南极罗斯海区域可培养微生物分离鉴定及产低温酶能力初步筛选

为了利用南极微生物资源、探索南极罗斯海区域可培养微生物多样性，利用传统平板培养法对中国第33次南极科学考察采自南极罗斯海6个站位的海洋沉积物样品进行了细菌、真菌的分离培养。经细菌16S rRNA、真菌ITS基因序列检测及系统发育分析，共获得5个属的36株细菌和6个属的29株真菌。其中嗜冷杆菌属为优势细菌类群，枝孢属为优势真菌类群。该结果表明南极罗斯海区域具有丰富的微生物多样性。细菌API 20 NE生理生化及真菌产胞外酶活性实验显示，分离得到的细菌和真菌绝大多数都有低温酶活性。研究结果为南极罗斯海区域可培养微生物的多样性认识和低温酶产生菌资源获取与开发利用提供了支撑。     

南极菲尔德斯半岛可培养土壤微生物多样性及理化性质鉴定

为了探索南极可培养土壤微生物的多样性,本研究对中国第31次南极科学考察采集自南极菲尔德斯半岛的5份土壤样品进行了细菌、真菌的分离培养。选择形态差异较大的细菌和真菌进行了16S rDNA和ITS鉴定及系统发育分析。最终共鉴定得到20个属的33株细菌和6个属的8株真菌,其中8株细菌,10株真菌序列相似性较低,可能是新种。该结果表明南极菲尔德斯半岛地区具有丰富的微生物多样性,其中假单胞菌属的细菌居多。对分离得到的细菌和真菌分别进行理化性质和胞外酶活性鉴定,实验结果显示,分离获得的细菌和真菌绝大多数可产生水解酶类并同化多种碳源。初步认定这些微生物在参与南极物质代谢、适应南极极端环境方面发挥作用。本研究丰富了对南极菲尔德斯半岛可培养土壤微生物多样性的认识,并筛选获得了一些产低温酶特性的菌株,这为极地微生物资源的利用研究奠定了基础。     

长江口可培养耐(嗜)盐细菌群落结构研究

针对长江口10个站位样品,采用HM培养基和ZMCA培养基分离培养耐(嗜)盐菌,通过PCR方法获取156株细菌的16S rDNA序列。其中,由HM培养基分离获得136株菌株,可划分为33个分类单元(OUT),分属α-变形菌纲(Alphaproteobacteria)、γ-变形菌纲(Gammaproteobacteria)和厚壁菌门(Firmicutes)的芽孢杆菌纲(Bacilli)3个类群,其中γ-变形菌纲为优势耐(嗜)盐细菌;靠近海岸的站点M5-1,N6-2和M4-1耐(嗜)盐细菌均属于厚壁菌门,而远离海岸的站点耐(嗜)盐细菌均属于变形菌门。以ZMCA培养基分离获得的20株菌株分属α-变形菌纲和放线菌纲(Actinobacteria),可划分为10个OTU。由HM培养基分离获得的菌株中,70.5%属于耐盐菌,29.5%属于中度嗜盐菌;由ZMCA培养基分离获得的菌株均为耐盐菌。HM培养基分离得到的136株细菌的硝酸盐和亚硝酸盐还原性分析表明,亚硝酸盐还原菌数量仅为硝酸盐还原菌的1/3左右。     

