云南阳宗海酵母菌种群结构及产胞外酶测试北大核心CSCD

【目的】研究阳宗海酵母菌种群结构,分析生物因子及非生物因子对酵母菌种群分布的影响;测试阳宗海酵母菌产胞外酶活性。【方法】水样用醋酸纤维素滤膜过滤,原位培养分离酵母菌;梯度稀释法分离土样和底泥样品;对分离得到的菌株进行DNA提取和测序,分析26S rDNA的D1/D2区域,并结合形态及生理生化指标进行鉴定;用产酶筛选培养基对分离得到的酵母菌进行产胞外酶活性测试。【结果】共分离得到201株酵母菌,鉴定分属于15个属48个种,其中包括10个潜在的新种;普鲁兰类酵母(Aureobasidium pullulans),库德里阿兹威氏毕赤酵母(Pichia kudriavzevii),胶红酵母(Rhodotorula mucilaginosa),Cryptococcus podzolicus是优势种;15.9%的酵母菌具有产胞外酶活性,主要是脂肪酶和淀粉酶。【结论】阳宗海酵母菌有较为丰富的多样性,人为活动对阳宗海酵母菌分布影响较大,其次浊度、电导率也是影响酵母菌种群分布的重要因素;阳宗海产胞外酶酵母菌可能参与湖泊生态系统的自然循环。     

汾酒酿造车间和车间外环境中酵母菌多样性研究

为解析汾酒厂区酵母菌资源的多样性，采用分离培养方法，从汾酒酿造车间和车间外环境中的微生物中分离纯化到300株酵母菌，经26S rDNA序列比对分析，检测到酵母菌包括6科15属21种。系统发育树结果表明，汾酒酿造车间和车间外环境中的酵母菌包括担子菌门（Basidiomycota）和子囊菌门（Ascomycota）两个分支。尽管汾酒酿造车间和车间外环境中各有特异的酵母菌，但是两者都存在与白酒酿造相关的酵母菌如酿酒酵母（Saccharomyces cerevisiae）、毕赤酵母（Pichia kudriavzevii）和扣囊复膜酵母（Saccharomycopsis fibuligera）等。汾酒环境中含有丰富的酵母菌资源，可能是大曲和酒醅发酵过程中酵母菌的主要来源。这些都为进一步研究指明了方向。     

150例酵母样真菌的菌种分布及其耐药性分析

目的分析引起医院酵母样真菌感染的菌种分布及其耐药性,以指导临床合理用药。方法对2007年10月-2008年10月的住院患者送至真菌室所分离的150株酵母样真菌,采用血清学鉴定及API 20C AUX酵母样真菌鉴定试剂条进行鉴定,采用ATB FUNGUS3进行药敏试验。结果分离到150株酵母样真菌,其中自假丝酵母菌94株（62．67％）,光滑假丝酵母菌14株（9．33％）,热带假丝酵母菌12株（8．00％）,无名假丝酵母菌10株（6．67％）,季也蒙假丝酵母菌6株（4．00％）,近平滑假丝酵母菌6株（4．00％）,其他酵母菌8株（5．33％）。以呼吸内科（54．67％）、老年科（9．33％）、皮肤性病科（9．33％）、血液科（8．67％）分离率最高。150株酵母菌对5氟胞嘧啶（5-FC）、两性霉素B（AmB）、氟康唑（FCA）、伊曲康唑（ITR）、伏立康唑（VRC）的敏感菌株分别为144、150、131、104、147株,敏感率分别为96．00％、100％、87．33％、69．33％、98．00％。结论真菌感染的因素有很多,临床分布科室广泛,临床上应重视真菌检测及真菌药敏试验,根据检测结果合理应用抗真菌药物。     

Asia1型口蹄疫病毒受体结合位点多样性

摘要：【目的】产甘油假丝酵母作为一株优良高产甘油菌株,已成功应用于工业生产15年。近年来由于产甘油假丝酵母染色体倍性尚不明确,限制了对其进行遗传改造的研究进展,因而我们对产甘油假丝酵母染色体倍性研究,分析确定其染色体倍性。【方法】选用酿酒酵母细胞进行生孢,制备酿酒酵母单倍体细胞作对照,并选用热带假丝酵母作为二倍体酵母细胞对照,利用血球计数板得到热带假丝酵母、产甘油假丝酵母、单倍体及二倍体酿酒酵母细胞数,提取染色体,通过二苯胺检测法测定DNA含量。由于在相同紫外照射条件下单倍体细胞比二倍体细胞更容易死亡,因     

