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Two yeast species, Papiliotrema leoncinii sp. nov. and Papiliotrema miconiae sp. nov., in the family Rhynchogastremataceae of the Tremellales are proposed. The two species are related to six species of the genus Papiliotrema: Papiliotrema aureus, P. flavescens, P. terrestris, P. baii, P. ruineniae and P. wisconsinensis. The novel species are proposed on the basis of the sequence-based phylogenetic species concept with analysis of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region. A total of 16 strains of Papiliotrema leoncinii sp. nov. were obtained from freshwater and bromeliad leaves collected in Brazil. Papiliotrema leoncinii sp. nov. differs by 11, 12, 16, 14, 11 and 13 substitutions in the D1/D2 domain from the related species P. aureus, P. flavescens, P. terrestris, P. baii, P. ruineniae and P. wisconsinensis, respectively. Differences of 11 substitutions and 21 or more substitutions in ITS regions were found when the sequences of Papiliotrema leoncinii sp. nov. were compared with P. wisconsinensis and its closest relatives. The type strain of Papiliotrema leoncinii sp. nov. is UFMG-CM-Y374T (=CBS 13918T). Papiliotrema miconiae sp. nov. is represented by two strains isolated from a flower of Miconia sp. and a water sample in Brazil. Papiliotrema miconiae sp. nov. differs from the related species P. aureus and P. ruineniae by eight substitutions, from P. flavescens and P. terrestris by 11 substitutions, from P. baii by 10 substitutions and from P. wisconsinensis by 6 substitutions in the D1/D2 domain, and by 7 substitutions from P. wisconsinensis and more than 19 substitutions in the ITS region from its closest relatives. The type strain of Papiliotrema miconiae sp. nov. is CBS 8358T (ML 3666T=DBVPG-4492T). The MycoBank numbers for Papiliotrema leoncinii sp. nov. and Papiliotrema miconiae sp. nov. are MB 813594 and MB 814882, respectively.

Aims: Research into the relationship between pathogens, faecal indicator microbes and environmental factors in beach sand has been limited, yet vital to the understanding of the microbial relationship between sand and the water column and to the improvement of criteria for better human health protection at beaches. The objectives of this study were to evaluate the presence and distribution of pathogens in various zones of beach sand (subtidal, intertidal and supratidal) and to assess their relationship with environmental parameters and indicator microbes at a non-point source subtropical marine beach. Methods and Results: In this exploratory study in subtropical Miami (Florida, USA), beach sand samples were collected and analysed over the course of 6 days for several pathogens, microbial source tracking markers and indicator microbes. An inverse correlation between moisture content and most indicator microbes was found. Significant associations were identified between some indicator microbes and pathogens (such as nematode larvae and yeasts in the genus Candida), which are from classes of microbes that are rarely evaluated in the context of recreational beach use. Conclusions: Results indicate that indicator microbes may predict the presence of some of the pathogens, in particular helminthes, yeasts and the bacterial pathogen Staphylococcus aureus including methicillin-resistant forms. Indicator microbes may thus be useful for monitoring beach sand and water quality at non-point source beaches. Significance and Impact of the Study: The presence of both indicator microbes and pathogens in beach sand provides one possible explanation for human health effects reported at non-point sources beaches.

Three novel species are described as Rhodotorula rosulata sp. nov. (type strain VKM Y-2962T =CBS 10977T), Rhodotorula silvestris sp. nov. (type strain VKM Y-2971T =CBS 11420T) and Rhodotorula straminea sp. nov. (type strain VKM Y-2964T =CBS 10976T) based on the study of eight isolates from needle litter. The new species, phylogenetically located within the Microbotryomycetes, are related to glucuronate-assimilating species of the genus Rhodotorula. Sequencing of the D1/D2 domains of the LSU rDNA gene and the internal transcribed spacer (ITS) region, as well as physiological characterization, revealed their distinct taxonomic positions.

