Yeast sexes: mating types do not determine the sexes in Metschnikowia species.

Although filamentous Ascomycetes may produce structures that are interpreted as male and female gametangia, ascomycetous yeasts are generally not considered to possess male and female sexes. In haplontic yeasts of the genus Metschnikowia, the sexual cycle begins with the fusion of two morphologically identical cells of complementary mating types. Soon after conjugation, a protuberance emerges from one of the conjugants, eventually maturing into an ascus. The originating cell can be regarded as an ascus mother cell, hence as female. We tested the hypothesis that the sexes, female or male, are determined by the mating types. There were good reasons to hypothesize further that mating type α cells are male. In a conceptually simple experiment, we observed the early stages of the mating reaction of mating types differentially labeled with fluorescent concanavalin A conjugates. Three large-spored Metschnikowia species, M. amazonensis, M. continentalis, and M. matae, were examined. In all three, the sexes were found to be independent of mating type, cautioning that the two terms should not be used interchangeably.

Metschnikowia: half tetrads, a regicide and the fountain of youth.

The purpose of this review is to introduce Metschnikowia to the yeast researcher community and to convince readers that the genus is a worthwhile object of study in developmental biology, genetics, ecology and biotechnology. Metschnikowia sits at the foundation of modern immunology, having been instrumental in the discovery of animal phagocytosis. Some 81 species form a monophyletic group within the Metschnikowiaceae, which also include the smaller genus Clavispora and a few clades of Candida species. The family stands out by the habit of forming, by meiosis, only two ascospores, which in Metschnikowia are needle shaped. In some cases, the spores can reach enormous proportions, exceeding 200 µm in length; in others, ascus formation is preceded by the development of chlamydospores. The adaptive value of such features remains to be elucidated. Extensive genetic studies are lacking, but attempts to apply methods developed for model species have been successful. Some species are found at the plant-insect interface whereas others are pathogens of aquatic animals and have served as model organisms in the exploration of host-parasite theory. Some species are globally distributed and others exhibit extreme endemism. Many species are remarkably easy to recover by sampling their known habitats. M. pulcherrima and close relatives may play an important role in wine quality and produce pulcherrimin, an iron-dipeptide complex that can interfere with the growth of other microorganisms. Some symbiotic species incapable of growth in culture media have been assigned to the genus, but their kinship with the group remains to be demonstrated. Copyright © 2016 John Wiley & Sons, Ltd.

Phylogenetic analysis of the angiosperm-floricolous insect-yeast association: have yeast and angiosperm lineages co-diversified?

Metschnikowia (Saccharomycetales, Metschnikowiaceae/Metschnikowia clade) is an ascomycetous yeast genus whose species are associated mostly with angiosperms and their insect pollinators over all continents. The wide distribution of the genus, its association with angiosperm flowers, and the fact that it includes some of the best-studied yeasts in terms of biogeography and ecology make Metschnikowia an excellent group to investigate a possible co-radiation with angiosperm lineages. We performed phylogenetic analyses implementing Bayesian inference and likelihood methods, using a concatenated matrix (≈2.6 Kbp) of nuclear DNA (ACT1, 1st and 2nd codon positions of EF2, Mcm7, and RPB2) sequences. We included 77 species representing approximately 90% of the species in the family. Bayesian and parsimony methods were used to perform ancestral character reconstructions within Metschnikowia in three key morphological characters. Patterns of evolution of yeast habitats and divergence times were explored in the Metschnikowia clade lineages with the purpose of inferring the time of origin of angiosperm-associated habitats within Metschnikowiaceae. This paper presents the first phylogenetic hypothesis to include nearly all known species in the family. The polyphyletic nature of Clavispora was confirmed and Metschnikowia species (and their anamorphs) were shown to form two groups: one that includes mostly floricolous, insect-associated species distributed in mostly tropical areas (the large-spored Metschnikowia clade and relatives) and another that comprises more heterogeneous species in terms of habitat and geographical distribution. Reconstruction of character evolution suggests that sexual characters (ascospore length, number of ascospores, and ascus formation) evolved multiple times within Metschnikowia. Complex and dynamic habitat transitions seem to have punctuated the course of evolution of the Metschnikowiaceae with repeated and independent origins of angiosperm-associated habitats. The origin of the family is placed in the Late Cretaceous (71.7 Ma) with most extant species arising from the Early Eocene. Therefore, the Metschnikowiaceae likely radiated long after the Mid-Cretaceous radiations of angiosperms and their diversification seems to be driven by repeated radiation on a pre-existing diverse resource.

Wickerhamiella pagnoccae sp. nov. and Candida tocantinsensis sp. nov., two ascomycetous yeasts from flower bracts of Heliconia psittacorum (Heliconiaceae).

Two novel yeast species were isolated from nectar of flower bracts of Heliconia psittacorum (Heliconiaceae) collected in a Cerrado ecosystem in the state of Tocantins, northern Brazil. Wickerhamiella pagnoccae sp. nov., which is closely related to Candida jalapaonensis, is heterothallic and produces one spheroid ascospore per ascus. Candida tocantinsensis sp. nov. belongs to the Metschnikowiaceae clade and its nearest relative is Candida ubatubensis, but the sequence identity (%) in the D1/D2 domains of the rRNA gene is low. The type strain of W. pagnoccae is UFMG-F18C1(T) ( = CBS 12178(T) = NRRL Y-48735(T)) and the type strain of C. tocantinsensis is UFMG-F16D1(T) ( = CBS 12177(T) = NRRL Y-48734(T)).

