Species Of Sporobolomyces Research Articles

High-Quality Genome Assembly of Oleaginous Red Yeast Sporobolomyces roseus CGMCC 2.4355.

Sporobolomyces roseus is an important oleaginous red yeast with critical biotechnological applications and has received significant recognition as a valuable source of industrial enzymes, carotenoids, and lipids. To reveal the genetic basis and functional components underlying its biotechnological applications, a high-quality genome assembly is required. Here, we present a novel genome assembly of S. roseus CGMCC 2.4355 using a combination of Illunima and Oxford Nanopore technologies. The genome has an assembly size of 21.4 Mb and consists of 15 scaffolds with an N50 length of 2,126,566 bp and GC content of 49.52%. The assembly is of high integrity, comprising 95.2% complete Benchmarking Universal Single-Copy Orthologs (BUSCOs) as evaluated by a genome completeness assessment. The genome was predicted to contain 8,124 protein-coding genes, 6,890 of which were functionally annotated. We believe that the combination of our analyses and high-quality genome assembly will promote the basic development of S. roseus as an agent for biotechnological applications and make a significant contribution to assess the evolutionary relationship of Sporobolomyces species.

Effects of Some Fungicides and Foliar Fertilizers on Epiphytic Fungal and Yeast Population of Citrus Leaves

In this study, the effects of some commonly used fungicides (Copper hydroxide, Diphenoconazole+Propiconazole, Iprodione, Mancozeb, Metiram and Propineb) and foliar fertilizers (Zinc, Phosphorus, Magnesium, Manganese, Potassium and Urea) were examined on phyllosphere microorganisms population on citrus leaves. As a results of the isolations from the leaves, mostly Cryptococcus spp. and Sporobolomyces spp. And Cladosporium spp. and Aureobasidium spp. were obtained in less rates. Mancozeb and Copper hydroxide had the greatest impact on mycoflora. During the period of these fungicide applications, fungal population decreased by approximately 50-fold. Effects of these fungicides continued up to 6 months after application. Diphenoconazole+Propiconazole and Iprodione had no significant effect on the fungal population. Effect of Metiram and Propineb fungicides disappeared after 2 months of applications. Although foliar fertilizers seemed to have a negative impact on fungal populations, repeated applications increased the mycoflora. Especially foliar fertilizers such as zinc and phosphorus remarkably increased Sporobolomyces species.

Phylogenetic classification of yeasts and related taxa within Pucciniomycotina.

Most small genera containing yeast species in the Pucciniomycotina (Basidiomycota, Fungi) are monophyletic, whereas larger genera including Bensingtonia, Rhodosporidium, Rhodotorula, Sporidiobolus and Sporobolomyces are polyphyletic. With the implementation of the “One Fungus = One Name” nomenclatural principle these polyphyletic genera were revised. Nine genera, namely Bannoa, Cystobasidiopsis, Colacogloea, Kondoa, Erythrobasidium, Rhodotorula, Sporobolomyces, Sakaguchia and Sterigmatomyces, were emended to include anamorphic and teleomorphic species based on the results obtained by a multi-gene phylogenetic analysis, phylogenetic network analyses, branch length-based methods, as well as morphological, physiological and biochemical comparisons. A new class Spiculogloeomycetes is proposed to accommodate the order Spiculogloeales. The new families Buckleyzymaceae with Buckleyzyma gen. nov., Chrysozymaceae with Chrysozyma gen. nov., Microsporomycetaceae with Microsporomyces gen. nov., Ruineniaceae with Ruinenia gen. nov., Symmetrosporaceae with Symmetrospora gen. nov., Colacogloeaceae and Sakaguchiaceae are proposed. The new genera Bannozyma, Buckleyzyma, Fellozyma, Hamamotoa, Hasegawazyma, Jianyunia, Rhodosporidiobolus, Oberwinklerozyma, Phenoliferia, Pseudobensingtonia, Pseudohyphozyma, Sampaiozyma, Slooffia, Spencerozyma, Trigonosporomyces, Udeniozyma, Vonarxula, Yamadamyces and Yunzhangia are proposed to accommodate species segregated from the genera Bensingtonia, Rhodosporidium, Rhodotorula, Sporidiobolus and Sporobolomyces. Ballistosporomyces is emended and reintroduced to include three Sporobolomyces species of the sasicola clade. A total of 111 new combinations are proposed in this study.