Three species of yeasts are taxonomically described for strains isolated from marine environments. Candida spencermartinsiae sp. nov. (type strain CBS 10894T =NRRL Y-48663T) and Candida taylorii sp. nov. (type strain CBS 8508T =NRRL Y-27213T) are anamorphic ascomycetous yeasts in a phylogenetic cluster of marine yeasts in the Debaryomyces/Lodderomyces clade of the Saccharomycetales. The two species were isolated from multiple locations among coral reefs and mangrove habitats. Pseudozyma abaconensis sp. nov. (type strain CBS 8380T =NRRL Y-17380T) is an anamorphic basidiomycete that is related to the smut fungi of the genus Ustilago in the Ustilaginales. P. abaconensis was collected from waters adjacent to a coral reef.

As an alternative to traditional, morphology-based methods, molecular techniques can provide detection of multiple species within the HAB community and, more widely, the phytoplankton community in a rapid, accurate and simultaneous qualitative analysis. These methods require detailed knowledge of the molecular diversity within taxa in order to design efficient specific primers and specific probes able to avoid cross-reaction with non-target sequences. Isolates from Florida coastal communities were sequence-analyzed and compared with the GenBank database. Almost 44% of the genotypes obtained did not match any sequence in GenBank, showing the existence of a large and still unexplored biodiversity among taxa. Based on these results and on the GenBank database, we designed 14 species-specific probes and 4 sets of specific primers. Multiple simultaneous detection was achieved with a bead array method based on the use of a flow cytometer and color-coded microspheres, which are conjugated to the developed probes. Following a parallel double PCR amplification, which employed universal primers in a singleplex reaction and a set of species-specific primers in multiplex, detection was performed in a cost effective and highly specific analysis. This multi-format assay, which required less than 4 h to complete from sample collection, can be expanded according to need. Up to 100 different species can be identified simultaneously in a single sample, which allows for additional use of this method in community analyses extended to all phytoplankton species. Our initial field trials, which were based on the 14 species-specific probes, showed the co-existence and dominance of two or more species of Karenia during toxic blooms in Florida waters.

Unicellular fungi, commonly referred to as yeasts, were found to be components of the culturable soil fungal population in Taylor Valley, Mt. Discovery, Wright Valley, and two mountain peaks of South Victoria Land, Antarctica. Samples were taken from sites spanning a diversity of soil habitats that were not directly associated with vertebrate activity. A large proportion of yeasts isolated in this study were basidiomycetous species (89%), of which 43% may represent undescribed species, demonstrating that culturable yeasts remain incompletely described in these polar desert soils. Cryptococcus species represented the most often isolated genus (33%) followed by Leucosporidium (22%). Principle component analysis and multiple linear regression using stepwise selection was used to model the relation between abiotic variables (principle component 1 and principle component 2 scores) and yeast biodiversity (the number of species present at a given site). These analyses identified soil pH and electrical conductivity as significant predictors of yeast biodiversity. Species-specific PCR primers were designed to rapidly discriminate among the Dioszegia and Leucosporidium species collected in this study.

Soils in the Dry Valleys of Antarctica are considered to be among the world's most extreme environments. These soils are old, cold and dry with low contents of organic carbon and nitrogen. Habitats adjacent to water (lakes and ice melts) have significant biological activity as demonstrated by the presence of algal mats, lichens and small invertebrates, particularly nematodes, tardigrades and rotifers. In contrast, there are extensive areas in the Dry Valleys that are extremely dry with less than 5% moisture content. These soils are often salty and appear to be barren of life as they have a coarse texture due to their lack of plant organic material. In contrast, molecular techniques (DNA extraction from soils, cloning and rDNA sequence analysis) demonstrated the presence of a complex micro-eukaryotic food web whose structure and composition varied with moisture content and location. Micro-eukaryotic communities in soils with 0.2-1.3% moisture were represented by species of the yeast genus Trichosporon and an unidentified clade of micro-eukaryotes, whereas levels from 3.1% to 4.9% contained complex food webs including primary producers (chlorophytes and stramenopiles), symbionts (lichen associated fungi), saprophytes (fungi), predators (alveolates and cercozoans) and fungal nematode parasite/pathogens. The soils had a diversity of species (80 species from 15 sites) with a restricted number (3-21 species) at each site. The sensitive and measurable community structure of the low moisture Dry Valley soils provides an unparalleled opportunity to examine local and global environmental effects on micro-eukaryotic community dynamics with multiple trophic levels.