产甘油假丝酵母(Candida glycerinogenes)染色体倍性分析

摘要：【目的】产甘油假丝酵母作为一株优良高产甘油菌株,已成功应用于工业生产15年。近年来由于产甘油假丝酵母染色体倍性尚不明确,限制了对其进行遗传改造的研究进展,因而我们对产甘油假丝酵母染色体倍性研究,分析确定其染色体倍性。【方法】选用酿酒酵母细胞进行生孢,制备酿酒酵母单倍体细胞作对照,并选用热带假丝酵母作为二倍体酵母细胞对照,利用血球计数板得到热带假丝酵母、产甘油假丝酵母、单倍体及二倍体酿酒酵母细胞数,提取染色体,通过二苯胺检测法测定DNA含量。由于在相同紫外照射条件下单倍体细胞比二倍体细胞更容易死亡,因     

一株耐盐酵母的分离、鉴定及其生物学特性

以洋河酒厂的酒曲为材料,首先利用盐浓度梯度富集培养和氯化三苯四唑(TTC)染色法初筛获得18株酵母菌株,再通过杜氏小管产气及摇瓶发酵实验,从18株初筛菌株中分离得到1株在1. 4 mol/L NaCl浓度下生长性能较好的菌株,命名为W58。经形态观察和26S rDNA基因序列比对分析,该酵母菌株被鉴定为1株库德里阿兹威氏毕赤酵母(Pichia kudriavzevii)。对菌株的生长性能及耐受性进行研究发现:该菌最适生长温度为30℃,最适pH为6. 0,在高盐条件下(1. 4 mol/L NaCl),其胞内海藻糖的积累显著增加,比对照菌株高28. 6%。此外,菌株W58在高盐条件下,其ATPase活性比对照菌显著提高(约3倍)。综上所述,菌株W58在高盐条件下能通过胞内积累海藻糖以及维持一定的ATPase活性抵抗外界环境压力。     

口腔黏膜真菌鉴定方法的比较

比较常见用于黏膜真菌菌种鉴别的多种方法,探寻最佳的鉴别方法。采集230例普通人群口腔黏膜样本,分别用玉米吐温-80培养观察厚膜孢子法、糖发酵生化反应法、CHROMagar假丝酵母菌显色培养基法、ITS基因的PCR-RFLP(聚合酶链反应-限制性片段长度多态性)法、ITS测序菌种鉴定法,鉴别真菌各菌株。结果显示:有56例菌株至少通过1种方法检出真菌;玉米吐温-80分离培养假丝酵母菌37株;50例菌株ITS基因测序共鉴定出8个菌种,白假丝酵母菌(C.albicans)29株,近平滑假丝酵母菌(C.parapsilosis)10株,热带假丝酵母菌(C.tropicalis)5株,Candida metapsilosis 1株,Lodderomyces elongisporus 1株,克柔假丝酵母菌(Candida krusei)1株,乙醇假丝酵母菌(C.ethanolica)1株,季也蒙毕赤酵母菌(Pichia guilliermondii)2株;CHROMagar假丝酵母菌显色培养基法鉴定出3种菌株,分别是白假丝酵母菌、热带假丝酵母菌、近平滑假丝酵母菌;PCR-RFLP法检出5种菌株,分别是白假丝酵母菌、热带假丝酵母菌、近平滑假丝酵母菌、季也蒙毕赤酵母菌、克柔假丝酵母菌,与基因的测序鉴定一致率为91%;糖发酵生化反应法阳性标本占被检出真菌例数的46.4%(26/56)。结果表明:ITS基因的测序法可以准确鉴定真菌各个菌种;PCR-RFLP法能鉴定常见的菌种,但操作繁琐;CHROMagar假丝酵母菌显色培养基法能快速准确鉴别3种常见假丝酵母菌菌种;玉米吐温-80可以准确培养鉴别白假丝酵母菌;糖发酵生化反应法,缺乏足够的敏感度和特异性,难以准确鉴别各个菌种。     

哈尔滨地区假丝酵母菌DNA异质性及药物敏感性分析

研究假丝酵母菌的DNA异质性及药物敏感性,为预防和监控院内假丝酵母菌感染奠定基础。将临床分离的假丝酵母菌菌株,用科玛嘉显色培养基鉴定菌种,经纸片扩散法进行药敏试验,应用随机扩增多态性DNA（RAPD）技术对这些菌株进行基因分型。结果显示：93株假丝酵母菌中白假丝酵母菌68株,非白假丝酵母菌25株,所有菌株对制霉菌素,两性霉素B两种药物的敏感率最高（100％）,酮康唑其次（70．9％）,氟康唑的敏感率最低（50．5％）,引物1和引物2将来源不同的68株白假丝酵母菌分别分成4型（A1、B1、C1、D1）和6型（A2、B2、C2、D2、E2、F2）。哈尔滨地区的假丝酵母菌感染以白假丝酵母菌为主,且主要为A1、B1型（引物1）或A2、B2型（引物2）;基因型与药敏谱无明显相关性。     

