title: Molecular phylogeny and ultrastructure of the lichen microalga Asterochloris mediterranea sp. nov. from Mediterranean and Canary Islands ecosystems.
abstract: 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.
authors.affiliation: Universitat de València, Dpto. Botánica, ICBIBE, Facultad Ciencias Biológicas, Dr Moliner 50, 46100 Burjassot, Valencia, Spain.
authors.forename: Patricia
authors.initials: P
authors.lastname: Moya
authors.affiliation: Charles University in Prague, Faculty of Sciences, Department of Botany, Benatska 2, 128 01 Praha 2, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Škaloud
authors.affiliation: Universitat de València, Dpto. Botánica, ICBIBE, Facultad Ciencias Biológicas, Dr Moliner 50, 46100 Burjassot, Valencia, Spain.
authors.forename: Salvador
authors.initials: S
authors.lastname: Chiva
authors.affiliation: Laboratorio de Anatomía Vegetal 'Julio Iranzo', Jardí Botànic de la Universitat de València, Quart 80, 46008 Valencia, Spain.|Dpto. Ecosistemas Agroforestales, Universidad Politécnica de Valencia. Camino de Vera, s/n., 46022 Valencia, Spain.
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: García-Breijo
authors.affiliation: Universitat de València, Dpto. Botánica, ICBIBE, Facultad Ciencias Biológicas, Dr Moliner 50, 46100 Burjassot, Valencia, Spain.|Laboratorio de Anatomía Vegetal 'Julio Iranzo', Jardí Botànic de la Universitat de València, Quart 80, 46008 Valencia, Spain.
authors.forename: José
authors.initials: J
authors.lastname: Reig-Armiñana
authors.affiliation: Charles University in Prague, Faculty of Sciences, Department of Botany, Benatska 2, 128 01 Praha 2, Czech Republic.
authors.forename: Lucie
authors.initials: L
authors.lastname: Vančurová
authors.affiliation: Universitat de València, Dpto. Botánica, ICBIBE, Facultad Ciencias Biológicas, Dr Moliner 50, 46100 Burjassot, Valencia, Spain.
authors.forename: Eva
authors.initials: E
authors.lastname: Barreno
mesh_terms: D001203:Ascomycota; D000460:Chlorophyta / Q000145:classification* / Q000235:genetics / Q000648:ultrastructure; D018744:DNA, Plant / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D014644:Genetic Variation; D008011:Lichens; D019083:Mediterranean Region; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D013030:Spain
chemical_list: D018744:DNA, Plant; D021903:DNA, Ribosomal Spacer