掷孢菌科的研究 Ⅱ.中国叶表掷孢酵母的初步分类及掷孢酵母属的种类

作者多年来从我国禾本科等植物叶表分离了大量掷孢酵母,本文重点报道70株掷孢酵母与类酵母的初步分类,共分为三属,即掷孢酵母属(Sporobolomyces),布勒掷孢酵母属(Bul-lera)与腥掷孢菌属(Tilletiopsis);其中掷孢酵母属占75％。至于53株掷孢酵母,共分为三种,即玫红掷孢酵母(Sporobolomyces roseus)、赭色掷孢酵母(Sporobolomyces salmonicolor)、Sporobolomyces shibatanus);经统计,后者最为常见,占此属的84％。这些种在我国尚未见报道,作为新记录。     

石河子不同树龄蟠桃土壤中可培养酵母多样性分析及功能酵母的筛选

为了探究新疆本土蟠桃园可培养酵母菌多样性,并挖掘功能酵母资源,本研究以新疆石河子蟠桃园3年、8年、15年树龄的根际和非根际土壤以及桃树叶片为材料,经过传统的分离培养方法获得可培养的酵母菌菌株,并进行形态学、生理生化以及26S r DNA的D1/D2区序列分析,共获得可培养酵母菌129株,从属于12个属17个种,其中子囊菌酵母为优势菌群,占分离属的88%,分布于威克汉姆酵母属(Wickerhamomyces),Vanrija属,Barnettozyma属和有孢圆酵母属(Torulaspora)等11个属的15个种。担子菌占分离属的12%,分布于隐球酵母属(Cryptococcus)的2个种。其中优势属威克汉姆酵母属,包括异常威克汉姆酵母(W. anomalus)和W. pijperi两个种,占总比例的33%,优势种异常威克汉姆酵母所占总株数比例为17%。从可培养酵母中共筛选出23株功能酵母,其中富硒酵母21株,优势种为白地霉(Galactomyces candidum),产蛋白酶酵母2株均属于隐球酵母属的Cryptococcus albidus。结果表明,新疆桃园中蕴含丰富的酵母菌资源,非根际土壤中的酵母多样性大于根际及叶片酵母多样性,且分离得到富硒酵母及产蛋白酶酵母。本研究挖掘了新疆本土可培养酵母菌资源,同时也为功能酵母的开发和利用提供理论指导。     

掷孢菌科的研究II．中国叶表掷孢酵母的初步分类及掷孢酵母属的种类

作者多年来从我国禾本科等植物叶表分离了大量掷孢酵母,本文重点报道70株掷孢酵母与类酵母的初步分类,共分为三属,即掷孢酵母属(Sporobolomyces),布勤掷孢酵母属(Bul-lera)与腥掷孢菌属(Tilletiopsis);其中掷孢酵母属占75％。至于53株掷孢酵母,共分为三种,即攻红掷孢酣母(Sporobolomyces roseus)、赭色掷孢酵母(Sporobolomyces salmonicolor)、Sporobolomyces shibatanus);经统计,后者最为常见,占此属的84％。这些种在我国尚未见报道,作为新记录。     

几株啤酒酵母的筛选及发酵性能比较*

啤酒酵母是啤酒酿造的灵魂,可以直接影响啤酒品质。在啤酒酿造过程中,由于啤酒酵母被多次传代和保藏,造成优良菌种发酵性能衰退等问题,导致发酵不彻底,影响最后啤酒的风味质量。为此以8株Lager型啤酒酵母为出发菌株,通过平板分离纯化获得80株分离菌株,再经过三角瓶发酵初筛和复筛、发酵罐中试发酵实验最终获得了8株发酵性能优良的啤酒酵母。其中,6株酵母可应用于酿造双乙酰含量低于0.1 mg/L的啤酒;3株酵母发酵度高于70%,适合酿造干啤酒;1株酵母发酵度低于50%,适合酿造低醇啤酒。在风味方面:1株酵母酿造的啤酒醇酯比为3.3,啤酒酯香味较突出;另1株酵母酿造的啤酒醇酯比为4.5,啤酒高级醇含量较高。8株经过选育的啤酒酵母发酵特征明显,便于精酿啤酒厂实际应用。     

