The taxonomy of yeasts found in exudates of certain trees and other natural breeding sites of some species ofDrosophila

Summary A survey was made of the yeasts occurring in slime fluxes ofQuercus kellogii (black oak),Abies concolor (red fir) and in certain mushrooms and dead logs in the mountains of the Yosemite region of California. Most of the 134 isolates identified were found to be non-fermentative or poorly fermentative (fermenting glucose only and usually weakly). The isolates were placed in the following genera:Pichia (34),Debaryomyces (35), imperfect forms ofHansenula (39),Endomyces (3),Saccharomyces (2),Candida (11),Trichosporon (7),Torulopsis (1),Cryptococcus (1) andSporobolomyces (1). Four new species have been described,Pichia silvestris, Pichia quercibus, Pichia carsonii andDebaryomyces fluxorum. P. quercibus andP. carsonii are unusual representatives of the genusPichia in that they form a very primitive pseudomycelium and lack pellicles on malt extract. To accommodate such species an amended diagnosis of the genusPichia has been proposed byPhaff in an accompanying paper.Saccharomyces pastori, of which 12 isolates were obtained, has been transferred to the amended genusPichia asP. pastori (Guilliermond) nov. comb.     

Synonomy of the yeast generaHansenula andPichia demonstrated through comparisons of deoxyribonucleic acid relatedness

The relationship between the generaHansenula andPichia was examined through comparisons of DNA relatedness among phenotypically similar species.Hansenula minuta andPichia lindneri showed 75% DNA base sequence complementarity. In other comparisons,H. nonfermentans was found to share nearly 50% of its DNA sequences with bothH. minuta andP. lindneri. Because of the high degree of relatedness observed, it is proposed that ability to assimilate nitrate, the sole distinction betweenHansenula andPichia, is of insufficient taxonomic value for the reliable separation of either species or genera. Hat-spored species ofHansenula H. et P. Sydow 1919 are being transferred toPichia Hansen 1904. Species ofHansenula andPichia with Saturn-shaped ascospores will be transferred to the genusWilliopsis. The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

The taxonomy of yeasts isolated fromDrosophila in the Yosemite region of California

Summary A survey was made of the yeasts occurring in the intestinal tract of wild species ofDrosophila occurring in the Yosemite Region of California. Two hundred and forty one yeasts, representing 42 species and varieties, were identified. Each isolate was obtained from a different fly. Almost half of the isolates belong toSaccharomyces. The most common species in this genus wereS. montanus (36 isolates),S. veronae (30 isolates),S. cerevisiae var.tetrasporus (22 isolates) andS. drosophilarum (13 isolates). Further species are listed in Table 1.Zygosaccharomyces fermentati Naganishi was shown to be a distinct species and not a synonym ofS. cerevisiae. In order to avoid confusion with another yeast of the same name, it has been proposed to change the nameZ. fermentati toS. montanus Naganishi. Two new species ofSaccharomyces were described,S. wickerhamii andS. kluyveri. S. mangini var.tetrasporus has been renamedS. cerevisiae var.tetrasporus. A non-cellobiose attacking strain ofS. drosophilarum has been designated tentativelyS. drosophilarum var.acellobiosa. A new species of the genusPichia was described asP. xylosa. Saccharomyces pastori andSaccharomyces pini were transferred to the genusPichia on the basis of arguments given in the preceding paper. A new species ofTrichosporon was described asTr. aculeatum on the basis of the presence of characteristic needlelike cells. Common species besides those mentioned inSaccharomyces wereHansenula angusta (19),Kloeckera apiculata (15),Kl. magna (13), andTorulopsis stellata (10). Other genera represented wereHanseniaspora, Cryptococus, Rhodotorula, Candida andOospora. Evidence was obtained that many species of imperfect genera consist of distinctly different physiological types.     

Characteristics delimiting the genusPichia Hansen

It is confirmed that asci with more than four spores are formed by strains ofPichia membranaefaciens, P. pseudopolymorpha andP. terricola. In view of this it is suggested that the diagnosis of the genusPichia should be amended. Tomato extract proved a more favourable medium for pellicle formation than malt extract.     

Genomic studies on killer yeasts belonging to the genusPichia

Twenty-four species belonging to the genusPichia were investigated using restriction fragment length polymorphism (RFLP) and Southern blot hybridization of their genomic DNA.Saccharomyces cerevisiae, Kluyveromyces lactis, Williopsis mrakii andCandida albicans were also included in this study. The RFLP patterns were obtained from digestion of yeast DNA with several restriction endonuclease enzymes, and showed various bands with different mobility; in most isolates, the more deeply stained bands were species-specific. This observation was confirmed by the results obtained from Southern blot hybridization of theEcoRI andXhoI RFLP patterns withP. anomala UCSC 25F DNA, digested with the same enzymes, used as probes. These bands are likely to be ribosomal DNA as shown by hybridization of digested DNA from unrelated yeast species (S. cerevisiae, K. lactis andC. albicans). However, one hybridized band, located at 3.9–4.1 Kb, seems to be peculiar to thePichia species. Our study confirms the usefulness of molecular tools in studying genetic relatedness among yeasts.     

