目的：我国冰川冻土资源丰富，但相关低温微生物的开发研究还比较落后，国内在低温酵母菌的研究基本为零。随着全球气候变暖逐渐加剧，冰川逐年退化，因此对低温微生物的研究迫在眉睫。在传统发酵技术和生物技术应用中，酵母菌是与其联系最为密切的真核微生物，并且与细菌相比酵母菌能够更好地适应低温环境。低温环境中的一些酵母菌特有的功能在生物、医药、洗涤、食品加工等行业有极好的应用前景。本课题在了解低温酵母菌表型、生理生化特性的基础上，研究低温酵母菌产酶特性并用分子生物学技术和统计学方法分析低温酵母菌在不同栖息环境中的生物多样性及生态格局分布。以期揭示低温酵母菌在我国天山1号冰川冷环境栖息地中的生态学意义，以及为低温酵母菌资源的开发、低温酶的研究及其在食品工业的应用提供基础资料。

方法：利用RB、DG、MEA、YEPG四种不同的培养基从新疆天山1号冰川不同生境的6种样品中，分离筛选出低温酵母菌；根据表性特征及生理生化特性，结合MSP-PCR指纹图谱及ITS rRNA基因序列测序分析确定低温酵母菌种属间进化关系，生物多样性及生态分布，并研究其产低温酶特性。

结论：利用4种培养基对天山1号冰川表面融水、底部融水、底部沉积物、冰川表面雪、冰川前沿积水及冰尘6种不同栖息环境中的低温酵母菌进行分离鉴定。共分离得到纯培养物629株，其中152株分离自冰川表面融水，96株分离自冰川底部沉积物，96株分离自冰川前沿积水，78株分离自冰尘；冰川表面雪中分离到的菌株最多共182株；冰川底部融水中最少，只分得25株。

ITSr RNA基因测序分析结果显示，629株酵母菌主要分属于Cryptococcus、Cystofilobasidium、Dioszegia、Filobasidium、Leucosporidium、Leucosporidiella、Microbotryum、Schizonella、Mrakia、Mrakiella、Rhodotorula、Sporidiobolus、Sporobolomyce、Tilletiopsi、Udeniomyce、Aureobasidium、Candida、Hanseniaspora、Pichia、Yarrowia和Lecythophora共21个属，51个种。所得菌株中Cryptococcus酵母菌最多，Candida次之，分别占总菌株数的46.5%和27.9%。Cryptococcus和Candida是各生境中的优势菌群。Cryptococcus victoriae和Candida akabanensis酵母菌数量最多且分布广泛，是冰川冷环境中的主要酵母种。冰川表面积雪中物种相对丰度最高，有较好的生物多样性。冰川底部沉积物中酵母菌物种相对丰度最低，生物多样性最差。

酵母菌生理生化实验结果表明，大部分酵母菌都是耐冷菌，生长温度范围在8-25℃之间，只有少数最适生长温度范围在12-16℃左右为嗜冷菌。耐盐性结果发现，大多数酵母菌在NaCl浓度达8%时停止生长，近60%的菌株最大耐盐度不超过6.5%，只有极个别酵母菌耐盐度较低，最大耐盐度不超过5%。产酶特性研究结果显示，共有产酶菌株共45株，其中产脂肪酶菌株37株，有24株产蛋白酶，31株产淀粉酶。Rhodotorula酵母菌都可以产脂肪酶和淀粉酶；Sporobolomyces几乎3种酶都产；Udeniomyces、Tilletiopsis、Hanseniaspora、Cystofilobasidium等属的酵母菌几乎都不产酶；Rhodotorula属酵母菌几乎都不产蛋白酶；Pichia属酵母菌只产蛋白酶；Yarrowia lipolytica、Aureobasidium pullulans 和Leucosporidiella fragaria三种酶都产。

Objective: Ice and frozen soil resources are very abundant in our country, but development research on cold-adapted microorganisms is still relatively backward, the domestic research in psychrophilic yeast is rare. As global warming, glaciers degraded with each passing year, study of psychrophile is around the corner. Yeasts are probably one of the most relevant microbial groups in both traditional fermentation technologies and biotechnological applications.And compared with bacteria, yeast can adapt to the low temperature environment much better. Some unique features of cold-adapted yeast have excellent application prospect in biology, medicine, washing and food industry. This paper aims to study the enzyme production features, biodiversity and ecological distribution pattern of the cold-adapted yeasts in different habitats by using molecular biology techniques and statistical methods, in basis of understanding the phenotype, physiological and biochemical characteristics of cold-adapted yeasts. In order to reveal the ecological significance of cold-adapted yeasts in cold habitats of Glacier No.1 in the Tianshan Mountains, and also provide basic information for bioprospecting in cold-adapted yeasts resources and their application in food industry.

