A proposal for amendment of the diagnosis of the genusPichia hansen

Summary An amended diagnosis has been given for the genusPichia Hansen to allow inclusion of species characterized by a lack of pellicle formation, by the absence or a strongly reduced pseudomycelium or by a combination of both properties. The following species fit the amended diagnosis ofPichia: P haplophila Shifrine et Phaff,P. carsonii Phaff et Knapp,P. quercibus Phaff et Knapp andP. xylosa Phaff, Miller et Shifrine. Furthermore, it has been proposed to transferSaccharomyces pastori Guilliermond,Saccharomyces pini Holst andDebaryomyces vini Zimmermann to the amended genusPichia, where they fit well rather than occupying and exceptional position in their present genera.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

A taxonomic study of soil yeasts

Summary Eighty-four samples of Minnesota soils were collected in the spring of the year. All samples yielded yeasts, and a total of 180 cultures were isolated of which 117 were studied taxonomically. Twenty-five cultures were black yeasts. Approximately one-third of the isolates were spore forming yeasts and members of the oxidative and film forming generaHansenula, Pichia, andDebaryomyces, of which a new species (D. mrakii) has been described. Not a single culture ofSaccharomyces was isolated. Perhaps later in the season when fruit was abundant members of this genus would have been present.The greatest number of imperfects belonged to the genusCandida (32 cultures), althoughPullularia (25 cultures),Rhodotorula (20 cultures), andTorulopsis (16 cultures) were well represented.The cultures ofPullularia darkened very slowly if at all, in the asbence of excess sugar. Under these circumstances they were very similar toT. pullulans.A single culture ofTrichisporon cerebriformis was isolated. The culture isolated formed both blastospores and arthrospores.I would like to express my appreciation to Dr.C. E. Skinner of Washington State College, for his help and guidance throughout this entire problem. I would also like to thank Dr.E. M. Mark, of the University of California, Berkeley, for the privilege of working under his direction, and in his laboratory during the study.     

Occurrence of keratinophilic chrysosporium corda species in indian soils

Summary Three species of the form genusChrysosporium Corda viz.C. stage ofCtenomyces serratus, C. keratinophilum andC. tropicum have been isolated during a search for keratinophilic fungi in Indian soils. An account of two atypical isolates ofC. tropicum is given in detail.     

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.     

The genusPicris (Asteraceae,Lactuceae) in Tropical Africa

Three species of the genusPicris L. are native in Tropical Africa:P. abyssinica Sch. Bip. (Ethiopia),P. xylopoda Lack, spec. nova (Nigeria, Ethiopia) andP. humilis DC. (Senegal, Mali). There are indications that the two perennial species,P. abyssinica andP. xylopoda, are related to and have evolved from a primitive Central Asiatic stock in a manner parallel to many African species ofCrepis L.P. humilis, on the other hand, is a small annual plant with a high number of derived characters. The introduced species of European origin growing south of the Sahara are briefly mentioned.     

Taxonomic studies of Choanephoraceae in India

Summary A taxonomic study of the members of the family Choanephoraceae was undertaken. Almost all the species of the family have been isolated from various sources in India. Two genera, viz.,Blakeslea andChoanephora have been recognized and a key to the genera has been given. The genusBlakeslea includes only one speciesB. trispora and the genusChoanephora five species. A key to the species has also been given. A new variety ofChoanephora circinans, viz.,C. circinans var.prolifera Mehrotra andMehrotra based on the proliferating vesicular apices of the sporangiophores has been described.     

A taxonomic analysis of seed proteins inPinus spp.(Pinaceae)

Seed storage proteins have proved to be a powerful biochemical marker for taxonomic research, but they have not been extensively employed in forest tree studies. In order to improve the understanding of the taxonomy of the genusPinus, total seed proteins of 12 pine species have been analyzed by means of SDS-PAGE (Sodium dodecylsulphate-polyacrylamide gel electrophoresis). The results showed the presence, in the genusPinus, of two main sub-taxa, corresponding to the subgeneraHaploxylon andDiploxylon. Differences and affinities between Mediterranean pine species were found in agreement with classification ofKlaus (1989).     

Accumulation of sugar alcohols by filamentous fungi

Five hundred isolates of different xerophilic and non-xerophilic fungi belonging to 10 genera and 74 species were screened for alditol (sugar alcohol) accumulation. Ninety-two of the isolates failed to grow on a salt medium, most of the isolates (408) produced alditols; 348,44 and 16 of them produced low, moderate and high levels of alditols, respectively. The high alditol producers belonged to five species ofAspergillus, six species ofEurotium andFennellia flavipes. Glycerol andd-mannitol were the main constituents of alditol pools of the 16 high alditol producers.d-Arabinitol andmeso-erythritol were also formed but at low concentrations by several of the tested isolates.     

Relevance of the ecology ofCitrus yeasts to the diet ofDrosophila

A study of the yeast flora of necrotic oranges and associatedDrosophila yielded a total of 221 isolates composed ofKloeckera apiculata (75),Pichia fermentans (75),Pichia kluyveri (50),Torulopsis stellata (17),Hanseniaspora uvarum (2),P. membranaefaciens (1), andCandida vini (1). The yeast species of all samples of oranges and adultDrosophila were very similar; however, the speciesof Drosophila contained a higher proportion ofP. fermentans and a lower proportion ofK. apiculata than was found in the rotting oranges.P. fermentans was subsequently found more frequently on the surface of the necrotic tissue, where the flies feed, than was found internally. SinceP. fermentans characteristically produces a pellicle and pseudomycelium andK. apiculata does not, it is concluded that the growth characteristics of the yeasts are an important factor determining adultDrosophila diets.     

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.     

Analyse numérique des 50 espèces classées dans le genre Pichia (Ascomycetes). Seconde application d'une méthode d'analyse factorielle

A second application of factor analysis among the 50 species of genusPichia corroborates the already identified three groups and underlines the non-appartenance of round-sporedPichia species to genusPichia sensu stricto. These results are compared with Campbell's numerical analysis.     

Pichia spartinae sp. n. from Louisiana marshland habitats

A new species ofPichia has been described.Pichia spartinae was one of the predominant yeasts isolated from the rhizosphere and tissue of oyster grass,Spartina alterniflora, in the Louisiana marshland. The species occurs in the environment in both the homo- and heterothallic state. This is the first species ofPichia in high densities in an estuarine locality.We wish to express our gratitude to Mr. J. C. Raadschelders for correcting the latin diagnosis.     

Observations on pollination inSarcostemma (Asclepiadaceae)

Field observations on pollination in New World species of the genusSarcostemma R. Br. sensuHolm are reported. Morphological and anatomical comparisons of pollinated flowers ofSarcostemma subg.Ceramanthus Kuntze (New World) andSarcostemma subg.Sarcostemma (Old World) are presented.