Seven phylloplane yeast strains that were collected in the Arrábida Natural Park, Portugal, and identified preliminarily as Bullera alba, the anamorphic stage of Bulleromyces albus, were investigated. In contrast to Bulleromyces albus, these isolates produced a brownish pigment when grown on potato dextrose agar. The pigment caused darkening of the cultures and diffused into the culture medium. Mating studies revealed that the Arrábida isolates did not react with the different mating types of Bulleromyces albus, but were sexually compatible with them and produced mycelium with clamp connections, haustoria and transversally septate basidia that ejected the basidiospores. Various taxonomic criteria that were evaluated during the present study and comparison with other sexual taxa of the Tremellales indicated that this teleomorph should be classified in a novel genus. Therefore, Auriculibuller fuscus gen. nov., sp. nov. (type strain, PYCC 5690T=CBS 9648T) is proposed. In addition, during the course of this investigation, a member of a novel Bullera species, Bullera japonica sp. nov. (type strain, PYCC 4534T=CBS 2013T), was found among collection isolates that were identified formerly as Bullera alba. In molecular phylogenetic analysis of the D1/D2 domains of the 26S rDNA and the internal transcribed spacer region, the two taxa were found to be closely related, but distinct at the species level.

Phylogenetic trees of the anamorphic basidiomycetous yeast genus Trichosporon Behrend, based on molecular sequence analysis of the internal transcribed spacer region and the D1/D2 region of the large subunit of ribosomal (26S) DNA, are presented. This study includes three novel species from soils, Trichosporon vadense sp. nov. (type strain, CBS 8901T), Trichosporon smithiae sp. nov. (type strain, CBS 8370T) and Trichosporon gamsii sp. nov. (type strain, CBS 8245T), one novel species from an insect, Trichosporon scarabaeorum sp. nov. (type strain, CBS 5601T) and one species of unknown origin, Trichosporon dehoogii sp. nov. (type strain, CBS 8686T). The phylogenetic positions and physiological characteristics that distinguish the new taxa from related species, based partly on growth tests that are not traditionally used in yeast taxonomy (uric acid, ethylamine, l-4-hydroxyproline, tyramine and l-phenylalanine as sources of carbon and nitrogen, and polygalacturonate, quinate, 4-ethylphenol, phloroglucinol, 2,3-dihydroxybenzoate and orcinol as sole carbon sources), are discussed. Assimilation of l-rhamnose and erythritol and maximum growth temperature were also used to delineate species.

A morphological and physiological characterization of yeast strains CBS 8908, CBS 8915, CBS 8920, CBS 8925(T) and CBS 8926, isolated from Antarctic soils, was performed. Phylogenetic analyses of the sequences of the D1/D2 regions and the adjacent internal transcribed spacer (ITS) regions of the large-subunit rDNA of these strains placed them into the Tremellales clade of the Hymenomycetes. The sequence data identified strains CBS 8908, CBS 8915 and CBS 8920 as belonging to the species Cryptococcus victoriae. Strains CBS 8925(T) and CBS 8926 were found to represent an unique clade within the Hymenomycetes, with Dioszegia crocea CBS 6714(T) being their closest phylogenetic relative. Fatty acid composition and proteome fingerprint data for these novel strains were also obtained. No sexual state was observed. A novel basidiomycetous species, Cryptococcus statzelliae, is proposed for strains CBS 8925(T) and CBS 8926.

