克鲁斯假丝酵母及其近似种的脉冲电泳核型分析

用钳位均匀电场脉冲电泳(CHEF)系统分析了克鲁斯假丝酵母(Candida krusei),郎比可假丝酵母(C. lambica)和粗状假丝酵母(C. valiad)的模式菌株的电泳核型,发现这三种表型相似的假丝酵母却具有互不相同的染色体DNA分子带型,为其分类学研究提供了可靠的鉴别依据。在常规分类学研究的基础上,测定了AS 2.75(原定种名为(C. incospicua),AS2.1182(原定种名为 C. lambica)和AS 2.1772(未定种)等三株假丝酵母的G+C含量和脉冲电泳核型。通过对已报道的C. inconspicu的G+C含量及上述三种假丝酵母模式菌株的脉冲电泳核型的比较分析证明,AS 2.75和AS 2.1772为粗状假丝酵母(C. valida),AS 2.1182为克鲁斯假丝酵母(C. krusei)。     

不同寄主来源白假丝酵母菌多态性与耐药性分析

为研究白色假丝酵母菌的分子多样性情况,探讨RAPD基因多态性与药物敏感性的关系,在获得病菌分离物的基础上,应用经筛选的10条10 bp随机引物,对18株病菌分离物进行RAPD分析,利用UPGMA对结果进行聚类.RAPD扩增的指纹图谱清晰,带型稳定,多态性丰富,可以作为白假丝酵母菌分型方法,18株不同来源的菌株可大致分为6种亲缘关系.药敏结果显示导致出现耐药的机制可能并不相同.所以利用RAPD标记技术在基因水平上对白色假丝酵母菌进行分子分型和鉴定是可行的.     

啤酒酵母和产朊假丝酵母属间原生质体融合子的筛选

利用不可逆生化抑制剂碘乙酸抑制一亲株细胞的生理活性和利用两亲株细胞间生理性状的差异性,进行啤酒酵母(Saccharomyces cerevisiae)和产朊假丝酵母(Candida utilis)原生质体融合子的筛选,属间原生质体融合率为3.47×10~(-6)。经菌落形态比较,碳化合物的同化和发酵,DNA含量测定,酯酶型分析,细胞核染色,产孢试验和自然的核分离实验证明,融合子分三种类型:87.21%产朊假丝醇母型;9.02%啤酒酵母型;3.77%真正核融合子型。     

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。     

48株临床分离假丝酵母菌DNA异质性分析

应用随机扩增多态性DNA技术(RAPD)对48株假丝酵母菌临床菌株进行PCR扩增,并对扩增产物的指纹图谱进行分析,结果显示:48株假丝酵母菌中白假丝酵母菌35株,非白假丝酵母菌13株,引物1和引物2将来源不同的35株白假丝酵母菌分成4型和6型,且RAPD技术可将白假丝酵母菌鉴定至种,并可分辩同种菌的不同型别,是假丝酵母菌致感染、追踪病原的分子流行病学研究有效方法。     

固囊酵母的一个新种

从中国保定槐茂甜面酱中分离到一株酵母。经鉴定,这株酵母菌属于固囊酵母属(Citeromyces),并为该属中的一个新种,命名为保定团囊酵母(Citeromyces baodingensis zhangsp.Nov.)。保定团囊酵母与固囊酵母生理生化特性有显著区别,保定固囊酵母(Citeromycesbaodingensis)不发酵蔗糖、棉子糖,同化半乳糖和纤维二糖,不同化海藻糖和棉子糖,G+C含量为48.5mol％。     

抑菌试验用于测定T-2毒素

本文从镰刀菌中提取T-2毒素,对不同的酵母菌进行抑菌试验,结果证明红酵母属(Rho-dotorula)的三个种,即红酵母(R.glutrnis)、深红酵母(R.ruber)和小红酵母(R.minuta)对T-2毒素最敏感,被定为测毒敏感菌。将抑菌试验与家兔皮肤反应试验、豌豆发芽抑制试验进行对比,结果有部分重合,但不能相互取代。由于抑菌试验具有快速、简便、准确的优点,可做为测定T-2毒素的方法之一。     

代谢木糖和葡萄糖的重组酿酒酵母的构建*

为使酿酒酵母(Saccharomyces cerevisiae)YS58代谢木糖产乙醇,采用PCR方法克隆得到树干毕赤酵母(Pichia stipitis)木糖醇脱氢酶基因xyl2,并将该基因和克隆得到的休哈塔假丝酵母(Candida shehatae)缺终止子的木糖还原酶基因xyt1一起连接到酵母表达载体pYES2的强启动子GAL下,得到融合表达载体pYES2-P12。通过醋酸锂转化的方法将pY- ES2-P12转入S．Cerevisiae YS5     