抑制尖刀镰孢菌(Fusarium oxysporum)南极细菌的筛选及Psychrobacter sp.P4-11-07-1的生长特性

以植物病原真菌尖刀镰孢菌(Fusarium oxysporum)为指示菌,采用平板扩散法从实验室极地微生物资源库中筛选到12株具有明显抑菌作用的活性菌株。分子鉴定与系统发育分析表明,7株活性菌株属于假交替单胞菌(Pseudoalteromonas)、3株属于嗜冷杆菌(Psychrobacter)、1株属于假单胞菌(Pseudomonas)、1株属于伦黑墨氏菌(Rheinheimera)。抑菌谱测定结果表明,不同活性菌株的抑菌谱也有所不同,12株活性菌株对尖刀镰孢菌、辣椒疫霉(Phytophthora copsici)、大丽轮枝菌(Verticillium dahliae)、瓜亡革菌(Thanatephorus cucumeris)均有一定的抑制作用,而对葡枝根霉(Rhizopus stolonifer)均没有抑制作用;除菌株1-Z11,1-Z18,4-Z11,4-Z18,IS-010-07-1,P3-11-10-1,Z18-3外,其余5株菌株均对水稻纹枯病菌(Rhizoctonia solani)有抑制作用。对一株具有较强抑菌活性的菌株Psychrobacter sp.P4-11-07-1生长条件的研究表明,当培养温度为5～30℃,培养基盐度为0～45,初始pH值为5.0～11.0时,菌株P4-11-07-1均可生长并产生抑菌活性物质,对应抑菌活性最高的培养条件分别是:培养温度为25℃,培养基盐度为0,初始pH为7.0。     

Community diversity of bacteria in digestive tract of mud snail (Bullacta exarata Philippi) and its rearing shoal

ＩＮＴＲＯＤＵＣＴＩＯＮＴｈｅｍｕｄｓｎａｉｌＢｕｌｌａｃｔａｅｘａｒａｔａＰｈｉｌｉｐｐｉｉｓｄｅｌｉｃｉｏｕｓａｎｄｎｕｔｒｉｔｉｏｕｓ ,ａｎｄｗｅｌｌ ａｃｃｅｐｔｅｄｂｙｐｅｏｐｌｅ,ｅｓｐｅｃｉａｌｌｙｉｎＺｈｅｊｉａｎｇＰｒｏｖｉｎｃｅ ,Ｃｈｉｎａ ,ｋｎｏｗｎａｓａｎｅｃｏｎｏｍｉｃａｌｌｙｉｍｐｏｒｔａｎｔｍａｒｉｃｕｌ ｔｕｒａｌｓｈｅｌｌｆｉｓｈｓｐｅｃｉｅｓ .Ｉｔｈａｓａｔｔｒａｃｔｅｄｃｏｎｓｉｄｅｒａｂｌｅｍａｒｉｃｕｌｔｕｒｅｉｎｔｅｒｅｓｔａｌｏｎｇｔｈｅｃｏａｓｔｏｆ…     

南沙深海沉积物中石油降解菌的分离鉴定和多样性分析

为了解我国南沙海域中石油降解菌的多样性,认识其在环境自净中作用并获得用于石油污染生物修复的菌种资源,以不同链长烷烃及支链烷烃对南沙深海沉积物样品进行富集与烃降解菌的分离鉴定,并结合16S rRNA基因文库和DGGE技术对降解菌群的结构进行了分析.结果表明：通过16S rRNA基因文库构建及PCR-DGGE分析发现,不同链长烷烃富集后的菌群中,食烷菌（Alcanivorax）在各降解菌群中都是绝对优势菌;另外,除烃海杆菌（Marinobacter hydrocarbonoclasticus）、海绵假单胞菌（Pseudomonas pachastrellae）,以及Idiomarina与Kangiella属的细菌也是菌群中较重要的优势菌.此外,通过2种平板培养基从烷烃富集的降解菌群中分离获得了8个不同属的细菌.本研究还首次发现Kangiella属（食烷菌科）的细菌可能参与了烷烃降解.南沙沉积物烷烃降解菌多为γ-Proteobacteria;其中,食烷菌科（Alcanivoraceae）（Alcanivorax与Kangiella属）的细菌在降解菌群中是绝对优势菌,假单胞菌与海杆菌在烷烃降解过程中也起着重要作用.     

