures.     

PHYLOGENETIC STUDIES OF THE GENUSARTHONIA

A phylogenetic analysis is presented, based on 57 morphological, anatomical, chemical and ecological characters, including 19 species from the generaArthonia,ArthotheliumandSyncesia. The aim of the study was to test the monophyly of some of the groups within the genusArthoniasuggested, for example, by Redinger (e.g. species with reddish ascomata, with grey–pruinose ascomata and with brown–black hypothecia). The results strongly support thatArthoniaandArthotheliumare paraphyletic genera. The best-supported node contains allArthoniaspecies together withA. crozalsianade Lesd.,A. ruanaA. Massal. (both hitherto placed inArthothelium) andSyncesia myrticola(Fée) Tehler. A well-supported clade is formed by a group of pioneer species, often non-lichenized or poorly lichenized, including the type species ofArthonia,A. radiata. The species with reddish and/or K+ reddish ascomata form one clade and the species with more or less brownish or blackish hypothecia form another clade withSyncesia myrticola, the sister group to theOpegraphaceaeandRoccellaceae. The results are discussed and compared with Redinger’s grouping. Relationships to other genera within Arthoniaceae are briefly discussed.Arthothelium scandinavicumTh. Fr.,Arthonia dispersa(Schrad.) Nyl.,A. punctiformisAch. andA. mediellaNyl. are lectotypified.     

Genetic evidence supports the new anatolian lupine accession,Lupinus anatolicus, as an old world “rough-seeded” lupine (sectionScabrispermae) related toL. pilosus

A noteworthy wild lupine accession was recently discovered in southwestern Turkey and was proposed as a new separate Old World “smooth-seeded” species close toL. micranthus and namedL. anatolicus. Its species status was controversial with respect to cytological and crossability data. In order to examine the position and the evolutionary relationships of this Anatolian accession relative to the Old World lupines, we investigated new data from seed coat micromorphology, and from internal transcribed spacer (ITS) nucleotide sequences of the nuclear ribosomal DNA repeat. The micromorphological seed coat pattern ofL. anatolicus, as revealed by scanning electron microscopy, is characterized by pluricellular tubercles, which represent the typical and unique pattern of the Old World “rough-seeded” lupines (sect.Scabrispermae). In accordance with the micromorphological results, the genetic distances and phylogenetic relationships among the Old World lupines, estimated from ITS data, unambiguously support the new Anatolian lupine accession as part of theL. pilosus-L. palaestinus lineage within the strongly monophyletic group containing all theScabrispermae. The results provided in this study, together with other lines of data available from the literature, are thus hardly compatible with the hypothesis that this new Anatolian lupine accession could be related to Old World “smooth-seeded” lupines (includingL. micranthus); instead, it appears closely related toL. pilosus.     

Characterization and Sequence Variation in the rDNA Region of Six Nematode Species of the Genus Longidorus (Nematoda)

Total DNA was isolated from individual nematodes of the species Longidorus helveticus, L. macrosoma, L. arthensis, L. profundorum, L. elongatus, and L. raskii collected in Switzerland. The ITS region and D1-D2 expansion segments of the 26S rDNA were amplified and cloned. The sequences obtained were aligned in order to investigate sequence diversity and to infer the phylogenetic relationships among the six Longidorus species. D1-D2 sequences were more conserved than the ITS sequences that varied widely in primary structure and length, and no consensus was observed. Phylogenetic analyses using the neighbor-joining, maximum parsimony and maximum likelihood methods were performed with three different sequence data sets: ITS1-ITS2, 5.8S-D1-D2, and combining ITS1-ITS2+5.8S-D1-D2 sequences. All multiple alignments yielded similar basic trees supporting the existence of the six species established using morphological characters. These sequence data also provided evidence that the different regions of the rDNA are characterized by different evolution rates and by different factors associated with the generation of extreme size variation.     

Molecular Phylogeny and Barcoding of Caulerpa (Bryopsidales) Based on the tufA,rbcL, 18S rDNA and ITS rDNA Genes