5株生物合成GABA酵母菌株的分离、筛选和鉴定

从水果表皮、果园土、酒曲、泡菜等场所分离筛选到5株产生1-氨基丁酸（GABA）酵母菌菌株,其中4#酵母菌株的GABA产量最高,为3．653g／L。通过形态特征观察和生理生化特征测定,5株酵母菌株分别鉴定为3#属于酿酒酵母（Saccharomyces cerevisiae）,4#和LJ3属于葡萄汁酵母（Saccharomyces uvarum）,MQ属于红冬孢酵母（Rhodosporidium toruloides）,JQ属于红酵母（Rhodotorula glutinis）。     

海南热带雨林腐木上酵母菌物种多样性研究

用含木糖为唯一碳源(培养基X)和含葡萄糖及7.6%乙醇(培养基E)的两种富集培养基分别从采自海南热带雨林的56和57份腐木样品中分离到酵母菌67和75株.依据26S rDNA D1/D2区域序列分析并结合形态学特征对这些菌株进行了分类学研究,探讨了该地区腐木上的酵母菌物种多样性及其分布.从分离的142株酵母菌中鉴定出14个属63个种,其中疑似新种25个,占总种数的近40%,说明在热带雨林腐木中尚存在大量酵母菌新分类群有待被发现.从用培养基X和E分离的酵母菌中分别鉴定出7属37种和11属33种,优势属均为假丝酵母属Candida Berkhout和毕赤酵母属Pichia Hansen,但种类组成基本不同.用培养基x富集分离的菌株以Candida quercitrusa S.A.Meyer＆Phaff的地理分布最广,用培养基E富集分离的菌株以异常毕赤酵母Pichia anomala(Hansen)Kurtzman、酿酒酵母Saccharomyces cerevisiae Meyen ex Hansen和亚膜毕赤酵母Pichiasubpelliculosa Kurtzman分布最广泛.同一样品用两种不同富集培养基分离的菌株大多数属于不同的种,在对比的23份样品中,只从2份样品中分离到了同一个种的菌株.用培养基X和E分离的菌株分别属于可利用木糖和可耐受乙醇的酵母菌,用两种培养基同时分离到的菌株属于具备利用木糖和耐受高浓度乙醇两种能力的菌株.这些酵母菌在木质纤维素物质的生物乙醇转化技术中的应用价值值得进一步研究.     

Physiological properties of some yeast strains

Twenty yeast strains have recently been isolated in pure cultures from natural and industrial sources and identified based mainly on physiological properties. The majority of the strains (15) are alcohologenic belonging to the genus Saccharomyces and comprise two brewer's (beer) yeast strains (S. carlsbergensis= S. uvarum A and B), two baker's yeast strains (S. cerevisiae CA and CP), one spirit yeast strain (S. cerevisiae CF) and ten wine yeast strains (S. cerevisiae var. ellipsoideus = S. ellipsoideus 1, 3, 4, 6, 8 and 9; S. oviformis 2, 5 and 7; and S. uvarum 10). The other 5 yeast strains belong to different species: Kloeckera apiculate, Candida mycoderma (Mycoderma vini), Pichia membranaefaciens, Rhodotorula glutinis and Torulopsis holmii, respectively.     

Isolation of the Candida tropicalis gene for P450 lanosterol demethylase and its expression in Saccharomyces cerevisiae

We have isolated the gene for cytochrome P450 lanosterol 14 alpha-demethylase (14DM) from the yeast Candida tropicalis. This was accomplished by screening genomic libraries of strain ATCC750 in E. coli using a DNA fragment containing the yeast Saccharomyces cerevisiae 14DM gene. Identity of this gene was confirmed by a) observing a heme binding region common to all P450s after sequencing the 3' portion of the gene, and b) based upon tests of its expression in strains of Saccharomyces cerevisiae.     

