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

University of Ljubljana, Biotechnical Faculty, Department of Food Science and Technology, Jamnikarjeva 101, 1000 Ljubljana, Slovenia.
International Journal of Systematic and Evolutionary Microbiology (Impact Factor: 2.51). 06/2006; 56(Pt 5):1157-65. DOI: 10.1099/ijs.0.64052-0
Source: PubMed


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.

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Available from: Maudy Th Smith, Apr 17, 2014
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    • "(5′-GCC GGT GAC GAC GCT CCA AGA GCT G-3′)/CA5r (5′-GTG AAC AAT GGA TGG ACC AGA TTC GTC-3′), CA21 (5′-ATT GAT AAC GGT TCC GGT ATG TG-3′)/CA22r (5′-TCG TCG TAT TCT TGC TTT GAG ATC CAC-3′) and Act1-f (5′-CTC GTG CTG TCT TCC CAT CT-3′)/CA22r as described by Daniel and Meyer (2003) and Cadez et al. (2006) "
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    ABSTRACT: The potential impact of aromatic and pectinolytic yeasts on cocoa flavour was investigated using two defined mixed starter cultures encompassing strains of Pichia kluyveri and Kluyveromyces marxianus for inoculating cocoa beans in small scale tray fermentations. Samples for microbial and metabolite analysis were collected at 12-24hour intervals during 120h of fermentation. Yeast isolates were grouped by (GTG)5-based rep-PCR fingerprinting and identified by sequencing of the D1/D2 region of the 26S rRNA gene and the actin gene. Pulsed Field Gel Electrophoresis (PFGE) was conducted on isolates belonging to the species P. kluyveri and K. marxianus to verify strain level identity with the inoculated strains. Furthermore, Denaturing Gradient Gel Electrophoresis (DGGE) was performed to follow yeast and bacterial dynamics over time including the presence of the bacterial inoculum consisting of Lactobacillus fermentum and Acetobacter pasteurianus. Yeast cell counts peaked after 12h of fermentation with the predominant species being identified as Hanseniaspora opuntiae and Hanseniaspora thailandica. P. kluyveri and K. marxianus were found to compose 9.3% and 13.5% of the yeast population, respectively, after 12h of fermentation whilst PFGE showed that ~88% of all P. kluyveri isolates and 100% of all K. marxianus isolates were identical to the inoculated strains. Despite never being the dominant yeast species at any stage of fermentation, the un-conched chocolates produced from the two inoculated fermentations were judged by sensory analysis to differ in flavour profile compared to the spontaneously fermented control. This could indicate that yeasts have a greater impact on the sensory qualities of cocoa than previously assumed.
    International journal of food microbiology 07/2013; 167(1). DOI:10.1016/j.ijfoodmicro.2013.06.024 · 3.08 Impact Factor
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    • "Although H. meyeri C ˇ adež, Poot, Raspor & M. T. Sm., H. guilliermondii Pijper, H. uvarum (Niehaus) Shehata, Mrak & Phaff ex M. T. Sm., and H. opuntiae C ˇ adež, Poot, Raspor & M. T. Sm. did not exhibit distinct physiological characteristics, they can be identified on the basis of differences in the internally transcribed spacer region (ITS) sequences or by polymerase chain reaction–restriction fragment-length polymorphism (PCR–RFLP) of ITS using two restriction enzymes (Cadez et al. 2003). Hanseniaspora pseudoguilliermondii originally described by Cadez et al. (2006) based on DNA relatedness resulted in DNA–DNA hybridization among Hanseniapora species, but physiological characteristics do not distinguish the two species (Cadez et al. 2003, 2006). "
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    ABSTRACT: In the course of a study on yeast diversity in Japan and Thailand, we isolated two yeast strains with bipolar budding patterns. Physiological and phylogenetic analysis suggested that these two strains were identical to Hanseniaspora pseudoguilliermondii. However, these strains produced hat-shaped ascospores and endospores, the latter of which was an unknown characteristic of the species. Endospores were produced on yeast extract–malt extract (YM) plates, though ascospores were produced on cornmeal agar of H. pseudoguilliermondii cultures. Endospores were formed in a twin-cell structure composed of a mother cell and a daughter cell, which did not separate after budding. Unlike the cell wall of the endospores, that of ascospore was stained with a chitin-specific stain. This was a feature distinguishing endospores and ascospores. Cell morphology of H. pseudoguilliermondii was compared with other species of the genus by observing their type strains. Other Hanseniaspora species did not show endospore formation under the same condition in which H. pseudoguilliermondii did. Therefore, the formation of endospores was considered to be a species-delimiting character of H. pseudoguilliermondii. KeywordsBipolar budding yeast-Cell morphology-Phenotypic characteristics
    Mycoscience 09/2010; 51(5):373-378. DOI:10.1007/s10267-010-0049-4 · 1.42 Impact Factor
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    • "pseudoguillermondii (Cadez et al., 2006). Unfortunately, ITS sequencing has proven unsuitable to discriminate species within this group, hence the unidentified Hanseniaspora isolates were subjected to sequencing of the actin gene (Cadez et al., 2006), which allowed their identification as H. opuntiae. This attribution was also confirmed by ITS restriction analysis with the enzymes HinfI and MboII (Cadez et al., 2003) and DraI (Nisiotou and Nychas, 2007) (data not shown). "
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    ABSTRACT: The composition and population dynamics of the yeast microflora of grape marcs were investigated during a pilot scale fermentation study using two white grape varieties, namely Moscato and Prosecco, from two distinct areas of the Veneto Region. Yeast counts were made at the beginning, after 4 and after 15 days of marc storage under anaerobic conditions. Seventy isolates from each sampling time were identified to species by RAPD-PCR analysis and subsequent ITS region sequencing. A good biodiversity of yeasts occurred in both marcs at the beginning of fermentation, with high presence of Hanseniaspora opuntiae, but without detectable presence of Saccharomyces strains, which instead became the dominant yeast after just 4 days of fermentation, remaining at that level until the end of fermentation. Colonization of Moscato marc by S. cerevisiae resulted better, in relation to its higher sugar content. Characterization of S. cerevisiae isolates by mitochondrial DNA restriction analysis revealed the presence of 66 different strains in the marc from the Moscato grapes, without the occurrence of a clearly dominant strain, while in the marc from the Prosecco grapes only 23 different profiles were scored, with a dominant strain that accounted for 62.7% of the Saccharomyces population after 4 days of fermentation.
    International journal of food microbiology 02/2009; 129(3):221-8. DOI:10.1016/j.ijfoodmicro.2008.11.025 · 3.08 Impact Factor
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