On four closely related hypotrichous ciliates(Protozoa,Ciliophora,Spirotrichea):molecular characters,interspecific relationships and phylogeny defined with multigene sequence information

In order to clarify the phylogeny and relationships of the most confused hypotrichous ciliates,Holosticha-complex,four closely related holostichids（five populations）,Holosticha bradburyae,H.diademata,Anteholosticha sp.,and A.manca,were compared and analyzed using ITS2 secondary structures,ITS1-5.8S-ITS2 region and SSrRNA gene sequences.The ITS1-5.8S-ITS2 region sequences of these four species were first sequenced,and they shared sequence identities ranging from 68.0% to 90.1%,while two populations of Anteholosticha sp.differed in three nucleotides（sequence identity 99.8%）.There were several minor differences among ITS2 secondary structures of these species,while two populations of Anteholosticha sp.had the identical secondary structure.Phylogenetic trees inferred from the ITS1-5.8S-ITS2 region sequences of stichotrichs using multiple algorithms（Neighbor-Joining,Maximum Parsimony and Bayesian） revealed similar topologies.The results show that：（1） Holosticha bradburyae and H.diademata firmly clustered together with strong bootstrap supports,forming a sister clade with Anteholosticha sp.,（2） Anteholosticha appeared to be a paraphyletic assemblage,in which the morphotype A.manca was more closely related to Diaxonella trimarginata than to its congener Anteholosticha sp.Phylogenetic analyses based on the SSrRNA gene and the combined sequences of SSrRNA gene and ITS1-5.8S-ITS2 region revealed the similar relationships between Holosticha and Anteholosticha,nevertheless their positions within the subclass Stichotrichia differed from each other inferred from different genes.

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.kluyverivar.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 kluyverivar.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%。

不同树龄葡萄自然发酵过程中酵母菌的研究

【目的】研究葡萄树龄与葡萄自然发酵过程中葡萄酒相关酵母菌之间的关系。【方法】利用WL营养培养基,对分离自华夏长城葡萄酒厂葡萄生产基地、不同树龄(2,3,5年)西拉葡萄自然发酵液的240株酵母菌进行了聚类分析。【结果】240株酵母菌共分为5个WL营养类型。结合酵母菌菌株5.8 S rDNA-ITS区基因序列分析,不同树龄西拉葡萄自然发酵液中的酵母菌共有5种,分别为酿酒酵母(Saccharomyces cerevisiae)、陆生伊萨酵母(Is-satchenkia terricola)、克鲁维毕赤酵母(Pichia kluyveri)、浅黄隐球酵母(Cryptococcus flavescens)、葡萄汁有孢汉逊酵母(Hanseniaspora uvarum)。不同树龄西拉葡萄自然发酵过程中酵母菌的种类及其比例是变化的。【结论】用树龄较高的葡萄发酵,可加速发酵的启动,缩短发酵周期,同时有利于性状优良野生酿酒酵母菌株的筛选。