Methods: In this experiment, four kinds of culture medium RB, DG, MEA, YEPG were used for isolating cold-adapted yeasts from 6 different habitats of Glacier No.1 in the Tianshan Mountains; The ecological distribution pattern, biodiversity and evolutionary relationship between yeast species were determinied according to the physiological and biochemical characteristics, MSP-PCR fingerprint and analysis of ITS rRNA sequences. The ability of yeasts synthesizing extracellular enzymes were tested using the culture mediums with different substrates.

Conclusion: A total of 629 strains were isolated by using four kinds of culture medium from 6 different habitats of Glacier No. 1 in the Tianshan Mountains. Among which, 152 from the supraglacial meltwater, 96 from the subglacial sediments, 96 from the water collected from puddles in the vicinity of the glacier, 78 from cryoconites; and the largest number of yeast strains were obtained from supraglacial snow, a total of 182 strains; yeasts in subglacial meltwater were the least, from which only 25 yeast strains were isolated.

Aalysis of the ITS rRNA gene sequences show that, the 629 yeast strains mainly belong to Cryptococcus, Cystofilobasidium, Dioszegia, Filobasidium, Leucosporidium, Leucosporidiella, Microbotryum, Schizonella, Mrakia, Mrakiella, Rhodotorula, Sporidiobolus, Sporobolomyce, Tilletiopsi, Udeniomyce, Aureobasidium, Candida, Hanseniaspora, Pichia, Yarrowia and Lecythophora, a total of 21 genus, 51 species. The number of Cryptococcus yeasts was the most, and for 46.5% of the yeasts were Cryptococcus. The number of Candida yeasts was less than the number of Cryptococcus. Species of Cryptococcus victoriae and Candida akabanensis were the most abundant yeast species, and they were widely distributed, and they were the dominant yeast species in glacial habitats. The species relative abundance of cold-adapted yeasts in snow was the highest, and in which it had a good biodiversity. And a very low species dominance was observed in subglacial meltwater.

The physiological and biochemical experiments show that most of the yeast strains were psychrotolerant. Their growth temperature ranged from 8 to 25℃; Only a minority of the yeasts optimum growth temperature were ranging from 12 to 16℃, and they were identified as psychrophilic yeasts. NaCl tolerance experiments found that most of the yeasts stopped growing when the concen

Objective: Ice and frozen soil resources are very abundant in our country, but development research on cold-adapted microorganisms is still relatively backward, the domestic research in psychrophilic yeast is rare. As global warming, glaciers degraded with each passing year, study of psychrophile is around the corner. Yeasts are probably one of the most relevant microbial groups in both traditional fermentation technologies and biotechnological applications.And compared with bacteria, yeast can adapt to the low temperature environment much better. Some unique features of cold-adapted yeast have excellent application prospect in biology, medicine, washing and food industry. This paper aims to study the enzyme production features, biodiversity and ecological distribution pattern of the cold-adapted yeasts in different habitats by using molecular biology techniques and statistical methods, in basis of

[1] Buzzini P, Branda E, Goretti M, et al. Psychrophilic yeasts from worldwide glacial habitats: diversity, adaptation strategies and biotechnological potential[J]. FEMS Microbiology Ecology, 2012, 82(2):217-41.

[2] Russell N, Cowan D A. 16 Handling of Psychrophilic Microorganisms[J]. Methods in Microbiology, 2006, 35(08):371-393.

[3] Liu YQ.Yao TD.Jiao NZ.Kang SC.Huang SJ .Li Q.Wang KJ. Liu XB. Culturable bacteria in glacial meliwater at 6,350 m on the East Rongbuk Glacier, Mount Everest[J]. Extremophiles, 2009, 13: 89-99.

[3] Sangorrín M P, Lopes C A, Vero S, et al. Cold-Adapted Yeasts as Biocontrol Agents: Biodiversity, Adaptation Strategies and Biocontrol Potential[M]//Cold-adapted Yeasts. Springer Berlin Heidelberg, 2014:441-464.

[4] Buford P, Todd S. Temperature dependence of metabolic rates for microbial growth, maintenance, and survival[J]. Proceedings of the National Academy of Sciences of the United States of America, 2004, 101(13):4631-6.

