摘要
对新疆地区和青海地区传统发酵乳制品中的酵母菌进行了分离及生理生化与分子生物学鉴定,同时对其中的酵母菌生物多样性进行了分析。本研究为进一步全面系统地了解新疆和青海地区传统发酵乳制品中的微生物组成提供了丰富的数据,以利于开发利用传统发酵乳制品中的酵母菌资源。
新疆地区28份传统发酵酸马奶中的酵母菌总数在3.12×104-2.23×106CFU/mL之间,主要集中分布于105和106CFU/mL,相对于同一样品中分离出的乳酸菌数量低1-2个对数级。青海地区11份酸山羊奶样品中的酵母菌总数在2.07×105-1.63×108 CFU/mL之间,比同一样品中的乳酸菌总数低2-3个对数级。青海地区25份酸牦牛奶样品中的酵母菌总数在2.60×106-2.00×109CFU/mL之间,主要集中在107和108 CFU/mL。其中青海省海北州酸牦牛奶样品中的酵母菌总数比乳酸菌总数低1-2个对数级;海南州酸牦牛奶样品中的酵母菌总数高于或接近乳酸菌总数。
新疆地区28份酸马奶样品中共分离到酵母菌87株,结合传统分类鉴定方法与26S rDNA序列分析方法对酵母菌进行鉴定。结果表明新疆地区发酵酸马奶中的酵母菌主要为Kazachstania unisporus 43株(占总分离株的49.4%)和Kluyveromyces marxianus 23株(26.4%),其他的酵母分离株分别是Saccharomyces cerevisiae 8株,Pichia membranaefaciens 2株,Pichia fermentans 6株,Pichia manshurica(Pichia galeiformis)2株和Pichia kudriavzevii(Issatchenkia orientalis)3株。将新疆酸马奶中酵母分离株的ITS扩增片段进行对比,通过菌株ITS区域之间的长度差异性对酸马奶中的酵母分离株进行分类,同时采用ITS序列分析的方法对酸马奶中部分酵母分离株进行鉴定。
新疆不同地区酸马奶中酵母菌的种类和数量呈现出多样性。K. unisporus在高达22个酸马奶样品中出现。K. unisporus - Kl. marxianus的发酵组合是酸马奶中酵母菌的优势组合。S. cerevisiae和Pichia属的菌株在酸马奶样品中没有单独出现的情况。对青海地区传统发酵乳制品中的酵母菌进行鉴定。青海2个地区的25份酸牦牛奶样品中共分离出74株酵母菌,其中S. cerevisiae 20株,Kl. marxianus 18株, K. unisporus 1株,Candida zeylanoides 2株,Rhodotorula mucilaginosa 1株,以及Pichia属的P. fermentans 31株和P. kudriavzevii 1株。青海地区酸牦牛奶中酵母菌的优势菌为P. fermentans (43.2%)、S. cerevisia(e27.0%)和Kl. marxianu(s24.3%)。P. fermentans - Kl. marxianus、P. fermentans - S. cerevisiae,以及Kl. marxianus - S. cerevisiae是青海传统发酵酸牦牛奶中酵母菌的优势组合。
青海海西州采集到的11份酸山羊奶样品中共分离出35株酵母菌。这些菌株经鉴定为Kl. marxianus 16株,P. fermentans 12株、S. cerevisiae 6株和Rh. mucilaginosa 1株。试验结果表明,青海地区酸山羊奶中酵母菌的优势菌为Kl. marxianus(45.7%)和P. fermentans (34.3%)。P. fermentans - S. cerevisiae和Kl. marxianus - S. cerevisiae是青海传统发酵酸山羊奶样品中的酵母菌优势组合。在酸山羊奶样品中没有分离出K. unisporus。
The yeast flora of traditional fermented milk products, such as koumiss, kurut and fermented goat milk in Xinjiang and Qinghai of China, were analyzed. The identification and biodiversity of the yeast involved in traditional fermented milk products were investigated. It forms an essential step towards the preservation and exploitation of the hidden potential of the wealth of yeast biodiversity. The results also provide the basic data for systematically understanding the characteristics of koumiss, kurut and fermented goat milk, give useful information for further utilization of yeast in traditional fermented milk products.
The counts of yeast in koumiss samples collected from Xinjiang were ranged from 3.12×104 CFU/mL to 2.23×106 CFU/mL. Generally, in koumiss samples, the counts of the yeasts were lower than those of lactic acid bacteria by one or two logarithm. A total of eighty-seven yeast strains were isolated from twenty-eight home-made koumiss samples. They were identified using standard conventional identification methods. And the results were combined with analysis of the large-subunit (26S) rDNA gene D1/D2 domain region sequences of them. The predominant strains proved to be mainly Kazachstania unisporus (49.4% of the total isolates) and Kluyveromyces marxianus (26.4%). Others were identified as Saccharomyces cerevisiae for eight strains, Pichia membranaefaciens for two strains, Pichia fermentans for six strains, Pichia manshurica (synonym Pichia galeiformis)for two strains and Pichia kudriavzevii (synonym Issatchenkia orientalis)for three strains. The combination of K. unisporus and Kl. marxianus was predominant group in koumiss samples in Xinjiang. And the strains of S. cerevisiae and Pichia sp. were never present alone.
Variation on the lengths of the ribosomal ITS region (ITS1, 5.8S rRNA and ITS2) of the different yeast strains in koumiss was observed. Three isolates and a type yeast strain were selected for analysis of ITS region sequences. The results confirmed that the ITS region sequencing was a reliable and useful method for yeast taxology.