A STUDY ON MICROBIAL DEGRADATION FOR OIL POLLUTANTS IN MARINE ENVIRONMENT

This paper presents the degradation of 10 strains of hydrocarbon-degrading bacteria isolated from Xiamen harbor for 8 kinds of hydrocarbon and their mixed hydrocarbons incubated for 7 days at 25℃. The results indicate that the degradation rates of different genera of bacteria, which are measured with gas chromatography, are different for 8 hydrocarbons, thus showing the differences among genera; that the degradation rates of most strains for alkane hydrocarbons are higher than those for aromatic hydrocarbons; that the mixed strains favour the degratation for mixed hydrocarbons; and that the concentration of hydrocarbon is an important factor affecting degradation rates.     

Hydrocarbon-utilising micro-organisms from Dona Paula Bay,Goa

Twenty-three hydrocarbon-utilising bacteria and one yeast were isolated, using enrichment techniques, from water and sediment samples. Vibrio and Pseudomonas were the predominant genera. Of the different organisms screened, Bacillus, Candida and Arthrobacter sp. exhibited the widest range of hydrocarbon-utilising profiles. Arabian Sea crude and kerosene supported the growth of most of the isolates.     

Phylogenetic analysis and biological characteristic tests of marine bacteria isolated from Southern Ocean (Indian sector) water

Fifty-seven bacteria were isolated from Southern Ocean(Indian sector) water samples which were collected from different latitude and longitude of the ocean. All the isolates were able to grow at 4°C, 20°C, 37°C and tolerable Na Cl concentration up to 13.5%(w/v). 29 out of 57 isolates were identified using 16 S rDNA amplification and the sequences were submitted to National Center for Biotechnology Information(NCBI). All the isolates were classified by using Ribosomal Database Project(RDP) and found that isolates belongs to Proteobacteria and Bacteriodes. The average G+C content was 56.4%. The isolates were screened for the presence of extracellular enzymes, viz. amylase, catalase, urease, esterase, lipase and protease. The disc diffusion method is used to screen antibiotic production by the isolates against four pathogenic bacteria, viz. Salmonella typhimurium(NCIM 2501),Staphylococcus aureus(NCIM 2122), Bacillus subtilis(NCIM 2193), and Pseudomonas aeruginosa(NCIM 2036).Nine out of 29 were found to be antibiotic producer.     

北极黄河站地区不同基底中真菌的分离培养及初步鉴定

为了探索北极陆域真菌的多样性,进一步揭示其生态学角色,本研究对中国第5次北极科学考察采集自黄河站区附近的24份样品进行了真菌的分离培养及鉴定,样品涂布划线后共分离得到60株真菌,其中腐殖质样品中分离到的真菌种类和数量最多;对选取的28株代表菌株进行了真菌ITS区序列鉴定,结果表明它们分属于4个纲,12个属,分别为散囊菌纲、粪壳菌纲、座囊菌纲和微藻菌纲,其中地丝霉属为优势类群。该结果表明北极地区具有丰富的真菌物种多样性。通过对代表土壤理化性质的测定,探究了真菌多样性和土壤理化参数的相关性,其中有机碳氮及可溶性营养盐含量是影响北极土壤真菌多样性的重要指标。     

厦门海区潮间带海洋动植物共附生微生物的抗菌活性

从厦门海区潮间带石纯、浒苔、江蓠、海兔、海葵及鲨鱼肠道分离得２８９株共附生微生物,以８种敏感菌为指示菌,从中获得拮抗活性的幼苗２８株,放线菌１７株,真菌８株。拮抗细菌以假单胞菌（３９％）、弧菌（２５％）和芽直菌（１４％）为主,主要为抗革兰氏阳性细菌,抗真菌的菌株比例较低。拮抗放线菌分属于链霉菌和小单孢菌,其抗革兰氏阳性菌、阴性菌和真菌的菌株比例分别为６５％、４７％和２３％。拮抗真菌分青霉属和芽枝霉     