尼罗红染色法筛选产油酵母及定量检测胞内油脂含量的研究

【目的】建立产油酵母筛选以及胞内油脂含量测定的简便方法。【方法】利用尼罗红与胞内油脂成分结合后在紫外光照射下发出荧光且荧光强弱与油脂含量相关的原理。通过在添加尼罗红的培养基中培养酵母,并观察菌落荧光的方法对385株深海酵母进行产油脂菌株筛选,利用26S rDNA D1/D2区序列分析方法对筛选获得的产油酵母菌株进行鉴定,并以其中的一株高产油脂酵母(2A00015)为试验菌株,建立了一套尼罗红染色快速测定油脂含量的方法。【结果】获得22株产油酵母,其中油脂含量最高可达62.9%,经分子鉴定后显示这22株酵母分别属于(Candida viswanathii)、近平滑假丝酵母(Candidaparapsilosis)、粘质红酵母(Rhodotorula mucilaginosa)、汉逊德巴利酵母(Debaryomyceshansenii)、季也蒙毕赤酵母(Pichia guilliermondii)以及Rhodosporidium paludigenum酵母。尼罗红染色快速测定油脂含量方法的最佳检测条件为:菌悬液OD600小于1.2,尼罗红浓度0.5 mg/L,染色时间5 min,激发波长488 nm,发射波长570 nm。该测定方法得到相对荧光强度与称重法得到油脂含量呈正相关性,R2=0.9637。     

假丝酵母一新种——北京假丝酵母

本文研究了一株从水果表皮分离到的假丝酵母,它与至今已发表的所有已知假丝酵母均不相同,定名为北京假丝酵母(Candida beijingensis)。     

食用营养酵母菌种的选育

通过对面包酵母、假丝酵母、啤酒酵母和奶酒酵母等四种酵母的核黄素、自溶浸出物比率进行研究,选定了核黄素及自溶浸出物比率较高的奶酒酵母(MY-1)作为出发菌株。经连续3次EMS诱变,依次选育富含核黄素的腺嘌呤营养缺陷型菌株(Ml-3)和腺嘌呤缺陷型的回复突变株(M2-8)。对所得的菌株进一步进行抗铵突变株的选育,最终得到富含核黄素的营养酵母突变株(M3-20),其核黄素含量与出发菌株相比提高了约215％。     

耐高温酵母菌株的分离、鉴定及其酒精发酵初步研究

从 38 1个自然样品中筛选得到 2株耐高温酵母THFY 4和THFY 1 6。THFY 4能够在51℃ ,含 30 %葡萄糖的培养基中生长 ;THFY 1 6能够在 45℃ ,30 %葡萄糖的培养基中生长。经初步鉴定 ,THFY 4为克鲁维属酵母 ,THFY 1 6为酵母属酵母。进一步的 37℃发酵实验证明 ,THFY 4在静置条件下的发酵性能很差 ,发酵 60h只能从 2 0 %的葡萄糖产生 4 88% (v v)的酒精 ;而THFY 1 6在相同的条件下从 2 0 %的葡萄糖中产生 1 1 44%     

芝麻香型白酒大曲中酵母菌的分离与鉴定

为了开发利用白酒大曲中的酵母菌资源,采用稀释涂布平板法,从芝麻香型白酒大曲中分离得到15株酵母菌株。利用26S rRNA基因序列分析技术对其进行分类鉴定。结果表明,其中6株酵母菌为酿酒酵母（Saccharomyces cerevisiae）,6株为库德里阿兹威氏毕赤酵母（Pichia kudriavzevii）,3株为热带假丝酵母（Candida tropicalis）;并对代表菌株Y1、Y2及Y4进行了形态观察;最后通过随机扩增多态性DNA标记（random amplified polymorphic DNA, RAPD）对分离得到的酵母菌进行分型,表明6株酿酒酵母分属于5个株型,6株库德里阿兹威氏毕赤酵母分属于5个株型,3株热带假丝酵母分属于3个株型。     

酵母样真菌感染的菌型分布及药敏试验分析

目的:调查酵母样真菌在临床标本中的菌型分布情况;了解临床常见真菌对常用抗真菌药物的敏感性,为临床使用抗真菌药提供依据.方法:按<临床检验操作规程>进行菌株分离和鉴定;对分离的菌株用法国生物梅里埃公司生产的ATBFUNGUS抗真菌药物敏感性试剂盒进行药敏试验.结果:共检出酵母样真菌256株,其构成比:白色念珠菌161株(62.88%)、热带念珠菌58株(22.8%)、高里念珠菌12株(4.69%)、酵母菌10株(3.91%),未定型7株(2.73%);91株酵母样真菌药敏结果:5-FC敏感率95.6%、AmB 100%、NYS 93.4%、MIZ 80.2%、ECO 81.3%、KET 84.6%.11株MIZ耐药株,2株同时耐ECO与KET,9株为ECO中介、8株为KET中介,15株耐药株有10株为热带念珠菌.结论:引起深部真菌感染的酵母样真菌种类日渐增多,其中以白色念珠菌为主;大部分酵母样真菌对抗真菌药敏感,咪唑类药物有交叉耐药,多数热带念珠菌有耐药性.因此,酵母样真菌的菌种鉴定及药敏试验对临床深部真菌感染的防治具有重要的意义.     