The counts of yeast in kurut samples collected from Qinghai were ranged from 2.60×106 CFU/mL to 2.00×109 CFU/mL. They were lower than those of lactic acid bacteria by one or two logarithm in the samples of Haibei, but were slightly more than those of lactic acid bacteria in the samples of Hainan. A total of seventy-four yeast strains were isolated from twenty-five samples. The predominant strains proved to be mainly P. fermentans (43.2%), S. cerevisiae(27.0%) and Kl. marxianus (24.3%). Others were identified as K. unisporus for one strain, P. kudriavzevii for one strain, Rhodotorula mucilaginosa for one strain and Candida zeylanoides for two strains. P. fermentans - Kl. marxianus, P. fermentans - S. cerevisiae and Kl. marxianus - S. cerevisiae were the predominant groups in kurut samples collected from Qinghai.
The counts of yeast in fermented goat milk samples collected from Qinghai were ranged from 2.07×105 CFU/mL to 1.63×108 CFU/mL. They were lower than those of lactic acid bacteria by two or three logarithm in the same sample. A total of thirty-five yeast strains were isolated from eleven samples. The predominant strains proved to be mainly Kl. marxianus (45.7%) and P. fermentans (34.3%). Others were identified as S. cerevisiae for six strains and Rh. mucilaginosa for one strain. Kl. marxianus - S. cerevisiae and P. fermentans - S. cerevisiae were the predominant groups in fermented goat milk samples collected from Qinghai.
引文
1 Watanabe K,Fujimoto J,Sasamoto M,et al. Diversity of lactic acid bacteria and yeasts in Airag and Tarag, traditional fermented milk products of Mongolia[J]. World J. Microbiol Biotechnol, 2008, 24:1313-1325
2杨印民.蒙元时期的葡萄酒和马奶酒[J].历史教学问题,2007,4:65-78
3杨文华,德慧,张珉.马乳及乳产品利用[J].内蒙古民族大学学报,2008,14(2):89-90
4王建光,孙玉江,芒来.马奶与几种奶营养成分的比较研究[J].食品研究与开发,2006,27(8):146-149.
5 Malacarne M, Martuzzi F, Summer A, et al. Protein and fat composition of mare’s milk: some nutritional remarks with reference to human and cow’s milk[J]. International Dairy Journal, 2002, 12(11): 869-877
6刘洪元、常江,高昆,等.鲜马奶对小鼠免疫功能的影响[J].中国乳品工业,2006,34(7):37-39
7刘洪元,高昆.马奶及酸马奶(马奶酒)的营养价值和医疗作用[J].中国食物与营养,2003,4:45-46
8关家怀.风味独特具有食疗作用的酸马奶酒[J].中国食品, 1999,15:33
9孙天松,孟和毕力格,王俊国,等.新疆维吾尔自治区酸马奶化学组成分与微生物学分析[J].中国乳品工业,2005 , 33(10) :9-13
10 Ishii S.“koumiss”,a treasure for nomad[J].Onko Chishin, 2004, 41:87-93
11贺银凤,李少英,母智深,等.酸马奶酒中微生物的分离鉴定及抗菌特性的研究[J].农业工程学报,2002,18(2):91-95
12 Kenji U, Masahiro H, Hidemasa M,et al. Microbiota of‘airag’,‘tarag’and other kinds of fermented dairy products from nomad in Mongolia[J]. Animal Science Journal, 2007, 78(6): 650-658(9)
13 Naersong,Tanaka Y,Mori N,et al.Microbial flora of“Airag”,a traditional fermented milk of Inner Mongolia in China[J].Anim.Sci.Technol,1996,67:78-83
14 Shuangquan, Burentegusi, Bai Y, et al. Microflora in traditional starter cultures for fermented milk, hurunge, from Inner Mongolia, China[J]. Animal Science Journal, 2006, 77(2):235-241(7)
15 Ishii S,Kikuchi M,Takao S.Isolation and Identification of lactic acid bacteria and yeasts from“Chigo”in Inner Mongolia[J].China Anim.Sci.Technol.,1997,68(3):325-329
16 Ishii S.Study on the kumiss(airag)of Mongolian nomads after severe cold in the winters of 2000 and 2001[J].Milk Science,2003,52(1):49-52
17殷文政,乌尼,钱建伟,等.锡林郭勒牧区马奶酒生物活性动态的研究(一)[J].内蒙古农业大学学报(自然科学版),2002, 23(1):9-16
18刘芳,都立辉,沙莎,等.利用16SrRNA基因同源性分析鉴定两株明串珠菌[J].中国微生态学杂志,2006,18(6):427-429
19 An Y, Adachi Y, Ogawa Y. Classification of lactic acid bacteria isolated from chigee and mare milk collected in Inner Mongolia[J].Animal Science Journal,2004,75 (3) :245-252
20孟和毕力格,乌日娜,王立平,等.不同地区酸马奶中乳杆菌的分离及其生物学特性的研究[J].中国乳品工业,2004,32(11):6-11
21 Wu R, Wang L, Wang J, et al. Isolation and preliminary probiotic selection of lactobacilli from koumiss in Inner Mongolia[J]. Journal of Basic Microbiology, 2009, Published Online
22 Wang J G,Chen X,Liu W J,et al. Identification of Lactobacillus from Koumiss by Conventional and Molecular Methods[J]. European Food Research and Technology, 2008,227:1555-1561