广西涠洲岛柳珊瑚共附生真菌多样性及其抑菌活性*

文章使用4种分离培养基, 从10个广西涠洲岛柳珊瑚样本中分离可培养真菌, 通过菌落形态观察和内转录间隔区序列系统发育分析, 得到涠洲岛柳珊瑚共附生真菌多样性信息, 并采用96孔板法测定了真菌发酵液乙酸乙酯提取物的抑菌活性和抗生物被膜形成活性。试验结果表明, 从涠洲岛柳珊瑚样本中分离得到191株共附生真菌, 鉴定为26个种。这些真菌均属于子囊菌门, 分布于6个属: 曲霉属(Aspergillus)、青霉属(Penicillium)、链格孢属(Alternaria)、枝孢菌属(Cladosporium)、黑孢霉属(Nigrospora)和Parengyodontium属。曲霉属(Aspergillus)为优势种属, 占总菌株种类的69.23%; 次优属为枝孢菌属(Cladosporium)。抑菌活性筛选发现有11种真菌至少对一种指示菌有抑制活性, 占菌种总数的42.31%; 有7种真菌对表皮葡萄球菌生物被膜形成具有抑制活性, 半数有效浓度(EC₅₀)范围为40.3~155μg·mL^-1, 菌株Penicillium cinnamopurpureum GXIMD00518、Aspergillus carneus GXIMD00519具有抗耐甲氧西林金黄色葡萄球菌生物被膜形成活性, EC₅₀分别为105.4μg·mL^-1和117.4μg·mL^-1。     

A culture-dependent survey of thermophilic bacteria from hot springs in Xiamen area in China

Microbes are believed to play important roles in ecosystem function in many environments. The hot springs of Xiamen Island are close to the Xiamen Sea, and may have some characteristics different from those of inland hot springs. Microbes living in the hot springs of Xiamen may have new characteristics. However, little is known about microbial communities of hot springs close to the Xiamen Sea. A culturedependent survey of microbial population in the Xiamen hot springs was performed by using an approach combining total cellular protein profile identification and 16S rRNA gene sequencing. A total of 328 isolates of bacteria were obtained from liquid and sediment samples from the Xiamen hot springs, including neutrophilic thermophilic bacteria and moderately thermophilic acidophiles. Neutrophilic thermophilic bacteria, which grow at a temperature range of 55—90℃ including Rhodothermus marinus (Strain 1), Thermus thermophilus (Strain 2), Thermus thiopara (Strain 3), Geobacillus stearothermophilus (Strain 4), Geobacillus thermoleovorans (Strain 5), and Pseudomonas pseudoalcaligenes (Strain 6), were recovered by 2216E plates. Moderately thermophilic acidophiles, which can grow at temperatures above 50℃ and a pH range of 1.8—3.5 such as Alicyclobacillus acidoterrestris (Strain 8), Sulfobacillus acidophilus (Strain 9), and Sulfobacillus thermosulfidooxidans (Strain 10), were isolated on selective solid medium containing sulfur and Fe2+. Among these strains, Rhodothermus marinus, Thermus thermophilus and Geobacillus stearothermophilus are not only thermophiles, but also halophiles. One bacterium strain (Strain 6) shared 99% nucleotide sequence homology with Pseudomonas pseudoalcaligenes on the 16S rRNA gene sequence, but was quite different from Pseudomonas pseudoalcaligenes in biological characteristics, suggesting that it may represent a novel thermophilic species. Results indicated that various species of neutrophilic thermophiles and moderately thermophilic acidophiles were widely distributed in the Xiamen hot springs and that Rhodothermus marinus and Thermus thermophilus dominated the cultivable microbial community.     