[5] Held B W, Jurgens J A, Duncan S M, et al. Assessment of fungal diversity and deterioration in a wooden structure at New Harbor, Antarctica[J]. Polar Biology, 2006, 29(6):526-531.

[6] Shivaji S, Prasad G S. Antarctic Yeasts: Biodiversity and Potential Applications[M]// Yeast Biotechnology: Diversity and Applications. Springer Netherlands, 2009:3-18.

[7] Vishniac H S. Yeast Biodiversity in the Antarctic[M]// Biodiversity and Ecophysiology of Yeasts. Springer Berlin Heidelberg, 2006:419-440.

[8] Connell L, Redman R, Craig S, et al. Diversity of Soil Yeasts Isolated from South Victoria Land, Antarctica[J]. Microbial Ecology, 2008, 56(3):448-59.

[9] Margesin R. Effect of temperature on growth parameters of psychrophilic bacteria and yeasts[J]. Extremophiles, 2009, 13(2):257-62.

[10] Vishniac H S. A multivariate analysis of soil yeasts isolated from a latitudinal gradient[J]. Microbial Ecology, 2006, 52(52):90-103.

[11] Goto S, Sugiyama J, Iizuka H. A taxonomic study of Antarctic yeasts[J]. Mycologia, 1969, 61(4):748-822.

[12] Panikov N S. Microbial Activity in Frozen Soils[J]. Permafrost Soils, 2008, 16:119-147.

[13] Gostincar C, Grube M, Hoog S D, et al. Extremotolerance in fungi: evolution on the edge[J]. FEMS Microbiology Ecology, 2009, 71(1):2-11.

[14] Bellemain E, Davey M L,Kauserud H,et al. Fungal palaeodiversity revealed using high-throughput metabarcoding of ancient DNA from arctic permafrost[J]. Environmental Microbiology, 2013, 15(4):1176-89.

[15] Uetake J, Kohshima S, Nakazawa F, et al. Evidence for propagation of cold-adapted yeast in an ice core from a Siberian Altai glacier[J]. Journal of Geophysical Research Biogeosciences, 2011, 116(G1):2144-2150.

[16] Shivaji S, Bhadra B, Rao R S, et al. Rhodotorula himalayensis sp. nov., a novel psychrophilic yeast isolated from Roopkund Lake of the Himalayan mountain ranges, India[J]. Extremophiles, 2008, 12(3):375-381.

[17] Hu W, Zhang Q, Tian T, et al. The microbial diversity, distribution, and ecology of permafrost in China: a review[J]. Extremophiles, 2015, 19(4):693-705.

[18] 姜远丽, 郑晓吉, 史学伟,等. 天山冻土中嗜冷酵母菌生物多样性[J]. 食品与生物技术学报, 2012, 31(12):1289-1294.

[19] 邵莎苑.青藏高原及其毗邻地区不同空间冰川雪坑中可培养酵母菌多样性研究[D]. 兰州大学, 2012.

[1] Buzzini P, Branda E, Goretti M, et al. Psychrophilic yeasts from worldwide glacial habitats: diversity, adaptation strategies and biotechnological potential[J]. FEMS Microbiology Ecology, 2012, 82(2):217-41.

[2] Russell N, Cowan D A. 16 Handling of Psychrophilic Microorganisms[J]. Methods in Microbiology, 2006, 35(08):371-393.

[3] Liu YQ.Yao TD.Jiao NZ.Kang SC.Huang SJ .Li Q.Wang KJ. Liu XB. Culturable bacteria in glacial meliwater at 6,350 m on the East Rongbuk Glacier, Mount Everest[J]. Extremophiles, 2009, 13: 89-99.

[3] Sangorrín M P, Lopes C A, Vero S, et al. Cold-Adapted Yeasts as Biocontrol Agents: Biodiversity, Adaptation Strategies and Biocontrol Potential[M]//Cold-adapted Yeasts. Springer Berlin Heidelberg, 2014:441-464.

[4] Buford P, Todd S. Temperature dependence of metabolic rates for microbial growth, maintenance, and survival[J]. Proceedings of the National Academy of Sciences of the United States of America, 2004, 101(13):4631-6.

[5] Held B W, Jurgens J A, Duncan S M, et al. Assessment of fungal diversity and deterioration in a wooden structure at New Harbor, Antarctica[J]. Polar Biology, 2006, 29(6):526-531.

[6] Shivaji S, Prasad G S. Antarctic Yeasts: Biodiversity and Potential Applications[M]// Yeast Biotechnology: Diversity and Applications. Springer Netherlands, 2009:3-18.