Phs1 and the synthesis of very long chain Fatty acids are required for ballistospore formation.

The production and dissemination of spores by members of the fungal kingdom is a major reason for the success of this eukaryotic lineage in colonizing most terrestrial ecosystems. Ballistospores are a type of spore produced by basidiomycete fungi, such as the mushrooms and plant pathogenic rusts. These spores are forcefully discharged through a unique liquid-drop fusion mechanism, enabling the aerosolization of these particles that can contribute to plant disease and human allergies. The genes responsible for this process are unknown due to technical challenges in studying many of the fungi that produce ballistospores. Here, we applied newly-developed techniques in a forward genetic screen to identify genes required for ballistospore formation or function in a tractable red yeast, a species of Sporobolomyces. One strain bearing a mutation in the PHS1 gene was identified as a mirror mutant. PHS1 encodes 3-hydroxyacyl-CoA dehydratase required for the third step in very long chain fatty acid biosynthesis. The Sporobolomyces PHS1 gene complements the essential functions of a S. cerevisiae phs1 mutant. The Sporobolomyces phs1 mutant strain has less dehydratase activity and a reduction in very long chain fatty acids compared to wild type. The mutant strain also exhibits sensitivity to cell wall stress agents and loss of shooting due to a delay in ballistospore formation, indicating that the role of Phs1 in spore dissemination may be primarily in cellular integrity.

Effect of selenium on lipid alternations in pigment-forming yeasts

The work deals with lipid modifications of pigment-forming yeasts Rhodotorula and Sporobolomyces growing under presence of selenium. This metal in the medium significantly prolonged lag-phase of all cultures and enlarged yeast cells. Total, neutral, and membrane yeast lipids (phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylinositol) consisted of predominantly palmitic, palmitoleic, stearic, oleic, linoleic, and linolenic acids. Selenium activated fatty acid unsaturation mainly in phosphatidylcholine due to elevated levels of linoleic and linolenic acids. Because biosynthesis of C18 unsaturated fatty acids in Rhodotorula and Sporobolomyces species may be associated with phosphatidylcholine moieties, selenium might be involved to the induction of membranebound fatty acid Δ12 and Δ15 desaturases in red yeasts. Oppositely, neutral lipids (primarily triacylglycerols) did not show such intensive changes in fatty acid composition as their polar counterparts. These observations could be applied for preparation of selenized red yeasts containing carotenoid pigments with enhanced accumulation of linoleic and linolenic acids.

Overcoming recalcitrant transformation and gene manipulation in Pucciniomycotina yeasts

The red yeasts of the Pucciniomycotina have rarely been transformed with DNA molecules. Transformation methods were recently developed for a species of Sporobolomyces, based on selection using uracil auxotrophs and plasmids carrying the wild-type copies of the URA3 and URA5 genes. However, these plasmids were ineffective in the transformation of closely related species. Using the genome-sequenced strain of Rhodotorula graminis as a starting point, the URA3 and URA5 genes were cloned and tested for the transformation ability into different Pucciniomycotina species by biolistic and Agrobacterium-mediated transformations. Transformation success depended on the red yeast species and the origin of the URA3 or URA5 genes, which may be related to the high G + C DNA content found in several species. A new vector was generated to confer resistance to nourseothricin, using a native promoter from R. graminis and the naturally high G + C nourseothricin acetyltransferease gene. This provides a second selectable marker in these species. Targeted gene disruption was tested in Sporobolomyces sp. IAM 13481 using different lengths of homologous DNA with biolistic and Agrobacterium transformation methods. Both DNA delivery methods were effective for targeted replacement of a gene required for carotenoid pigment biosynthesis. The constructs also triggered transgene silencing. These developments open the way to identify and manipulate gene functions in a large group of basidiomycete fungi.

