新疆地区酸马奶中酵母菌的鉴定及其生物多样性分析

从新疆少数民族牧民家庭采集的28份传统工艺酿造酸马奶样品中分离出87株酵母菌,并对其进行了生理生化鉴定、分子生物学鉴定和生物多样性分析。生化试验结果表明,新疆地区酸马奶中的酵母菌为Saccharomyces unisporus(占总分离株的48.3%),Kluyveromyces marxianus(27.6%),Pichia membranaefaciens(15.0%)和Saccharomyces cerevisiae(9.2%)。选取其中的6株酵母菌和1株参考菌株,进行大亚基(26S)rDNA D1/D2区域(600bp左右)碱基序列分析,并通过GenBank进行同源序列搜索以确定各菌株的归属,进一步验证生理生化方法的正确性。从得到的结果中可以看出,S.unisporus和K.marxianus为新疆地区酸马奶中的优势菌。     

中国吐鲁番两株产木聚糖酶的极端耐碱Bacillus halodurans的分类鉴定

通过生理生化实验、16S rDNA序列分析和同源性杂交,将分离到的XJU-1和XJU-80菌种进行了分类鉴定。XJU-1和XJU-80具有较宽的pH生长范围(分别是pH4·5 ~12·6和pH3·8 ~12·6) ,其G C mol%含量分别是40·5mol%和42·2mol%。16S rDNA序列分析和DNA-DNA同源杂交结果表明,XJU-1和XJU-80与Bacillushalodurans C-125和Bacillus halodurans DSM497T具有较高的同源性(99 %) ;两者之间也具有85 %的相关性,但其与Bacillus halodurans C-125和Bacillus halodurans DSM497T分别具有81·3 %和71·5 %的相关性。基于以上结果,将两株分离菌株分类为Bacillus halodurans的两个新品系。     

26S rDNA单链构象多态性分析在临床酵母菌菌种鉴定中的应用

摘要： 【目的】探讨酵母菌临床分离株26S rDNA D1/D2区序列种内相似性和种间差异性的快速检测方法,为临床酵母菌菌种鉴定方法的改进奠定基础。调查北京地区临床酵母菌的种群多样性,为国内酵母菌感染的流行病学研究提供新的基础数据。【方法】用5种常见临床酵母菌种的模式和权威菌株作为标准参考菌株,从北京四家综合性医院收集临床酵母菌260余株,PCR扩增其26S rDNA D1/D2区,对扩增产物进行单链构象多态性（Single-Strand Conformation Polymorphism,SSCP）分析和序列测定分析。【结果】常见病原酵母菌26S rDNA D1/D2区的SSCP图谱具有明显的种间差异性和种内相似性,可以通过该方法对菌株进行初步的菌种鉴定。D1/D2-SSCP和序列分析相结合,对260余株临床酵母菌进行了菌种鉴定,共鉴定有10个属20个种,优势属为念珠菌属(Candida),优势种及其所占比例分别是：C. albicans (57.7%), C. parapsilosis (10.0%), C. tropicalis (9.2%), C. glabrata (6.7%)和C. krusei (5.8%),并发现过去从未或很少报道致病的酵母菌种,愈来愈多地出现在临床分离菌株中。【结论】 26S rDNA D1/D2区的SSCP图谱分析为临床酵母菌株的快速鉴定提供了新的方法;北京地区酵母菌临床分离株呈种群多样性分布,C. albicans虽然仍占优势,但其它念珠菌种的比例已达42%。     

26S rDNA-RFLP分析在非酿酒酵母菌分类研究中的应用

使用26S rDNA-RFLP分析,分析了分离自甘肃莫高葡萄酒厂的29株非酿酒酵母菌,被测菌株被分为10个类型。通过26S rDNA D1/D2区序列分析验证,证明此方法在葡萄酿酒酵母菌种多样性研究中良好的应用价值。     

