Species delimitation in the genus Lipomyces by nuclear genome comparison.

Species delimitation in Lipomyces was attempted by nuclear genome comparison in conjunction with the re-evaluation of 48 physiological characters of 65 strains. High intraspecific (> 75%) and low interspecific (< 28%) similarity values established that L. japonicus, L. lipofer and L. tetrasporus are genetically isolated, and also distinct from L. kononenkoae and L. starkeyi. Ambiguous similarity values were obtained with L. kononenkoae and L. starkeyi. Strains previously assigned to L. kononenkoae constitute two related clusters. While similarity values within each cluster range from 76-99%, representatives of the two clusters reassociate for only 47%. Since these clusters are differentiated by their ecologically relevant maximum growth temperature, L. kononenkoae is subdivided. Strains previously assigned to L. starkeyi resolve into four closely related clusters. While similarity values within each cluster range from 78-100%, representatives of the four clusters reassociate for only 59-69%. Since these four clusters are poorly differentiated, the subdivision of L. starkeyi does not appear possible without recourse to other criteria. Four unassigned strains constitute a further two clusters. Reassociation within these clusters is of the order of 91-100%, while reassociation between them occurs only at 59%. Reassociation of representatives of these clusters with those of the L. kononenkoae and L. starkeyi complexes is around 40% and 31%, respectively. These two clusters consequently appear to be intermediate between L. kononenkoae and L. starkeyi, and will, as such, have to be considered in any delimitation of these two species. A key to the taxa of Lipomyces and related genera of the Lipomycetaceae is given.

Babjevia gen. nov.--a new genus of the Lipomycetaceae.

The species described as Lipomyces anomalus Babjeva & Gorin shows significant genetic and phenotypic divergence from the type species Lipomyces starkeyi Lodder & Kreger-van Rij in terms of rRNA base sequence substitution and ascosporal and septal ultrastructure. The species is consequently reclassified in the new, unispecific genus Babjevia, as Babjevia anomala.

Involvement of mitochondria in the assimilatory metabolism of anaerobic Saccharomyces cerevisiae cultures.

The possible physiological role of mitochondria in anaerobically grown Saccharomyces cerevisiae was investigated via enzyme localization and inhibitor studies. Almost all of the activity of citrate synthase (EC 4.1.3.7) was recovered in the mitochondrial fraction after differential centrifugation of spheroplast lysates. The enzyme exhibited a high degree of latency which was demonstrated by sonication of the mitochondrial fractions. Since citrate synthase is an important enzyme in anabolic reactions, a consequence of this localization is the requirement for transport of metabolites across the mitochondrial membranes. Such transport is likely to require energy which, as a result of anaerobiosis, cannot be supplied by respiration. It was therefore investigated whether ATP translocation into the mitochondria by an ADP/ATP translocase might be involved in anaerobic mitochondrial energy metabolism. It was shown that addition of the ADP/ATP translocase inhibitor bongkrekic acid to anaerobic cultures indeed inhibited growth, although only partially. It is concluded that mitochondria of S. cerevisiae fulfil a vital role in anaerobic sugar metabolism.

Botryozyma nematodophila gen. nov., spec. nov. (Candidaceae).

The new genus Botryozyma with a single species, B. nematodophila is proposed for two isolates from nematodes (Panagrellus zymosiphilus) occurring in grapes with sour-rot. The new genus has typical ascomycetous characteristics and, being unable to produce ascospores, is placed in the family Candidaceae.

Contributions to a revision of the genus Trichosporon.

The genus Trichosporon was revised using characters of morphology, ultrastructure, physiology, ubiquinone systems, mol% G + C of DNA, DNA/DNA reassociations and 26S ribosomal RNA partial sequences. A total of 101 strains was used, including all available type and authentic cultures of previously described taxa. Nineteen taxa could be distinguished, 15 of which having Q-9 coenzyme systems and 4 having Q-10. Sixteen previously described names were reduced to synonymy. One new species was described. The genus is characterized by the presence of arthroconidia. Few species possess further diagnostic morphological characters, such as the presence of appressoria, macroconidia or meristematic conidiation. The septa of two species were found to be non-perforate, while those of the remaining species contained dolipores at variable degrees of differentiation, with or without vesicular or tubular parenthesomes. All species were able to assimilate a large number of carbon compounds; visible CO2 production was absent. The genus was found to be fairly homogeneous on the basis of a phylogenetic analysis of partial 26S rRNA sequences, with the exception of T. pullulans which proved to be unrelated. Most taxa were found to occupy well-defined ecological niches. Within the group of taxa isolated from humans, a distinction could be made between those involved in systemic mycoses and those which mainly caused pubic or non-pubic white piedras, respectively. One species was consistently associated with animals, while others came mainly from soil or water. One species was mesophilic and another psychrophilic.

Septal micropores in Zygozyma and their taxonomic significance.

Septal micropores or plasmadesmal canals have been observed in two species of the lipomycetaceous genus Zygozyma. The presence of these canals is considered as further evidence for the connexion between the Lipomycetaceae and the Dipodascaceae. The genus Zygozyma has been emended.

