Psychrophilic yeasts in glacial environments of Alpine glaciers

The presence of psychrophilic yeasts in supra- and subglacial sediments, ice and meltwater collected from two glaciers of the Italian Alps (Forni and Sforzellina-Ortles-Cevedale group) was investigated. After incubation at 4 degrees C, subglacial sediments contained from 1.3 x 10(3) to 9.6 x 10(3) CFU of yeasts g(-1). The number of yeast cells in supraglacial sediments was c. 10-100-fold lower. A significant proportion of isolated yeasts exhibited one or more extracellular enzymatic activities (starch-degrading, lipolytic, esterolytic, proteolytic and pectinolytic activity) at 4 degrees C. Selected isolates were able to grow at 2 degrees C under laboratory-simulated in situ conditions. In all, 106 isolated yeasts were identified by MSP-PCR fingerprinting and 26S rRNA gene sequencing of the D1/D2 region as belonging to 10 species: Aureobasidium pullulans, Cryptococcus gilvescens (over 50% of the total), Cryptococcus terricolus, Mrakia gelida, Naganishia globosa, Rhodotorula glacialis, Rhodotorula psychrophenolica, Rhodotorula bacarum, Rhodotorula creatinivora and Rhodotorula laryngis. Four strains, all belonging to a new yeast species, yet to be described, were also isolated.     

Biodiversity of cold-adapted yeasts from glacial meltwater rivers in Patagonia, Argentina

The occurrence of culturable yeasts in glacial meltwater from the Frías, Casta?o Overo and Río Manso glaciers, located on Mount Tronador in the Nahuel Huapi National Park (Northwestern Patagonia, Argentina) is presented. Subsurface water samples were filtered for colony counting and yeast isolation. The total yeast count ranged between 6 and 360 CFU L(-1). Physiologic and molecular methods were employed to identify 86 yeast isolates. In agreement with yeast diversity data from studies for Antarctic and Alpine glaciers, the genera Cryptococcus, Leucosporidiella, Dioszegia, Rhodotorula, Rhodosporidium, Mrakia, Sporobolomyces, Udeniomyces and Candida were found. Cryptococcus and Leucosporidiella accounted for 50% and 20% of the total number of strains, respectively. Among 21 identified yeast species, Cryptococcus sp. 1 and Leucosporidiella fragaria were the most frequent. The typically psychrophilic Mrakia yeast strain and three new yeast species, yet to be described, were also isolated. All yeast strains were able to grow at 5, 10, and 15 degrees C. Among yeast strains expressing extracellular enzymatic activity, higher proteolytic and lipolytic activities were obtained at 4 degrees C than at 20 degrees C.     

Rhodotorula svalbardensis sp. nov., a novel yeast species isolated from cryoconite holes of Ny-Ålesund,Arctic

A psychrophilic yeast species was isolated from glacier cryoconite holes of Svalbard. Nucleotide sequences of the strains were studied using D1/D2 domain, ITS region and partial sequences of mitochondrial cytochrome b gene. The strains belonged to a clade of psychrophilic yeasts, but showed marked differences from related species in the D1/D2 domain and biochemical characters. Effects of temperature, salt and media on growth of the cultures were also studied. Screening of the cultures for amylase, cellulase, protease, lipase, urease and catalase activities was carried out. The strains expressed high amylase and lipase activities. Freeze tolerance ability of the isolates indicated the formation of unique hexagonal ice crystal structures due to presence of ‘antifreeze proteins’ (AFPs). FAME analysis of cultures showed a unique trend of increase in unsaturated fatty acids with decrease in temperature. The major fatty acids recorded were oleic acid, linoleic acid, linolenic acid, palmitic acid, stearic acid, myristic acid and pentadecanoic acid. Based on sequence data and, physiological and morphological properties of the strains, we propose a novel species, Rhodotorula svalbardensis and designate strains MLB-I (CCP-II) and CRY-YB-1 (CBS 12863, JCM 19699, JCM 19700, MTCC 10952) as its type strains (Etymology: sval.bar.den′sis. N.L. fem. adj. svalbardensis pertaining to Svalbard).     