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。     

稷酸米汁中新耐碱酵母Trichosporon asahii var.asahii XJU-1的分离鉴定

目的:从山西偏关县地方饮食稷米汁中分到一株酵母,命名为XJU-1,并进行分类学鉴定及pH耐受检测。方法:XJU-1的形态学观察、生理学测试及脂肪酸分析、测定基因的ITS区、26S大亚基D1/D2区以及IGS区,测序序列用邻近法建树推算XJU-1的归属。结果:XJU-1生长谱为pH 2.0～13.0,最适为8.0。仅检测到8种脂肪酸,3种优势脂肪酸总量为91.24%。基因ITS区和26S大亚基D1/D2区显示与Trichosporon asahii(Strain 2479T)最为相近,分别仅有1bp差异。基因IGS区表明该菌是T.asahii的一个变种T.asahii var.asahii。结论:已知的T.asahii无耐碱性报道,XJU-1(=CGMCC AS 2.3199=CCTCC AF207026)pH生长谱宽泛,是一株新的兼性耐碱菌株。     

一株碱性淀粉酶产生菌Bacillus flexus XJU-3的分离鉴定及酶学特性分析

[目的]从新疆石河子市一处碱性工业污水(pH11.0)分离﹑鉴定高产碱性淀粉酶菌株,并对其所产酶酶学特性进行研究.[方法]在碱性淀粉酶分离培养基上对所分离菌株进行筛选,分离到一株高产碱性淀粉酶菌株,并将其编号XJU-3.应用生理生化试验,脂肪酸含量,16S rDNA序列以及(G C) mol%含量等方法对菌株进行鉴定,同时对XJU-3所产碱性淀粉酶的生物学特性进行研究.[结果] XJU-3可在pH4.0～12.5的LB培养基上生长,最适生长温度37℃.16S rDNA序列构建的系统进化树表明XJU-3与Bacillus flexus类聚在一起,且序列同源性为99%.该菌产生的淀粉酶最适pH10.0,最适温度40℃,且在pH9.0～13.0内有较高活性和稳定性.Co2 和Mg2 能明显提高酶的活性.[结论] XJU-3被鉴定为Bacillus flexus,由于XJU-3与B. flexus DSM 1320T在尿素水解和优势脂肪酸含量上有差异,且具有宽范围pH耐受性,因此XJU-3被认为是B.flexus的一个新菌株.XJU-3所产的碱性淀粉酶酶学特性良好,具有极大的工业应用潜力.     

一株分离自酸梅酱的酵母菌的分离鉴定

目的:分离及鉴定来自于新疆塔城民间自制酸梅酱中的酵母菌。方法:用NL1/NL4引物对扩增酵母菌株的26S rDNAD1/D2区,测序结果进行序列分析,用Neighbour-joining(N-J)方法构建系统发育进化树,同时结合酵母的传统形态学鉴定对菌株进行鉴定。结果:26S rDNA D1/D2区序列分析表明菌株KKS与Metschnikowia aff.fructicola D3895相近,相似率为99.2%。在N-J法构建的系统发育进化树中,菌株KKS与Metschnikowia aff.fructicola聚类在同一分枝上。结论:从新疆塔城民间自制酸梅酱中分离得到一株酵母菌并将该菌株鉴定为Metschnikowia aff.fructicola。     

