黑比诺葡萄接种发酵过程酵母菌的变化监控

采用WL营养琼脂培养基和Interdella指纹图谱分析以及UPGMA聚类分析,对接种工业酵母RC212的黑比诺葡萄酒发酵过程中分离的63株酵母进行种类区分与鉴定,并对其中的49株酿酒酵母单菌落进行菌株区分,以监控发酵过程中酵母菌的种类和动态变化,明确工业酵母是否主导发酵过程,探讨野生酿酒酵母与工业酵母之间的竞争性,为葡萄酒发酵过程中的微生物控制提供依据.结果表明,接种发酵过程中共分离得4种不同培养类型的酵母菌,分别是葡萄汁有孢汉逊酵母(Hanseniaspora uvarum)、美极梅奇酵母(Metschnikowia pulcherrima)、红冬孢酵母(Rhodotorula mucilaqinosa)和酿酒酵母(Soccharomyces cerevisiae).用Interdelta指纹图谱将酿酒酵母区分为3种基因型,包括2种野生酿酒酵母和已接种的工业酵母RC212.工业酵母在发酵初、中、后期分别占酿酒酵母的6.25%、18.75%、100%.工业酵母在发酵末期才成为发酵优势菌,野生酿酒酵母在发酵初期和中期都位据优势地位,表现出很强的竞争力.     

祁连葡萄酒产区葡萄酒相关野生酵母菌株的分离及初步分类

取祁连葡萄酒产区葡萄园原料自然发酵筛选葡萄酒相关野生酵母菌株68株,利用YEPD培养基的形态观察以及WL营养培养基的聚类分析,并结合生理生化实验对其进行初步分类,结果表明,与葡萄酒相关的野生酵母菌株有假丝酵母、有孢汉逊酵母、酒香酵母、毕赤氏酵母、红酵母以及酿酒酵母,其中有孢汉逊酵母属和酒香酵母属为优势菌株.     

接种不同嗜杀特性的酿酒酵母对赤霞珠发酵中酵母多样性的影响

以赤霞珠葡萄为原料,分别接种不同嗜杀特性的酿酒酵母(Saccharomyces cerevisiae)菌株NXU17-26(中性)、UCD522(敏感菌株)和UCD2610(嗜杀菌株),并以自然发酵为对照,研究各菌株对赤霞珠葡萄酒的发酵特征及发酵中酵母菌多样性的影响。结果表明,接种发酵在启酵和发酵速度上显著快于自然发酵。WLN培养基将分离到的480株酵母菌鉴定为7种类型,26S rDNA D1/D2序列分析进一步将其鉴定为4属5种:葡萄汁有孢汉逊酵母(Hanseniaspora uvarum)、克鲁维毕赤酵母(Pichia kluyveri)、伯顿丝孢毕赤酵母(Hyphopichia burtonii)、S.cerevisiae、库徳毕赤酵母(Pichia kudriavzevii)。这4属5种的酵母均存在于自然发酵中,而接种发酵中仅有H.uvarum和S.cerevisiae两种酵母,接种发酵中酵母菌多样性较低。Interdelta指纹图谱分析表明,所接种的酿酒酵母菌株是相应发酵中的优势菌株:接种中性酵母NXU17-26的发酵中,NXU17-26的基因型占比为63.46%;接种敏感菌株UCD522中,UCD522的基因型占比为44.68%,野生酿酒酵母NXU18-15表现出较强的竞争力,基因型占比为34.04%;接种嗜杀酵母UCD2610的发酵中,UCD2610的基因型占比为62.74%。非加权算术平均数法聚类分析表明,分离自同一发酵中的不同酿酒酵母菌株间的遗传差异性较小;分离自不同发酵中的酿酒酵母菌株间遗传差异性较大。     

桓仁产区优良冰酒酵母菌株的筛选及酿造特性评价（英文）

为获得可用于东北桓仁地区威代尔冰酒生产的酿酒酵母菌株,采用一种快速的酵母菌株筛选方法,通过测定菌株乙醇和二氧化硫耐受性,从威代尔冰酒自然发酵过程中筛选到9株酿酒酵母菌株。进一步酿造实验结果显示,所筛选的酿酒酵母可以顺利完成冰酒发酵,产生的香气轮廓与商业酵母DV10相比也不同(主成分分析结果),最终获得了2株具有高发酵活力且香气特征与商业酵母差异显著的酵母菌株SC42和SC45,其中SC42能够高产高级醇和酯类物质,并且低产乙酸,而SC45能够产生高含量的甘油、酯类物质以及反式玫瑰醚和β-大马士酮。结果表明,采用本研究的筛选方法能够快速有效地筛选到具有应用潜力的冰酒生产菌株,同时也证明了使用本土野生酵母菌株能够有效地改善冰酒香气品质,生产出与接种商业酵母不同风格的冰酒产品。     

