title: Phylogenetic placement of Hanseniaspora-Kloeckera species using multigene sequence analysis with taxonomic implications: descriptions of Hanseniaspora pseudoguilliermondii sp. nov. and Hanseniaspora occidentalis var. citrica var. nov.
abstract: Two protein-coding genes, actin and translation elongation factor-1alpha (EF-1alpha), as well as two ribosomal gene regions, D1/D2 domains of the large subunit and both internal transcribed spacers including the 5.8S gene region, were evaluated regarding their usefulness for reconstruction of phylogenetic relationships in the Hanseniaspora-Kloeckera species group. This included analyses of sequence divergence values, heterogeneity of evolutionary rates and the reliability of the inferred trees. Both protein-coding genes showed greater capacities to resolve at the strain level and between the closely related species of Hanseniaspora-Kloeckera, compared with the ribosomal gene regions. However, to obtain a fully resolved and reliable phylogenetic tree that reflected the biological relationships it was necessary to combine three congruent sequence datasets. The novel species Hanseniaspora pseudoguilliermondii sp. nov. (type strain CBS 8772T) is described as a result of the application of various molecular approaches to delimit species. Furthermore, incongruent gene genealogies of genetically divergent strains of Hanseniaspora occidentalis, as determined by amplified fragment length polymorphism analysis and DNA-DNA reassociation measurements, indicated the presence of two novel varieties, H. occidentalis var. occidentalis (type strain CBS 2592T) and H. occidentalis var. citrica var. nov. (type strain CBS 6783T), which could be distinguished by habitat preference.
authors.affiliation: University of Ljubljana, Biotechnical Faculty, Department of Food Science and Technology, Jamnikarjeva 101, 1000 Ljubljana, Slovenia.
authors.forename: Neza
authors.initials: N
authors.lastname: Cadez
authors.affiliation: University of Ljubljana, Biotechnical Faculty, Department of Food Science and Technology, Jamnikarjeva 101, 1000 Ljubljana, Slovenia.
authors.forename: Peter
authors.initials: P
authors.lastname: Raspor
authors.affiliation: Centraalbureau voor Schimmelcultures, PO Box 85167, 3508 AD Utrecht, The Netherlands.
authors.forename: Maudy Th
authors.initials: MT
authors.lastname: Smith
mesh_terms: D000199:Actins / Q000235:genetics; D016172:DNA Fingerprinting; D004271:DNA, Fungal / Q000737:chemistry / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000737:chemistry / Q000302:isolation & purification; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D020459:Genes, rRNA; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length; D012331:RNA, Fungal / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics; D012340:RNA, Ribosomal, 5.8S / Q000235:genetics; D004718:Saccharomycetales / Q000145:classification* / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D000199:Actins; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer; D005656:Fungal Proteins; D020648:Peptide Elongation Factor 1; D012331:RNA, Fungal; D012335:RNA, Ribosomal; D012340:RNA, Ribosomal, 5.8S; C065096:RNA, ribosomal, 26S