[7] Vishniac H S. Yeast Biodiversity in the Antarctic[M]// Biodiversity and Ecophysiology of Yeasts. Springer Berlin Heidelberg, 2006:419-440.

[8] Connell L, Redman R, Craig S, et al. Diversity of Soil Yeasts Isolated from South Victoria Land, Antarctica[J]. Microbial Ecology, 2008, 56(3):448-59.

[9] Margesin R. Effect of temperature on growth parameters of psychrophilic bacteria and yeasts[J]. Extremophiles, 2009, 13(2):257-62.

[10] Vishniac H S. A multivariate analysis of soil yeasts isolated from a latitudinal gradient[J]. Microbial Ecology, 2006, 52(52):90-103.

[11] Goto S, Sugiyama J, Iizuka H. A taxonomic study of Antarctic yeasts[J]. Mycologia, 1969, 61(4):748-822.

[12] Panikov N S. Microbial Activity in Frozen Soils[J]. Permafrost Soils, 2008, 16:119-147.

[13] Gostincar C, Grube M, Hoog S D, et al. Extremotolerance in fungi: evolution on the edge[J]. FEMS Microbiology Ecology, 2009, 71(1):2-11.

[14] Bellemain E, Davey M L,Kauserud H,et al. Fungal palaeodiversity revealed using high-throughput metabarcoding of ancient DNA from arctic permafrost[J]. Environmental Microbiology, 2013, 15(4):1176-89.

[15] Uetake J, Kohshima S, Nakazawa F, et al. Evidence for propagation of cold-adapted yeast in an ice core from a Siberian Altai glacier[J]. Journal of Geophysical Research Biogeosciences, 2011, 116(G1):2144-2150.

[16] Shivaji S, Bhadra B, Rao R S, et al. Rhodotorula himalayensis sp. nov., a novel psychrophilic yeast isolated from Roopkund Lake of the Himalayan mountain ranges, India[J]. Extremophiles, 2008, 12(3):375-381.

[17] Hu W, Zhang Q, Tian T, et al. The microbial diversity, distribution, and ecology of permafrost in China: a review[J]. Extremophiles, 2015, 19(4):693-705.

[18] 姜远丽, 郑晓吉, 史学伟,等. 天山冻土中嗜冷酵母菌生物多样性[J]. 食品与生物技术学报, 2012, 31(12):1289-1294.

[19] 邵莎苑.青藏高原及其毗邻地区不同空间冰川雪坑中可培养酵母菌多样性研究[D]. 兰州大学, 2012.

[20] 郑晓吉,孙海龙, 关波,等.天山一号冰川融水中耐冷酵母菌多样性及系统发育[J]. 食品与发酵工业, 2015, 41(7):45-50.

[21] Michael Z, Wilfried H, Martin H, et al. Alpine Glaciers to Disappear within Decades?[J]. Geophysical Research Letters, 2006, 331(13):1-4.

[22] Turchetti B, Buzzini P, Goretti M, et al. Psychrophilic yeasts in glacial environments of Alpine glaciers[J]. FEMS Microbiology Ecology, 2008, 63(1):73-83.

[23] Eva B, Benedetta T, Guglielmina D, et al. Yeast and yeast-like diversity in the southernmost glacier of Europe (Calderone Glacier, Apennines, Italy) [J]. FEMS Microbiology Ecology, 2010, 72(3):354-69.

[24] Skvarca P, Raup B, De Angelis H. Recent behaviour of Glaciar Upsala, a fast-flowing calving glacier in Lago Argentino, southern Patagonia[J]. Annals of Glaciology, 2003, 36(1):184-188.

[25] Margesin R, Miteva V. Diversity and ecology of psychrophilic microorganisms[J]. Research in Microbiology, 2011, 162(3):346-361.

[26] Libkind D, Brizzio S, Ruffini A, et al. Molecular characterization of carotenogenic yeasts from aquatic environments in Patagonia, Argentina[J]. Antonie Van Leeuwenhoek, 2003, 84(4):313-22.

[27] Frengova G I, Beshkova D M. Carotenoids from Rhodotorula and Phaffia: yeasts of biotechnological importance[J]. Journal of Industrial Microbiology and Biotechnology, 2009, 36(2):163-180.

[28] Chen M. Study on Carotenoid Pigments Produced by Red Yeasts[J]. Amino Acids and Biotic Resources, 1998.

[29] Gostin?ar C, Gunde-Cimerman N, Turk M. Environmental Impacts on Fatty Acid Composition of Fungal Membranes[J]. Fungi from Different Environments, 2009:278-327.