New data on the distribution of certain psychrophilic yeasts in Moscow oblast

Several rare species and varieties of psychrophilic yeasts were isolated from the Sphagnum mosses and paludal vascular plants of Moscow oblast. Based on their 26S rDNA D1/D2 nucleotide sequences, they were assigned to the species Sterigmatosporidium polymorphum, Rhodotorula psychrophenolica, and Aureobasidium pullulans var. subglaciale. Thus, a new habitat of S. polymorphum was found and yeasts known previously only for the Alpine environments and the Arctic region have been isolated in the central regions of Russia for the first time.     

Alkalitolerant yeasts from natural biotopes

Using a solid nutrient medium containing alkaline buffer (pH 10) and an antibiotic, alkalitolerant yeasts were isolated from samples of soda-rich saline soils (solonchaks) of Armenia (Arazdayan) and the Transbaikal Region (the Kungur Steppe). The species diversity of the yeast populations of the tested soda-rich soils was relatively insignificant. They only contained alkalitolerant representatives of asporogenic capsulated yeasts belonging to the species Cryptococcus laurentii, C. albidus, Rhodotorula glutinis, R. mucilaginosa, and Sporobolomyces roseus. C. laurentii representatives clearly dominated the isolates obtained, their number exceeding that of the other species by 2-3 orders of magnitude. All of the isolates grew on acidic wort agar, suggesting that they did not include obligate alkaliphiles.     

Diversity of Yeasts from Puddles in the Vicinity of Midre Lovénbreen Glacier, Arctic and Bioprospecting for Enzymes and Fatty Acids

A total of 132 yeast strains were characterised from 4 sediment samples collected from small puddles in the vicinity of Midre Lovénbreen glacier, Arctic. Based on the D1/D2 domain sequence similarity, the isolates could be categorised into 6 groups. The nearest phylogenetic neighbour of groups I to VI were identified as Cryptococcus gastricus, Cryptococcus terricolus, Rhodotorula muscorum, Mrakia psychrophila, Mrakia gelida and Rhodotorula glacialis, respectively. Strains representative of the six groups were psychrophilic and salt tolerant but varied in their ability to produce cold-active extracellular enzymes such as lipase, protease, pectinase, cellulase and amylase. C_{18:1 (w9C)} and C_{18:2 (w9,12C)} were the only two fatty acids common to all the yeasts and branched and (or) unsaturated fatty acids increased in yeasts growing at 8°C compared to 22°C, probably as an adaptation to low temperature. The present study establishes that psychrophilic yeasts are predominant in Arctic and could be used as work horses to produce cold-active enzymes and poly unsaturated fatty acids which have been implicated in low temperature adaptation and also for their use in biotechnology.     

Relative Incidence of Ascomycetous Yeasts in Arctic Coastal Environments

Previous studies of fungi in polar environments have revealed a prevalence of basidiomycetous yeasts in soil and in subglacial environments of polythermal glaciers. Ascomycetous yeasts have rarely been reported from extremely cold natural environments, even though they are known contaminants of frozen foods. Using media with low water activity, we have isolated various yeast species from the subglacial ice of four glaciers from the coastal Arctic environment of Kongsfjorden, Spitzbergen, including Debaryomyces hansenii and Pichia guillermondii, with counts reaching 10⁴ CFU L⁻¹. Together with the basidiomycetes Cryptococcus liquefaciens and Rhodotorula mucilaginosa, these yeasts represent the stable core of the subglacial yeast communities. Other glacial ascomycetous species isolated included Candida parapsilosis and a putative new species that resembles Candida pseudorugosa. The archiascomycete Protomyces inouyei has seldom been detected anywhere in the world but was here recovered from ice in a glacier cave. The glacier meltwater contained only D. hansenii, whereas the seawater contained D. hansenii, Debaryomyces maramus, Pichia guilliermondii, what appears to represent a novel species resembling Candida galli and Metschnikowia bicuspidata. Only P. guilliermondii was isolated from sea ice, while snow/ice in the fjord tidal zone included C. parapsilosis, D. hansenii, P. guilliermondii and Metschnikowia zobellii. All of these isolated strains were characterized as psychrotolerant and xero/halotolerant, with the exception of P. inouyei.     

