Savitreea siamensis sp. nov., an ascomycetous yeast species in the family Saccharomycetaceae discovered in Thailand.

Four yeast strains, representing a novel anamorphic species, were isolated in Thailand. The two strains (ST-3660T and ST-3647) were obtained from two different estuarine water samples in a mangrove forest. Strain DMKU-FW1-37 was derived from a grease sample, and another strain (TSU57) was isolated from a fruiting body of Phallus sp. Pairwise sequence analysis showed that the four strains had identical or differed by only one nucleotide substitution in the D1/D2 domains of the large subunit (LSU) rRNA gene, and differed by one to three nucleotide substitutions in the internal transcribed spacer (ITS) regions. Savitreea pentosicarens is the most closely related species to the four strains, but with 9-10 (1.57-1.72 %) nucleotide substitutions in the D1/D2 domains of the LSU rRNA gene and 29-31 (4.22-4.45 %) nucleotide substitutions in the ITS regions. Phylogenetic analyses based on the concatenated sequences of the ITS regions and the D1/D2 domains of the LSU rRNA gene showed that the four strains form a well-separated lineage from S. pentosicarens with high bootstrap support, confirming that they represent a distinct species. Therefore, the four strains are assigned as representives of a novel species of the genus Savitreea, for which the name Savitreea siamensis sp. nov. is proposed. The holotype is TBRC 4481T and the ex-type is PYCC 9794T (=ST-3660T). The MycoBank number of the novel species is MB 851951.

Phylogenomic delineation of two new species of ascomycetous yeasts, Wickerhamiella koratensis sp. nov. and Wickerhamiella limtongiae sp. nov., and proposal of two synonyms, Wickerhamiella infanticola and Wickerhamiella tropicalis.

Two novel ascomycetous yeast species of the genus Wickerhamiella are proposed based on isolates obtained in Thailand from food waste and the fruiting body of a polypore fungus, and on a combination of conventional DNA-barcode sequence analyses and whole-genome phylogenies. We focus on a particular subclade of the genus Wickerhamiella that contains species found in anthropic environments and describe Wickerhamiella limtongiae sp. nov. (DMKU-FW31-5T=PYCC 9022T=TBRC 15055T), found on food waste samples. In an adjacent clade, we describe Wickerhamiella koratensis sp. nov. (DMKU-KO16T=PYCC 8908T=TBRC 14869T), which represents the closest relative of Wickerhamiella slavikovae and was isolated from the fruiting body of Bjerkandera sp. In the subclade of W. limtongiae sp. nov., we propose that Wickerhamiella infanticola should be regarded as a synonym of Wickerhamiella sorbophila and that Wickerhamiella tropicalis should be regarded as a synonym of Wickerhamiella verensis.

Yeasts from tropical forests: Biodiversity, ecological interactions, and as sources of bioinnovation.

Tropical rainforests and related biomes are found in Asia, Australia, Africa, Central and South America, Mexico, and many Pacific Islands. These biomes encompass less than 20% of Earth's terrestrial area, may contain about 50% of the planet's biodiversity, and are endangered regions vulnerable to deforestation. Tropical rainforests have a great diversity of substrates that can be colonized by yeasts. These unicellular fungi contribute to the recycling of organic matter, may serve as a food source for other organisms, or have ecological interactions that benefit or harm plants, animals, and other fungi. In this review, we summarize the most important studies of yeast biodiversity carried out in these biomes, as well as new data, and discuss the ecology of yeast genera frequently isolated from tropical forests and the potential of these microorganisms as a source of bioinnovation. We show that tropical forest biomes represent a tremendous source of new yeast species. Although many studies, most using culture-dependent methods, have already been carried out in Central America, South America, and Asia, the tropical forest biomes of Africa and Australasia remain an underexplored source of novel yeasts. We hope that this review will encourage new researchers to study yeasts in unexplored tropical forest habitats.

Yamadazyma sisaketensis f.a., sp. nov. and Yamadazyma koratensis f.a., sp. nov., two novel ascomycetous yeast species from mushrooms and cocoa leaves in Thailand, and reassignment of Candida andamanensis, Candida jaroonii and Candida songkhlaensis to the genus Yamadazyma.