基于可培养法分析新疆葡萄与葡萄酒相关酵母菌菌群结构

从葡萄、葡萄发酵物、土壤及酒厂设备分离到3 991株新疆本土葡萄酒酵母菌,并通过Wallerstein Laboratory(WL)营养琼脂培养形态分类、代表菌株26S rDNA Dl/D2和5.8S-ITS区域序列鉴定,对新疆葡萄酒酵母菌株进行系统分类和多样性分析,得到22个WL培养类型,7个属13个种。伊犁和于田地区酵母菌种类最多,达10个种;叶城地区和中信国安、乡都酒业、野果林(伊犁)酒厂酵母菌种类最少,仅有1种。新疆优势种为葡萄汁有孢汉逊酵母(Hanseniaspora uvarum)和酿酒酵母(Saccharomyces cerevisiae)。陆生毕赤酵母(Pichia terricola)、Metschnikowia aff、核果梅奇酵母(Metschnikwoia fructicola)、韩国梅奇酵母(Metschnikwoia koreensis)在新疆地区首次分离鉴定到。对新疆葡萄酒相关的酵母菌群落结构进行全面分析,结果显示新疆与葡萄酒相关的酵母菌丰富多样。为新疆葡萄酒酵母资源的开发利用和葡萄酒的质量控制等奠定了菌种基础。     

甘肃莫高葡萄酒厂酵母种群的生态分布

葡萄酿酒酵母的菌群组成和多样性对葡萄酒的化学成分和感观特性具有重要贡献,因此葡萄产区酵母菌的生态研究成为国外近年来的研究热点.应用WL营养琼脂培养基聚类分析、26S rRNA基因序列和5.8S-ITS区序列分析对甘肃莫高葡萄酒厂酵母菌群进行研究,共分离到葡萄酒相关酵母9属10个种:Saccharomyces cerevisiae、Issatchenkia orientalis、Rhodosporidium kratochvilovae、Pichia kluyveri var. kluyveri、Metschnikowia pulcherrima、Hanseniaspora uvarum、Cryptococcus magnus、Pichia fermentans、Rhodotorula mucilaginosa、Cryptococcus uzbekistanensis.不同生态条件下的菌群分布各有不同,在自然发酵过程中出现的菌种有M.pulcherrima, H.uvarum, P. kluyveri var. kluyveri和I. orientalis.发酵初期Metschnikowia pulcherrima和Hanseniaspora uvarum为启动发酵的主要菌群,3～4d后至发酵中期Saccharomyces cerevisiae开始出现并逐渐取代non-Saccharomyces,随后主导发酵结束.在酒厂设备表面分离到的菌种以Rhodotorula mucilaginosa和Hanseniaspora uvarum为主,分别占到34%和26%,其次C. magnus占16%,I. orientalis占12%,Pichia kluyveri var. kluyveri占12%.葡萄园土壤中分离到的菌群R. mucilaginosa占62.5%、C.magnus占21%,S. cerevisiae和Rhodosporidium kratochvilovae分别占12.5%和4%.     

Cr(VI) reduction in a chromate-resistant strain of Candida maltosa isolated from the leather industry

A Cr(VI)-resistant yeast was isolated from tanning liquors from a leather factory in Leon, Guanajuato, Mexico. Based on morphological and physiological analyses and the D1/D2 domain sequence of the 26S rDNA, the yeast was identified as Candida maltosa. Resistance of the strain to high Cr(VI) concentrations and its ability to chemically reduce chromium was studied. When compared to the three laboratory yeasts Candida albicans, Saccharomyces cerevisiae and Yarrowia lipolytica, the C. maltosa strain was found to tolerate chromate concentrations as high as 100 micro g/ml. In addition to this phenotypic trait, the C. maltosa strain showed ability to reduce Cr(VI). Chromate reduction occurred both in intact cells (grown in culture medium or in soil containing chromate) as well as in cell-free extracts. NADH-dependent chromate reductase activity was found associated with soluble protein and, to a lesser extent, with the membrane fraction.     