Isolation and phylogenetic classification of culturable psychrophilic prokaryotes from the Collins glacier in the Antarctica

Culturable psychrophilic prokaryotes were obtained of samples of glacier sediment, seaside mud, glacier melted ice, and Deschampsia antarctica rhizosphere from Collins glacier, Antarctica. The taxonomic classification was done by a culture-dependent molecular approach involving the Amplified Ribosomal DNA Restriction Analysis. Two hundred sixty colonies were successfully isolated and sub-cultivated under laboratory conditions. The analysis showed a bacterial profile dominated by Beta-proteobacteria (35.2%) followed by Gamma-proteobacteria (18.5%), Alpha-proteobacteria (16.6%), Gram-positive with high GC content (13%), Cytophaga–Flavobacterium–Bacteroides (13%) and Gram-positive with low GC content (3.7%). Eleven of the isolates have been reported previously and the others microorganisms remain uncharacterized. The isolated microorganisms here could be a potential source for biotechnological products, such as cold-active enzymes and secondary metabolites.     

异龙湖湿地修复后可培养酵母菌的多样性及其与理化因子的相关性

【目的】研究湿地修复后异龙湖中可培养酵母菌多样性及其与理化因子的相关性。【方法】对异龙湖湖水中可培养酵母菌进行DNA提取和测序,通过分析26SrRNA基因的D1/D2区域和形态以及生理生化特征对酵母菌进行鉴定,测定各个水样点的总有机碳(TOC)、总氮(TN)、总磷(TP)、总硬度(TH)和电导率(Cond),运用R 4.0.5、Canoco 5软件来分析异龙湖湖水中可培养酵母菌多样性和与理化因子的相关性。【结果】从湿地修复后的异龙湖湖水中分离获得519株可培养酵母菌,鉴定为24个属42个种和1个潜在的新种,胶红酵母(Rhodotorula mucilaginosa)、皮肤皮状新丝孢酵母(Cutaneotrichosporon dermatis)以及产黑色素短梗霉(Aureobasidium melanogenum)是异龙湖中的优势种,分别分离到了320株(52.29%)、40株(7.71%)和37株(7.13%)。【结论】湿地修复后的异龙湖具有丰富的可培养酵母菌资源,其群落结构较湿地修复前有了明显的变化,湖区西北部的可培养酵母菌多样性要比湖区东南部丰富。TN与异龙湖中可培养酵母菌多样性...     

Occurrence and dominance of yeast species in naturally fermented milk from the Tibetan Plateau of China

To determine which yeasts are present in the naturally fermented milks of China, 69 samples made by the nomads of Tibet were collected from the Tibetan Plateau in China. From these samples, 225 strains of yeast were isolated and identified using conventional microbiological analysis and gene sequencing analysis of the D1/D2 domain of the large subunit (26S) ribosomal DNA. The results showed that the total concentration of yeasts in these samples ranged from 5.01 to 8.97 log10 colony-forming units (CFU)/mL (6.91?± 1.02 log10 CFU/mL; mean?± SD). The number of cultivable yeasts was higher in the samples from Qinghai (7.55?± 0.75 log10 CFU/mL) than those from Tibet (6.21?± 0.79 log10 CFU/mL, P?< 0.05). Moreover, there were 15 phylotypes in these 69 samples. Among these phylotypes, Kluyveromyces marxianus (49.3%, frequency percentage), Saccharomyces cerevisiae (62.3%), and Pichia fermentans (46.4%) appeared frequently and can be considered the most common culturable species in naturally fermented milk products. Traditional fermented Mongolian cow milk featured a wide diversity of yeast species, including Issatchenkia orientalis, Kazachstania unisporus, Rhodotorula mucilaginosa, Candida pararugosa, Torulaspora delbrueckii, Geotrichum sp., Kazachstania unisporus, Geotrichum fragrans, Debaryomyces hansenii, Yarrowia lipolytica, Trichosporon gracile, and Pichia membranifaciens. This study provides new data on yeast composition in naturally fermented milk and shows the yeast biodiversity of fermented milk products from the Tibetan Plateau of China.     