两株野生沙棘酵母菌株的分子生物学鉴定及其发酵特性

应用26SrDNAD1／D2区域序列测定和系统发育分析,对从野生沙棘果中分离出的两株酵母菌JHZ3、WKZl-2进行了鉴定。结果表明,其分别为葡萄汁有孢汉逊酵母（Hanseniaspora uvarum）和戴尔凯氏有孢圆酵母（Torulaspora delbrueckii）。对这两株茵发酵特性的研究结果表明,JHZ3和WKZl—2在较强酸性条件下发酵能力强。     

树莓酒自然发酵过程中酵母菌的分离与鉴定

目的:从树莓酒自然发酵过程中分离和鉴定酵母菌,目的是筛选适宜树莓酒发酵特点的专用酵母菌株。方法:采用传统微生物分离纯化技术与和26S r DNA D1/D2区序列分析相结合的方法,从树莓酒自然发酵的不同时期分离纯化酵母菌,结合菌落和细胞显微形态特征进行区分,随机选择代表菌株进行26S r DNA D1/D2区序列分析。结果:从树莓酒自然发酵过程中共分离出323株酵母菌,形态学初步鉴定为12类,分子鉴定为分属于5个属的7种酵母菌,分别为葡萄汁有孢汉逊酵母(Hanseniaspora uvarum)、东方伊萨酵母(Issatchenkia terricola)、酿酒酵母(Saccharomyces cerevisiae)、覆膜孢酵母属亚种(Saccharomycopsis crataegensis)、叉开假丝酵母(Candida diversa)、东方拟无枝酸菌(Candida sorboxylosa)和假丝酵母属亚种(Candida stellimalicola)。结论:传统的微生物培养形态区分与分子鉴定技术结合,对筛选树莓酒自然发酵过程中的酵母菌准确有效。     

种植方式对‘赤霞珠’葡萄酒发酵中酵母菌多样性的影响

为明确种植方式对‘赤霞珠’葡萄酒发酵中酵母多样性的影响,以有机种植、生物动力种植及其常规种植的葡萄为原料进行自然发酵,并于破碎的第0、2、4、6、8天（发酵结束）分别取样,采用WLN培养基形态学鉴定和26SrDNAD1/D2区序列分析,调查不同种植方式下不同发酵阶段的酵母群落结构差异。结果表明,共分离出1075株酵母,鉴定为16种形态类型,分为8属9种,分别为葡萄汁有孢汉逊酵母（Hanseniaspora uvarum）、假丝酵母（Candida zemplinina）、拜尔接合酵母（Zygosaccharomyces bailii）、美极梅奇酵母（Metschnikowiapulcherrima）、桃梅奇酵母（Metschnikowiafructicola）、酿酒酵母（Saccharomyces cerevisae）、隐球酵母（Cryptococcus saitoi）、库德毕赤酵母（Pichia kudriavzevii）、单孢酿酒酵母（Kazachstania bulderi）。不同发酵时期酵母种属及占比在各种植方式下具有差异,破碎当天均以非酿酒酵母为主;破碎第2天有机种植组以酿酒...     

葡萄自然发酵过程中酵母菌的研究

目的:筛选性状优良的野生酿酒酵母,对葡萄自然发酵过程中的酵母菌进行分离鉴定,探讨葡萄自然发酵期间酵母菌群的变化.方法:利用WL培养基,对分离自吉林松源的"双红"、"双优"、"赤霞珠"和"黑塞比尔"4个葡萄品种自然发酵液的245株酵母进行初步鉴定,并对其中的120株进行分子鉴定.结果:利用WL培养基可将245株酵母分为6个营养类型.结合酵母菌菌株5.8S-ITS区域的RFLP分析,4个葡萄品种的自然发酵液中共有5种酵母,分别是葡萄汁有孢汉逊酵母(Hanseniaspora uvarum)、酿酒酵母(Saccharomyces cerevisiae)、粘红酵(Rhodutorula glutinis)、陆生伊萨酵母(Issatchenkia terricola)和假丝酵母属的Candida sorbosa.结论:自然发酵过程中酵母菌群的比例是不断变化的,"双优"品种的自然发酵液中可能存在性状优良的野生酿酒酵母.     