对从面包中筛选酵母菌及其发酵特性的研究

目的 从野生酵母筛选出对可溶性糖产生既可氧化又可还原、并发生反式反应的酵母菌株,使其能应用于相关物质的生物合成.方法 对从面包中滋生出多种的野生菌落采用依形态、使易糖氧化为主要特征筛选出野生酵母菌落.结果 从3株野生菌株(JM1、JM2、JM3)中筛选出JM1,该菌株能使体内发生反式催化的氧化反应.结论 该菌株适宜的生...     

CARDIOLIPIN CONTENT OF WILD TYPE AND MUTANT YEASTS IN RELATION TO MITOCHONDRIAL FUNCTION AND DEVELOPMENT

The phospholipid composition of various strains of the yeast, Saccharomyces cerevisiae, and several of their derived mitochondrial mutants grown under conditions designed to induce variations in the complement of mitochondrial membranes has been examined. Wild type and petite (cytoplasmic respiratory deficient) yeasts were fractionated into various subcellular fractions, which were monitored by electron microscopy and analyzed for cytochrome oxidase (in wild type) and phospholipid composition. 90% or more of the phospholipid, cardiolipin was found in the mitochondrial membranes of wild type and petite yeast. Cardiolipin content differed markedly under various growth conditions. Stationary yeast grown in glucose had better developed mitochondria and more cardiolipin than repressed log phase yeast. Aerobic yeast contained more cardiolipin than anaerobic yeast. Respiration-deficient cytoplasmic mitochondrial mutants, both suppressive and neutral, contained less cardiolipin than corresponding wild types. A chromosomal mutant lacking respiratory function had normal cardiolipin content. Log phase cells grown in galactose and lactate, which do not readily repress the development of mitochondrial membranes, contained as much cardiolipin as stationary phase cells grown in glucose. Cytoplasmic mitochondrial mutants respond to changes in the glucose concentration of the growth medium by variations in their cardiolipin content in the same way as wild type yeast does under similar growth conditions. It is concluded that cardiolipin content of yeast is correlated with, and is a good indicator of, the state of development of mitochondrial membrane.     

Comparison of fermentative capacities of industrial baking and wild-type yeasts of the species Saccharomyces cerevisiae in different sugar media

AIMS: To compare the fermentative capacity of wild and domesticated isolates of the genus Saccharomyces. METHODS AND RESULTS: The fermentative capacity of yeasts from a variety of wild and domesticated sources was tested in synthetic dough media that mimic major bread dough types. Domesticated yeast strains were found to have better maltose-utilizing capacity than wild yeast strains. The capacity to ferment sugars under high osmotic stress was randomly distributed amongst wild and baking strains of Saccharomyces. CONCLUSION: The domestication of bakers' yeast has enhanced the ability of yeasts to ferment maltose, without a similar impact on the fermentative capacity under high osmotic conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This study, combined with molecular studies of both wild and domesticated yeast, showed that domestication of bakers' yeast has resulted in improved maltose utilization, apparently via the duplication and mutation of the MAL genes.     

Karyotype rearrangements in a wine yeast strain by rad52-dependent and rad52-independent mechanisms

Yeast strains isolated from the wild may undergo karyotype changes during vegetative growth, a characteristic that compromises their utility in genetic improvement projects for industrial purposes. Karyotype instability is a dominant trait, segregating among meiotic derivatives as if it depended upon only a few genetic elements. We show that disrupting the RAD52 gene in a hypervariable strain partially stabilizes its karyotype. Specifically, RAD52 disruption eliminated recombination at telomeric and subtelomeric sequences, had no influence on ribosomal DNA rearrangement rates, and reduced to 30% the rate of changes in chromosomal size. Thus, there are at least three mechanisms related to karyotype instability in wild yeast strains, two of them not requiring RAD52-mediated homologous recombination. When utilized for a standard sparkling-wine second fermentation, Deltarad52 strains retained the enological properties of the parental strain, specifically its vigorous fermentation capability. These data increase our understanding of the mechanisms of karyotype instability in yeast strains isolated from the wild and illustrate the feasibility and limitations of genetic remediation to increase the suitability of natural strains for industrial processes.     