Alkalitolerant Yeasts from Natural Biotopes

Using a solid nutrient medium containing alkaline buffer (pH 10) and an antibiotic, alkalitolerant yeasts were isolated from samples of soda-rich saline soils (solonchaks) of Armenia (Arazdayan) and the Transbaikal region (the Kungur Steppe). The species diversity of the yeast populations of the tested soda-rich soils was relatively insignificant. They only contained alkalitolerant representatives of asporogenic capsulated yeasts belonging to the species Cryptococcus laurentii, C. albidus, Rhodotorula glutinis, R. mucilaginosa,and Sporobolomyces roseus. C. laurentii representatives clearly dominated the isolates obtained, their number exceeding that of the other species by two to three orders of magnitude. All of the isolates grew on acidic wort agar, suggesting that they did not include obligate alkaliphiles.     

Phylogenetic Diversity of Numerically Important Arctic Sea-Ice Bacteria Cultured at Subzero Temperature

Heterotrophic bacteria in sea ice play a key role in carbon cycling, but little is known about the predominant players at the phylogenetic level. In a study of both algal bands and clear ice habitats within summertime Arctic pack ice from the Chukchi Sea, we determined the abundance of total bacteria and actively respiring cells in melted ice samples using epifluorescence microscopy and the stains 4', 6'-diamidino-2-phenylindole 2HCl (DAPI) and 5-cyano-2,3-ditolyl tetrazolium chloride (CTC), respectively. Organic-rich and -poor culturing media were used to determine culturable members by plating (at 0 degrees C and 5 degrees C) and most-probable-number (MPN) analyses (at -1 degrees C). Total bacterial counts ranged from 5.44 x 10(4) ml(-1) in clear ice to 2.41 x 10(6) ml(-1) in algal-band ice samples, with 2-27% metabolically active by CTC stain. Plating and MPN results revealed a high degree of culturability in both types of media, but greater success in oligotrophic media (to 62% of total abundance) and from clear ice samples. The bacterial enumeration anomaly, commonly held to mean 相似文献   

Sources and export fluxes of inorganic and organic carbon and nutrient species from the seasonally ice-covered Yukon River

Climate and environmental changes are having profound impacts on Arctic river basins, but the biogeochemical response remains poorly understood. To examine the effect of ice formation on temporal variations in composition and fluxes of carbon and nutrient species, monthly water and particulate samples collected from the lower Yukon River between July 2004 and September 2005 were measured for concentrations of organic and inorganic C, N, and P, dissolved silicate (Si(OH)₄), and stable isotope composition (δD and δ¹⁸O). All organic carbon and nutrient species had the highest concentration during spring freshet and the lowest during the winter season under the ice, indicating dominant sources from snowmelt and flushing of soils in the drainage basin. In contrast, inorganic species such as dissolved inorganic carbon (DIC) and Si(OH)₄ had the highest concentrations in winter and the lowest during spring freshet, suggesting dilution during snowmelt and sources from groundwater and leaching/weathering of mineral layer. The contrasting relation with discharge between organic, such as dissolved organic carbon (DOC), and inorganic, such as DIC and Si(OH)₄, indicates hydrological control of solute concentration but different sources and transport mechanisms for organic and inorganic carbon and nutrient species. Concentration of DOC also shows an inter-annual variability with higher DOC in 2005 (higher stream flow) than 2004 (lower stream flow). Average inorganic N/P molar ratio was 110?±?124, with up to 442 under the ice and 38–70 during the ice-open season. While dissolved organic matter had a higher C/N ratio under the ice (45–62), the particulate C/N ratio was lower during winter (21–26) and spring freshet (19). Apparent fractionation factors of C, N, P, Si and δD and δ¹⁸O between ice and river water varied considerably, with high values for inorganic species such as DIC and Si(OH)₄ (45 and 9550, respectively) but lower values for DOC (4.7). River ice formation may result in fractionation of inorganic and organic solutes and the repartitioning of seasonal flux of carbon and nutrient species. Annual export flux from the Yukon River basin was 1.6?×?10¹² g-DOC, 4.4?×?10¹² g-DIC, and 0.89?×?10¹² g-POC during 2004–2005. Flux estimation without spring freshet sampling results in considerable underestimation for organic species but significant overestimation for inorganic species regardless of the flux estimation methods used. Without time-series sampling that includes frozen season, an over- or under-estimation in carbon and nutrient fluxes will occur depending on chemical species. Large differences in carbon export fluxes between studies and sampling years indicate that intensive sampling together with long-term observations are needed to determine the response of the Yukon River to a changing climate.     

