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Isolation and phylogenetic assignation of actinomycetes in the marine sediments from the Arctic Ocean

Acta Oceanologica Sinica 2005年第6期24卷 135-142页

作者： yu yong li huirong zeng yinxin chen bo State Oceanic Administration Key Laboratory for Polar Science Polar Research Institute of China Shanghai 200136 China State Oceanic Administration Key Laboratory for Polar Science Polar Research Institute of China Shanghai 200136 China State Oceanic Administration Key Laboratory for Polar Science Polar Research Institute of China Shanghai 200136 China State Oceanic Administration Key Laboratory for Polar Science Polar Research Institute of China Shanghai 200136 China

Actinomycetes in five marine sediments collected from the Arctic Ocean atdepths of 43 to 3 050 m were cultivated using a variety of media. A total of 61 actinomycetecolonies with substrate mycelia only were observed, and no colonies with aerial mycelia wereobserved under aerobic conditions at 15℃. From these colonies, 28 were selected to representdifferent morphological types. Denaturing gradient gel electrophoresis (DGGE) was used to check thepurity of isolates and select representatives for subsequent sequencing. Phylogentic analyses basedon nearly full-length 16S ribosomal RNA gene (rDNA) sequences indicated that the actinomycetesisolated were accommodated within genus Rhodococcus of family Nocardiaceae, genus Dietzia of familyDietziaceae, genera Janibacter and Terrabacter of family Instrasporangiaceae and genera Kocuria andA nhrobacter of family Micrococcaceae. One of the strains (P27-24) from the deep-sea sediment atdepth of 3 050 m was found to be identical in 16S rDNA sequence(1474/1474) with theradiation-resistant Kocuria rosea ATCC 187~T isolated from air. More than half of the isolatesshowed the similarities ranging from 99.5% to 99.9% in 16S rDNA sequence to dibenzofran-degrading,butyl 2-ethylhexanoate-hydrolysising and nitrile-metabolizing actinomycetes. All the strainsisolated were psychrotolerant bacteria and grew better on the media prepared with natural seawaterthan on the media prepared with deionized water. Three of them (Dietzia sp. P27-10, Rhodococcus sp.S11-3 and Rhodococcus sp.P11-5) had an obligate growth requirement for salt, confirming that thesestrains are indigenous marine actinomycetes.

关键词： marine actinomycetes genetic diversity arctic ocean

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Phylogenetic diversity and phenotypic characterization of cultivable bacterioplankton isolated from polar oceans

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Acta Oceanologica Sinica 2007年第4期26卷 93-103页

作者： zeng yinxin li huirong yu yong chen bo ZHENG Tianling School of life Sciences Xiamen University Xiamen 361005 China Key Laboratory for Polar Science of State Oceanic Administration Polar Research Institute of China State Oceanic Administration Shanghai 200136 China

A set of 27 marine planktonic bacteria isolated from the polar regions was characterized by 16S rDNA sequencing and physiological and biochemical testing. More than half of these bacteria were positive for caseinase, gelatinase and 13-glucosidase, and could utilize glucose, maltose or malic acid as carbon source for cell growth. Twelve isolates expressed nitrate reduction activities. Except for one antarctic isolate BSwlO175 belonging to Actinobacteria phylum, these isolates were classified as γ-Proteobacteria, suggesting that γ-Proteobacteria dominated in cultivable marine bacterioplankton at both poles. Genus Pseudoalteromonas was the predominant group in the Chukchi Sea and the Bering Sea, and genus ShewaneUa dominated in cultivable bacterioplankton in the Prydz Bay. With sequence similarities above 97%, genus Psychrobacter was found at both poles. These 27 isolates were psychrotolerant, and significant 16S rDNA sequence similarities were found not only between arctic and antarctic marine bacteria （〉 99% ）, but also between polar marine bacteria and bacteria from other aquatic environments （ ≥ 98.8% ）, including temperate ocean, deep sea, pond and lake, suggesting that in the polar oceans less temperature-sensitive bacteria may be cosmopolitan and have a bipolar, even global, distribution at the species level.