[30] Carman G M, Kersting M C. Phospholipid synthesis in yeast: regulation by phosphorylation[J]. Biochemistry and Cell Biology, 2004, 82(1):62-70.

[31] Rossi M, Buzzini P, Cordisco L, et al. Growth, lipid accumulation, and fatty acid composition in obligate psychrophilic, facultative psychrophilic, and mesophilic yeasts[J]. FEMS Microbiology Ecology, 2009, 69(3):363–372.

[32] Phadtare S, Alsina J, Inouye M, et al. Cold-shock response and cold-shock proteins[J].Current Opinion in Microbiology, 1999, 2(2):175–180.

[33] Poli A, Anzelmo G, Tommonaro G, et al. Production and chemical characterization of an exopolysaccharide synthesized by psychrophilic yeast strain Sporobolomyces salmonicolor AL? isolated from Livingston Island, Antarctica[J]. Folia Microbiologica, 2010, 55(6):576-81.

[34] Kawahara H. Cryoprotectants and Ice-Binding Proteins[M]. Springer Berlin Heidelberg, 2007.

[35] Pavlova K, Panchev I, Krachanova M, et al. Production of an exopolysaccharide by Antarctic yeast[J]. Folia Microbiologica, 2009, 54(4):343-8.

[36] Lee J K, Park K S, Park S, et al. An extracellular ice-binding glycoprotein from an Arctic psychrophilic yeast[J]. Cryobiology, 2010, 60(2):222-8.

[37] Richter K, Haslbeck M, Buchner J. The Heat Shock Response: Life on the Verge of Death[J]. Molecular Cell, 2010, 40(2):253–266.

[38] Atomi H, Sato T, Kanai T. Application of hyperthermophiles and their enzymes[J]. Current Opinion in Biotechnology, 2011, 22(5):618-26.

[39] Barbuddhe S, Chakraborty T. Biotechnological applications of psychrophiles[M]. Microbial Biotechnology, 2011, 4(5):685-686(2).

[40] María P D, Carboni-Oerlemans C, Tuin B, et al. Biotechnological applications of Candida antarctica lipase A: State-of-the-art[J]. Journal of Molecular Catalysis B Enzymatic, 2005, 37(1–6):36-46.

[41] Cavicchioli R, Charlton T, Ertan H, et al. Biotechnological uses of enzymes from psychrophiles[J]. Microbial Biotechnology, 2011, 4(4):449-60.

[42] Ray M K, Devi K U, Kumar G S, et al. Extracellular protease from the antarctic yeast Candida humicola[J]. Applied and Environmental Microbiology, 1992, 58(6): 1918–1923.

[43] Zlatanov M, Pavlova K, Antova G, et al. Biomass Production by Antarctic Yeast Strains: An investigation on the Lipid Composition[J]. Biotechnology and Biotechnological Equipment, 2014, 24(4):2096-2101.

[44] Krishnan A, Convey P, Gonzalez-Rocha G, et al. Production of extracellular hydrolase enzymes by fungi from King George Island[J]. Polar Biology, 2014, 39(1):1-12.

[45] Bergauer P, Fonteyne P A, Nolard N, et al. Biodegradation of phenol and phenol-related compounds by psychrophilic and cold-tolerant alpine yeasts[J].Chemosphere, 2005, 59(7):909-18.

[46] Silvia B, Benedetta T, Virginia D G, et al. Extracellular enzymatic activities of basidiomycetous yeasts isolated from glacial and subglacial waters of northwest Patagonia (Argentina)[J]. Canadian Journal of Microbiology, 2007, 53(4):519-525(7).

[47] Sitepu I R, Garay L A, Sestric R, et al. Oleaginous yeasts for biodiesel: Current and future trends in biology and production[J]. Biotechnology Advances, 2014, 32(7):1336-1360.

[48] Mishra V, Srivastava I, Hasija Y. A Review on Bio-hydrogen production: Current trends and Advances[C]// Indian Society for Technical Education (ISTE) Delhi Section Convention on “Technological Universities and Institutions in New Knowledge Age: Future Perspectives and Action Plan”. 2013.

[49] José I. Rovati , Hipólito F. Pajot, Ruberto L, et al. Polyphenolic substrates and dyes degradation by yeasts from 25 de Mayo/King George Island (Antarctica)[J]. Yeast, 2013, 30(11):459-70.

[50] Atlas R M. Handbook of Microbiological Media, Fourth Edition[J]. CRC Press, 2010.