Several independent surveys of yeasts associated with different plant materials and soil led to the proposal of a novel yeast species belonging to the Tremellales clade (Agaricomycotina, Basidiomycota). Analysis of the sequences of the D1/D2 domains and internal transcribed spacer region of the large subunit of the rRNA gene suggested affinity to a phylogenetic lineage that includes Hannaella coprosmaensis, Hannaella oryzae and Hannaella sinensis. Thirty-two isolates were obtained from different sources, including bromeliads, nectar of Heliconia psittacorum (Heliconiaceae), flowers of Pimenta dioica (Myrtaceae), roots and leaves of sugar cane (Saccharum spp.) in Brazil, leaves of Cratoxylum maingayi, Arundinaria pusilla and Vitis vinifera in Thailand, soil samples in Taiwan, and prairie soil in the USA. Sequence analysis of the D1/D2 domains of the large subunit of the rRNA gene showed that the novel species differs from Hannaella coprosmaensis and Hannaella oryzae by 36 and 46 nt substitutions, respectively. A novel species is suggested to accommodate these isolates, for which the name Hannaella pagnoccae sp. nov. is proposed. The type strain is BI118T ( = CBS 11142T = ATCC MYA-4530T).

Several isolates representing the genus Trichosporon were collected over a 6-year period from soils in The Netherlands. Based on classical growth tests with carbon and nitrogen compounds these were identical. Three of these (CBS 8396, CBS 8397 and CBS 8522) were subjected to molecular analysis of the D1/D2 region of the large subunit of rDNA. This confirmed that the three strains were identical, yet distinct from other members of the genus. Conspecificity was demonstrated with the type strain (CBS 2040) of Apiotrichum porosum Stautz (1931), with the exception that A. porosum, which had been isolated from exudate of a yew tree, differed morphologically from the soil strains. Based on the identity of DNA base sequences, morphology was not considered to be an adequate parameter to separate otherwise identical strains into two genera. Therefore, the new combination Trichosporon porosum is presented. Based on molecular sequence analysis, T. porosum may be related to T. sporotrichoides, within a weakly related clade that includes species such as Trichosporon laibachii and Trichosporon loubieri. The strains of T. porosum degrade phenolic compounds and hemicelluloses, which are characteristics with potential ecological importance in soil habitats. Characters distinguishing the nine species of the laibachii/loubieri group of species were listed. These include traditionally used tests as well as assimilation patterns of some aliphatic and phenolic compounds. Based on these tests, species such as Trichosporon multisporum and T. laibachii could be separated.

Cryptococcus adeliensis sp. nov. (CBS 8351) is described based on phenotypic characteristics and molecular sequence analysis of the D1/D2 large subunit and internal transcribed spacer regions of the ribosomal DNA. Molecular comparisons include species closely related to Cryptococcus albidus and several species isolated from the Antarctic. C. adeliensis, which has a cold-adapted xylanase, was isolated from Terre Adelie, Antarctica. ATCC 34633, which has a mesophilic xylanase, was identified as Cryptococcus albidosimilis.

A morphological and physiological description of an alkane-assimilating anamorphic basidiomycetous yeast species, named Trichosporon veenhuisii, is presented. The ability to assimilate several aliphatic and aromatic compounds as sole source of carbon and energy is reported. The phylogenetic position within the genus, based on nuclear base sequencing of the D1/D2 region of the large subunit of rDNA, is discussed. The type strain is CBS 7136T.

Evidence accumulated from studies based on physiological, biochemical, and molecular characteristics has pointed to the heterogeneity of the ubiquitous anamorphic basidiomycetous yeast species Cryptococcus albidus (Saito) Skinner, with its current varieties and synonyms. The taxonomic status of this species has not been reappraised because different studies, mostly involving limited numbers of strains, have not been integrated. To assess species diversity within the clade containing Cryptococcus albidus and other phylogenetically related Cryptococcus and Filobasidium species, we determined ribosomal DNA (rDNA) sequences of 69 strains from the 5prime end of the 26S gene, D1/D2 region, and in some cases, the non-coding ITS2 region. Analysis of the sequence data together with available physiological, biochemical, and molecular characteristics, showed the segregation of C. albidus into at least 12 species, leading to the elevation of former varieties to the rank of species (C. aerius, C. diffluens), the reins...