23 Zhang W Y, Yu D L, Sun Z H, et al. Complete nucleotide sequence of plasmid plca36 isolated from Lactobacillus casei Zhang[J]. Plasmid,2008,60(2):131-135
24 F M德赖森等著,顾瑞霞等译.发酵乳科学与技术[M].东南大学出版社,1991:71-74,185-189
25 Koroleva N S.Starters for fermented milks. Section 4: Kefir and kumys starters[C]. Bulletin International Dairy Federation,1988,96-100
26 Montanari G, Zambonelli C, Grazia L, et al. Saccharomyces unisporus as the principal alcoholic fermentation microorganism of traditional koumiss[J]. Journal of Dairy Research, 1996, 63(2):327-331
27 Lozovich S.Medical uses of whole and fermented mare milk in Russia[J].Cultured Dairy Products Journal,1995,30(1):18-19(21)
28 Mitchell G E,Fedrick I A,Rogers S. The relationship between somatk cell count,composition and manufacturing propereies of bulk milk[J]. Australlan Journal of Dariy Technology,1986,41(3):12-14
29李星科,李开雄,邹圣东.酸马奶酒[J].乳品加工,2006,07:58-60
30乌·扎木苏.酸马奶疗法[M].呼和浩特:内蒙古人民出版社,1986
31呼春,达古拉.蒙古族酸马奶疗法[J],中国民族医药杂志,1996,2(1):32
32敖道夫,布日额.酸马奶酒的研究进展[J].中国民族医药杂志,2008,4:65-67
33张和平,那日苏.蒙古族的酸马奶[J].中国乳品工业,1993,21(4):165-168
34 Barbano D M,Rasmussen R R,Lynch J M. Influence of milk somatic cell count and milk age on cheese yield[J]. Journal of Dairy Science,1991,74(2):369-388
35查哈达.酸马奶治疗肺结核56例临床观察[J].中国民族民间医药杂志,1999,36:13-14
36苏雅拉吉日嘎,胡毕斯哈拉图.用酸马奶治疗肺结核及结核性胸膜炎58例临床疗效观察[J].中国民族民间医药杂志,2005,74:142
37刘洪元,高昆.马奶及酸马奶(马奶酒)的营养价值和医疗作用[J].中国食物与营养,2003,4:45-46
38包德必理格,吴江媛,马凤珍,等.策格(酸马奶)霜倒模面膜治疗黄褐斑44例疗效观察[J].中国皮肤性病学杂志,1998,12(2):110
39哈斯苏荣,阿木古楞,芒来.酸马奶及其医学价值[J].中国中药杂志,2003,28(1):11-14
40刘忠贤,马丽亚汉.马奶酒治疗便秘[J].中国民间疗法,2000,8(6):27-28
41孙健,贺银凤,田建军.酸马奶酒中有机酸的抑菌作用[J].内蒙古农业大学学报,2003,24(1):94-97
42贺银凤,李少英,母智深,等.酸马奶酒中微生物的分离鉴定及抗菌特性的研究[J].农业工程学报,2002,18(2):91-95
43殷文政,乌尼,钱建伟,等.锡林郭勒牧区马奶酒生物活性动态的研究(一)[J].内蒙古农业大学学报,2002,23(1):9-16
44殷文政,马玉玲,胡炜东,等.马奶酒抑菌活性的研究[J].中国乳品工业,2005,33 (7):15-17
45 Khrisanfova L P, Antimicrobial proterties of kumys from cow and mare milk[J]. Moloch.Prom.,1969,30(10):16-19
46王锂韫,贺银凤,李少英,等.酸马奶酒中粪肠球菌抑菌特性的研究[J].食品科技,2006,15-18
47贺银凤,靳烨,田建军,等.酸马奶酒中一株产抑菌物质酵母菌的分离鉴定及系统发育分析[J].食品研究与开发,2008,29(12):56-60
48母智森,赵美霞,白英,等.酸马奶中酵母菌的分离、鉴定及抗菌特性研究[J].内蒙古农业大学学报,2002,4(7):35-38
49惠丰立,冯金荣,杨柯金,等.产几丁质酶酵母菌的筛选与鉴定[J].东北林业大学学报,2007,35(8):66-67(70)
50冯金荣,惠丰立,文祯中.拮抗性酵母几丁质酶的纯化、性质及抗菌活性[J].食品与生物技术学报,2007,26(1):90-94
51齐木德道尔吉.蒙古族传统饮食文化[J].蒙古社会科学(汉文版) ,2002 ,23(4):37-39
52原琪,赵淑萍,乌云高娃,等.酸马奶治疗冠心病62例临床观察[J].医学论坛,2006,12:111-112
53李蓓,杨虹,芒来.用酸马奶中降胆固醇有效成分的研究[J].畜牧与饲料科学,2008,29(2):56-59
54扎木苏,刘月英,金柱.酸马奶降脂及抗凝作用50例临床观察[J].内蒙古中医药,1994,13(4):16
55扎木苏.酸马奶治疗冠心病临床总结[J].蒙医药,1991,COO:19-23
56李蓓,杨虹,芒来.用酸马奶中降胆固醇有效成分的研究[J].畜牧与饲料科学2008,(2):56-59
57张和平,等.分离自内蒙古传统发酵酸马奶中L. casei.Zhang潜在益性特性的研究[J].中国乳品工业,2006,4(34):4-10
58陆晓娜,杨海燕,阿布力米提.新疆马奶酒中降胆固醇乳酸菌的筛选及性状研究[J].农产品加工,2006,1:31-32(37)
59董全民,李青云.世界牦牛的分布及生产现状[J].青海草业,2003,12(4):32-35
60欧阳熙,王杰,王永,等.藏山羊的生态地理分布及其生态类型[J].西南民族学院学报·自然科学版,1995,21(3):264-271
61李慧,邵建新,范志涛,等.乳汁中主要成分的含量测定[J].中国卫生检验杂志,2002,12(4):441
62于莉,罗章,孟云,等.西藏地区牦牛乳理化和微生物指标的检测分析[J].中国乳品工业,2006,34(9):8-11
63胡小成,郑玉才,邝光荣.牦牛乳中Ca、K、Mg、P的含量[J].西南民族学院学报·自然科学版,1996,22(4):425-427
64罗章,史有国,于莉,等.西藏发酵牦牛乳微生物菌相与理化指标初探[J].中国酿造,2005,10:40-41
65 Zhang H, Xu J, Wang J, et al. A survey on chemical and microbiological composition of kurut,naturally fermented yak milk from Qinghai in China[J]. Food Control, 2008, 19: 578-586
66庞晓娜,韩北忠,陈晶瑜,等.西藏牦牛发酵乳中乳酸菌及酵母菌的特性与相互作用[J].中国酿造,2007,11:23-26
67 Park Y W,Chukwu H I. Macro-mineral concentrations in milk of two goat breeds at different stages of lactation[J]. Small Ruminant Research,1988,1(2):157-166
68陈明汝,林庆文,叶又慈.台湾羊乳克弗尔之研究[J].台湾农业化学与食品科学,2005,43(2):97-102
69 Duitschaever C L. Yogurt from goat milk[J]. Cult. Dairy Prod.,1978,11(11):20-28
70 Samolada M,Litopoulou-Tzanetaki E,Xanthopoulos V,et al. Changes in microbial flora during manufacture of a traditional fermented milk from ewe’s milk[J]. Food Microbiology,1998,15:43-50
71李洪臣,杨秀艳.四大类群微生物菌落的比较[J].生物学教学,2007,32(11):79.