[51] Fredricks D N, Smith C, Meier A. Comparison of six DNA extraction methods for recovery of fungal DNA as assessed by quantitative PCR[J]. Journal of Clinical Microbiology, 2005, 43(10):5122-5128.

[52] Garcia V, Brizzio S, Broock M R. Yeasts from glacial ice of Patagonian Andes, Argentina[J]. FEMS Microbiology Ecology, 2012, 82(2):540-50.

[53] Purnima S, Masaharu T, Shiv Mohan S, et al. Taxonomic characterization, adaptation strategies and biotechnological potential of cryophilic yeasts from ice cores of Midre Lovénbreen glacier, Svalbard, Arctic[J]. Cryobiology, 2013, 66(2):167-175.

[54] Brandão L R, Libkind D, Vaz A B M, et al. Yeasts from an oligotrophic lake in Patagonia (Argentina): diversity, distribution and synthesis of photoprotective compounds and extracellular enzymes[J]. FEMS Microbiology Ecology, 2011, 76(1):1-13.

[55] Selbmann L, Zucconi L, Onofri S, et al. Taxonomic and phenotypic characterization of yeasts isolated from worldwide cold rock-associated habitats[J]. Fungal Biology, 2014, 118(1):61-71.

[56] Silva-Bedoya L M, Ramírez-Castrillón M, Osorio-Cadavid E.Yeast diversity associated to sediments and water from two Colombian artificial lakes[J]. Brazilian journal of microbiology: publication of the Brazilian Society for Microbiology, 2014, 45(1):135-142.

[57] Li L, Singh P, Ying L, et al. Diversity and biochemical features of culturable fungi from the coastal waters of Southern China[J]. Amb Express, 2014, 4(1):1-11.

[58] Zhang W, Sun Z. Random local neighbor joining: A new method for reconstructing phylogenetic trees[J]. Molecular Phylogenetics and Evolution, 2008, 47(1):117-128.

[59] Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees[J]. Molecular Biology and Evolution, 1987, 4(6):406-425.

[60] Buzzini P, Martini A. Extracellular enzymatic activity profiles in yeast and yeast-like strains isolated from tropical environments[J]. Journal of Applied Microbiology, 2002, 93(6):1020-5.

[61] Pathan A A K, Bhadra B, Begum Z, et al. Diversity of Yeasts from Puddles in the Vicinity of Midre Lovénbreen Glacier, Arctic and Bioprospecting for Enzymes and Fatty Acids[J]. Current Microbiology, 2010, 60(4):307-314.

[62] Buzzini P, Vaughan-Martini A. Yeast Biodiversity and Biotechnology[M]. Yeast Handbook, 1970:533-559.

[63] Butinar L, Spencer-Martins I, Gunde-Cimerman N. Yeasts in high Arctic glaciers: the discovery of a new habitat for eukaryotic microorganisms[J]. Antonie Van Leeuwenhoek, 2007, 91(3):277-89.

[64] Turchetti B, Skye R. Thomas Hall, Laurie B. Connell, Eva Branda, Pietro Buzzini, Bart Theelen, Wally H. Muller, Teun Boekhout. Psychrophilic yeasts from Antarctica and European glaciers: description of Glaciozyma gen. nov., Glaciozyma martinii sp. nov. and Glaciozyma watsonii sp. nov[J]. Extremophiles, 2011, 15:573–586.

[65] Thevenieau F, Nicaud J M, Gaillardin C. Yeast Biotechnology: Diversity and Applications[J]. Yeast Biotechnology Diversity and Applications, 2009:590-613.

[66] Pathan A A K, Bhadra B, Begum Z, et al. Diversity of Yeasts from Puddles in the Vicinity of Midre Lovénbreen Glacier, Arctic and Bioprospecting for Enzymes and Fatty Acids[J]. Current Microbiology, 2010, 60(4):307-314.

[67] Eva B, Benedetta T, Guglielmina D, et al. Yeast and yeast-like diversity in the southernmost glacier of Europe (Calderone Glacier, Apennines, Italy)[J]. FEMS Microbiology Ecology, 2010, 72(3):354-69.

[68] Turchetti B, Buzzini P, Goretti M, et al. Psychrophilic yeasts in glacial environments of Alpine glaciers[J]. FEMS Microbiology Ecology, 2008, 63(1):73-83.

[69] García V D, Brizzio S, Libkind D, et al. Biodiversity of cold-adapted yeasts from glacial meltwater rivers in Patagonia,