Development of resources for the analysis of gene function in Pucciniomycotina red yeasts

The Pucciniomycotina is an important subphylum of basidiomycete fungi but with limited tools to analyze gene functions. Transformation protocols were established for a Sporobolomyces species (strain IAM 13481), the first Pucciniomycotina species with a completed draft genome sequence, to enable assessment of gene function through phenotypic characterization of mutant strains. Transformation markers were the URA3 and URA5 genes that enable selection and counter-selection based on uracil auxotrophy and resistance to 5-fluoroorotic acid. The wild type copies of these genes were cloned into plasmids that were used for transformation of Sporobolomyces sp. by both biolistic and Agrobacterium-mediated approaches. These resources have been deposited to be available from the Fungal Genetics Stock Center. To show that these techniques could be used to elucidate gene functions, the LEU1 gene was targeted for specific homologous replacement, and also demonstrating that this gene is required for the biosynthesis of leucine in basidiomycete fungi. T-DNA insertional mutants were isolated and further characterized, revealing insertions in genes that encode the homologs of Chs7, Erg3, Kre6, Kex1, Pik1, Sad1, Ssu1 and Tlg1. Phenotypic analysis of these mutants reveals both conserved and divergent functions compared with other fungi. Some of these strains exhibit reduced resistance to detergents, the antifungal agent fluconazole or sodium sulfite, or lower recovery from heat stress. While there are current experimental limitations for Sporobolomyces sp. such as the lack of Mendelian genetics for conventional mating, these findings demonstrate the facile nature of at least one Pucciniomycotina species for genetic manipulation and the potential to develop these organisms into new models for understanding gene function and evolution in the fungi.

Immunodetection and Characterization of Antigens Expressed by Uncinula necator

AbstractConidia from four genetically distinct isolates of Uncinula necator (Schw.) Burr. were used to raise a polyclonal antiserum. Immunofluorescent detection of the fungus on Vitis vinifera (cv. Chardonnay) indicated that the antiserum bound specifically to fungal antigens present on both conidia and hyphae, with no detection of underlying berry tissues. The antibody reacted with three antigens present on the conidia (Mr 21, 29 and > 250 kDa). Immunoreactivity with the 21 kDa antigen was dependent upon the preservation of at least one disulphide bond linkage. Evidence from immunoblot staining and enzyme immunoassay indicated that only a small proportion of the recognized epitopes contained carbohydrate moieties. The antibody detected homologous U. necator conidial antigens in a plate trapped antigen‐enzyme‐linked immunoabsorbent assay (PTA‐ELISA), with a linear range of detection extending from 1000 to 9000 conidia/ml at a 1/5000 dilution of serum. Under these conditions the immunoassay also detected antigens from pooled heterologous U. necator isolates. The antiserum exhibited cross‐reactivity with antigens present on Aspergillus, Pithomyces and Sporobolomyces species co‐isolated from powdery mildew‐infected grapes, which could not be removed by fractionation of the antiserum on antigen affinity columns. Monoclonal antibodies were subsequently produced to avoid the problems of cross‐reactivity associated with the polyclonal antibody. Antibodies produced from two clonal lines exhibited specificity for U. necator and were shown to detect a 21 kDa conidial antigen. Use of either of these antibodies enabled the differentiation of grapes grouped on the basis of powdery mildew disease levels.

Sporobolomyces odoratus sp. nov., a new species in the Sporidiobolus ruineniae clade.