72 Guarro J,GenéJ,Stchigel A M. Developments in Fungal Taxonomy[J].Clinical Microbiology Reviews,1999,12(3):454-500
73白逢彦.酿酒酵母属的分类学研究进展[J].微生物学通报,2007,27(2):139-142
74 Lodder J,Kreger-van Rij. The Yeasts, a Taxonomic Study, Amaterdam: North-Holland Publishing Company, 1952
75 van der Walt J P,Genus.16 Saccharomyces Meyen emend. In Lodder J ed. The Yeasts,a Taxonomic Study,2rd. edn. Amaterdam: North-Holland Publishing Company, 1970,555-718
76 van der Walt J P, Johannsen E. Trichosporon terrestre sp. Nov[J]. Antonie van Leeuwenhoek,1975,41:361-365
77 Yarrow D. Saccharomyces Meyen ex Reess. In Kreger-van Rij N J W ed. The Yeasts, a Taxonomic Study, 3rd. edn. Amaterdam: Elsevier Science Publishers B V,1984
78 Kurtzman C P, Fell J W. The Yeasts, a Taxonomic Study. 4th edn. Amsterdam:Elsevier, Science B V, Amsterdam 1998
79 Kurtzman C P, Robnett C J. Phylogenetic relationships among yeasts of the‘Saccharomyces complex’determined from multigene sequence analyses[J]. FEMS Yeast Res,2003,3(4): 417-432
80 Kurtzman C P. Phylogenetic circumscription of Saccharomyces, Kluyveromyces and other members of the Saccharomycetaceae, and the proposal of the new genera Lachancea, Nakaseomyces,Naumovia,Vanderwaltozyma and Zygotorulaspora[J].FEMS Yeast Res,2003,4: 233-245
81 Esteve-Zarzoso B,Belloch C,Uruburu F, et al. Identification of yeasts by RFLP analysis of the 5.8S rRNA gene and the two ribosomal internal transcribed spacers[J]. International Journal of Systematic Bacteriology,1999(49):329-337
82 Gomes L H,Duarte K M R,Argueso J L,et al. Methods for yeast characterization from industrial products[J]. Food Microbiology,2000,17(2):217-223
83 Zubkova R D. Genus novum Saccharomycetacearum e Kazachstania[J]. Bot Mater Gerb Inst Bot Akad Nauk Kazakh SSR,1971,7:53-56
84 Kurtzman C P. Phylogenetic circumscription of Saccharomyces,Kluyveromyces and other members of the Saccharomycetaceae,and the proposal of the new generaLachancea,Nakaseomyces,Naumovia,Vanderwaltozyma and Zygotorulaspora[J].FEMS Yeast Res,2003,4: 233-245
85 Lu H Z,Cai Y,Wu Z W,et al. Kazachstania aerobia sp. nov.,an ascomycetous yeast species from aerobically deteriorating corn silage[J].International Journal of Systematic and Evolutionary Microbiology,2004,54:2431-2435
86 Wu Z W,Bai F Y. Kazachstania aquatica sp. nov. and Kazachstania solicola sp. nov., novel ascomycetous yeast species[J].International Journal of Systematic and Evolutionary Microbiology, 2005,55:2219-2224
87 Imanishi Y,Ueda-Nishimura K,Mikata K. Two new species of Kazachstania that form ascospores connected by a belt-like intersporal body: Kazachstania zonata and Kazachstania gamospora[J]. FEMS Yeast Res,2007,7(2):330-338
88 Limtong S,Yongmanitchai W,Tun M M,et al. Kazachstania siamensis sp. nov., an ascomycetous yeast species from forest soil in Thailand[J].Int J Syst Evol Microbiol,2007,57:419-422
89 Lee C F,Liu C H,Young S S,et al. Kazachstania jiainicus sp. nov.,an ascomycetous yeast species isolated from soil in Taiwan[J]. FEMS Yeast Res,2008,8(1):114-118
90 Liu Y R,Liu C H, Young S S,et al. New Records of Kazachstania Species in Taiwan[J]. Taiwania,2008,53(3): 293-300
91 Nisiotou A A,Nychas G J. Kazachstania hellenica sp. nov., a novel ascomycetous yeast from a Botrytis-affected grape must fermentation[J]. Int. J. Syst. Evol. Microbiol.,2008,58: 1263-1267
92 Urubschurov V,Janczyk P,Pieper R,et al. Biological diversity of yeasts in the gastrointestinal tract of weaned piglets kept under different farm conditions[J]. FEMS Yeast Res., 2008, 8(8): 1349-1356
93 Lachance M A. (1998) Kluyveromyces van der Walt emend. van der Walt. In: The Yeasts, A Taxonomic Study, Kurtzman C P and Fell J W,Eds.4th edn. Elsevier Science, Amsterdam:227-247
94 Naumov G I. Nomenclature of a yeast genus, Zygofabospora Kudriavzev emend. G. Naumov[J]. Mikologiya i Fitopatologiya, 1987,21:134-140
95 Naumov G I,Naumova E S. Five new combinations in the yeast genus Zygofabospora Kudriavzev emend. G. Naumov. (pro parte Kluyveromyces) based on genetic data[J].FEMS Yeast Res.,2002,2(1):39-46
96 Kurtzman C P,Lachance M A,Nguyen H V,et al. Proposal to conserve Kluyveromyceswith a conserved type (Ascomycota: Hemiascomycetes, Saccharomycetaceae) [J]. Taxon,2001,50(3):907-908