Six strains representing two novel ascomycetous yeast species were isolated from mushroom fruiting bodies and cocoa leaves collected in Thailand. Analysis of the internal transcribed spacer (ITS) regions and the D1/D2 domains of the large subunit rRNA gene sequences showed that the six strains were divided into two groups. The first group consisted of four strains (DMKU-SSK46, DMKU-SK1, SCCL3-5 and SCCL19-3), that were closely related to the type strains of Candida conglobata, Candida insectorum, Yamadazyma dushanensis, Yamadazyma mexicana and Yamadazyma riverae, but with 12-14 (2.5-2.9 %) and 28-50 (5.4-8.8 %) nucleotide substitutions in the D1/D2 domains and the ITS regions, respectively. However, two strains (DMKU-KMY40 and DMKU-KO18) of the second group differed from a group of described species, Candida diddensiae, Candida dendronema, Candida germanica, Candida kanchanaburiensis, Candida naeodendra, Candida vaughaniae and Yamadazyma siamensis by 8-15 (1.5-2.8 %) and 45-53 (8.2-9.6 %) nucleotide substitutions in the D1/D2 domains and the ITS regions, respectively. Phylogenetic analysis based on the concatenated sequences of the ITS regions and D1/D2 domains showed that these strains represented two species of the Yamadazyma clade that were distinct from the other related species. Based on the phylogenetic analysis and phenotypic characteristics, these six strains were assigned to two novel species of the genus Yamadazyma, although formation of ascospores was not observed. Yamadazyma sisaketensis f.a., sp. nov., is proposed for the first group (four strains). The holotype is TBRC 17139T (ex-type culture: PYCC 9797). The MycoBank number is MB 849637. Yamadazyma koratensis f.a., sp. nov. is proposed for the second group (two strains). The holotype is TBRC 14868T (ex-type culture: PYCC 8907). The MycoBank number is MB 849638. In addition, it is proposed that Candida andamanensis, Candida jaroonii and Candida songkhlaensis are reassigned to the genus Yamadazyma as Yamadazyma andamanensis comb. nov., Y. jaroonii comb. nov. and Y. songkhlaensis comb. nov., respectively.

Kodamaea samutsakhonensis f.a., sp. nov., a novel ascomycetous yeast species isolated from wild mushrooms in Thailand.

Two strains of genus Kodamaea, representing a novel anamorphic yeast species, were isolated from two samples of Marasmiellus sp. collected in Thailand. Analysis of the sequences of the internal transcribed spacer (ITS) regions and the D1/D2 domains of the large subunit (LSU) rRNA gene showed that the two strains differed by 27-42 nucleotide substitutions in the D1/D2 domains of the LSU rRNA gene and 7-34 nucleotide substitutions in the ITS region of a group of related species, Kodamaea smagusa CBS 11430T, Kodamaea fungicola JCM 10142T, Kodamaea plutei ATCC MYA-4329T, Kodamaea lidongshanica SD5S01T and Kodamaea jinghongensis NYNU 167162T. Phylogenetic analysis based on the concatenated sequences of the ITS and the D1/D2 domains of the LSU rRNA gene showed that the two strains were placed in the Kodamaea clade and clearly separated from other recognized species of the genus. Therefore, the two strains were assigned as a novel species of the genus Kodamaea, for which we propose the name Kodamaea samutsakhonensis f.a., sp. nov. The holotype is TBRC 16043T (=DMKU-BP19T) and the isotype is PYCC 9354. The MycoBank number of the novel species is MB 846490.

Distinct Metabolic Flow in Response to Temperature in Thermotolerant Kluyveromyces marxianus.

The intrinsic mechanism of the thermotolerance of Kluyveromyces marxianus was investigated by comparison of its physiological and metabolic properties at high and low temperatures. After glucose consumption, the conversion of ethanol to acetic acid became gradually prominent only at a high temperature (45°C) and eventually caused a decline in viability, which was prevented by exogenous glutathione. Distinct levels of reactive oxygen species (ROS), glutathione, and NADPH suggest a greater accumulation of ROS and enhanced ROS-scavenging activity at a high temperature. Fusion and fission forms of mitochondria were dominantly observed at 30°C and 45°C, respectively. Consistent results were obtained by temperature upshift experiments, including transcriptomic and enzymatic analyses, suggesting a change of metabolic flow from glycolysis to the pentose phosphate pathway. The results of this study suggest that K. marxianus survives at a high temperature by scavenging ROS via metabolic change for a period until a critical concentration of acetate is reached. IMPORTANCE Kluyveromyces marxianus, a thermotolerant yeast, can grow well at temperatures over 45°C, unlike Kluyveromyces lactis, which belongs to the same genus, or Saccharomyces cerevisiae, which is a closely related yeast. K. marxianus may thus bear an intrinsic mechanism to survive at high temperatures. This study revealed the thermotolerant mechanism of the yeast, including ROS scavenging with NADPH, which is generated by changes in metabolic flow.