A morphological and physiological description of an anamorphic basidiomycetous yeast species, named Trichosporon guehoae (CBS 8521T), is presented. The ability to assimilate several aliphatic and aromatic compounds as sole source of carbon and energy is reported. The phylogenetic position within the genus, based on nuclear base sequencing of the D1/D2 region of the large subunit of rDNA is discussed.Key words: basidiomycetes, taxonomy, Trichosporon guehoae sp.nov., yeasts.

We describe Candida aquaetextoris, a new yeast species isolated from sludge that accumulates at the main wastewater treatment facility which processes discharges from textile factories located in the Prato metropolitan district, northern Tuscany, Italy. This yeast degrades 4-(1-nonyl)phenol, a toxic intermediate originating from the microbial attack of nonylphenol polyethoxylates, which are nonionic surfactants largely used in leather and textile industries. In the investigation we employed conventional and molecular taxonomy techniques to compare the new isolate to strains of physiologically similar species, such as Candida maltosa and Candida tropicalis, as well as strains of quite phenotypically different species, such as Candida haemulonii. The results demonstrate that the yeast which we identified represents a separate taxon. The type strain of C. aquaetextoris is strain Lmar1, which has been deposited in the Industrial Yeast Collection of the Division of Applied Microbiology, Department of Plant Biology, University of Perugia, Perugia, Italy, as strain DBVPG 6732.

Made available in DSpace on 2013-08-28T14:12:28Z (GMT). No. of bitstreams: 1 WOS000309889400009.pdf: 1706691 bytes, checksum: b7c188ac88fe5caf97006c61887b511f (MD5) Made available in DSpace on 2013-09-30T18:47:42Z (GMT). No. of bitstreams: 2 WOS000309889400009.pdf: 1706691 bytes, checksum: b7c188ac88fe5caf97006c61887b511f (MD5) WOS000309889400009.pdf.txt: 53174 bytes, checksum: 68557318676cf691b3300cc209ab8da7 (MD5) Previous issue date: 2012-10-09 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:56:33Z No. of bitstreams: 2 WOS000309889400009.pdf: 1706691 bytes, checksum: b7c188ac88fe5caf97006c61887b511f (MD5) WOS000309889400009.pdf.txt: 53174 bytes, checksum: 68557318676cf691b3300cc209ab8da7 (MD5) Made available in DSpace on 2014-05-20T13:56:33Z (GMT). No. of bitstreams: 2 WOS000309889400009.pdf: 1706691 bytes, checksum: b7c188ac88fe5caf97006c61887b511f (MD5) WOS000309889400009.pdf.txt: 53174 bytes, checksum: 68557318676cf691b3300cc209ab8da7 (MD5) Previous issue date: 2012-10-09 Fundação de Amparo à Pesquisa no Estado do Rio Grande do Sul (FAPERGS) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Brazilian Electricity Regulatory Agency BANDEIRANTE Energy S. A. National Science Foundation Background: Independent surveys across the globe led to the proposal of a new basidiomycetous yeast genus within the Bulleromyces clade of the Tremellales, Bandoniozyma gen. nov., with seven new species.Methodology/Principal Findings: The species were characterized by multiple methods, including the analysis of D1/D2 and ITS nucleotide sequences, and morphological and physiological/biochemical traits. Most species can ferment glucose, which is an unusual trait among basidiomycetous yeasts.Conclusions/Significance: In this study we propose the new yeast genus Bandoniozyma, with seven species Bandoniozyma noutii sp. nov. (type species of genus; CBS 8364(T) = DBVPG 4489(T)), Bandoniozyma aquatica sp. nov. (UFMG-DH4.20(T) = CBS 12527(T) = ATCC MYA-4876(T)), Bandoniozyma complexa sp. nov. (CBS 11570(T) = ATCC MYA-4603(T) = MA28a(T)), Bandoniozyma fermentans sp. nov. (CBS 12399(T) = NU7M71(T) = BCRC 23267(T)), Bandoniozyma glucofermentans sp. nov. (CBS 10381(T) = NRRL Y-48076(T) = ATCC MYA-4760(T) = BG 02-7-15-015A-1-1(T)), Bandoniozyma tunnelae sp. nov. (CBS 8024(T) = DBVPG 7000(T)), and Bandoniozyma visegradensis sp. nov. (CBS 12505(T) = NRRL Y-48783(T) = NCAIM Y.01952(T)). Univ Fed Rio Grande do Sul, Dept Microbiol Imunol & Parasitol, Porto Alegre, RS, Brazil Univ Fed Rio Grande do Sul, Ctr Biotecnol, Porto Alegre, RS, Brazil Cent Bur Schimmelcultures Fungal Biodivers Ctr, Utrecht, Netherlands São Paulo State Univ, Dept Bioquim & Microbiol, Rio Claro, SP, Brazil Univ Estadual Campinas, Ctr Pluridisciplinar Pesquisas Quim Biol & Agr, Div Recursos Microbianos, Campinas, SP, Brazil Univ Fed Minas Gerais, Dept Microbiol, Belo Horizonte, MG, Brazil Univ Fed Tocantins, Lab Microbiol Ambiental & Biotecnol, Palmas, TO, Brazil Univ Fed Rio de Janeiro, Inst Microbiol Prof Paulo de Goes, Rio de Janeiro, RJ, Brazil Natl Hsinchu Univ Educ, Dept Appl Sci, Hsinchu, Taiwan Amer Type Culture Collect, Mycol & Bot Program, Manassas, VA USA Corvinus Univ Budapest, Fac Food Sci, Budapest, Hungary Univ Miami, Rosenstiel Sch Marine & Atmospher Sci, Key Biscayne, FL USA São Paulo State Univ, Dept Bioquim & Microbiol, Rio Claro, SP, Brazil NSF: DEB-0206521 NSF: DEB-0072741 NSF: REU NSF: DEB-0417180 NSF: TAMOP 4.2.1/B-09/1/KMR-2010-0005