葡萄酒病害相关酵母的分离与鉴定

利用3种培养基和3种分离方法,从贺兰山东麓葡萄酒产区发生葡萄酒病害的5款酒样中分离获得23株酵母菌株。经形态学鉴定,初步将所分离的酵母菌归为4种培养类型;26S rDNA D1/D2区序列分析结果表明,4种培养类型的酵母分属于3个属3个种,分别为酿酒酵母（Saccharomyces cerevisiae）、葡萄汁有孢汉逊酵母（Hanseniaspora uvarum）和乳酪隐球酵母（Cryptococcus friedmannii）。     

宁夏玉泉营地区酿酒葡萄酵母菌的分离筛选及分子鉴定

为进一步探究宁夏玉泉营地区酿酒葡萄酵母菌的多样性,更好地开发利用本土葡萄酒产区酵母菌资源,从宁夏玉泉营地区的葡萄园土壤、葡萄果实表皮以及葡萄自然发酵过程中进行酵母菌的分离筛选,对得到的22株酵母菌株运用WL营养琼脂培养基进行初步聚类分类,并采用26S rDNA D1/D2区序列分析法进行菌株鉴定。结果表明,供试菌株共鉴定为6种,分别为大隐球酵母（Cryptococcus magnus）、美极梅奇酵母（Metschnikowia pulcherrima）、葡萄酒有孢汉生酵母（Hanseniaspora vineae）、酿酒酵母（Saccharomyces cerevisiae）、克鲁维毕赤酵母（Pichia kluyveri）、东方伊萨酵母（Issatchenkia orientalis）。     

Dynamic study of yeast species and Saccharomyces cerevisiae strains during the spontaneous fermentations of Muscat blanc in Jingyang,China

The evolution of yeast species and Saccharomyces cerevisiae genotypes during spontaneous fermentations of Muscat blanc planted in 1957 in Jingyang region of China was followed in this study. Using a combination of colony morphology on Wallerstein Nutrient (WLN) medium, sequence analysis of the 26S rDNA D1/D2 domain and 5.8S-ITS-RFLP analysis, a total of 686 isolates were identified at the species level. The six species identified were S. cerevisiae, Hanseniaspora uvarum, Hanseniaspora opuntiae, Issatchenkia terricola, Pichia kudriavzevii (Issatchenkia orientalis) and Trichosporon coremiiforme. This is the first report of T. coremiiforme as an inhabitant of grape must. Three new colony morphologies on WLN medium and one new 5.8S-ITS-RFLP profile are described. Species of non-Saccharomyces, predominantly H. opuntiae, were found in early stages of fermentation. Subsequently, S. cerevisiae prevailed followed by large numbers of P. kudriavzevii that dominated at the end of fermentations. Six native genotypes of S. cerevisiae were determined by interdelta sequence analysis. Genotypes III and IV were predominant. As a first step in exploring untapped yeast resources of the region, this study is important for monitoring the yeast ecology in native fermentations and screening indigenous yeasts that will produce wines with regional characteristics.     

赤霞珠葡萄自然发酵酵母菌的动态变化研究

葡萄酒发酵是一个复杂的生化过程,包含了不同酵母属种此消彼长的变化。利用WLN琼脂培养基,对陕西杨凌赤霞珠自然发酵过程中,不同阶段分离的403株酵母菌进行了归类鉴定。结果表明,其分属于9属10种:酿酒酵母(Saccharomyces cerevisiae)、葡萄汁有孢汉逊酵母(Hanseniaspora uvarum)、东方伊萨酵母(Issatchenkia orientalis)、毕赤克鲁维酵母(Pichia kluyver)、膜璞毕赤酵母(Pichia membranefaciens)、美极梅奇酵母(Metschnikowia pulcherrima)、戴尔有孢圆酵母(Torulaspora delbrueckii)、红冬孢酵母(Rho-dosporidium mucilaginos)、假丝酵母(Candida species)和佛罗伦萨接合酵母(Zygosaccharomyces florentinus),分别占到了分离总菌株的53.00%、35.00%、1.00%、1.75%、0.75%、6.75%、0.25%、0.25%、1.00%和1.00%。     