粤北一种虫草分离菌的抗酵母现象

CR 95 1 2是一株分离自粤北一种虫草的无孢丝状真菌 ,经培养初步鉴定为无孢菌目(^{Agonomycetales})的丝核菌属 (^Rhizoctonia)。对包括革兰氏阳性及阴性细菌、放线菌、丝状真菌和酵母菌在内的 2 6株致敏菌株的抗性测定表明 ,CR 95 1 2对包括红酵母属 (^Rhodotorula)、酵母属 (^{Saccharomyces})、毕赤氏酵母属 (Pichia)和隐球酵母属 (^Cryptoc     

不同类群酵母胞壁甘露聚糖核磁共振氢谱的比较研究*

运用核磁共振氢谱(PMR谱) 对各类酵母的细胞壁甘露聚糖进行比较研究,在我国尚无报道,其中某些酵母也尚无文献记载。本文结果表明：1．同菌株的胞壁甘露聚糖PMR谱型的重复性很好。2．同种不同株的酿酒酵母(Saccharomyces cerevisiae)的多糖谱型也相同。3．所测的二端芽殖酵母中完全型与不完全型菌株的谱型很相似, 如柠檬形克勒克酵母(Kloeokera apiculata)与葡萄有孢汉逊酵母(Hanseniaspora uvarum。4．某些分类系统上来源较杂的子囊菌酵母如单宁管囊酵母(Packysolen tanophilus)、萤光威克酵母(Wickerkamiaflurescens) 与高糖固囊酵母(Citeromyses matritensis) 则体现了各不相同的谱型。 5．二株分自西双版纳的极为相近的类酵母(Saccharomycodes sp.)其多糖的(PMR)谱型与多糖的组分都彼此相同,有助于对它们的适当归类。这一切证明酵母胞壁多糖PMR谱型相似程度的比较是分类上较有意义的性状,有助于探讨亲缘关系,核实完全型与不完全型,也有助于对疑难菌株的分析     

Prospects of using the electrophoresis technique for identification of the taxonomic status of earthworms (Lumbricidae)

The polyacrylamide gel electrophoresis technique was used for comparison of common protein of body wall tissues in five species of Lumbricidae (Eisenia nordenskioldi, E. fetida, Lumbricus rubellus, Aporrectodea caliginosa, A. longa). The statistic processing of indexes of electrophoretic similarity revealed three levels of similarity: intraspecies, interspecies, and intergenus. It was discovered that the similarity of E. nordenskioldi and E. fetida proteins corresponded to the intergenus level. The use of this method for determination of the earthworm’s taxonomic status is discussed.     

Identification of medically important yeasts by sequence analysis of the internal transcribed spacer and D1/D2 region of the large ribosomal subunit

BackgroundThe prevalence of opportunistic yeast infections has increased in recent decades as the result of an increasing immunocompromised patient population.AimsTo evaluate ribosomal RNA (rRNA) gene sequence to identify medically important yeast species, to investigate the performance of both the rRNA gene internal transcribed spacer (ITS) and D1/D2 region in identifying clinically relevant yeasts, and to compare these results with those of a standard phenotypic method.MethodsBoth regions from 50 yeast strains, comprising 45 clinical isolates and 5 reference strains, were amplified using PCR and then sequenced. The sequences were compared to reference data available from the GenBank database of the National Center for Biotechnology Information using the BLASTn tool.ResultsUsing ID32C, 88% (44/50) of all strains were identified accurately at the species level, although 6% were misidentified; two Candida eremophila isolates were identified as Candida glabrata and Candida tropicalis, and one Saprochaete clavata isolate was identified as Saprochaete capitata. Two of the four isolates identified by phenotypic methods as Trichosporon asahii were defined so by analyzing the ITS region, but the remaining two were not distinguishable from closely related species. Based on the D1/D2 region, these four isolates had 100% sequence identity with T. asahii, Trichosporon japonicum, and Trichosporon asteroides. The isolate identified as Trichosporon inkin using ID32C could not be distinguished from Trichosporon ovoides by analyzing the ITS and D1/D2 regions.ConclusionsIdentifying medically important yeasts by sequencing the ITS and D1/D2 region is a rapid and reliable alternative to conventional identification methods. For a diagnostic algorithm, we suggest a two-step procedure integrating conventional methods (e.g. microscopic morphology on corn meal agar with Tween® 80 and API ID32C®) and sequence analysis of the ITS and D1/D2 region.     