97 Steensma H Y, de Jongh F C M, Linnekamp M. The use of electrophoretic karyotypes in the classification of yeasts: Kluyveromyces marxianus and K. lactis[J]. Curr Genet,1988, 14:311-317
98 Algeri A,Tassi F,Ferrero I,et al. Different phenotypes for the lactose utilization system in Kluyveromyces and Saccharomyces species[J]. Antonie van Leeuwenhoek, 1978, 44: 177-182
99 Gomez-ruiz L, Garcia-garibay M, Barzana E. Utilization of Endo- Polygalacturonase from Kluyveromyces fragilis in the Clarification of Apple Juice[J]. Journal of Food Science, 2006, 53(4):1236-1238
100 Johnson D A, Rogers J D, Regner K M. A soft rot of onion caused by the yeast Kluyveromyces marxianus var. marxianus[J]. Plant Disease,1988,72(4):359-361
101 Schroeder B K,Rogers J D,Johnson D A,et al. Occurrence of Kluyveromyces marxianus var. marxianus Causing Onion Soft Rot in the Columbia Basin of Washington State[J]. Plant Disease, 2007,91(8):1059-1061
102 Kurtzman C P, Robnett C J, Basehoar-Powers E. Phylogenetic relationships among species of Pichia, Issatchenkia and Williopsis determined from multigene sequence analysis, and the proposal of Barnettozyma gen.nov., Lindnera gen.nov. and Wickerhamomyces gen.nov.[J]. FEMS Yeast Res,2008,8:939-954.
103 Ueda-Nishimura K, Mikata K. Reclassification of Pichia scaptomyzae and Pichia galeiformis[J]. Antonie van Leeuwenhoek,2001,79:371-375
104 Kurtzman C P,Robnell C J.Identification and phylogeny of Ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences. Antonie van Lee,uwenhoek,1998,73(4):33l-371
105 Mikata K,Ueda-Nishimura K. Reclassification of Pichia membranifaciens sensu Kurtzman[J]. Antonie van Leeuwenhoek, 2000,77:159-171
106 Kudrjawzew W I. Die Systematik der Hefen[J]. Berlin:Akademie Verlag, 1960,88:366-373
107唐玲,刘平,黄瑛,等.酵母的分子生物学鉴定[J].生物技术通报,2008,5:84-87.
108张传博,苏晓庆.几种基于基因组DNA的真菌分类技术研究进展[J].贵州师范大学学报(自然科学版),2006,24(1):113-118
109熊庆,刘作易.几种分子生物学方法在真菌中的应用[J].贵州农业科学,2002,30 (3) : 69-73
110徐超德,李绍兰.核酸分子系统学方法在酵母菌分类中的应用进展[J].微生物学通报,2004,31(3):126-129
111毛志群,张伟,马雯.分子生物技术在酵母菌分类中的应用进展[J].河北农业大学学报,2002,25:230-233
112周春艳,张秀玲,王冠蕾.酵母菌的5种鉴定方法[J].中国酿造,2006,161(8):51-54
113王国庆,刘天明.酵母菌分类学方法研究进展[J].微生物学杂志,2007,27(3):96-101
114 Fenn J P,Segal H,Barland B,et al. Comparison of updaed Vitek yeast biochemical card and API 20C yeast identification systems[J]. J. Clin. Microbiol.,1994,32(5):1184-1187
115贺玉梅,琚志昌,董葵,等.北京市售食品污染酵母菌鉴定[J].卫生研究,2004,33(4):497-498
116 Gündes S G, Gulenc S, Bingol R. Comparative performance of fungichrom I, Candifast and API 20C AUX systems in the identification of clinically significant yeasts [J]. J. Med. Microbiol.,2001,50:1105-1110
117 Peterson S W,Kurtzman C P.Ribosomal RNA sequence divergence among sibling species of yeasts[J]. Systematic Applied Microbiology,1990,14(2):124-129
118 Kurtzman C P,Robnell C J.Identification and phylogeny of Ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences. Antonie van Leeuwenhoek,1998,73(4):33l-371
119 Kurtzman C P,Robnell C J.Identification of clinically Important Ascomycetous yeasts based on nucleotide divergence in the 5’end of the Large-Subunit(26S) ribosomal DNA Gene[J]. Journal of Clinical Microbiology,1997,35(5):1216-1223
120 Fell J W, Boekhout T, Fonseca A , et al. Biodiveristy and systematics of basidiomycetous yeasts as determined by large-subunit rDNA D1/D2 domain sequence analysis[J]. Int. J. Syst. Evol. Microbiol.,2000,50:1351-1371
121 Cappa F, Cocconcelli P S. Identification of fungi from dairy products by means of 18S rRNA analysis[J]. International Journal of Food Microbiology,2001, 69(1-2): 157-160
122 Bock M,Maiwald M,Kappe R,et al. Polymerase chain reaction-based detection of dermatophyte DNA with a fungus-specific primer system[J]. Mycoses.,1994,37(3-4): 79-84
123 Frutos R L,Fernández-Espinar M T, Querol A. Identification of species of the genus Candida by analysis of the 5.8S rRNA gene and the two ribosomal internal transcribed spacers[J]. Antonie van Leeuwenhoek,2004,85:175-185
124 Schwartz D C,Cantor C R. Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis[J]. Cell,1984,37(1):67-75