Publisher Summary The genus studied in this chapter is Xanthophyllomyces. In the determination of the asexual reproduction it is seen that cells are ellipsoidal, reproduce by enteroblastic budding, and occur singly, in pairs, and occasionally in short chains. Carotenoid pigments are synthesized (mainly astaxanthin and a minor proportion of β-carotene), which gives the colonies an orange to salmon-red color. In the sexual reproduction it is found that slender cylindrical holobasidia arise mainly after conjugation between a cell and its bud. Cell to cell conjugation, although rare, may take place. Basidiospores are produced terminally on minute pegs on an elongate holobasidium. The chapter also discusses physiology/biochemistry and phylogenetic placement of the genus. The type species taken is Xanthophyllomyces dendrorhous. In the systematic discussion of the species growth on yeast morphology agar, growth in glucose–yeast extract–peptone water, slide cultures on corn meal agar, formation of basidiospores, origin of the strains studied, cell carbohydrates, and ecology are determined. The present view that Xanthophyllomyces is limited to one species is due to lack of information on the genetic variability within the genus and the difficulty of examining cross-strain mating studies, due to the primary cell/bud mode of sexual reproduction.

Yeast populations in the Shark River Slough of the Florida Everglades, USA, were examined during a 3-year period (2002–2005) at six locations ranging from fresh water marshes to marine mangroves. Seventy-four described species (33 ascomycetes and 41 basidiomycetes) and an approximately equal number of undescribed species were isolated during the course of the investigation. Serious human pathogens, such as Candida tropicalis, were not observed, which indicates that their presence in coastal waters is due to sources of pollution. Some of the observed species were widespread throughout the fresh water and marine habitats, whereas others appeared to be habitat restricted. Species occurrence ranged from prevalent to rare. Five representative unknown species were selected for formal description. The five species comprise two ascomycetes: Candida sharkiensis sp. nov. (CBS 11368T) and Candida rhizophoriensis sp. nov. (CBS 11402T) (Saccharomycetales, Metschnikowiaceae), and three basidiomycetes: Rhodotorula cladiensis sp. nov. (CBS 10878T) in the Sakaguchia clade (Cystobasidiomycetes), Rhodotorula evergladiensis sp. nov. (CBS 10880T) in the Rhodosporidium toruloides clade (Microbotryomycetes, Sporidiobolales) and Cryptococcus mangaliensis sp. nov. (CBS 10870T) in the Bulleromyces clade (Agaricomycotina, Tremellales).

Electronic cell sorting for isolation and culture of dinoflagellates and other marine eukaryotic phytoplankton was compared to the traditional method of manually picking cells using a micropipette. Trauma to electronically sorted cells was not a limiting factor, as fragile dinoflagellates, such as Karenia brevis (Dinophyceae), survived electronic cell sorting to yield viable cells. The rate of successful isolation of large-scale (> 4 litres) cultures was higher for manual picking than for electronic cell sorting (2% vs 0.5%, respectively). However, manual picking of cells is more labor intensive and time consuming. Most manually isolated cells required repicking, as the cultures were determined not to be unialgal after a single round of isolation; whereas, no cultures obtained in this study from electronic single-cell sorting required resorting. A broad flow cytometric gating logic was employed to enhance species diversity. The percentages of unique genotypes produced by manual picking or electronic cell sorting were similar (57% vs 54%, respectively), and each approach produced a variety of dinoflagellate or raphidophyte genera. Alternatively, a highly restrictive gating logic was successfully used to target K. brevis from a natural bloom sample. Direct electronic single-cell sorting was more successful than utilizing a pre-enrichment sort followed by electronic single-cell sorting. The appropriate recovery medium may enhance the rate of successful isolations. Seventy percent of isolated cells were recovered in a new medium (RE) reported here, which was optimized for axenic dinoflagellate cultures. The greatest limiting factor to the throughput of electronic cell sorting is the need for manual postsort culture maintenance and assessment of the large number of isolated cells. However, when combined with newly developed automated methods for growth screening, electronic single-cell sorting has the potential to accelerate the discovery of new algal strains.

Extracts of fifty-seven newly isolated strains of dinoflagellates and raphidophytes were screened for protein phosphatase (PP2A) inhibition. Five strains, identified by rDNA sequence analysis as Prorocentrum rhathymum, tested positive and the presence of okadaic acid was confirmed in one strain by HPLC-MS/MS and by HPLC with fluorescence detection and HPLC-MS of the okadaic acid ADAM derivative. Quantitation of the ADAM derivative indicated that the concentration of okadaic acid in the culture medium is 0.153 microg/L.

The teleomorphic basidiomycetous yeast Xanthophyllomyces dendrorhous is important as a commercial source of astaxanthin, which is a component of feeds for mariculture. Phaffia rhodozyma is the anamorphic state of Xanthophyllomyces; however, there are conflicting reports in the literature concerning the presence of a sexual cycle in P. rhodozyma. The current study attempted to explain this enigma. Strains were obtained from the Phaff Yeast Culture Collection (University of California, Davis) and other sources in the northern hemisphere. Molecular sequences of three nuclear rDNA regions were examined: the internal transcribed spacers (ITS), intergenic spacer (IGS1) and the D1D2 region at the 5' end of the 26S gene. Different levels of genetic variability were observed in the three regions. The D1D2 differentiated major groups of strains, while an increased variability in the ITS suggested that the ITS region could be employed as an ecological marker. The greatest variability was in the IGS1 region, where strains can be defined by the presence and location of indels. Intragenomic sequence heterogeneity in the ITS and IGS1 regions led to the hypothesis that the type strain of P. rhodozyma (CBS 5905 T , UCD 67-210 T ) was derived as a mating-deficient basidiospore from the parent teleomorphic strain CBS 9090.