The biodiversity assessment of different taxa of the genus Caulerpa is of interest from the context of morphological plasticity, invasive potential of some species and biotechnological and pharmacological applications. The present study investigated the identification and molecular phylogeny of different species of Caulerpa occurring along the Indian coast inferred from tufA, rbcL, 18S rDNA and ITS rDNA nucleotide sequences. Molecular data confirmed the identification of 10 distinct Caulerpa species: C. veravalensis, C. verticillata, C. racemosa, C. microphysa, C. taxifolia, C. sertularioides, C. scalpelliformis, C. serrulata, C. peltata and C. mexicana. All datasets significantly supported the sister relationship between C. veravalensis and C. racemosa var. cylindracea. It was also concluded from the results that the specimen identified previously as C. microphysa and C. lentillifera could not be considered as separate species. The molecular data revealed the presence of multiple lineages for C. racemosa which can be resolved into separate species. All four markers were used to ascertain their utility for DNA barcoding. The tufA gene proved a better marker with monophyletic association as the main criteria for identification at the species level. The results also support the use of 18S rDNA insertion sequences to delineate the Caulerpa species through character-based barcoding. The ITS rDNA (5.8S-ITS2) phylogenetic analysis also served as another supporting tool. Further, more sequences from additional Caulerpa specimens will need to be analysed in order to support the role of these two markers (ITS rDNA and 18S insertion sequence) in identification of Caulerpa species. The present study revealed the phylogeny of Caulerpa as complete as possible using the currently available data, which is the first comprehensive report illustrating the molecular phylogeny and barcoding of the genus Caulerpa from Indian waters.     

DNA Barcoding Green Microalgae Isolated from Neotropical Inland Waters

This study evaluated the feasibility of using the Ribulose Bisphosphate Carboxylase Large subunit gene (rbcL) and the Internal Transcribed Spacers 1 and 2 of the nuclear rDNA (nuITS1 and nuITS2) markers for identifying a very diverse, albeit poorly known group, of green microalgae from neotropical inland waters. Fifty-one freshwater green microalgae strains isolated from Brazil, the largest biodiversity reservoir in the neotropics, were submitted to DNA barcoding. Currently available universal primers for ITS1-5.8S-ITS2 region amplification were sufficient to successfully amplify and sequence 47 (92%) of the samples. On the other hand, new sets of primers had to be designed for rbcL, which allowed 96% of the samples to be sequenced. Thirty-five percent of the strains could be unambiguously identified to the species level based either on nuITS1 or nuITS2 sequences’ using barcode gap calculations. nuITS2 Compensatory Base Change (CBC) and ITS1-5.8S-ITS2 region phylogenetic analysis, together with morphological inspection, confirmed the identification accuracy. In contrast, only 6% of the strains could be assigned to the correct species based solely on rbcL sequences. In conclusion, the data presented here indicates that either nuITS1 or nuITS2 are useful markers for DNA barcoding of freshwater green microalgae, with advantage for nuITS2 due to the larger availability of analytical tools and reference barcodes deposited at databases for this marker.     

Morphological and molecular study of <Emphasis Type="Italic">Longicollum pagrosomi</Emphasis> Yamaguti, 1935 (Acanthocephala: Pomphorhynchidae) from the barred knifejaw <Emphasis Type="Italic">Oplegnathus fasciatus</Emphasis> (Temminck &amp; Schlegel) (Perciformes: Oplegnathidae) in the East China Sea

Longicollum pagrosomi Yamaguti, 1935 (Acanthocephala: Pomphorhynchidae) collected from the barred knifejaw Oplegnathus fasciatus (Temminck & Schlegel) (Perciformes: Oplegnathidae) in the East China Sea (off Zhoushan Islands) was studied using light and scanning electron microscopy. The SEM observations revealed for the first time the presence of about 28 well-developed sensory papillae arranged in a circle on the copulatory bursa. In addition, L. pagrosomi was characterised using molecular methods by sequencing of the internal transcribed spacer (ITS) of the ribosomal DNA based on the newly collected material. Longicollum pagrosomi is the first species of the genus with the ITS region sequenced for the purpose of species identification. These new morphological and molecular data contributed to a reliable and accurate specific identification and differentiation of species.     

A Middle Llandovery shelly fauna from Motala in Östergötland,south central Sweden. 1.Trilobita

A temporary exposure of Middle Llandovery (Aeronian) shales in the Swedish town of Motala has yielded a rich collection of shelly fauna and graptolites. The shelly fauna is restricted to a thin mudstone band in a graptolitic shale sequence, and is within theconvolutus Biozone. This mudstone band may be correlated with a similar mudstone in the Kolka core from northwestern Latvia. The faunas, including the trilobites, from the two occurrences represent the same species. In the collection from Motala, nine species of trilobites are present, of whichAcernaspis dispersa sp. nov. is new. The material represents the first trilobite fauna described from the Middle Llandovery of Sweden.     

Additional data for a new Theileria sp. from China based on the sequences of ribosomal RNA internal transcribed spacers

Theileria sinensis was recently isolated and named as an independent Theileria species that infects cattle in China. To date, this parasite has been described based on its morphology, transmission and molecular studies, indicating that it should be classified as a distinct species. To test the validity of this taxon, the two internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA gene were cloned and sequenced from three T. sinensis isolates. The complete ITS sequences were compared with those of other Theileria sp. available in GenBank. Phylogenetic analyses based on sequence data for the complete ITS sequences indicate that T. sinensis lies in a distinct clade that is separate from that of T. buffeli/orientalis and T. annulata. Sequence comparisons indicate that different T. sinensis isolates possess unique sizes of ITS1 and ITS2 as well as species-specific nucleotide sequences. This analysis provides new molecular data to support the classification of T. sinensis as a distinct species from other known Theileria spp. based on ITS sequences.     