The genusSaccharomyces (Meyen) Reess

Twenty-nine different strains of several fermentation type II species of the genusSaccharomyces, whose main common characteristic was complete fermentation of raffinose, were studied. These can be regarded as the most highly developed species of the genusSaccharomyces. Their common and differential morphological, physiological, biochemical and serological characteristics were studied. In addition to the usual criteria, factors which played a role in the history and formation of these species were taken into account. Large species were compared with typical strains chosen on the basis of statistical typing, e.g.Saccharomyces carlsbergensis Hansen, and small species were compared with type strains, e.g.Saccharomyces logos van Laer et Den. andSaccharomyces uvarum Beijer, as nomenelatural types. It was found that the whole group could be divided into two categories, one grouped roundSaccharomyces carlsbergensis Hansen and the other roundSaccharomyces uvarum Beijer., which overlapped at the place occupied bySaccharomyces logos van Laer et Den. Another group of intermediate strains isolated from grapes came betweenSaccharomyces logos andSaccharomyces uvarum. In future, closer attention will have to be paid to strains which do not give a serological reaction with serum against the typical speciesSaccharomyces carlsbergensis, i. e. to the speciesSaccharomyces monacensis Hansen andSaccharomyces mandshuricus Saito.     

Differential toxinogenesis in the genusPichia detected by an anti-yeast killer toxin monoclonal antibody

The differential toxinogenesis of 25 isolates belonging to species of the potential yeast killer genusPichia that were previously classified in the genusHansenula was comparatively demonstrated by two serologic techniques (indirect immunofluorescence and double immunodiffusion) by using a monoclonal antibody against a yeast killer toxin produced by a selected strain ofPichia anomala (UCSC 25F). The killer phenotypes of thePichia isolates were evaluated by their ability to kill each other. The results, although of insufficient taxonomic value for a reliable separation of either species or genera, attest to the genomic heterogeneity for the killer character in the genusPichia as well as the presumptive dual killer/sensitive identity for each single isolate.     

Pichia robertsii nov. spec. A new haploid homothallic yeast

Summary A new haploid, homothallic yeast isolated from the larval feed ofXylocopa caffra is described. A study of its properties shows that the evolution of its sexual and biochemical characteristics has not taken place at the same rate as in the genusHansenula and the proposed genusDekkeromyces. This led the author to the conclusion that in the yeast domain phylogenetic lines may exist in which a unilateral evolution of biochemical characteristics has taken place. At least thre groups of yeasts exist where such discrepancies exist between the evolution of their sexual and biochemical characteristics. The inclusion of the new species in these groups is discussed. The new species has provisionally been classified in the genusPichia (in sensu Phaff). A Latin diagnosis of the species is given.     

Immunochemical studies on mannans of the genusSaccharomyces

Structural and immunochemical studies were carried out on mannans isolated from various species of the genusSaccharomyces. It has been found that mannans ofSaccharomyces cerevisiae, Saccharomyces bayanus, Saccharomyces chevalieri, Saccharomyces italicus, Saccharomyces logos, Saccharomyces oviformis, Saccharomyces uvarum andSaccharomyces willianus are structurally and antigenically closely related, they gave high cross-reactions with anti-Saccharomyces cerevisiae serum. Mannotetraosa was found to be the immunodominant group of these mannans. Mannans ofSaccharomyces rouxii andSaccharomyces rosei showed different structural features and immunochemical properties.     

Rhodotorula fujisanensis,a new taxonomic combination

The demonstration of carotenoid pigments in the type strain and one additional strain ofTorulopsis fujisanensis Soneda necessitates the transfer of this species to the genusRhodotorula asRhodotorula fujisanensis (Soneda) Johnson et Phaff comb. nov.     

DNA relatedness among saturn-spored yeasts assigned to the generaWilliopsis andPichia

Saturn-spored species assigned to the generaWilliopsis andPichia were compared from extent of nuclear DNA complementarity. Of thePichia spp., four were recognized as distinct taxa:P. dispora, P. saitoi, P. zaruensis andPichia sp. nov. AmongWilliopsis spp., the following were accepted:W. californica, W. mucosa comb. nov.,W. pratensis, W. saturnus var.saturnus, W. saturnus var.mrakii comb. nov.,W. saturnus var.sargentensis comb. nov.,W. saturnus var.subsufficiens comb. nov. andWilliopsis sp. nov. The newPichia andWilliopsis species are described elsewhere. Moderate (36–68%) DNA relatedness was detected between the formerPichia sargentensis and varieties ofW. saturnus again demonstrating that nitrate assimilation is not a reliable criterion for separating yeast species.     

Reevaluation of the taxonomy ofVibrio,beneckea, andPhotobacterium: Abolition of the genusBeneckea

As a result of studies on the evolution of glutamine synthetase and superoxide dismutase, the genusBeneckea has been abolished and its constituent species, along withPhotobacterium fischeri andP. logei, assigned to the genusVibrio. The definitions ofVibrio andPhotobacterium have been modified accordingly.     