Metahyphopichia suwanaadthiae sp. nov., an anamorphic yeast species in the order Saccharomycetales and reassignment of Candida silvanorum to the genus Metahyphopichia.

Seven yeast strains, representing a single novel anamorphic species, were isolated in Thailand. They consisted of five strains (DMKU-MRY16T, DMKU-SK18, DMKU-SK25, DMKU-SK30 and DMKU-SK32) obtained from five different mushrooms, and two strains (ST-224 and 11-14.2) derived from insect frass and soil, respectively. The pairwise sequence analysis indicated that all seven strains had identical sequences in the D1/D2 domains of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region. Metahyphopichia silvanorum was the most closely related species, but with 11.9-12.4% nucleotide substitutions in the D1/D2 domains of the LSU rRNA gene and 13.1-13.3% nucleotide substitutions in the ITS region. The phylogenetic analyses based on the concatenated sequences of the ITS region and the D1/D2 domains of the LSU rRNA gene showed that the seven strains form a well-separated subclade in a clade containing M. silvanorum and Metahyphopichia laotica with high bootstrap support. A phylogenetic analysis of a multilocus dataset including the small subunit (SSU) rRNA gene, the ITS region, the D1/D2 domains of the LSU rRNA gene, translation elongation factor 1-alpha gene, actin gene and the RNA polymerase II subunit 2 gene, confirmed the presence of the monophyletic clade that also includes M. silvanorum and M. laotica, and strongly supported the phylogenetic isolation of the seven strains from its neighbouring species. Therefore, the seven strains were assigned as a single novel species of the genus Metahyphopichia, according to their phylogenetic relationships. The name Metahyphopichia suwanaadthiae sp. nov. is proposed to accommodate the seven strains. The holotype is DMKU-MRY16T (TBRC 11775T=NBRC 114386T=PYCC 8655T). The MycoBank number of the novel species is MB 841280. In addition, Candida silvanorum is reassigned to the genus Metahyphopichia. The MycoBank number of M. silvanorum comb. nov. is MB 841279.

Pichia bovicola sp. nov., a yeast species associated with small-intestinal content of cattle.

Three yeast strains, DMKU-MP6-4T, DMKU-MP2-6 and DMKU-MP5-1, were isolated from the small-intestinal content or Pia of cattle in Thailand during the investigation of yeast diversity in this habitat. According to the D1/D2 domain of the large subunit (LSU) rRNA gene and internal transcribed spacer (ITS) region sequence analysis, these strains represent a novel yeast species in the genus Pichia. The species produced one to four ascospores per ascus with spherical to ellipsoidal shape and heterogenous in terms of size. These three strains were identical and differed from their closely related species, Pichia exigua NRRL Y-10920T by 2% (six nucleotide substitutions and five gaps) in the D1/D2 domain of the large subunit rRNA gene, while ITS region sequences differed by 3.1% (16 nucleotide substitutions and 27 gaps), 3.7% (19 nucleotide substitutions and 28 gaps) and 3.1% (16 nucleotide substitutions and 27 gaps) for DMKU-MP6-4T, DMKU-MP2-6 and DMKU-MP5-1, respectively. The name, Pichia bovicola, is proposed to accommodate these species. The holotype is DMKU-MP6-4T (TBRC 15616T=PYCC 8905T).

Wickerhamiella nakhonpathomensis f.a. sp. nov., a novel ascomycetous yeast species isolated from a mushroom and a flower in Thailand.