Yeast contaminants of micropropagated plant cultures

L eifert , C., W aites , W.M., N icholas , J.R. & K eetley , J.W. 1990. Yeast contaminants of micropropagated plant cultures. Journal of Applied Bacteriology 69 , 471–476.
Of 36 yeast strains isolated from contaminated plant cultures 78% were Candida, 20% Rhodotorula and 2% were Cryptococcus species. Strains of Candida guilliermon-dii represented 45% of all yeasts isolated. Yeasts grew rapidly on plant growth media with a pH of between 2.5 and 6.0 and decreased the medium pH to values of between 2.0 and 3.0. Candida yeasts growing on plant medium for 28 d metabolized about 75% of the sucrose and produced fermentation products such as ethanol and acetic acid. In contrast, Rhodotorula did not produce ethanol or acetic acid and only metabolized about 20% of the sucrose. Possible sources of contamination are discussed.     

Seasonal and altitudinal changes of culturable bacterial and yeast diversity in Alpine forest soils

The effect of altitude and season on abundance and diversity of the culturable heterotrophic bacterial and yeast community was examined at four forest sites in the Italian Alps along an altitude gradient (545–2000 m). Independently of altitude, bacteria isolated at 0 °C (psychrophiles) were less numerous than those recovered at 20 °C. In autumn, psychrophilic bacterial population increased with altitude. The 1194 bacterial strains were primarily affiliated with the classes Alpha-, Beta-, Gammaproteobacteria, Spingobacteriia and Flavobacteriia. Fifty-seven of 112 operational taxonomic units represented potential novel species. Strains isolated at 20 °C had a higher diversity and showed similarities in taxa composition and abundance, regardless of altitude or season, while strains isolated at 0 °C showed differences in community composition at lower and higher altitudes. In contrast to bacteria, yeast diversity was season-dependent: site- and altitude-specific effects on yeast diversity were only detected in spring. Isolation temperature affected the relative proportions of yeast genera. Isolations recovered 719 strains, belonging to the classes Dothideomycetes, Saccharomycetes, Tremellomycetes and Mycrobotryomycetes. The presence of few dominant bacterial OTUs and yeast species indicated a resilient microbial population that is not affected by season or altitude. Soil nutrient contents influenced significantly abundance and diversity of culturable bacteria, but not of culturable yeasts.     

Polyphasic identification of yeasts isolated from bark of cork oak during the manufacturing process of cork stoppers

A two-step protocol was used for the identification of 52 yeasts isolated from bark of cork oak at initial stages of the manufacturing process of cork stoppers. The first step in the identification was the separation of the isolates into groups by their physiological properties and RFLPs of the ITS-5.8S rRNA gene. The second step was the sequencing of the D1/D2 domains of the 26S rRNA gene of selected isolates representing the different groups. The results revealed a predominance of basidiomycetous yeasts (11 species), while only two species represented the ascomycetous yeasts. Among the basidiomycetous yeasts, members representing the species Rhodosporidium kratochvilovae and Rhodotorula nothofagi, that have been previously isolated from plant material, were the most abundant. Yeasts pertaining to the species Debaryomyces hansenii var. fabryii, Rhodotorula mucilaginosa and Trichosporon mucoides were isolated in small numbers.     