Yeast communities of the cactusPilosocereus arrabidae and associated insects in the Sandy Coastal Plains of Southeastern Brazil

The yeast communities from necrotic tissues, decaying flowers and fruits, and from larval feeding sites of the mothSigelgaita sp. in the cactusPilosocereus arrabidae were surveyed in three restinga ecosystems in Southeastern Brazil. Insects associated with these substrates were sampled to verify the vectoring of yeasts. The cactusPilosocereus arrabidae was shown to have four different yeast communities associated with it. Necrotic stems had a diverse yeast community with the prevalent speciesPichia barkeri, Candida sonorensis, Pichia cactophila, Geotrichum sp.,Myxozyma mucilagina andSporopachydermia sp. A, representing about 80% of the total isolates.Pichia sp. A and aCandida domercqii-like species represented more than 90% of the yeast isolates from decaying flowers. Fruits had a heterogeneous yeast community with typical fruit yeasts of the genusKloeckera, basidiomicetous anamorphs of the genusCryptococcus, the black yeastAureobasidium pullulans, Pichia sp. A, aCandida domercqii-like species, and some cactophilic yeasts, especiallyClavispora opuntiae. The feeding site ofSigelgaita sp. larvae hadClavispora opuntiae as the prevalent species. Insect vectors are suggested as one the most important factors influencing the composition of these yeast communities.     

Systematics of the generaDebaryomyces andMetschnikowia: Proton magnetic resonance spectra of their mannans as an aid in classification

The proton magnetic resonance spectra of the mannans of a number ofDebaryomyces andMetschnikowia species (Endomycetales) were determined. The spectra of all of the mannans had several similar characteristics, regardless of the species from which they originated. Pichia vini, a species originally classified asDebaryomyces vini, formed a mannan with a spectrum almost identical with those ofMetschnikowia (Candida) reukaufii andPichia haplophila. Debaryomyces vanrijii, originally placed in the genusPichia, formed a mannan with a spectrum identical with those ofPichia robertsii andCandida (Pichia) guilliermondii mannans.The authors wish to thank Mr. M. Mazurek for the determination of the p.m.r. spectra, and Mr. N. R. Gardner and Mr. R. J. Magus for valuable technical assistance.     

Saccharomyces Marxianus Hansen

III Conclusion and Summary Zygosaccharomyces Marxianus andSaccharomyces macedoniensis belong to the same species. This species is met with in the haplophase (Z. Marxianus) as well as in the diplophase (S. macedoniensis). It was possible to bring this yeast from the haplophase into the diplophase and vice versa. By keeping this yeast during long times on maltagar it showed a tendency to change from the haplophase into the diplophase, but not into the opposite direction.It seems quite possible thatHansen, who did not describe a conjugation in this yeast, had met with the diplophase.It has been once more emphasized — at whichWinge andLaustsen and alsoLindegren andLindegren have pointed —, that the genusZygosaccharomyces is no valid genus.The yeast studied here belongs to the genusSaccharomyces and must be designated with the original name given to it byHansen:Saccharomyces Marxianus.For the sake of completeness it is mentioned here that also an imperfect stage ofS. Marxianus has been describedviz., Candida macedoniensis (A. Castellani) Berkhout (I).Saccharomyces fragrans Beijerinck has to be considered as its synonym.     

Two new species ofPichia isolated from slime fluxes of deciduous trees

Two new homothallic species ofPichia, associated with exudates ofPopulus trichocarpa andSalix sp. have been described.Pichia trehalophila sp.n. was isolated from slime exudates in two widely separatedPopulus trichocarpa trees andPichia salictaria sp.n. was isolated from two exudates ofSalix sp. The latter species was previously present as an unidentified strain ofPichia in the culture collection of the Centraalbureau voor Schimmelcultures and had been isolated from a patient in Germany. The natural habitat ofP. salictaria is considered to be in association with willow trees.     

Phylogenetic relationships among species ofWilliopsis andSaturnospora gen. nov. as determined from partial rRNA sequences

Phylogenetic relationships among those yeast species that form saturn-shaped ascospores and which are assigned to the generaWilliopsis andPichia were estimated from their extent of nucleotide sequence divergence in three regions of ribosomal RNA. ThePichia species (P. dispora, P. saitoi, P. zaruensis andP. sp. nov.) are a closely clustered group only distantly related toWilliopsis, and it is proposed that they be reassigned toSaturnospora gen. nov. The extent of divergence amongWilliopsis species (W. californica, W. mucosa, W. pratensis, W. saturnus andW. sp. nov.) is greater than that previously observed within other ascomycetous yeast genera.     

Cytological and chemical changes in cell walls ofRhodotorula gracilis

The phenotype and genotype of six strains of the genusRhodotorula Harrison and of one strain of the genusCryptococcus Phaff et Fell, with anomalous thickening of cell walls were investigated. The present studies showed that the strains investigated represent different stages of the life cycles of the genusRhodosporidium Banno. The anomalous thickening of the cell walls can be explained by extreme conditions resulting in the formation of surviving forms (teliospores, chlamydospores).