Strains SU22T (TBRC 14875T) and FLA11.5, representing a novel anamorphic yeast species, were respectively isolated from a fruiting body of a Coprinus species and an inflorescence of a Coffea species collected in Thailand. Analysis of the sequences of the D1/D2 domains of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) regions showed that the two strains differed by two nucleotide substitutions in the D1/D2 domains of the LSU rRNA gene and were identical in the ITS regions. Wickerhamiella drosophilae CBS 8459T was the most closely related species, but with 24-26 nucleotide substitutions in the D1/D2 domains of the LSU rRNA gene and 24 nucleotide substitutions in the ITS regions. A phylogenetic analysis, based on the sequences of the D1/D2 domains, indicated that the two strains represented a species in the genus Wickerhamiella which was distinct from other recognized species of the genus. Therefore, the two strains were assigned as a novel species, for which we propose the name Wickerhamiella nakhonpathomensis f.a. sp. nov. The holotype is TBRC 14875T (isotype PYCC 8914T). The MycoBank number of the novel species is MB 840833.

Kazachstania surinensis f.a., sp. nov., a novel yeast species isolated from Thai traditional fermented food.

Seven yeast strains (UBIF12-1, UBFB13-1, SRFS56-3, SRFS57-2, SKFS62-1, SKFS66-1 and SKFS67-1) representing a single anamorphic novel yeast species were isolated from traditional Thai fermented foods in Ubon Ratchathani, Surin and Sisaket in the northeast part of Thailand. The results of analysis of the sequences of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region indicated that the seven strains showed zero to one nucleotide substitutions in the sequences of the D1/D2 region of the LSU rRNA gene, and zero to four nucleotide substitutions in the ITS region. Kazachstania humilis CBS 5658T was the most closely-related species, but with 0.7-0.9% nucleotide substitutions in the D1/D2 region of the LSU rRNA gene, and 2.0-2.2% nucleotide substitution in the ITS region. The results of a phylogenetic analysis based on the concatenated ITS and D1/D2 regions confirmed that the seven strains represented a single species of the genus Kazachstania distinct from the other recognized species of the genus. Furthermore, the morphological, biochemical and physiological properties of the seven strains not only indicated that they represented members of the genus Kazachstania, but that they were separated from K. humilis and K. pseudohumilis, the two most closely related species in the phylogenetic tree. Therefore, the seven strains were identified as representing a novel species, for which we propose the name Kazachstania surinensis f.a., sp. nov. The holotype is TBRC 15053T (isotype: SRFS57-2 and PYCC 9021). The MycoBank number of the novel species is 841892.

Cultivable yeasts associated with marine sponges in the Gulf of Thailand, South China Sea.

Marine sponges harbor numerous microorganisms, among which sponge-associated yeasts are the least explored. To gain greater knowledge of sponge-associated yeasts, an investigation was therefore performed on marine sponges in Sattahip Bay, Gulf of Thailand, South China Sea. Seventy-one (71) marine sponge samples were collected at sites near Samae-san, Mu, and Khram islands, and were subsequently identified as 17 sponge species in 14 genera. Eighty-seven (87) yeast strains were isolated from 42 samples. The identification of yeasts by similarity analysis of the D1/D2 domain sequences of the large subunit rRNA gene revealed that 64% of the yeast strains obtained belonged to the phylum Basidiomycota, while the remaining strains belonged to the phylum Ascomycota. The strains that belonged to Ascomycota comprised 11 known yeast species in five genera (Candida, Kodamaea, Magnusiomyces, Meyerozyma, and Pichia). The strains belonging to the phylum Basidiomycota comprised 14 known yeast species in eight genera (Cutaneotrichosporon, Cystobasidium, Naganishia, Papiliotrema, Rhodosporidiobolus, Rhodotorula, Trichosporon, and Vishniacozyma). In addition, three strains represented a potential novel species closest to Cys. slooffiae; one strain represented a potential novel species closest to R. toruloides; and one strain represented a potential novel species closest to V. foliicola. The species with the highest occurrence was Rhodotorula mucilaginosa. No marked difference was found in the principal coordinates analysis of the ordinations of yeast communities from the three sampling sites. The estimation using EstimateS software showed that the expected species richness was higher than the observed species richness. As the marine sponge-yeast association remains unclear, more systematic investigations should be carried out.

Ethanol and H2O2 stresses enhance lipid production in an oleaginous Rhodotorula toruloides thermotolerant mutant L1-1.