A culture-dependent survey of thermophilic bacteria from hot springs in Xiamen area in China

Microbes are believed to play important roles in ecosystem function in many environments. The hot springs of Xiamen Island are close to the Xiamen Sea, and may have some characteristics different from those of inland hot springs. Microbes living in the hot springs of Xiamen may have new characteristics. However, little is known about microbial communities of hot springs close to the Xiamen Sea. A cuhure-dependent survey of microbial population in the Xiamen hot springs was pcrformed by using an approach combining total cellular protein profile identification and 16S rRNA gene sequencing. A total of 328 isolates of bacteria were obtained from liquid and sediment samples from the Xiamen hot springs, including neutrophilie thermophilic bacteria and moderately thermophilic acidophiles. Neutrophilic thermophilic bacteria, which grow at a temperature range of 55-90℃ including Rhodothermus marinus (Strain 1) , Thermus thermophilus (Strain 2), Thermus thiopara (Strain 3) , Geobacillus stearothermophilus(Strain 4) , Geobacillus thermoleovorans (Strain 5) , and Pseudomonas pseudoal-caligenes (Strain 6), were recovered by 2216E plates. Moderately thermophilic acidophiles, which can grow at temperatures above 50℃ and a pH range of 1. 8-3.5 such as Alicyclobacillus acidoterrestris (Strain 8) , Sul-fobacillus acidophilus (Strain 9), and Sulfobacillus thermosulfidooxidans (Strain 10), were isolated on selective solid medium containing sulfur and Fe2+. Among these strains, Rhodothermus marinus, Thermus thermophilus and Geobacillus stearothermophilus are not only thermophilcs, but also halophiles. One bacterium strain (Strain 6) shared 99% nucleotide sequence homology with Pseudomonas pseudoalcaligenes on the 16S rRNA gene se-quence, but was quite different from Pseudomonas pseudoalcaligenes in biological characteristics, suggesting that it may represent a novel thermophilic species. Results indicated that various species of neutrophilic thermophiles and moderately thermophilic acidophiles were widely distributed in the Xiamen hot springs and that Rhodother-mus marinus and Thermus thermophilus dominated the cultivable microbial community.     

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

Actinomycetes in five marine sediments collected from the Arctic Ocean at depths of 43 to 3 050 m were cultivated using a variety of media. A total of 61 actinomycete colonies with substrate mycelia only were observed, and no colonies with aerial mycelia were observed under aerobic conditions at 15 ℃. From these colonies, 28 were selected to represent different morphological types.Denaturing gradient gel electrophoresis (DGGE) was used to check the purity of isolates and select representatives for subsequent sequencing. Phylogentic analyses based on nearly full-length 16S ribosomal RNA gene (rDNA) sequences indicated that the actinomycetes isolated were accommodated within genus Rhodococcus of family Nocardiaceae, genus Dietzia of family Dietziaceae,genera Janibacter and Terrabacter of family Instrasporangiaceae and genera Kocuria and Arthrobacter of family Micrococcaceae. One of the strains (P27-24) from the deep-sea sediment at depth of 3 050 m was found to be identical in 16S rDNA sequence(1474/1474)with the radiation-resistant Kocuria rosea ATCC 187T isolated from air. More than halfofthe isolates showed 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 strains isolated were psychrotolerant bacteria and grew better on the media prepared with natural seawater than 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 these strains are indigenous marine actinomycetes.     

Fungi in deep-sea sediments of the Central Indian Basin

Although a great amount of information is available on bacteria inhabiting deep-sea sediments, the occurrence of fungi in this environment has been poorly studied and documented. We report here the occurrence of fungi in deep-sea sediments from ∼5000 m depth in the Central Indian Basin (9–16°S and 73–76°E). A total of 181 cultures of fungi, most of which belong to terrestrial sporulating species, were isolated by a variety of isolation techniques. Species of Aspergillus and non-sporulating fungi were the most common. Several yeasts were also isolated. Maximum species diversity was observed in 0–2 cm sections of the sediment cores. Direct staining of the sediments with Calcofluor, a fluorescent optical brightener, revealed the presence of fungal hyphae in the sediments. Immunofluorescence using polyclonal antibodies raised against a deep-sea isolate of Aspergillus terreus (# A 4634) confirmed its presence in the form of hyphae in the sub-section from which it was isolated. A total of 25 representative species of fungi produced substantial biomass at 200 bar pressure at 30° as well as at 5 °C. Many fungi showed abnormal morphology at 200 bar/5 °C. A comparison of terrestrial isolates with several deep-sea isolates indicated that the former could grow at 200 bar pressure when growth was initiated with mycelial inocula. However, spores of a deep-sea isolate A. terreus (# A 4634), but not the terrestrial ones, showed germination at 200 bar pressure and 30 °C. Our results suggest that terrestrial species of fungi transported to the deep sea are initially stressed but may gradually adapt themselves for growth under these conditions.     