80份甘蔗种质RAMP标记遗传多样性分析

采用RAMP分子标记技术对80份甘蔗种质(32份祖亲种、48份栽培品种或品系)的遗传基础进行了分析。从30对引物组合中筛选出4对多态性较强引物,构建了甘蔗80份种质的RAMP指纹图谱,这四对引物组合共扩增出84条带,其多态性为91.7%。80份种质的遗传相似系数变化范围在0.433～0.988,平均0.710。聚类分析表明,随着相似系数结合线的不同,可分别将参试的甘蔗种质从属间(甘蔗属与斑茅种)、野生种(割手密种、大茎野生种、印度种、中国种)与栽培种(热带种)间、栽培种与杂交栽培品种(或品系)间区别开来。各祖亲种与杂交栽培品种(或品系)的遗传相似性由大到小依次为热带种>印度种和中国种>大茎野生种＞云南割手密种＞其它割手密种＞斑茅。另外,本试验首次利用RAMP标记,获得了部分热带种、野生种及斑茅种特异片段,并发现这些特异片段能不同程度地在具有其血缘的栽培种中得到传递。     

鸢尾属植物遗传多样性的 RAPD和ISSR分析

应用RAPD和ISSR标记技术,对来自不同产地的鸢尾属(IrisL.)4个野生种的遗传多样性进行了分析。结果表明,12个RAPD和ISSR引物分别扩增出225和196条带,多态性条带分别为215和196条,多态性条带百分率分别为95.56%和100.00%;种间总基因多样度分别为0.368 9和0.357 5、种内基因多样度分别为0.107 7和0.138 0,表明鸢尾属种间遗传多样性较高,且种间变异大于种内变异。4个野生种中,蝴蝶花(I.japonicaThunb.)的遗传组成较为丰富。此外,种内遗传关系与地理分布和环境差异有一定的相关性。     

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。     

Genetic variation of the genus Kengyilia by ISSR markers

We investigated the genetic variation within 32 accessions distributed to 14 species and one variety by using ISSR (inter-simple sequence repeat) markers.The results showed that genetic variation was relatively higher among the accessions.A total of 593 bands were amplified by 12 ISSR primers,of which 535 bands (90.2%) were polymorphic.Eleven to 80 polymorphie bands were amplified from each prime,with an average of 44.6 bands.The interspecies GS (genetic similarity)value ranged from 0.430 to 0.866,and the average was 0.620.Cluster analysis showed that all accessions could be classified into 4 groups by ISSR markers.The different accessions in a species were clustered together,but they had genetic variation in molecular levels.There was obvious interspecies genetic variation.Species with similar morphological characteristics and from the same areas or neighboring geographical regions were clustered together and had close relationships.ISSR markers are useful in analyzing interspecies variation in Kengyilia.     

基于RAPD标记的薄荷属(Mentha L.)植物亲缘关系分析

应用RAPD标记方法分析了薄荷属(Mentha L. )7个种38个种源间的遗传多样性,并采用UPGMA聚类分析方法探讨了38个种源间的亲缘关系.结果表明,用20条随机引物从38个种源的总DNA中共扩增出111条带,其中多态性条带91条,多态性条带百分率达81.98%,表明薄荷属植物种间和种内存在丰富的遗传多样性.聚类分析结果表明,在遗传相似系数0.43 处,供试的38个种源被分为2大类,其中第1大类包含日本薄荷(M. arvensis L. )、灰薄荷(M. vagans Boriss. )、留兰香(M. spicata L. )、皱叶留兰香(M. crispata Schrad. ex Willd. )、椒样薄荷(M.×piperita L. )和薄荷(M. haplocalyx Briq. )的37个种源,第2大类仅包含唇萼薄荷(M. pulegium L. )1个种源.在遗传相似系数0.74处,38个种源被分为6组:A组仅包含日本薄荷1个种源;B组包含灰薄荷的4个种源;C组包含留兰香的2个种源和皱叶留兰香的6个种源;D组包含椒样薄荷的5个种源和留兰香的2个种源;E组包含薄荷的17个种源;F组仅包含唇萼薄荷1个种源.在遗传相似系数0.83处,B组、C组、D组和E组可各自进一步划分为不同的亚组.研究结果显示,基于RAPD标记分析的聚类分析结果与传统形态学分类结果基本相吻合;同一种类来源相同或相近的种源聚在一起,说明薄荷属植物种内的遗传关系与地理分布和环境差异有一定的相关性.     