关键词： bacterioplankton diversity arctic antarctic

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Molecular genetic diversity of bacteria in the bottom section of arctic sea ice from the Canada Basin

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Acta Oceanologica Sinica 2005年第6期24卷 153-161页

作者： li huirong yu yong chen bo zeng yinxin REN Daming State Oceanic Administration Key Laboratory for Polar Science Polar Research Institute of ChinaShanghai 200136China State Key Laboratory of Genetic Engineering Institute of GeneticsFudan UniversityShanghai 200433China

PCR-DGGE approach was used to analyze bacterial diversity in the bottomsection of seven arctic sea ice samples colleted from the Canada ***-two 16S rDNAsequences were obtained from prominent DGGE *** closest relatives of these sequences arefound to be those of cultivated or uncultured bacteria from antarctic or arctic sea *** analysis clustered these sequences or phylotypes withinα-proteobacteria,γ-proteobacteria and CFB(cytophaga-flexibacter-bacteroides)*** belonging toγ-proteobacteria were dominant and members of the CFB group were highly *** was suggestedthat the CFB group was the representative of the bottom section of sea ice samples.

关键词： sea ice arctic ocean bacteria 16S rDNA phylogenetic analysis

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Phylogenetic relationship and phenotypic comparison of Psychrobacter species isolated from polar oceans

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Progress in Natural Science:Materials International 2007年第1期17卷 44-48页

作者： zeng yinxin, li huirong, yu yong and chen bo (Key Laboratory for Polar Science of State Oceanic Administration, Polar Research Institute of China, Shanghai 200136, China) Key Laboratory for Polar Science of State Oceanic Administration Polar Research Institute of China Shanghai 200136 China

To investigate the phylogenetic relationship and biogeography of bacterioplankton in polar oceans, four Psychrobacter strains, BSw10170, BSw20352, BSw20370, and BSw20461, isolated from seawater of the Beijing Sea, the Chukchi Sea, and the Prydz Bay, were characterized by 16S rDNA sequencing and physiological and biochemical testing. Results demonstrated that close relationships existed between the Arctic and Antarctic strains with sequence similarities higher than 97 %. These four Psychrobacter strains not only showed almost identical phenotypic characteristics among them, but also shared a lot of similarities with those related Psychrobacter species, indicating that psychrotolerance and halotolerance of Psychrobacter strains may be among the reasons for their bipolar, even global distribution in marine environments at the genus level.

关键词： Psychrobacter phylogenetic phenotypic Arctic Antarctic.

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低温纤维素酶产生菌的筛选、鉴定、生长特性及酶学性质

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高技术通讯 2005年第4期15卷 58-62页

作者：曾胤新俞勇陈波李会荣中国极地研究中心国家海洋局极地科学重点实验室上海200129

从北极楚科奇海分离出一株产纤维素酶的耐冷假交替单胞菌BSw20308.研究表明:该菌最适生长温度10℃,35℃不生长;在pH7.0～8.0、含2.0%～3.0% NaCl的培养基条件下最适宜菌株生长与产酶;可溶性淀粉、酵母浸出液是有利于菌株生长及产酶的碳... 详细信息

从北极楚科奇海分离出一株产纤维素酶的耐冷假交替单胞菌BSw20308.研究表明:该菌最适生长温度10℃,35℃不生长;在pH7.0～8.0、含2.0%～3.0% NaCl的培养基条件下最适宜菌株生长与产酶;可溶性淀粉、酵母浸出液是有利于菌株生长及产酶的碳源、氮源物质;蔗糖、可溶性淀粉、麦芽糖及麸皮对CMCase的合成具有明显诱导作用;CMC、麸皮、可溶性淀粉及麦芽糖对滤纸酶的合成有一定诱导作用,单糖与部分双糖(蔗糖、纤维二糖及乳糖)则起阻遏作用;在指数生长后期至稳定早期,大量产CMCase;发酵液含CMCase、滤纸酶及葡萄糖苷酶活性,以CMCase活力最高;胞外CMCase活力占总CMCase活力的74.1%左右;酶最适作用温度35℃,5℃时酶活保留50%左右;酶对热敏感,35℃半衰期为3 h,25℃下酶活稳定;酶最适pH8.0.