125白逢彦,贾建华.克鲁斯假丝酵母及其近似种的脉冲电泳核型分析[J].微生物学报,1996,36(5):329-334
126白逢彦,贾建华.具有不同碳源利用方式的酿酒酵母菌株的脉冲电泳核型分析[J].菌物系统,2000,19(1):65-71
127白逢彦,贾建华.脉冲电泳核型分析在酿酒酵母菌分类学研究中的应用[J].微生物学报,2000,40(1):9-13
128白逢彦,贾建华.四种假丝酵母与其有性型的分子核型比较[J].菌物系统,2001,20(4):471-474
129吴作为,白逢彦.大亚基rRNA基因D1/D2区序列相同或相似的子囊菌酵母ITS序列比较和核型分析[J].菌物学报,2005,24(2):193-198
130 Bai F Y,Liang H Y,Jia J H. Taxonomic relationships among the taxa in the Candida guilliermondii complex,as revealed by comparative electrophoretic karyotyping[J]. International Journal of Systematic and Evolutionary Microbiology,2000,50:417-422
131 Wu Z W,Robert V,Bai F Y. Geneticdiversityof thePichiamembranifaciens strains revealed fromrRNAgenesequencingand electrophoretic karyotyping,and theproposal ofCandida californica comb.nov.[J]. FEMS Yeast Res,2006,6:305-311
132夏青,谢田,文红梅,等. 8株国外引进的酿酒酵母菌的分类鉴定[J].贵州农业科学,2007,35(2):13-17
133 Forster H, Oudemans P, Coffy M D. Mitochondrial and nuclear DNA diversity within six species of Phytophthora[J]. Experimental Mycology,1990,14(1):18-31
134 Su C S, Meyer S A. Characterization of Mitochondrial DNA in Various Candida Species: Isolation, Restriction Endonuclease Analysis, Size, and Base Composition[J]. International Journal of Systematic Bacteriology,1991,41(1):6-14
135 Hennequin C, Thierry G F, Richard G, et al. Microsatellite typing as a new tool for identification of Saccharomyces cerevisiae strains [J]. J. Clin. Microbiol., 2001,39:551-559
136 Welsh J,Michael M. Fingerprinting genomes using PCR with arbitrary primers[J]. Nuc. Aci. Res.,1990,18:7213-7218
137汪小全,邹喻苹. RAPD应用于遗传多样性和系统学研究中的问题[J].植物学报,1996,38(12): 954-962
138 Gomes L H,Duarte K M R,Argueso J L,et al. Methods for yeast characterization from industrial products[J]. Food Microbiology,2000,17(2):217-223
139 Baleiras Couto M M, Vogels J T W E,Hofstra H,et al. Random amplified polymorphic DNA and restriction enzyme analysis of PCR amplified rDNA in taxonomy: two identification techniques for food-borne yeasts[J]. Journal of Applied Microbiology,1995,79(5):525-535
140 Vos P, Hogers R, Bleeker M, et al. AFLP: a new technique for DNA fi ngerprinting[J]. Nucleic Acids Research, 1995,23(21): 4407-4414
141 Thomas C M,Vos P,Zabeau M,et al. Identification of amplified restriction fragment length polymorphism (AFLP) markers lightly linked to the tomato Cf-9 gene for resistance to Cladosporium fulvum[J]. The Plant Jour.,1995,8(5): 785-794
142郑先云,郭亚平,马恩波. AFLP分子标记技术的发展[J].生命的化学,2003,23(1): 65-67
143 Flores Berrios E P,Alba González J F,Arrizon Gavi?o J P,et al. The uses of AFLP for detecting DNA polymorphism, genotype identification and genetic diversity between yeasts isolated from Mexican agave-distilled beverages and from grape musts[J]. Letters in Applied Microbiology,2005,41:147-152
144 Capece A, Sciancalepore A, Sunseri F, et al. Molecular Tools for Assessing Genetic Diversity in Saccharomyces cerevisiae and in the Grapevine Cultivar Aglianico del Vulture Typical of South Italy[J]. Journal of Wine Research, 2004, 15(3):179–188
145 Gallego F J,Pérez M A,Nú?ez Y,et al. Comparison of RAPDs,AFLPs and SSR markers for the genetic analysis of yeast strains of Saccharomyces cerevisiae[J]. Food Microbiology,2005,22:561–568
146 Charlesworth B, Sniegowski P, Stephan W. The evolutionary dynamics of repetitive DNA in eukaryotes[J]. Nature,1994,371:215-220
147 Fleet G H. Yeast in dairy products[J]. J. Appl. Bacterial.,1990,68: 199-211
148 Jakobsen M,Larsen M D,Jespersen L. Production of bread, cheese and meat. In: Osiewacz HD (ed) The Mycota. A comprehensive freatise on fungi as experimental systems and applied research, vol X. 2002,Industrial applications. Springer, Berlin Heidelberg New York, 3-22
149 Beresford T P, Fitzsimons N A, Brennan N L, et al. Recent advances in cheese microbiology[J]. Int. Dairy J.,2001,11:259-274
150 Petersen K M,Moller P L,Jespersen L. DNA typing methods for differentiationof Debaryomyces hansenii strains and yeasts related to surface ripened cheeses[J]. Int. J. Food Microbiol.,2001,69(1-2): 11-24
151 Leclercq-Perlat M N, Oumer A, Bergère J L,et al. Growth of Debaryomyces hansenii on a bacterial surface-ripened soft cheese[J]. J. Dairy Res.,1999,66:271-281