Phylogenetic Relationships of American Willows (Salix L., Salicaceae)

Salix L. is the largest genus in the family Salicaceae (450 species). Several classifications have been published, but taxonomic subdivision has been under continuous revision. Our goal is to establish the phylogenetic structure of the genus using molecular data on all American willows, using three DNA markers. This complete phylogeny of American willows allows us to propose a biogeographic framework for the evolution of the genus. Material was obtained for the 122 native and introduced willow species of America. Sequences were obtained from the ITS (ribosomal nuclear DNA) and two plastid regions, matK and rbcL. Phylogenetic analyses (parsimony, maximum likelihood, Bayesian inference) were performed on the data. Geographic distribution was mapped onto the tree. The species tree provides strong support for a division of the genus into two subgenera, Salix and Vetrix. Subgenus Salix comprises temperate species from the Americas and Asia, and their disjunction may result from Tertiary events. Subgenus Vetrix is composed of boreo-arctic species of the Northern Hemisphere and their radiation may coincide with the Quaternary glaciations. Sixteen species have ambiguous positions; genetic diversity is lower in subg. Vetrix. A molecular phylogeny of all species of American willows has been inferred. It needs to be tested and further resolved using other molecular data. Nonetheless, the genus clearly has two clades that have distinct biogeographic patterns.     

Systematics of Fraxinus (Oleaceae) and evolution of dioecy

Phylogenetic relationships among 40 of the 43 recognized species of Fraxinus L. (Oleaceae) were estimated on the basis of 106 nuclear ribosomal ITS sequences. ITS trees resulting from maximum likelihood (ML), maximum parsimony (MP) and Bayesian inference (BI) are congruent and identify six distinct lineages. These clades allow establishing sections with high molecular and morphological support. The basal resolution generally has low ML bootstrap and MP jackknife support, but the Bayesian posterior probabilities are high for certain relationships. An independent data set of combined sequences from the chloroplast rps16 and trnL-F regions contains few informative sites but corroborate most of the relationships in the ITS tree. The molecular phylogeny is discussed in the light of morphological and other data and a revised infrageneric classification with six sections are presented. The subgenera and subsections are abandoned and the section Pauciflorae is a new combination. Fraxinus quadrangulata and Fraxinus anomala are united with Fraxinus dipetala in the section Dipetalae and Fraxinus platypoda is transferred to the section Fraxinus. Fraxinus chiisanensis, Fraxinus spaethiana and Fraxinus cuspidata are treated as incertae sedis. A sectional key is given, together with a systematic list of the 43 recognized species, with common synonyms and distribution. Breeding system and other traits mapped on the phylogeny show that dioecy has three separate origins, and in each case followed after the transition from insect to wind pollination. In one instance dioecy evolved from hermaphroditism via androdioecy and twice via polygamy.     

Study on Sequences of Ribosomal DNA Internal Transcribed Spacers of Clams Belonging to the Veneridae Family (Mollusca: Bivalvia)

The first and second internal transcribed spacer (ITS1 and ITS2) regions of the ribosomal DNA from four species, Meretrix meretrix L., Cyclina sinensis G., Mercenaria mercenaria L., and Protothaca jedoensis L., belonging to the family Veneridae were amplified by PCR and sequenced. The size of the ITS1 PCR amplification product ranged from 663 bp to 978 bp, with GC contents ranging from 60.78% to 64.97%. The size of the ITS1 sequence ranged from 585 bp to 900 bp, which is the largest range reported thus far in bivalve species, with GC contents ranging from 61.03% to 65.62%. The size of the ITS2 PCR amplification product ranged from 513 bp to 644 bp, with GC contents ranging from 61.29% to 62.73%. The size of the ITS2 sequence ranged from 281 bp to 412 bp, with GC contents ranging from 65.21% to 67.87%. Extensive sequence variation and obvious length polymorphisms were noted for both regions in these species, and sequence similarity of ITS2 was higher than that of ITS1 across species. The complete sequences of 5.8S ribosomal RNA gene were obtained by assembling ITS1 and ITS2 sequences, and the sequence length in all species was 157 bp. The phylogenetic tree of Veneridae clams was reconstructed using ITS2-containing partial sequences of both 5.8S and 28S ribosomal DNA as markers and the corresponding sequence information in Arctica islandica as the outgroup. Tree topologies indicated that P. jedoensis shared a close relationship with M. mercenaria and C. sinensis, a distant relationship with other species.