慕萨莱思酵母菌YPD表型多样性及其酿酒酵母δ序列遗传多样性分析

利用YPD稀释涂布法,从新疆阿瓦提地区6个不同采样点46份样液(原料、预处理液,不同时期发酵液)中分离出700株酵母菌,具有15种表型特征。挑选141株酿酒酵母代表株,与7株不同年份酿酒酵母,采用δ序列分析,得到41种基因型,显示阿瓦提慕萨莱思相关酵母菌,在不同采样点之间及同一采样点的原料批次,不同发酵期,不同发酵容器,不同年份之间均存在多样性。     

Yeast species associated with the spontaneous fermentation of cider

This paper reports the influence of cider-making technology (pneumatic and traditional pressing) on the dynamics of wild yeast populations. Yeast colonies isolated from apple juice before and throughout fermentation at a cider cellar of Asturias (Spain), during two consecutive years were studied. The yeast strains were identified by restriction fragment length polymorphism analysis of the 5.8S rRNA gene and the two flanking internal transcribed sequences (ITS). The musts obtained by pneumatic pressing were dominated by non-Saccharomyces yeasts (Hanseniaspora genus and Metschnikowia pulcherrima) whereas in the apple juices obtained by traditional pressing Saccharomyces together with non-Saccharomyces, were always present. The species Saccharomyces present were S. cerevisiae and S. bayanus. Apparently S. bayanus, was the predominant species at the beginning and the middle fermentation steps of the fermentation process, reaching a percentage of isolation between 33% and 41%, whereas S. cerevisiae took over the process in the final stages of fermentation. During the 2001 harvest, with independence of cider-making technology, the species Hanseniaspora valbyensis was always isolated at the end of fermentations.     

贺兰山东麓产区葡萄自然发酵液中酵母的多样性研究

为了分析宁夏贺兰山东麓葡萄自然发酵液中酵母多样性,从贺兰山东麓不同产区（广夏、立兰、银川和红寺堡）葡萄自然发酵的不同阶段分离酵母菌,并采用WL培养基与26S rDNA D1/D2序列分析法进行鉴定。结果表明,共分离到316株酵母,经鉴定被归为6个属9个种,其中在各产区均分离到有孢汉逊酵母（Hanseniaspora uvarum）、美极梅奇酵母（Metschnikowia pulcherrima）、Candida zemplinina和酿酒酵母（Saccharomyces cerevisiae）;在立兰、银川和红寺堡均分离到克鲁维毕赤酵母（Pichia kluyveri）;在红寺堡还分离到加利福尼亚假丝酵母（Candida californica）和伽状毕赤酵母（Pichia galeiformis）;库德里阿兹威氏毕赤酵母（Pichia kudriavzevii）仅存在于立兰和银川;戴尔有孢圆酵母（Torulaspora delbrueckii）仅存在于银川。宁夏贺兰山东麓葡萄自然发酵汁中酵母的多样性分析不仅为该产区酵母的开发利用奠定基础,也将为其他产区酵母筛选提供参考依据。     

Indigenous wine killer yeasts and their application as a starter culture in wine fermentation

Wine yeast strains were isolated from seven fermentations of the red wines ‘Refošk’ and ‘Teran’, produced in the southwestern part of Slovenia. Among 613 isolated yeast strains, 22 expressed killer activity against the supersensitive strain Saccharomyces cerevisiae. Killer strains were isolated at different stages of wine fermentation but did not dominate in any of them. Species identification was based on the combination of RFLP analysis of an amplified rDNA region and biochemical–physiological tests. Killer isolates were identified as S. cerevisiae, Pichia anomala, Pichia kluyveri, Pichia pijperi, Hanseniaspora uvarum and Candida rugosa. Electrophoretic karyotyping was used to differentiate strains of the same species. Fermentation properties of four S. cerevisiae strains that possessed stable killer activity were characterized in fermentations of Malvasia must by studying their population dynamics and chemical composition and by sensory analysis of the wines produced. In order to compare the results, spontaneous fermentations and fermentations induced by commercial yeast starters were performed concomitantly. The local killer strain S_S12/10 showed the best fermentation properties and produced wine with favourable characteristics.     

产地葡萄酒优良酵母菌株的筛选及鉴定

从山东、甘肃、宁夏、陕西、新疆酿酒葡萄产区采集的99份葡萄果粒中分离到酵母258株,通过抗性和发酵特性评价,筛选出3株发酵性能优良的菌株.经菌落特征、细胞形态、生理生化特性鉴定,结合26S rDNA D1/D2序列,菌株ZYFJQ和QLMLZ2被鉴定为酿酒酵母,菌株ZYY73-2被鉴定为葡萄酒有孢汉逊氏酵母.