酵母鉴定程序在酵母分类鉴定中的应用

本文介绍了Barnett等1985年编制并由剑桥大学发行的计算机软件《酵母鉴定程序》,以及如何使用该程序在IBM PC DOS操作系统上进行酵母菌的分类鉴定。我们使用该程序对从云南鸡足山和紫金山两地森林土壤中分离到的82株酵母菌进行了分类鉴定,其中:鉴定到种的有47株,占总株数的57.3％;鉴定到属但未能直接定到种的有21株,占总株数的25.6％;暂未定名的有14株,占总株数的17.1％。     

Isolation of alcohol tolerant,osmotolerant and thermotolerant yeast strains and improvement of their alcohol tolerance by UV mutagenesis

In this study, the yeast strains were isolated from grapes by serial dilution technique to determine their alcohol-, sugar- and thermotolerance. 34 wild type yeast strains were isolated and alcohol-, sugar- and thermotolerance of these strains were determined. The maximum alcohol tolerance was found to be 9% (v/v) in yeast strain which is named Y2. Thermotolerance behavior of 6 strains were investigated. The strains were treated with UV light with intervals of 20, 30, 40 and 50 seconds. Selected resistant colonies were investigated for alcohol tolerance. It was found that alcohol tolerance increased from 9% (v/v) to 12% (v/v) on Y2 strain.     

S-nitrosoglutathione induced nitrosative stress in yeast: modifying role of catalases

Possible role of catalases in modification of yeast Saccharomyces cerevisiae response to nitrosative stress was studied in the work. Yeast cell of a wild strain and catalase-defective strains were treated with nitric oxide donor S-nitrosoglutathione, then the cell survival rate, and the levels of protein carbonyls and oxidized glutathione were measured. It was shown that S-nitrosoglutathione decreased cells viability of wild and catalase-defective strains. Unexpected, yeast cells defective by both catalases survived successfully as compared with the cells of the wild strain. The intensity of stress was evaluated by measures of oxidative protein modifications and glutathione oxidation. Treatment with S-nitrosoglutathione did not affect the level of protein carbonyls and was lower by about 14 i 22 % in the cells of double mutant strains after treatments with S-nitrosoglutathione in concentrations of 10 and 20 mM. S-nitrosoglutathione induced a strong increase of the level of oxidized glutathione in yeast cells of the wild strain. This stress slightly increased the level of oxidized glutathione in the yeast cells defective by peroxisomal or both catalases. It is interesting, that an increase of oxidized glutathione level was not observed in the yeast cells defective by cytosolic catalase. The obtained results prove that catalases can modify yeast cell response to the nitrosative stress.     

Karyotype Rearrangements in a Wine Yeast Strain by rad52-Dependent and rad52-Independent Mechanisms

Yeast strains isolated from the wild may undergo karyotype changes during vegetative growth, a characteristic that compromises their utility in genetic improvement projects for industrial purposes. Karyotype instability is a dominant trait, segregating among meiotic derivatives as if it depended upon only a few genetic elements. We show that disrupting the RAD52 gene in a hypervariable strain partially stabilizes its karyotype. Specifically, RAD52 disruption eliminated recombination at telomeric and subtelomeric sequences, had no influence on ribosomal DNA rearrangement rates, and reduced to 30% the rate of changes in chromosomal size. Thus, there are at least three mechanisms related to karyotype instability in wild yeast strains, two of them not requiring RAD52-mediated homologous recombination. When utilized for a standard sparkling-wine second fermentation, Δrad52 strains retained the enological properties of the parental strain, specifically its vigorous fermentation capability. These data increase our understanding of the mechanisms of karyotype instability in yeast strains isolated from the wild and illustrate the feasibility and limitations of genetic remediation to increase the suitability of natural strains for industrial processes.     

脱落酸产生菌的遗传育种

从南京蔬菜研究所植物致病株中分离到3株脱落酸（ABA）产量较高菌株,经初步鉴定均为灰葡萄孢（BolrytiscinereaPers．）。分别命名为A23,A101,A160通过紫外诱变及菌株生长状况比较,选出一株菌A101作为出发菌,经紫外,硫酸二乙酯多次诱变,在ANA合成代谢关键酶抑制剂平板上定向选育出一株ANA高产菌UUD－1,该菌固体发酵可产生ABA374μg／ml,是出发菌株产量的14倍,且其产量连续培养5代稳定。     

降解三硝基甲苯的酵母和类酵母菌的研究

从受三硝基甲苯（TNT）严重污染的土壤和废水中分离筛选到17株可降解TNT的酵母菌和白地霉。其中6株为克鲁斯假丝酵母（Candidakrusei）,4株为橡树假丝酵母（C.quercitrusa）,一株为无名假丝酵母（C.famata）,一株为伯杰汉逊酵母（Hansenulabeijerinckii）,一株为亚膜汉逊酵母（H.subpelliculosa）,4株为白地霉（Geotrichumcandidum）。对其中6株菌进行了降解TNT的条件实验,发现降解TNT的适宜pH为7,温度为37～40℃。在含75～80mg／LTNT的培养基中,40h内能降解TNT56～74mg／L,去除率达71％～93％。在培养基中加入0．01％～0．05％的葡萄糖作碳源,或加入0.01％～0．1％的酵母膏对6株菌降解TNT的能力略有促进作用。加入铵盐作为氮源则明显抑制这些菌对TNT的降解。