Stress tolerance is a desired characteristic of yeast strains for industrial applications. Stress tolerance has been well described in Saccharomyces yeasts but has not yet been characterized in oleaginous Rhodotorula yeasts even though they are considered promising platforms for lipid production owing to their outstanding lipogenicity. In a previous study, the thermotolerant strain L1-1 was isolated from R. toruloides DMKU3-TK16 (formerly Rhodosporidium toruloides). In this study, we aimed to further examine the ability of this strain to tolerate other stresses and its lipid productivity under various stress conditions. We found that the L1-1 strain could tolerate not only thermal stress but also oxidative stress (ethanol and H2O2), osmotic stress (glucose) and a cell membrane disturbing reagent (DMSO). Our results also showed that the L1-1 strain exhibited enhanced ability to maintain ROS homeostasis, stronger cell wall strength and increased levels of unsaturated membrane lipids under various stresses. Moreover, we also demonstrated that ethanol-induced stress significantly increased the lipid productivity of the thermotolerant L1-1. The thermotolerant L1-1 was also found to produce a higher lipid titer under the dual ethanol-H2O2 stress than under non-stress conditions. This is the first report to indicate that ethanol stress can induce lipid production in an R. toruloides thermotolerant strain.

Teunia siamensis f.a., sp. nov., a novel tremellaceous yeast species isolated from soil in a secondary peat swamp forest area.

The strain DMKU-XD44, representing an anamorphic novel yeast species, was isolated from soil collected in a peat swamp forest (PSF) area in Rayong Botanical Garden in eastern Thailand. On the basis of sequence analysis of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region, Teunia cuniculi CBS 10309T was the most closely related species. The novel species differed from the T. cuniculi type by 2.5 % (14 nucleotide substitutions) in the D1/D2 region of the LSU rRNA gene and by 8.0 % (40 nucleotide substitutions) in the ITS region. The results of a phylogenetic analysis, based on the combined sequences of the ITS region and the D1/D2 region, indicated that DMKU-XD44 represents a member of the Teunia clade in the Cryptococcaceae (Tremellales, Tremellomycetes, Agaricomycotina and Basidiomycota) and is phylogenetically distinct from other species of the genus Teunia in the clade. Therefore, DMKU-XD44 represents a novel species of the genus Teunia. The name Teunia siamensis f.a., sp. nov. is proposed. The holotype is DMKU-XD44, while the MycoBank number is MB 832816.

Torulaspora nypae sp. nov., a novel yeast species isolated from nipa (Nypa fruticans Wurmb.) inflorescence sap in southern Thailand.

Two strains (YSP-384 and YSP-399), representing a novel Torulaspora species, were isolated from two nipa inflorescence sap samples collected in Trang province in the southern part of Thailand. The two strains had identical sequences of the D1/D2 domains of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) regions. The two strains were closest to Torulaspora maleeae CBS 10694T, but with 1.1 % nucleotide substitutions in the D1/D2 domains of the LSU rRNA gene and 5.2 % nucleotide substitutions in the ITS regions. Phylogenetic analysis based on the concatenated sequences of the ITS regions and the D1/D2 domains of the LSU rRNA gene supported that the two strains represented a distinct species in the genus Torulaspora. Some phenotypic characteristics of the two strains differed from T. maleeae including the two strains have ability to assimilate d-xylose, d-glucono-δ-lactone and melizitose, and inability to ferment maltose and raffinose, whereas T. maleeae has opposite results. Therefore, the two strains are described as representing a novel species, for which the name Torulaspora nypae sp. nov. was proposed.

Wickerhamiella osmotolerans sp. nov. and Wickerhamiella tropicalis sp. nov., novel ascomycetous yeast in the family Wickerhamiellaceae.