This report presents the description of a new Sporobolomyces species, Sp. odoratus sp. nov. The new species was characterized by DNA fingerprinting using the micro/minisatellite-primed PCR approach (MSP-PCR). A phylogenetic analysis using the D1/D2 region of the 26S rDNA revealed that Sp. odoratus belongs to a clade that includes Rhodosporidium fluviale and Sporidiobolus ruineniae. An integrated comparison with related species is presented. A number of presumptive strains of S. ruineniae were also investigated using the MSP-PCR method and 26S rDNA sequence analysis. Mating experiments revealed, for the first time, that S. ruineniae includes heterothallic strains, besides those already known to be self-fertile.

Distribution and identification of red yeasts in deep-sea environments around the northwest Pacific Ocean.

We isolated 99 yeast strains, including 40 red yeasts, from benthic animals and sediments collected from the deep-sea floor in various areas in the northwest Pacific Ocean. Comparing the yeast isolates from animals and sediments collected from shallow locations, the proportion of red yeasts differed considerably, comprising 81.5% and 10.6% of the isolates from animals and sediments, respectively. All of the red yeast isolates belonged to the genera Rhodotorula and Sporobolomyces. On the basis of morphological and physiological characteristics, the isolates were identified as R. aurantiaca, R. glutinis, R. minuta and R. mucilaginosa of the genus Rhodotorula, and S. salmonicolor and S. shibatanus of the genus Sporobolomyces. Only R. glutinis and R. mucilaginosa were isolated from sediments. All of the others were isolated from animal sources. Phylogenetic analyses based on internal transcribed spacer (ITS) regions and 5.8S rRNA gene sequences allowed us to establish the precise taxonomic placement of each of the isolates and thereby investigate the intraspecific relationships among the isolates. Twenty-two strains identified as members of R. glutinis, which showed a wide distribution in the deep-sea, and five isolates identified as R. minuta, which were isolated only from benthic animals, showed substantial heterogeneity within the species. The isolates phenotypically identified as Sporobolomyces species and R. mucilaginosa phylogenetically occupied the placements corresponding to these species. Some strains assigned to known species on the basis of phenotypic features should be regarded as new species as suggested by the results of molecular analysis.

Phylogenetic analysis of the ballistoconidium-forming yeast genus Sporobolomyces based on 18S rDNA sequences.

The 18S rDNA nucleotide sequences of 25 Sporobolomyces species and five Sporidiobolus species were determined. Those of Sporobolomyces dimmenae JCM 8762T, Sporobolomyces ruber JCM 6884T, Sporobolomyces sasicola JCM 5979T and Sporobolomyces taupoensis JCM 8770T showed the presence of intron-like regions with lengths of 1586, 324, 322 and 293 nucleotides, respectively, which were presumed to be group I introns. A total of 63 18S rDNA nucleotide sequences was analysed, including 33 published reference sequences. Sporobolomyces species and the other basidiomycetes species were distributed throughout the phylogenetic tree. The resulting phylogeny indicated that Sporobolomyces is polyphyletic. Sporobolomyces species were mainly divided into four groups within the Urediniomycetes. The groups are designated as the Sporidiales, Agaricostilbum/Bensingtonia, Erythrobasidium and subbrunneus clusters. The last group, comprising four species, Sporobolomyces coprosmicola, Sporobolomyces dimmenae, Sporobolomyces linderae and Sporobolomyces subbrunneus, forms a new and distinct cluster in the phylogenetic tree in this study.

Ballistosporous yeasts found on the surface of plant materials collected in New Zealand. 1. Six new species in the genus Sporobolomyces.

Six new species of ballistosporous yeast, the genus Sporobolomyces, were isolated from dead leaves and fruit of plants collected in New Zealand; Sp. novazealandicus, Sp. dimmenae, Sp. coprosmicola, Sp. coprosmae, Sp. dracophyllus, and Sp. taupoensis. These species differ from any hitherto known species of Sporobolomyces based on chemotaxonomic characteristics.

Molecular approaches to the taxonomy of ballistosporous yeasts based on the analysis of the partial nucleotide sequences of 18S ribosomal ribonucleic acids.