Wickerhamomyces queroliae sp. nov. and Candida jalapaonensis sp. nov., two yeast species isolated from Cerrado ecosystem in North Brazil.

Two novel yeast species, Wickerhamomyces queroliae sp. nov. and Candida jalapaonensis sp. nov., were isolated, respectively, from larvae of Anastrepha mucronata (Diptera: Tephritidae) collected from ripe fruit of Peritassa campestris ('Bacupari', Hippocrateaceae) and from flowers of Centropogon cornutus (Campanulaceae) in the Cerrado ecosystem of the state of Tocantins, Brazil. Analysis of the D1/D2 large-subunit rRNA gene sequences placed W. queroliae in the Wickerhamomyces clade near Wickerhamomyces ciferri and Candida silvicultrix. Candida jalapaonensis belongs to the Wickerhamiella clade and is related to Candida drosophilae. The type strain of Wickerhamomyces queroliae is UFMG-05-T200.1(T) (=CBS 10936(T)=NRRL Y-48478(T)) and the type strain of Candida jalapaonensis is UFMG-03-T210(T) (=CBS 10935(T)=NRRL Y-48477(T)).

Kurtzmaniella gen. nov. and description of the heterothallic, haplontic yeast species Kurtzmaniella cleridarum sp. nov., the teleomorph of Candida cleridarum.

The teleomorph of Candida cleridarum was discovered through the detection of conjugation between isolates of a large collection from the nitidulid beetles of the genus Carpophilus found in the flowers of various cacti in Arizona, USA. The previous oversight of the sexual cycle of this yeast is attributed to the inequality (ca. 5 : 1) of the two mating types. Extensive conjugation between compatible mating types is observed after overnight incubation on 5 % malt agar, followed after 3-5 days by the formation of mature asci. The hat-shaped ascospores are reminiscent of those seen in Kodamaea species, which are members of the same guild. However, published analyses of D1/D2 large subunit rDNA sequences indicate an affinity with the genus Debaryomyces. As the latter is polyphyletic and morphologically heterogeneous, and in view of the distinct life cycle of the new teleomorph, the new genus Kurtzmaniella is described with a novel species, Kurtzmaniella cleridarum sp. nov. Given the close relatedness of Kurtzmaniella cleridarum sp. nov. to Candida quercitrusa, Candida oleophila and Candida railenensis, for which several natural isolates were available, strains of these species were mixed in pairs under the conditions found favourable for the former. Conjugation was not detected in those species. The type strain of Kurtzmaniella cleridarum sp. nov. is UWOPS 99-101.1(T) (=CBS 8793(T)=NRRL Y-48386(T), h(+)), type of Candida cleridarum. The allotype is UWOPS 07-123.1 (=CBS 10688=NRRL Y-48387, h(-)).

Speciation in the large-spored Metschnikowia clade and establishment of a new species, Metschnikowia borealis comb. nov.

The reproductive boundaries among species in the large-spored Metschnikowia clade were studied by prototrophic recombinant selection, electrophoretic karyotyping, mitochondrial DNA restriction analysis, and DNA sequence analysis. Inviable ascospores arose from crosses between the two varieties of Metschnikowia continentalis, indicating that they should be recognized as separate species. Prototrophic recombinants were recovered from crosses between auxotrophic mutants of Metschnikowia borealis, M. continentalis, Metschnikowia lochheadii, Metschnikowia sp. UWO(PS)00-154.1, and Candida ipomoeae, showing that some genetic exchange is possible in spite of the sterility of the asci formed in interspecific crosses. Metschnikowia hawaiiensis, although capable of ascus formation when its h(-) mating type is crossed with the h(+) mating type of the other species, did not give rise to recombinants. In the other species, some recombinants acquired the ability to form asci directly from single cells. These often contained the chromosomes of both parents, suggesting formation of allodiploid hybrids. Other recombinants behaved as haploids and were similar to one parent except for having inherited the selectable wild-type allele from the other parent. In most, but not all cases, inheritance of the mitochondrial genome was uniparental and correlated with the inheritance of the nuclear chromosome complement. In some cases, what appeared to be a recombinant mitochondrial genome was observed. Phylogenies derived from the sequences of various DNA regions were not congruent, indicating that hybridization may have taken place in nature as the large-spored species diverged from their common ancestor. Further evidence that C. ipomoeae arose from a natural recombination event was obtained, but a pair of Metschnikowia species that might represent derived forms of the parents could not be identified conclusively. C. ipomoeae and most of its closely related Metschnikowia species contained a group-II intron in the mitochondrial small-subunit ribosomal gene. The intron was absent in M. borealis, M. hawaiiensis, and other species in the genus Metschnikowia.

The origin of the cactus-yeast community.

The yeast community found in decaying cactus stems and cladodes is stable in terms of species membership and is similar in composition over space and time. The ecological origins of the three core and four common species in the assemblage were inferred by mapping yeast habitats onto a phylogeny of yeasts reconstructed from rDNA sequences. The members of the community belong to distinct clades and consequently have independent origins. The inferred evolutionary pathways of the taxa originate in either tree-flux or decaying fruit habitats and lead to decaying Opuntia cladode and columnar stem habitats. The reasons for the polyphyletic origins of the cactus-yeast community could be due to unique aspects of cactus chemistry, environmental extremes, vector association and interactions among the members.