Bulleromyces genus novum (Tremellales), a teleomorph for Bullera alba, and the occurrence of mating in Bullera variabilis.

Mating is observed in Bullera alba and B. variabilis, resulting in the formation of dikaryotic mycelium with clamps, haustorial branches, and lateral and terminal dikaryotic, clavate, lageniform or subglobose cells. These cells develop in B. alba into tremellaceous phragmobasidia. Karyogamy has been observed in young non-divided basidia. Germination of the phragmobasidia occurred by acropetal chains of yeast cells, ballistospores or hyphae. Septal pores are dolipores with parenthesomes made up of U-shaped vesicles (Tremellales type). For the teleomorph of B. alba a new genus, Bulleromyces, is proposed, with only one species, viz. Bulleromyces albus.

Oxygen requirements of yeasts.

Type species of 75 yeast genera were examined for their ability to grow anaerobically in complex and mineral media. To define anaerobic conditions, we added a redox indicator, resazurin, to the media to determine low redox potentials. All strains tested were capable of fermenting glucose to ethanol in oxygen-limited shake-flask cultures, even those of species generally regarded as nonfermentative. However, only 23% of the yeast species tested grew under anaerobic conditions. A comparative study with a number of selected strains revealed that Saccharomyces cerevisiae stands out as a yeast capable of rapid growth at low redox potentials. Other yeasts, such as Torulaspora delbrueckii and Candida tropicalis, grew poorly mu max, 0.03 and 0.05 h-1, respectively) under anaerobic conditions in mineral medium supplemented with Tween 80 and ergosterol. The latter organisms grew rapidly under oxygen limitation and then displayed a high rate of alcoholic fermentation. It can be concluded that these yeasts have hitherto-unidentified oxygen requirements for growth.

Mixotrophic and Autotrophic Growth of Thiobacillus acidophilus on Glucose and Thiosulfate.

Mixotrophic growth of the facultatively autotrophic acidophile Thiobacillus acidophilus on mixtures of glucose and thiosulfate or tetrathionate was studied in substrate-limited chemostat cultures. Growth yields in mixotrophic cultures were higher than the sum of the heterotrophic and autotrophic growth yields. Pulse experiments with thiosulfate indicated that tetrathionate is an intermediate during thiosulfate oxidation by cell suspensions of T. acidophilus. From mixotrophic growth studies, the energetic value of thiosulfate and tetrathionate redox equivalents was estimated to be 50% of that of redox equivalents derived from glucose oxidation. Ribulose 1,5-bisphosphate carboxylase (RuBPCase) activities in cell extracts and rates of sulfur compound oxidation by cell suspensions increased with increasing thiosulfate/glucose ratios in the influent medium of the mixotrophic cultures. Significant RuBPCase and sulfur compound-oxidizing activities were detected in heterotrophically grown T. acidophilus. Polyhedral inclusion bodies (carboxysomes) could be observed at low frequencies in thin sections of cells grown in heterotrophic, glucose-limited chemostat cultures. Highest RuBPCase activities and carboxysome abundancy were observed in cells from autotrophic, CO(2)-limited chemostat cultures. The maximum growth rate at which thiosulfate was still completely oxidized was increased when glucose was utilized simultaneously. This, together with the fact that even during heterotrophic growth the organism exhibited significant activities of enzymes involved in autotrophic metabolism, indicates that T. acidophilus is well adapted to a mixotrophic lifestyle. In this respect, T. acidophilus may have a competitive advantage over autotrophic acidophiles with respect to the sulfur compound oxidation in environments in which organic compounds are present.

Genome comparison among species of the genus Arthroascus von Arx.

Genome comparison in strains of the genus Arthroascus indicates that two species, A. javanensis (CBS 2555, Type) and A. schoenii (CBS 7223, Type), can be recognized.

Zygozyma suomiensis, sp. nov. (Lipomycetaceae), a new species from Finland.

A new species of the genus Zygozyma, Z. suomiensis, is described, based on the study of a single strain, derived from a bovine skin lesion.

Pichia triangularis sp. nov., the teleomorph of Candida polymorpha Ohara et Nonomura, nom. nud.

The type strain of Candida polymorpha Ohara et Nonomura, nom. nud. was found to produce hat-shaped ascospores. On the basis of its morphology and physiology, it is considered a new species of the genus Pichia and is described as Pichia triangularis sp. nov.

Ultrastructure in septa in Blastobotrys and Sporothrix.

Transmission electron micrographs of septa in Blastobotrys species invariably showed central micropores. Septa of species of Sporothrix, however, exhibited three types of pores: micropores which were central if single, or scattered; central simple pores with Woronin bodies; dolipores. The results confirm the heterogeneity of the genus Sporothrix.

Cryptococcus mollis Kützing, type species of the genus Cryptococcus: investigation of the type material.

The legitimacy of the name Cryptococcus mollis for the type of the genus Cryptococcus is shown. The type material, available in the Rijksherbarium at Leiden University was investigated by transmission electron microscopy. The cell wall of most of the cells showed a structure, similar to that of basidiomycetous yeasts. Bud scars were also found.