一株分离于酸性矿坑水的异养菌的特性与分子鉴定

采用平板划线分离法,从江西某铜矿酸性矿坑水中分离出一株极端嗜酸、又能耐受中度碱性条件的异养微生物,命名为HJM菌株。该菌株能在pH1.5~10.0的范围内生长。形态学以及18SrDNA和26S rDNAD1/D2区序列分析表明,HJM菌株属于P.guilliermondii这个种。金属抗性试验表明,该菌株对重金属铜离子的抗性可高达45mmol/L,因此,它的分离为研究极端环境中微生物的抗铜机制提供了材料。     

西藏曲拉和云南乳饼中酵母菌的鉴定及其生物多样性

【目的】探讨西藏曲拉和云南乳饼中酵母菌的生物多样性及其分布特征,为我国传统乳制品中酵母菌资源的利用提供基础数据。【方法】从西藏和云南分别采集的5份曲拉样品和8份乳饼样品中分离出41株酵母菌,利用26SrDNAD1/D2区域序列分析对这些菌株进行了分类鉴定。【结果】曲拉和乳饼样品中酵母菌的总数分别在106-107cfu/g和102-106cfu/g之间,曲拉样品的酵母菌平均数比乳饼样品中的高34倍。共鉴定出10属12种,其中西藏曲拉的优势菌株为发酵毕赤氏酵母(Pichia fermentans)和酿酒酵母(Saccharomyces cerevisiae);云南乳饼的优势菌株为类筒假丝酵母(Candida zeylanoides)和喜仙人掌毕赤氏酵母(Pichia cactophila)。毕赤氏酵母属(Pichia)是曲拉和乳饼的共同优势属。【结论】西藏曲拉和云南乳饼中的酵母菌都具有丰富的生物多样性,但其差异性很大。     

Molecular genetic identification of yeast strains isolated from egyptian soils for solubilization of inorganic phosphates and growth promotion of corn plants

Forty yeast strains isolated from soils taken from different locations in Egypt were tested for their P-solubilizing activities on the basis of analyzing the clear zone around colonies growing on a tricalcium phosphate medium after incubation for 5 days at 25degreesC, denoted as the solubilization index (SI). Nine isolates that exhibited P-solubilization potential with an SI ranging from 1.19 to 2.76 were genetically characterized as five yeasts belonging to the genus Saccharomyces cerevisiae and four non-Saccharomyces, based on a PCR analysis of the ITS1-26S region amplied by SC1/SC2 species-specific primers. The highest Psolubilization efficiency was demonstrated by isolate PSY- 4, which was identified as Saccharomyces cerevisiae by a sequence analysis of the variable D1/D2 domain of the 26S rDNA. The effects of single and mixed inoculations with yeast PSY-4 and Bacillus polymyxa on the P-uptake and growth of corn were tested in a greenhouse experiment using different levels of a phosphorus chemical fertilizer (50, 100, and 200 kg/ha super phosphate 15.5% P2O5). The results showed that inoculating the corn with yeast PSY-4 or B. polymyxa caused significant increases in the shoot and root dry weights and P-uptake in the shoots and roots. The P-fertilization level also had a significant influence on the shoot and root dry weights and P-uptake in the shoots and roots when increasing the P-level from 50 up to 200 kg/ha. Dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 200 kg/ha gave higher values for the shoot and root dry weights and P-uptake in the shoots and roots, yet these increases were nonsignificant when compared with dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 100 kg/ha. The best increases were obtained from dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 100 kg/ha, which induced the following percentage increases in the shoot and root dry weights, and P-uptake in the shoots and roots; 16.22%, 46.92%, 10.09%, and 31.07%, respectively, when compared with the uninoculated control (fertilized with 100 kg/ha).     

不对称还原苯乙酮的酵母菌株筛选与分子鉴定

利用苯乙酮作为模式底物,对145株菌株进行初筛和复筛,获得一株具有高效立体选择性的酵母菌株YS6-2,能够不对称还原苯乙酮生成(S)-1-苯基乙醇.在苯乙酮浓度为70mmol/L时,底物的初始转化率达26.8%,产物(S)-1-苯基乙醇的对映体过量值为98.8%.基于形态学、生理生化特征、18S rDNA和26S rDNA D1/D2区域的分析表明,YS6-2为胶红酵母(Rhodotorula muci-laginosa).