内蒙古西部区酸粥中酵母菌的分离鉴定及优势菌分析

从内蒙古地区采集28份酸粥样品,从中分离出40株酵母菌,并对其进行了分子生物学鉴定和生物多样性分析。26S rDNA D1/D2区域 (600bp左右)碱基序列分析结果表明,酸粥中的酵母菌有Issatchenkia orientalis、Saccharomyces cerevisiae、Geotrichum sp.、Candida pararugosa、 Candida parapsilosis、Trichosporon asahii、Trichosporon coremiiforme、Clavispora lusitaniae和Candida tropicalis。经过分析,Issatchenkia orientalis(75%,Frequency percentage)为酸粥中的优势菌。     

秦岭地区子囊菌酵母物种多样性研究

从陕西秦岭地区的果实和叶等不同基物上分离得到子囊菌酵母262株,利用26SrDNA的D1/D2区域序列分析并结合形态学特征对这些菌株进行了分类学研究,探讨了该地区子囊菌酵母的物种多样性及其分布。共鉴定出10属31种,其中优势属为假丝酵母属Candida,12种,(其中新种2个,另文发表)、酿酒酵母属Saccharomyces,5种、毕赤酵母属Pichia,5种和有孢汉逊酵母属Hanseniaspora,3种。对分离自陕西秦岭不同地区与海拔的同一个种的不同菌株进行了D1/D2序列分析比较,以探讨子囊菌酵母种内序列稳定性和变异幅度。     

清香型白酒固态酿造过程中酵母种群结构和多样性分析

【目的】探索清香型白酒固态酿造过程中酵母的种群结构和生态多样性变化规律,为科学认识白酒酿造的过程与机制提供理论依据。【方法】运用WL鉴别培养基和26S rRNA D1/D2序列分析方法对清香型白酒3种典型大曲和酒醅发酵过程的酵母进行分类学研究。【结果】从清香型白酒固态酿造过程中共鉴定出10种酵母,分别为Saccharomyces cerevisiae、Issatchenkia orientalis、Pichia anomala、Saccharomycopsis fibuligera、Pichia fermentans、Trichosporon asahii、Hanseniaspora osmophila、Pichia farinosa、Pichia membranifaciens和Clavispora lusitaniae。其中T.asahii、C.lusitaniae、H.osmophila、P.membranifaciens、P.farinose和P.fermentans为首次从清香型白酒酿造过程中分离获得的酵母种类。考察3种典型大曲(清茬、红心、后火曲)和大茬、二茬酒醅发酵过程的酵母种群结构变化规律显示,3种大曲具有相同的优势菌种S.fibuligera,但三者酵母结构组成差异较大,且清茬曲含有最多的酵母数量和种类。酒醅发酵过程中的酵母种群结构与3种大曲均明显不同,大茬和二茬酒醅酵母结构也不同,两种酒醅发酵后期的优势酵母均为S.cerevisiae,而发酵前期优势酵母则分别是H.osmophila和P.membranifaciens。【结论】深入研究了清香型白酒酿造过程中微生物的分布特征和规律,对认识清香型白酒酿造过程和群体微生物的发酵机制,以及丰富我国传统酿造食品微生物的研究,具有重要的理论和实践价值。     

Identification of yeast species from orange fruit and juice by RFLP and sequence analysis of the 5.8S rRNA gene and the two internal transcribed spacers

Yeast isolates from orange fruit and juice in a spontaneous fermentation were identified and classified by two molecular techniques. The first was analysis of the restriction pattern generated from the polymerase chain reaction (PCR)-amplified 5.8S rRNA gene and the two internal transcribed spacers (ITS) using specific primers. The second technique was sequence analysis of the ITS regions using the same two primers. Nine different restriction profiles were obtained from the size of the PCR products and the restriction analyses with three endonucleases (CfoI, HaeIII and HinfI). These groups were identified as Candida tropicalis, Clavispora lusitaniae, Hanseniaspora uvarum, Pichia anomala, Pichia fermentans, Rhodotorula mucilaginosa, Saccharomyces cerevisiae, Saccharomyces unisporus, and Trichosporon asahii. Checking against identification according to morphological, physiological and biochemical traits corroborated this molecular identification. A total concordance was found in the identification with PCR-restriction fragment length polymorphism of the ITS region after analysing certified yeast strains from two different culture collections. Consequently, a rapid and reliable identification of the yeast populations was achieved by using molecular techniques.     

