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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.

¹ University of Ljubljana, Biotechnical Faculty, Department of Food Science and Technology, Jamnikarjeva 101, 1000 Ljubljana, Slovenia
² Centraalbureau voor Schimmelcultures, PO Box 85167, 3508 AD Utrecht, The Netherlands

Correspondence
Neza Cadez
neza.cadez{at}bf.uni-lj.si

Two protein-coding genes, actin and translation elongation factor-1 (EF-1), 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 8772^T) 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 2592^T) and H. occidentalis var. citrica var. nov. (type strain CBS 6783^T), which could be distinguished by habitat preference.

The GenBank/EMBL/DDBJ accession numbers for the gene sequences determined in this study are listed in Table 1.

Figures showing saturation plots and phylogenetic analyses of sequences of actin and EF-1 genes and the D1/D2 regions of the LSU rDNA and ITS regions of Hanseniaspora–Kloeckera strains, and tables comparing dataset characteristics and characteristics inferred from the datasets using maximum-parsimony analyses and DNA–DNA relatedness values among strains of Hanseniaspora occidentalis are available as supplementary material in IJSEM Online.