152 Eliskases-Lechner F, Ginizinger W. The yeast flora of surface ripened cheeses[J]. Milchwissenschaft,1995,50(8):458-462
153 Addis E, Fleet G H, Cox J M,et al. The growth, properties and interactions of yeasts and bacteria associated with the maturation of Camembert and blue-veined cheeses[J].Int. J. Food Microbiol.,69(1-2):25-36
154 Corsetti A, Rossi J, Gobbetti M. Interactions between yeasts and bacteria in the smear surface-ripened cheeses[J]. Int. J. Food Microbiol.,2001,69(1-2):1-10
155 Petersen K M,Westall S,Jespersen L. Microbial succession of Debaryomyces hansenii strains during the production of Danish surface ripened cheeses[J]. J. Dairy. Sci.,2002,85:478-486
156 Beresford T P,Fitzsimons N A,Brennan N L,et al. Recent advances in cheese microbiology[J]. Int. Dairy J.,2001,11:259-274
157 Hansen T K, Jakobsen M. Taxonomical and technological characteristics of Saccharomyces spp. associated with blue veined cheese[J]. Int. J. Food Microbiol.,2001,69(1-2):59-68
158 Hansen T K,van der Tempel T,Cantor M D,et al. Saccharomyces cerevisiae as a starter culture in Mycella[J]. Int J Food Microbiol,2001,69(1-2):101-111
159 Molimard P,Vassal L,Bouvier I,et al. Growth of Penicillium camemberti and Geotrichum candidum in pure and mixed cultures on experimental mold ripened cheese of camembert-type[J]. Food processing and preservation,1995,75(1):3-16
160 Jakobsen M, Larsen M D, Jespersen L. Production of bread, cheese and meat. In: Osiewacz HD (ed) The Mycota. A comprehensive freatise on fungi as experimental systems and applied research, vol X. 2002,Industrial applications. Springer, Berlin Heidelberg New York:3-22
161 Guerra J B, Araújo R A C, Patago C, et al. Genetic diversity of Saccharomyces cerevisiae strains during the 24 h fermentative cycle for the production of the artisanal Brazilian cachaca[J]. Lett. Appl. Microbiol.,2001,33(2):106-111
162 Wyder M T, Bachmann H P, Puhan Z. Role of selected yeasts in cheese ripening: an evaluation in foil wrapped Raclette cheese[J]. Lebensm Wiss Technol.,1999,32(6):333-343
163 Oberman H, Libudzisz Z. Fermented milk. In: Wood BJB (ed) Microbiology of fermented foods, vol 1. 1998,Blackie, London:308-350
164 Abdelgadir W S, Hamad SH, M?ller P L,et al. Characterisation of the dominant microbiota of Sudanese fermented milk Rob[J]. Int. Dairy J.,2001,11: 63-70
165 Okagbue R N, Bankole N O. Use of starter cultures obtaining Streptococcus diacetilactis, Lactobacillus brevis and Saccharomyces cerevisiae for fermenting milk for production of Nigerian nono[J]. World J. Microbiol. Biotechnol.,1992,8(3):251-253
166 Gadaga T H, Mutukumira A N, Narvhus J A. The growth and interaction of yeasts and lactic acid bacteria isolated from Zimbabwean naturally fermented milk in UHT milk[J]. Int. J. Food Microbiol.,2001,68(1-2):21-23
167 Jespersen L. Occurrence and taxonomic characteristics of strains of Saccharomyces cerevisiae predominant in African indigenous fermented foods and beverages[J]. FEMS Yeast Res,2003,3(2):191-200
168 Burentegusi,Miyamoto T,Nakamura S,et aL. Identification of lactic acid bacteria isolated from fermented mare’s milk“chigee”in Inner Mongolia ,China[J].Anim.Sci.Technol,2002,73(3):441-448
169 Ishii S, Kikuchi M, Takao S. Isolation and identification of lactic acid bacteria and yeasts from“Chigo”in Inner Mongolia, China[J]. China Animal Science and Technology, 1997, 68(3):325-329
170孟和毕力格,张和平,陈永福,等. Lactobacillus acidophilus MG2-1对大鼠血清脂质代谢的影响研究[J].微生物学报,2005,45(6):52-54
171 Bevan E A, Makover M. The physiological basis of the killer character in yeast[J]. Proc. Int. Congr. Genet, 1963,1:202-203
172 White T J, Bruns S, Lee S, Taylor J. Amplidication and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ, Eds. PCR Protocols: a guide to methods and applications. Academic Press, San Diego,1990
173 F.奥斯伯,R.布伦特编.精编分子生物学实验指南[M].颜子颖,王海林译.北京:科学出版社,1998
174巴尼特J A.酵母菌的特征与鉴定手册.胡瑞卿译.青岛:青岛海洋大学出版社, 1990
175 Thompson J D, Gibson T J, Plewniak. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools[J].Nucleic Acids Research, 1997, 25(24): 4876-4882
176 Saitou N, Nei M. The Neighbor-joining Method: A New Method for Reconstructing Phylogenetic Trees[J]. Mol. Biol. Evol.,1987,4(4):406-425