胶州湾沉积物中石油降解菌的降解能力及多样性研究

2013年青岛输油管道爆炸，大量石油污染了附近海岸。课题组采集了污染的沉积物样品，以原油为唯一碳源和能源，富集了四个石油降解菌群。生物多样性和群落分析表明，Luteibacter、Parvibaculum 和属于食烷菌科的一个属是降解菌群的主要优势菌，都属于变形菌门。从石油降解菌群中分离筛选，获得了9株具有不同16S rRNA基因序列的降解菌，分别属于8个属。重量法测定降解菌的石油降解率，其中5株的石油降解率大于30%。GC-MS分析结果表明，石油降解菌多倾向于降解烷烃，对多环芳烃的降解能力较差，其中5株细菌的烷烃降解率较大，仅1株菌D2对多环芳烃的降解率较大，其降解率在34.9%到77.5%。通过对高通量数据的分析，表明食烷菌属是菌群A和菌群E的主要降解菌群，其中筛选获得的菌株E4可能是菌群E的一株优势降解菌。本研究所筛选菌株证明了其石油降解潜力，为油污染海滩生物修复提供了菌株资源。     

北极海洋沉积物中可培养细菌及其多样性分析

利用Zobell 2216E培养基和涂布平板法对北极海洋沉积物中可培养细菌进行分离纯化,并利用16SrRNA基因进行分子鉴定与系统发育分析。根据菌落形态学特征,从59个站点的沉积物样品中共分离纯化获得570株细菌;基于16SrRNA基因的分子鉴定与系统发育分析表明,分离到的可培养细菌分别属于细菌域的4个门,5个纲,12个目,23个科,47个属,102个种,其中γ-Proteobactria占绝大多数;有14株菌株与模式菌株的16SrRNA基因序列相似性小于97%,为6个潜在的新种。北极海域的海洋沉积物中存在着丰富的微生物种质资源,为开发新型生物活性物质和特殊功能基因打下了基础。     

地芽孢杆菌属DYth03 DNA聚合酶基因的克隆、表达及性质分析

从东太平洋热液区E53站位的深海沉积物样品中分离出1株能在65℃生长的嗜热菌（DYth03）.该菌的16S rDNA序列与地芽孢杆菌属（Geobacillus）内各种之间的同源性为98%以上.克隆得到DYth03的DNA聚合酶基因（DYth-pol）,序列分析表明该基因全长为2 631 bp,G＋C含量为55.5%,推测编码为876个氨基酸,与BstDNApolI的同源性最高（达98%）.将该聚合酶基因克隆到pTTQ-h表达载体上,并在大肠杆菌DH1中进行表达.对纯化到的表达产物进行酶活性测定,结果表明该酶具有聚合酶活性和5’-3’外切酶活性.     

光合细菌对褶皱臂尾轮虫的饵料价值

我们自青岛栈桥浅海海泥中分离到一株光合细菌,经鉴定为球形红假单胞菌(Rhodopseudomonas spheroides)。对其生化特性、生长条件以及做为褶皱臂尾轮虫(Brachionus plicatilis)的饵料价值,进行了研究。结果表明,以球形红假单胞菌的新鲜培养物,混以青岛大扁藻(Platymonas helgolandica var.tsingtaoensis)喂养轮虫,轮虫的增殖率明显高于单独使用光合细菌、扁藻和海洋酵母,也高于光合细菌与海洋酵母的混合。提示在轮虫的生产性培养中,以一定浓度的光合细菌与单胞藻混合投喂轮虫,可能取得较高产量。