Seven yeast strains, DMKU VGT1-14T, DMKU VGT1-19T, DMKU-JMGT1-28, DMKU-JMGT1-32, DMKU VGT2-06, DMKU VGT2-19 and DMKU VGT6-14, were isolated from a grease trap in Thailand and two strains, SJ-1 and SN-102 were isolated from the sea surface microlayer in Taiwan. On the basis of phenotypic characteristics and sequence analysis of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) region, these strains represented two novel yeast species of the genus Wickerhamiella. In terms of pairwise sequence similarity, four strains, DMKU VGT1-14, DMKU-JMGT1-32, DMKU VGT6-14 and SN-102, were closely related to Wickerhamiella infanticola NRRL Y-17858T but differed by 13 nucleotide substitutions with one gap (2.46 %) in the D1/D2 domain of the LSU rRNA gene and 15 nucleotide substitutions with 23 gaps (4.2 %) in the ITS region. The strains DMKU VGT1-19T, DMKU-JMGT1-28, DMKU VGT2-06, DMKU VGT2-19 and SJ-1, differed from the type strain of the most closely related species, Wickerhamiella sorbophila NRRL Y-7921T, by nine nucleotide substitutions with one gap (1.66 %) in the D1/D2 domain of the LSU rRNA gene and nine nucleotide substitutions with 17 gaps (2.52%) in the ITS region. Hence, the names Wickerhamiella osmotolerans sp. nov. and Wickerhamiella tropicalis sp. nov. are proposed to accommodate these species in the genus Wickerhamiella. The holotypes are W. osmotolerans DMKU VGT1-14T (ex-type strain TBRC 11425=PYCC 8359=CGMCC 2.6179; Mycobank number 833394) and W. tropicalis DMKU VGT1-19T (ex-type strain TBRC 11426=PYCC 8360=CGMCC 2.6180; Mycobank number 833393).

Yeast communities of primary and secondary peat swamp forests in southern Thailand.

Khanthuli peat swamp forest (PSF) is one of a few fertile peat swamp forests that remain in Thailand. It is composed of primary PSF and some areas which have been degraded to secondary PSF due to drought, wildfires and land conversion, which have resulted in a decrease in peat layers and change in the species of the plant community. In this study, diversity of yeasts in peat from both primary and secondary PSF areas of the Khanthuli PSF was determined based on culture-dependent approaches, using dilution plate and enrichment techniques. A total of 66 yeast isolates were identified by the analysis of sequence similarity of the D1/D2 region of the large subunit rRNA gene or the combined analysis of sequence of the D1/D2 region and internal transcribed spacer region and confirmed by phylogenetic analysis of the D1/D2 region to belong to 22 known yeast species and six potential new species in the genera Candida (Kurtzmaniella, Lodderomyces, Ogataea, Pichia and Yamadazyma clades), Clavispora, Cyberlindnera, Galactomyces, Hanseniaspora, Metschnikowia, Saturnispora, Schwanniomyces, Cryptotrichosporon, Pichia, Curvibasidium, Papiliotrema, Rhodotorula, and Saitozyma. The most prevalent yeasts in the primary PSF were Cyberlindnera subsufficiens and Galactomyces candidus, while Saitozyma podzolica was the most frequently found in peat from the secondary PSF. Common yeast species in both, primary and secondary PSF, were Cy. subsufficiens, G. candidus and Rhodotorula mucilaginosa.

Exploration of yeast communities in fresh coconut, palmyra, and nipa palm saps and ethanol-fermenting ability of isolated yeasts.

This study aimed to explore communities and the ethanol-fermenting ability of yeasts in fresh coconut, palmyra, and nipa palm saps. From the 90 samples of coconut, palmyra, and nipa palm saps, 204 yeast isolates were isolated and identified as 15 species in the phylum Ascomycota and a species (one strain) in Basidiomycota. Saccharomyces cerevisiae, Hanseniaspora guilliermondii, and Lachancea thermotolerans were found in the saps of all three palm species. Candida tropicalis and Pichia kudriavzevii were obtained from the coconut and palmyra palm saps, Hanseniaspora vineae, Lachancea fermentati, and Pichia manshurica were present in the coconut and nipa palm saps, whereas Torulaspora delbrueckii was found in the palmyra and nipa palm saps. The species with the highest occurrence in the saps of coconut, palmyra, and nipa palms was S. cerevisiae with 76.67%, 86.70%, and 100% frequency of occurrence, respectively. Using principal coordinates analysis for ordination, no marked difference was observed in the yeast communities from the saps of the three palm species. A total of 199 isolates were found to possess ethanol-fermentation ability when cultivated using shake flask in 160 g/L of glucose medium at 28°C for 48 h. Lachancea fermentati YSP-383, isolated from nipa palm sap, produced the highest amount of ethanol (76.74 g/L). Twenty-six isolates of Candida sanyaensis (1), C. tropicalis (1), H. guilliermondii (7), L. fermentati (8), L. thermotolerans (1), Pichia kudriavzevii (2), and S. cerevisiae (6) produced high amounts of ethanol ranging from 69.57 to 76.74 g/L. The result demonstrated that yeasts in the palm saps could play roles in the natural fermentation of palm saps.