Point prevalence, microbiology and antifungal susceptibility patterns of oral Candida isolates colonizing or infecting Mexican HIV/AIDS patients and healthy persons

We have conducted a longitudinal study over a 3-year period to address the point prevalence, microbiological characteristics and antifungal susceptibility patterns of yeast isolates colonizing or infecting the oral cavities of 111 HIV-infected (51 adults, 60 children) and 201 non HIV-infected (109 adults, 92 children) Mexican persons. Regarding the epidemiology of oral candidiasis, Candida albicans was the most frequent species isolated. Seventy-one out of 85 isolates from colonized persons were C. albicans (83.5%), 27 isolates of them were from HIV-infected children and 44 from non HIV-infected patients. Sixty-two isolates belonged to serotype A which was the most prevalent serotype of C. albicans. Non-albicans species (Candida glabrata, Candida tropicalis and Candida parapsilosis, and Saccharomyces cerevisiae) were isolated from 16.5% of colonized patients and from 38.5% patients with candidiasis or Candida-related lesions. There were nine episodes of infection or colonization by at least 2 different yeast species. In the case of HIV/AIDS patients, it was determined that yeast carriage was not associated with the number of CD4+ cells or the viral load, but HAART reduced the prevalence of oral candidiasis. Overall, most patients harbored strains in vitro susceptible to fluconazole, however 10.8% of the yeasts were resistant to one or more azole antifungal agents and 29% were intermediate susceptible to them. On the contrary, 5-fluorocytosine was very active against all isolates tested, and amphotericin B was active against 97.9% of them.     

渤海海水中酵母菌的种类

本文报道了渤海海水中酵母菌种类的调查结果,从31个站位中,共分离鉴定了228株酵母菌,这些酵母菌分别属于8属40种:红酵母属(Rhodotorula)5个种,隐球酵母属(Cryptococcus)8个种,德巴利酵母属(Debaryomyces)2个种,毕赤酵母属(Pichia)8个种,酵母属(Saccharomyces)2个种,短梗霉属(Aureobasidium)1个种,球孢酵母属(Torulaspora)1个种,假丝酵母属(candida)13个种。40个种中,有13种是中国新记录,即:黑隐球酵母(Cryptococcus ater),大型隐球酵母(Cr.magnus),多型德巴利酵母(Debaryomyces polymorphus),克鲁维酵母(Saccharomyces kluyveri),伯顿毕赤酵母(Pichia burtonii),卡森毕赤酵母(P.carsonii),埃切毕赤酵母(P.etchellsii),季也蒙毕赤酵母(P.guilliermondii),海梅尔毕赤酵母(P.heimii),嗜土毕赤酵母(P.philogaea),奥默毕赤酵母(P.ohmeri),棘胫小蠹毕赤酵母(P.scolyti),和戴尔有孢圆酵母(Torulaspora delbrueckii),在这13种中,除埃切毕赤酵母和奥默毕赤酵母外,其余11种在海水里也是首次报道。     

Development of an integrative DNA transformation system for the yeast Candida tropicalis.

We developed the alkane and fatty-acid utilizing yeast Candida tropicalis as a host for DNA transformations. The system is based on an auxotrophic mutant host of C. tropicalis which is defective in orotidine monophosphate decarboxylase (ura3). The ura3 host was isolated by mutagenesis and a double-selection procedure that combined nystatin enrichment selection and 5-fluoro-orotic acid resistance selection. As a selectable marker, we isolated and characterized the C. tropicalis URA3 gene. Plasmid vectors that contained the C. tropicalis URA3 gene transformed the C. tropicalis mutant host at a frequency of 10(3) to 10(4) transformants per micrograms of plasmid DNA. Vectors that contained the Saccharomyces cerevisiae URA3 gene could not transform C. tropicalis. DNA transfer was accomplished by modified versions of either spheroplast generation (CaCl2-polyethylene glycol)-fusion or cation (LiCl) procedures developed for S. cerevisiae. Plasmid vectors that had been cut within the C. tropicalis URA3 fragment integrated by homologous recombination at the URA3 locus.     

Heat Resistance Studies on Yeast spp. Causing Spoilage in Soft Drinks

A test method to investigate the heat resistance of yeasts has been developed. The method was used to study the heat resistance of 120 yeast strains, representative of the fungal flora in soft drinks and certain acid food products: 35 asporogenous yeast strains ( Brettanomyces, Candida, Kloeckera, Rhodotorula and Torulopsis ) and 85 ascomycetous strains ( Debaryomyces, Hansenula, Kluyveromyces, Lodderomyces, Pichia, Saccharomyces and Saccharomycopsis ) were tested. Generally, asporogenous yeasts were found to be less heat resistant than ascomycetous types. The genus Saccharomyces showed the highest heat resistance, especially strains of the species Sacch. cerevisiae and Sacch. chevalieri . For an evaluation of the practical implications of these results additional studies on environmental factors influencing the heat resistance of ascomycetous yeast species are required.