关键词：纤维素酶产生菌酶学性质生长特性可溶性淀粉筛选低温鉴定最适生长温度菌株生长诱导作用酵母浸出液葡萄糖苷酶楚科奇海 NaCl 阻遏作用纤维二糖生长后期作用温度麦芽糖滤纸酶单胞菌培养基源物质 CMC

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北极耐冷假交替单胞菌的培养温度对其胞外蛋白酶合成的影响

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高技术通讯 2007年第5期17卷 535-539页

作者：曾胤新李会荣俞勇陈波中国极地研究中心国家海洋局极地科学重点实验室上海200136

从北极楚科奇海水中分离筛选到2株产蛋白酶耐冷细菌BSw20308及BSw20353,经分子鉴定为假交替单胞菌属。这2株菌虽存在较近亲缘关系,16SrDNA序列相似性为98%,但属于不同种,表型方面也存在差异。酶谱分析显示,此2株菌的胞外产物中存在至少... 详细信息

从北极楚科奇海水中分离筛选到2株产蛋白酶耐冷细菌BSw20308及BSw20353,经分子鉴定为假交替单胞菌属。这2株菌虽存在较近亲缘关系,16SrDNA序列相似性为98%,但属于不同种,表型方面也存在差异。酶谱分析显示,此2株菌的胞外产物中存在至少3种蛋白水解活性组分。培养温度的变化对这2株菌胞外蛋白水解活性组分的生成具有不同的影响效果,表明极地海洋细菌可能在酶学水平上通过酶的种类与活力的调整和采取不同的策略来适应环境温度的变化。

关键词：耐冷假交替单胞菌北极蛋白酶温度

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Archaeal diversity and abundance within different layers of summer sea-ice and seawater from Prydz Bay,Antarctica

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Advances in Polar Science 2014年第1期25卷 54-60页

作者： MA Jifei DU Zongjun LUO Wei yu yong zeng Yixin chen bo li huirong Polar Research Institute of China College of Marine Science Shandong University

Fluorescence in-situ hybridization （FISH） and 16S rRNA gene clone library analyses were used to determine the abundance and diversity of archaea in Prydz Bay, Antarctica. Correlation analysis was also performed to assess links between physicochemical parameters and archaeal abundance and diversity within the sea-ice. Samples of sea-ice and seawater were collected during the 26th Chinese National Antarctic Research Expedition. The results of FISH showed that archaea were relatively abundant within the top layer of the sea-ice, and correlation analysis suggested that the concentration of 4NH＋ might be one of the main factors underlying this distribution pattern. However, using 16S rRNA gene libraries, archaea were not detected in the top and middle layers of the sea-ice. All archaeal clones obtained from the bottom layer of the sea-ice were grouped into the Marine Group I Crenarchaeota while the archaeal clones from seawater were assigned to Marine Group I Crenarchaeota, Marine Group II Euryarchaeota, and Marine Group III Euryarchaeota. Overall, the ifndings of this study showed that the diversity of archaea in the sea-ice in Prydz Bay was low.