Nucleotide sequence analyses of the hymenomycetous yeasts demonstrated that Hyalodendron lignicola should be considered as a member of the genus Trichosporon within the Trichosporonales and that Trichosporon pullulans is associated closely with the Cystofilobasidiales, rather than the Trichosporonales. Accordingly, the following proposals are made: Trichosporon lignicola comb. nov. and Guehomyces gen. nov., to accommodate Guehomyces pullulans comb. nov. in the Cystofilobasidiales.

Three novel mite-associated basidiomycetous species are described in two new anamorph genera as Meira geulakonigii gen. nov., sp. nov. (type CBS 110052(T)=NRRL Y-27483(T)=AS 004(T)), Meira argovae sp. nov. (type CBS 110053(T)=NRRL Y-27482(T)=AS 005(T)) and Acaromyces ingoldii gen. nov., sp. nov. (type CBS 110050(T)=NRRL Y-27484(T)=AS 001(T)). Morphologically, these fungi are similar to the yeast-like fungi classified in the Ustilaginales, such as Pseudozyma species. However, analysis of the D1/D2 domain of the LSU rDNA suggests that they belong to two different lineages within the Exobasidiomycetidae of the Ustilaginomycetes (Basidiomycota). Furthermore, these fungi may be of interest for the biocontrol of mites, as they reduced mite numbers by approximately 80 % after inoculation.

Sporobolomyces ruberrimus Yamasaki and Fujii nom. inval. is established as a distinct species by ribosomal-DNA base composition in the D1, D2 and ITS regions and by morphology. A Latin description is given to validate the name with CBS 7500 as the type strain.

Basidiomycetous yeasts in the Urediniomycetes and Hymenomycetes were examined by sequence analysis in two ribosomal DNA regions: the D1/D2 variable domains at the 5' end of the large subunit rRNA gene (D1/D2) and the internal transcribed spacers (ITS) 1 and 2. Four major lineages were recognized in each class: Microbotryum, Sporidiobolus, Erythrobasidium and Agaricostilbum in the Urediniomycetes; Tremellales, Trichosporonales, Filobasidiales and Cystofilobasidiales in the Hymenomycetes. Bootstrap support for many of the clades within those lineages is weak; however, phylogenetic analysis provides a focal point for in-depth study of biological relationships. Combined sequence analysis of the D1/D2 and ITS regions is recommended for species identification, while species definition requires classical biological information such as life cycles and phenotypic characterization.

The molecular systematics of 337 strains of basidiomycetous yeasts and yeast-like fungi, representing 230 species in 18 anamorphic and 24 teleomorphic genera, was determined by sequence analysis of the D1/D2 region of the large-subunit rDNA. The data were compared with published sequences of other basidiomycetous fungi. The results demonstrated that the yeast species and genera are phylogenetically distributed among the Microbotryum, Sporidiobolus, Agaricostilbum and Erythrobasidium clades of the Urediniomycetes; the Tremellales, Trichosporonales ord. nov., Filobasidiales and Cystofilobasidiales clades of the Hymenomycetes; and the Ustilaginales, Microstromatales and Malasseziales clades of the Ustilaginomycetes. Genera such as Bensingtonia, Cryptococcus, Rhodotorula and Sporobolomyces are polyphyletic, i.e. they occur in two or more clades. In contrast, other genera, e.g. Bullera, Cystofilobasidium, Fellomyces, Filobasidiella, Filobasidium, Kondoa, Kurtzmanomyces, Leucosporidium, Rhodosporidium, Sporidiobolus and Udeniomyces, are monophyletic. The majority of the species can be identified using D1/D2 analyses, although the internal transcribed spacer region is required to distinguish closely related species. The intergenic spacer region is recommended for additional differentiation of species and strains.