The partial nucleotide sequences of 18S ribosomal RNA (positions 1451 to 1618 in Saccharomyces cerevisiae) of 48 strains representing 46 species of ballistosporous yeasts and a related genus were determined by dideoxy sequencing with reverse transcriptase. The sequence data were analyzed by neighbor-joining method and several unrooted phylogenetic trees were drawn. Ballistosporous yeast species were separated into two groups: a of xylose-containing species and a of xylose-lacking species. All of the species of Bullera and Kockovaella are included in the former group, and all of the species of Ballistosporomyces, Bensingtonia, Sporobolomyces, and Sporidiobolus are included in the latter group. Species of the genus Bullera are scattered on several branches of the phylogenetic trees and species of Cryptococcus are located at several positions close to those of species in several groups of Bullera. Species of piricola group of Bullera, Bullera megalospora, Bullera punicea and Bullera piricola, are considered to be closely related and are far removed from where the remaining species of the genus are located. These three species are characterized by the production of extremely large asymmetrical (bilaterally symmetrical) ballistospores and are considered to be included in a different genus from Bullera. A close relationship of Kockovaella to Fellomyces is suggested. Species of Sporobolomyces are scattered on several branches of the phylogenetic trees. The same phenomenon is observed in the case of Bensingtonia and Ballistosporomyces. These observations clearly suggest that the mode of vegetative reproduction such as the productivity of ballistospores, stalked conidia and budding yeast cells, does not reflect the phylogenetic relationship among basidiomycetous yeasts and is considered to be a less significant taxonomic criterion than hitherto believed.

An electrophoretic comparison of enzymes in strains of the Q9-equipped species in the genus Sporobolomyces Kluyver et van Niel(Sporobolomycetaceae).

A taxonomic study below the generic or at the specific level was made of electrophoretic patterns of seven enzymes in 18 strains of the Q9-equipped Sporobolomyces and Bensingtonia species. The seven enzymes were glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, isocitrate dehydrogenase, malate dehydrogenase, hexokinase, phosphoglucomutase, and fumarase. Seven species, S. intermedius, S. miscanthi, S. subroseus, S. yamatoanus, S. naganoensis, S. yuccicola, and S. phylladus, constituted clusters separate from each other (s=0 or 14%). The two strains of S. miscanthi showing identical electrophoretic enzyme patterns (s=100%) were linked to Sporobolomyces species NZ-3 with a similarity value of 29%. Within the five strains of S. yamatoanus examined, the type strain had a similarity value of 86% with one other strain. The remaining three strains whose similarity values were calculated as 100% and 57% were linked to the two strains including the type strain with a similarity value of 43%. The three strains of S. phylladus had identical electrophoretic enzyme patterns (s=100%). Sporobolomyces species NZ-24 had a separate cluster (s=0%). Sporobolomyces weijmanii was synonymous with S. intermedius (s=100%). Bensingtonia ciliata constituted its own cluster (s=0%). These data are discussed from the taxonomic point of view.

The distribution of xylose in the cells of ballistosporous yeasts - Application of high performance liquid chromatography without derivatization to the analysis of xylose in whole cell hydrolysates.

The presence or absence of xylose in the whole cell hydrolysates from all type strains of 41 known species of the genera Bullera, Sporidiobolus, and Sporobolomyces, and three strains of Itersonilia perplexans was examined using high performance liquid chromatography (HPLC) which makes itpossible to analyze the carbohydrates without derivatization. Of 16 species of the genus Bullera, 14 had xylose in the cells. The other two species, Bullera salicina and Bullera tsugae, had no xylose in the cells. None of the 25 species of the genera Sporidiobolus and Sporobolomyces had xylose in the cells. The lack of xylose in the cells of seven Sporobolomyces species with Q-9 as the major ubiquinone proves that these species are not yeast phases of certain species of the genus Itersonilia with Q-9, since both the yeast and filamentous phases of Itersonilia perplexans had xylose in the cells. These results show that the presence or absence of xylose in the cells is very useful as one criterion for the generic differentiation of Bullera and Sporobolomyces. The lack of xylose in the cells of B. salicina and B. tsugae indicates that these two species may be closely related to the genus Sporobolomyces. Our HPLC method, by which xylose in the whole cell hydrolysates can be analyzed without derivatization, is a useful way to differentiate taxa by this criterion.