北极高维度海域海冰嗜冷菌系统发育多样性及其低温水解酶分析

应用样品直接稀释涂布平板、-1℃富集培养和-20℃冷冻24h后富集培养等3种方法,从北极加拿大海盆和格陵兰海的高纬度海域(77°30′N~81°12′N)海冰中分离到37株嗜冷菌。根据其16S rDNA全长序列所进行的系统发育分析表明,分离菌株分属于γ_变形细菌群(γ_Proteobacteria)的Colwellia、Marinobacter、Shewanella、Thalassomonas、Glaciecola、Marinomonas、Pseudoalteromonas和嗜纤维菌_曲挠杆菌_拟杆菌群(Cytophaga_Flexibacter_Bacteroide,CFB)的Flavobacterium、Psychroflexus等9个属。其中有9株菌的16S rDNA序列与已明确鉴定种的相似性在93.4%~96.9%,为潜在的新种。北极加拿大海盆海冰细菌BSi20002与南极威德尔海海冰细菌Marinobactersp.ANT8277的16S rDNA序列相似性为100%,表明在种水平上南、北两极也存在相同的细菌。分离的嗜冷菌在4℃条件下能产生多种大分子物质水解酶类,其中62.6%、51.4%和40.5%的菌株分别能水解Tween_80、明胶和淀粉。     

The presence of some yeasts species in occupational dermatitis

The samples of skin lesions were collected from forearms, palms and phalanges of ten females working in a manufacturing section (clothes tailoring and assemblage) using imported textiles. The samples were examined for bacteriological and mycotic flora by using classical methods. The textile fragments of 1-2 square centimeters from four samples were investigated using the same classical methods. The gauze fragments were used as toxic negative control samples. In 6 out of 10 samples from cutaneous lesions, Candida albicans was isolated and in 5 samples Rhodotorula species was identified. The yeasts species were identified by cultural and biochemical characteristics. All C. albicans strains were positive for the germ tube test and produced spherical chlamydospores on corn meal agar. The Rhodotorula strains grew on Sabouraud agar pure orange-red pigmented and mucoid colonies of 2-3 mm diameter. The textile samples incubated on blood--agar (10% sheep blood) demonstrated a high toxic activity producing a surrounding beta-hemolysis area of 3-5 cm. This lysis of the sheep red cell was caused by an unidentified chemical agent. In our cases, dermatites were associated with chemical and yeasts agents. The chemical agents irritated and made the derma sensitive, favouring C. albicans and Rhodotorula species multiplication.     

Diversity and association of psychrophilic bacteria in Antarctic sea ice.

The bacterial populations associated with sea ice sampled from Antarctic coastal areas were investigated by use of a phenotypic approach and a phylogenetic approach based on genes encoding 16S rRNA (16S rDNA). The diversity of bacteria associated with sea ice was also compared with the bacterial diversity of seawater underlying sea ice. Psychrophilic (optimal growth temperature, < or = 15 degrees C; no growth occurring at 20 degrees C) bacterial diversity was found to be significantly enriched in sea ice samples possessing platelet and bottom ice diatom assemblages, with 2 to 9 distinct (average, 5.6 +/- 1.8) psychrophilic taxa isolated per sample. Substantially fewer psychrophilic isolates were recovered from ice cores with a low or negligible population of ice diatoms or from under-ice seawater samples (less than one distinct taxon isolated per sample). In addition, psychrophilic taxa that were isolated from under-ice seawater samples were in general phylogenetically distinct from psychrophilic taxa isolated from sea ice cores. The taxonomic distributions of psychrotrophic bacterial isolates (optimal growth temperature, > 20 degrees C; growth can occur at approximately 4 degrees C) isolated from sea ice cores and under-ice seawater were quite similar. Overall, bacterial isolates from Antarctic sea ice were found to belong to four phylogenetic groups, the alpha and gamma subdivisions of the Proteobacteria, the gram-positive branch, and the Flexibacter-Bacteroides-Cytophaga phylum. Most of the sea ice strains examined appeared to be novel taxa based on phylogenetic comparisons, with 45% of the strains being psychrophilic. 16S rDNA sequence analysis revealed that psychrophilic strains belonged to the genera Colwellia, Shewanella, Marinobacter, Planococcus, and novel phylogenetic lineages adjacent to Colwellia and Alteromonas and within the Flexibacter-Bacteroides-Cytophaga phylum. Psychrotrophic strains were found to be members of the genera Pseudoalteromonas, Psychrobacter, Halomonas, Pseudomonas, Hyphomonas, Sphingomonas, Arthrobacter, Planococcus, and Halobacillus. From this survey, it is proposed that ice diatom assemblages provide niches conducive to the proliferation of a diverse array of psychrophilic bacterial species.