177孙天松.中国新疆地区传统发酵酸马奶的化学组成及乳酸菌生物多样性研究[D].内蒙古农业大学,博士论文,2006
178徐杰.青海部分地区自然发酵酸牦牛奶的化学与微生物组成分析及乳酸菌的分离鉴定[D].内蒙古农业大学,硕士论文,2006
179云月英.青海海西州传统发酵酸山羊奶的化学与微生物组成分析及乳酸菌的分离鉴定[D].内蒙古农业大学,硕士论文,2006
180 Berlin P J. Koumiss[M].In: Annual Bulletin,PartⅣ, Section A[C],International Dairy Federation, Brussels,1962:4-16
181颜子颖,王海林.精编分子生物学实验[M].北京:科学出版社, 1999
182萨姆布鲁克J,拉塞尔D W著.分子克隆实验指南(第三版)[M].北京:科学出版社, 2002
183 Buzzini P, Martini A. Extracellular enzymatic activity profiles in yeast and yeast-like strains isolated from tropical environments. Journal of Applied Microbiology, 2002, 93(6):1020-1025
184刘云鹏,倪慧娟,孙天松,等.适于AFLP分析的酵母菌DNA提取方法[J].微生物学通报,2008,35(3):1-4
185朱衡,瞿峰,朱立煌.利用氯化苄提取适于分子生物学分析的真菌DNA[J].真菌学报, 1994, 13(1): 34-40
186 Oberman H.Fermented milks,In:Microbiology of Fermented Foods(Edited by Wood B J B)[M].Elsevier Applied Science Publishers,London,1985,167-195
187 Montanari G, Zambonelli C, Fiobi G. The koumiss, milk beverage[J].Industrie Alimentari, 1997,36(355):5-9
188 Montanari G,Grazia L. Galactose-fermenting yeasts as fermentation microorganisms in traditional koumiss[J]. Food Technology and Biotechnology, 1997, 35 (4):305-308
189 Fernández-Espinar M T, Esteve-Zarzoso B, Querol A,et al. RFLP analysis of the ribosomal internal transcribed spacers and the 5.8S rRNA gene region of the genus Saccharomyces: a fast method for species identification and the differentiation of flor yeasts[J]. Antonie Van Leeuwenhoek,2000,78(1): 87-97
190 Petersen K M, Moller P L, Jespersen L. DNA typing methods for differentiation of Debaryomyces hansenii strains and yeasts related to surface ripened cheeses[J]. Int. J. Food Microbiol.,2001,69(1-2): 11-24
191 Scorzetti G, Fell J W, Fonseca A, et al. Systematics of basidiomycetous yeasts: a comparison of large subunit D1/D2 and internal transcribed spacer rDNA regions[J]. FEMS Yeast Res.,2002,2(4): 495-517
192 James S A, Collins M D, Roberts I N. Use of an rRNA internal transcribed spacer region to distinguish phylogenetically closely related species of the genera Zygosaccharomyces and Torulaspora[J]. Int. J. Sys. Bacteriol.,1996,46(1): 189-194
193 Leaw S N, Chang H C, Sun H F, et al. Identification of medically important yeast species by sequence analysis of the Internal Transcribed Spacer regions[J]. J. Clin. Microbiol.,2006,44(3): 693-699
194 Valles B S, Bedri?ana R P, Tascón N F, et al. Yeast species associated with the spontaneous fermentation of cider[J]. Food Microbiol.,2007,24: 25-31
195 Liu Z,Wang C,Chen Y. (1989). Yak resources and qualified populations in China.In:Chinese Yakology.Sichuan Scientific and Technology Press,Chengdu,China:36-77
196 Vasdinyeri R, Deák T. Characterization of yeast isolates originating from Hungarian dairy products using traditional and molecular identification techniques[J]. Int. J. Food Microbiol.,2003,86(1-2): 123-130
197 Andrighetto C, Psomas E, Tzanetakis N, et al. Randomly amplified polymorphic DNA (RAPD) PCR for the identification of yeasts isolated from dairy products[J]. Lett. Appl. Microbiol.,2000,30: 5-9
198 Sirnova E, Beshkova D, Angelov A, et al. Lactic acid bacteria and yeasts in kefir grains and kefir made from them[J]. J. Ind. Microbiol. Biot.,2002,28(1):1-6
199 Latorre-García L, Castillo-Agudo L, Polaina J. Taxonomical classification of yeasts isolated from kefir based on the sequence of their ribosomal RNA genes[J]. World J. Microbiol. Biotechnol.,2007,23:785-791
200 Watabe J, Ikeda N,Mizutani J,et al. Comparison of microbiological and chemical characteristics among types of traditionally fermented milk in Inner Mongolia in China and Calpis sour milk[J]. Milk Science,1998,47(1):1-8
201 Kenji U, Masahiro H, Hidemasa M,et al. Microbiota of‘airag’,‘tarag’and other kinds of fermented dairy products from nomad in Mongolia[J]. Animal Science Journal, 2007, 78(6): 650-658(9)
202许美鑫,刘天明,相茂功,等.青海东部土壤中酵母物种多样性研究[J].微生物学通报,2009,36(3):360-364
203 Martini A V,Rosini D,Martini A. Killer sensitivity patterns as a tool for the fingerprinting of strains within the yeast species Kluyveromyces lactis andK. Marxianus[J]. Biotechnology Techniques,1988,2(4):293-296