Sporobolomyces inositophilus, a new species of ballistosporous yeast isolated from a dead leaf of Sasa sp. in Japan.

A new species of ballistosporous yeast, Sporobolomyces inositophilus was isolated from a dead leaf of Sasa sp. collected at a mountain in Japan. This yeast is assumed to have a close relationship to Sporobolomyces singularis since it produces bilaterally symmetrical ballistospores and pale-coloured colonies and resembles Sporobolomyces singularis in these respects. These two species differ from typical species of Sporobolomyces and seem to constitute a natural taxonomic group.

Mating among isolates of three species of Sporobolomyces

Mating tests were made with the type strains of Sporobolomyces hispanicus Peláez & Ramirez, S. odorus Derx, and S. salmonicolor (Fischer & Brebeck) Kluyver & van Niel, and with 19 additional strains. These tests show that 21 of the strains are of two, intercompatible mating types. Since the three type strains are linked by compatibility, the names that have been applied are synonymous. Additionally, since mycelial and strictly yeast-like strains are compatible, the genus Prosporobolomyces Novák & Zolt appears to be unacceptable.

Sporobolomycetaceae in Kansas

Isolations of Sporobolomycetaceae in Kansas from air and leaf surfaces yielded four species of Sporobolomyces, S. pararoseus, S. gracilis, S. alborubescens, and S. odorus; two species of Tilletiops...

Sporobolomycetaceae in Kansas

SUMMARYIsolations of Sporobolomycetaceae in Kansas from air and leaf surfaces yielded four species of Sporobolomyces, S. pararoseus, S. gracilis, S. alborubescens, and S. odorus; two species of Tilletiopsis, T. washingtonensis and T. minor; and Bullera alba. Sporobolomyces was isolated throughout the year, Tilletiopsis only in the summer, and Bullera in fall, winter, and spring. Optimum temperature for ballistospore production was 16 C.

Asparaginase and glutaminase activities of micro-organisms.

Summary: l-Asparaginase and l-glutaminase activities were detected in many microorganisms and the distribution of these activities was found to be related to the classification of micro-organisms. Among 464 bacteria, the activities occurred in many Gram-negative bacteria and in a few Gram-positive bacteria. Most members of the family Enterobacteri-aceae possessed l-asparaginase. l-Asparaginase and l-glutaminase occurred together in a large proportion of pseudomonads. Among Gram-positive bacteria many strains of Bacillus pumilus showed strong l-asparaginase activity. Amidase activities were also observed in several strains in other families. l-Asparaginase activity was not detected in culture filtrates of 261 strains of species of the genera Streptomyces and Nocardia, but l-asparaginase and l-glutaminase were detected when these organisms were sonicated. The amidase activities in culture filtrates of 4158 fungal strains were tested. All the strains of Fusarium species formed l-asparaginase. Organisms of the genera Hypomyces and Nectria, which are regarded as the perfect stage of the genus Fusarium, also formed l-asparaginase. Several Penicillium species formed l-asparaginase. Two organisms of the family Moniliaceae formed l-glutaminase together with l-asparaginase, and a fewascomycetous fungi formed l-asparaginase or l-glutaminase. Among 1326 yeasts, l-asparaginase or l-glutaminase occurred frequently in certain serological groups of yeasts: VI (Hansenula) group, Cryptococcus group and Rhodotorula group. Many strains of Sporobolomyces species also showed l-asparaginase activity. Several strains of Cryptococcus and Rhodotorula group possessed l-glutaminase and l-asparaginase. l-Glutaminase alone was formed in many strains of Candida scottii and Cryptococcus albidus, both of which are related to Basidiomycetes.