Candida xylosifermentans sp. nov., a d-xylose-fermenting yeast species isolated in Thailand.

Three strains, representing a novel anamorphic and d-xylose-fermenting yeast species, were isolated from moss (ST-302T), seawater (ST-1169) and peat (DMKU-XE12) collected from the southern part of Thailand. The three strains had identical sequences of the D1/D2 regions of the large subunit (LSU) rRNA gene and the internal transcribed spacer (ITS) regions. Candida flosculorum CBS 10566T and Candida sharkiensis CBS 11368T were the most closely related species with 7.9 % nucleotide substitutions in the D1/D2 regions of the LSU rRNA gene, and 10.3 and 12.6% nucleotide substitutions in the ITS regions, respectively. Phylogenetic analysis based on the concatenated sequences of the ITS and the D1/D2 regions confirmed that the three strains represented a distinct anamorphic species in the Clavispora clade. Therefore, the three strains were described as a novel species, for which we propose the name Candida xylosifermentans sp. nov.

Tremella saccharicola f.a., sp. nov., a novel tremellaceous yeast species isolated from tropical regions.

Ten strains representing a single anamorphic novel yeast species were isolated from the external surface (DMKU-SP23 and DMKU-SP40) and tissue (DMKU-SE89, DMKU-SE99, DMKU-SE100 and DMKU-SE147) of sugarcane leaves in Thailand, and phylloplane (IMUFRJR 52034) and rhizoplane (IMUFRJ 52036 and 52037) of sugarcane and associated soil (IMUFRJ 52035) in Brazil. These strains showed zero to two nucleotide substitutions in the sequences of the D1/D2 region of the LSU rRNA gene and zero to three nucleotide substitutions in the internal transcribed spacer (ITS) region. Tremella globispora was the most closely related species, but with 1.7-2.1 % nucleotide substitutions in the D1/D2 region of the LSU rRNA gene, and 5.3-6.0 % nucleotide substitutions in the ITS region. Phylogenetic analysis based on the concatenated sequences of the ITS and the D1/D2 regions showed that these 10 strains represented a single species belonging to the genus Tremella (class Tremellomycetes, subphylum Agaricomycotina) that was distinct from related species. They therefore represented a novel species of the genus Tremella although the formation of basidia and basidiocarp were not observed. The name Tremella saccharicola f.a., sp. nov. is proposed. The type strain is DMKU-SP23T (=NBRC 109698T=BCC 61186T).

Functional analysis of Mig1 and Rag5 as expressional regulators in thermotolerant yeast Kluyveromyces marxianus.

To analyze the glucose repression mechanism in the thermotolerant yeast Kluyveromyces marxianus, disrupted mutants of genes for Mig1 and Rag5 as orthologs of Mig1 and Hxk2, respectively, in Saccharomyces cerevisiae were constructed, and their characteristics were compared with those of the corresponding mutants of S. cerevisiae. MIG1 mutants of both yeasts exhibited more resistance than the corresponding parental strains to 2-deoxyglucose (2-DOG). Histidine was found to be essential for the growth of Kmmig1, but not that of Kmrag5, suggesting that MIG1 is required for histidine biosynthesis in K. marxianus. Moreover, Kmrag5 and Schxk2 were more resistant than the corresponding MIG1 mutant to 2-DOG, and only the latter increased the utilization speed of sucrose in the presence of glucose. Kmrag5 exhibited very low activities for gluco-hexokinase and hexokinase and, unlike Schxk2, showed very slow growth and a low level of ethanol production in a glucose medium. Furthermore, Kmrag5, but not Kmmig1, exhibited high inulinase activity in a glucose medium and exhibited greatly delayed utilization of accumulated fructose in the medium containing both glucose and sucrose. Transcription analysis revealed that the expression levels of INU1 for inulinase and GLK1 for glucokinase in Kmrag5 were higher than those in the parental strain; the expression level of INU1 in Kmmig1 was higher, but the expression levels of RAG1 for a low-affinity glucose transporter in Kmmig1 and Kmrag5 were lower. These findings suggest that except for regulation of histidine biosynthesis, Mig1 and Rag5 of K. marxianus play similar roles in the regulation of gene expression and share some functions with Mig1 and Hxk2, respectively, in S. cerevisiae.