RAPD分析与ITS序列分析在拟茎点霉分类鉴定上的应用

采用随机扩增多态性DNA(RAPD)技术和核糖体RNA基因(rDNA)转录间隔区(ITS)序列分析对22种拟茎点霉共34个菌株进行了系统发育研究。RAPD分析构建的UPGMA聚类图所反映的种间、种内关系与形态学分类结果基本一致,可以清楚地将分自7科寄主植物上的不同的种分别区分开来,但分自同科或同属寄主植物上的不同的种并不具有相近的亲缘关系。ITS序列分析结果不支持将Phomopsis mangiferae Ahmad和P. cytosporella Penz. et Sacc. 合并为同一个种的观点,同时还显示出P. mangiferae与P. psidii de Camara的亲缘关系非常近, 可能是异名同物;而为害木棉叶的拟茎点霉与杨梅枝枯病菌P. myricae Y.J.Huang et P.K.Chi之间的碱基差异亦属于种下不同菌株间的正常差别范围,很可能就是同一个种。对相同的供试菌株两技术所反映的亲缘关系趋势相同,表明两技术用于拟茎点霉的亲缘关系分析和种类鉴定均是可行的。     

RAPD分析与ITS序列分析在拟茎点霉分类鉴定上的应用*

采用随机扩增多态性DNA(RAPD)技术和核糖体RNA基因(rDNA)转录间隔区(ITS)序列分析对22种拟茎点霉共34个菌株进行了系统发育研究。RAPD分析构建的UPGMA聚类图所反映的种间、种内关系与形态学分类结果基本一致,可以清楚地将分自7科寄主植物上的不同的种分别区分开来,但分自同科或同属寄主植物上的不同的种并不具有相近的亲缘关系。ITS序列分析结果不支持将Phomopsis mangiferae Ahmad和P. cytosporella Penz. et Sacc. 合并为同一个种的观点,同时还显示出P. mangiferae与P. psidii de Camara的亲缘关系非常近, 可能是异名同物;而为害木棉叶的拟茎点霉与杨梅枝枯病菌P. myricae Y.J.Huang et P.K.Chi之间的碱基差异亦属于种下不同菌株间的正常差别范围,很可能就是同一个种。对相同的供试菌株两技术所反映的亲缘关系趋势相同,表明两技术用于拟茎点霉的亲缘关系分析和种类鉴定均是可行的。     

The microbiology of Bandji,palm wine of Borassus akeassii from Burkina Faso: identification and genotypic diversity of yeasts,lactic acid and acetic acid bacteria

Aim

To investigate physicochemical characteristics and especially genotypic diversity of the main culturable micro‐organisms involved in fermentation of sap from Borassus akeassii, a newly identified palm tree from West Africa.

Methods and Results

Physicochemical characterization was performed using conventional methods. Identification of micro‐organisms included phenotyping and sequencing of: 26S rRNA gene for yeasts, 16S rRNA and gyrB genes for lactic acid bacteria (LAB) and acetic acid bacteria (AAB). Interspecies and intraspecies genotypic diversities of the micro‐organisms were screened respectively by amplification of the ITS1‐5.8S rDNA‐ITS2/16S‐23S rDNA ITS regions and repetitive sequence‐based PCR (rep‐PCR). The physicochemical characteristics of samples were: pH: 3·48–4·12, titratable acidity: 1·67–3·50 mg KOH g^?1, acetic acid: 0·16–0·37%, alcohol content: 0·30–2·73%, sugars (degrees Brix): 2·70–8·50. Yeast included mainly Saccharomyces cerevisiae and species of the genera Arthroascus, Issatchenkia, Candida, Trichosporon, Hanseniaspora, Kodamaea, Schizosaccharomyces, Trigonopsis and Galactomyces. Lactobacillus plantarum was the predominant LAB species. Three other species of Lactobacillus were also identified as well as isolates of Leuconostoc mesenteroides, Fructobacillus durionis and Streptococcus mitis. Acetic acid bacteria included nine species of the genus Acetobacter with Acetobacter indonesiensis as predominant species. In addition, isolates of Gluconobacter oxydans and Gluconacetobacter saccharivorans were also identified. Intraspecies diversity was observed for some species of micro‐organisms including four genotypes for Acet. indonesiensis, three for Candida tropicalis and Lactobacillus fermentum and two each for S. cerevisiae, Trichosporon asahii, Candida pararugosa and Acetobacter tropicalis.

Conclusion

fermentation of palm sap from B. akeassii involved multi‐yeast‐LAB‐AAB cultures at genus, species and intraspecies level.

Significance and Impact of the Study

First study describing microbiological and physicochemical characteristics of palm wine from B. akeassii. Genotypic diversity of palm wine LAB and AAB not reported before is demonstrated and this constitutes valuable information for better understanding of the fermentation which can be used to improve the product quality and develop added value by‐products.