关键词： Antarctica summer sea-ice archaea diversity abundance

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北极海洋细菌BSw20353的胞外蛋白酶性质

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极地研究 2006年第3期18卷 206-214页

作者：曾胤新俞勇李会荣陈波中国极地研究中心

从北极楚科奇海水中分离到一株产蛋白酶耐冷河豚毒素假交替单胞菌BSw20353。该菌可合成多种胞外蛋白酶,而培养温度影响蛋白酶的合成。分离纯化出其中一种在15℃、25℃培养时均有生成的酶:该酶最适作用温度55℃;最适pH8;金属离子Hg2+、S... 详细信息

从北极楚科奇海水中分离到一株产蛋白酶耐冷河豚毒素假交替单胞菌BSw20353。该菌可合成多种胞外蛋白酶,而培养温度影响蛋白酶的合成。分离纯化出其中一种在15℃、25℃培养时均有生成的酶:该酶最适作用温度55℃;最适pH8;金属离子Hg2+、Sr2+抑制酶活,Fe2+、Mn2+则具有激活作用;该酶属于金属蛋白酶,受螯合剂邻二氮杂菲、EDTA及EGTA抑制,基本不受丝氨酸蛋白酶抑制剂PMSF、TPCK及TLCK抑制,半胱氨酸蛋白酶抑制剂E-64对该酶无抑制作用,天冬氨酸蛋白酶抑制剂pepstatin则具有一定抑制作用;SDS、二硫苏糖醇、碘乙酸可抑制酶活;而在变性剂尿素、盐酸胍作用下,酶活反而有所增高。

关键词：河豚毒素假交替单胞菌北极金属蛋白酶

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一株产海藻糖合成酶南极海洋低温细菌的鉴定

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极地研究 2005年第2期17卷 127-133页

作者：俞勇李会荣陈波曾胤新中国极地研究中心国家海洋局极地科学重点实验室上海200136

从南极普里兹湾海域海水中分离到一株产海藻糖合成酶的海洋低温细菌BSw10041,革兰氏阴性,杆状,有极生鞭毛,能运动,菌落半透明。进行了常规生理生化和BIOLOG GN2细菌鉴定系统测试,结果表明菌株BSw10041与恶臭假单胞菌(Pseudomonas putida... 详细信息

从南极普里兹湾海域海水中分离到一株产海藻糖合成酶的海洋低温细菌BSw10041,革兰氏阴性,杆状,有极生鞭毛,能运动,菌落半透明。进行了常规生理生化和BIOLOG GN2细菌鉴定系统测试,结果表明菌株BSw10041与恶臭假单胞菌(Pseudomonas putida)的表型特征非常相似。为了进一步确定菌株BSw10041的分类学地位,测定了其16S rDNA序列,分析了相关细菌相应序列的同源性,构建了系统发育树,结果表明BSw10041与***的亲缘关系最近。综合上述结果,菌株BSw10041可鉴定为Pseudomonas putida,定名为Pseudomonas putida BSw10041。

关键词：海藻糖合成酶低温微生物 Pseudomonas putida 南极细菌鉴定低温海洋株产 rDNA序列

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海冰细菌研究进展

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极地研究 2005年第1期17卷 75-85页

作者：俞勇李会荣陈波曾胤新何剑锋中国极地研究中心国家海洋局极地科学重点实验室上海200136

地球两极海洋上覆盖的海冰为微生物提供了一个极端的生存环境。微生物在这环境中生存需要生理和代谢上的复杂适应机制。现有的研究表明,细菌在海冰中旺盛地生长繁殖,并在极区生态系统中占有重要地位。同时海冰细菌也已成为生命现象多样... 详细信息

地球两极海洋上覆盖的海冰为微生物提供了一个极端的生存环境。微生物在这环境中生存需要生理和代谢上的复杂适应机制。现有的研究表明,细菌在海冰中旺盛地生长繁殖,并在极区生态系统中占有重要地位。同时海冰细菌也已成为生命现象多样性等基础研究领域和生物工程领域的新热点。本文就近年来国内外在海冰细菌系统发育组成、生态生理学、生态作用和环境适应等方面所取得的研究进展进行简述,并初步探讨了今后海冰细菌研究中需要解决的一些问题。

关键词：海冰微生物生存环境复杂适应机制细菌生态系统发育组成生态生理学生态作用环境适应

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