一株石油烃降解菌分子鉴定及特性分析

从受污海水中分离筛选了具有石油烃降解能力的降解菌Bac1020,经PCR扩增得到1 497 bp长16S rDNA序列,通过Blast比对,与主要石油烃降解菌属16S rDNA序列构建系统发生树,鉴定其为不动杆菌(Acinetobactersp.)。降解菌(Acinetobactersp.)在72 h内生长稳定,对石油烃的降解率随时间的延长而不断增加。建立了快速筛选及鉴定石油烃降解菌的方法,应用于海洋石油烃污染的生物降解。     

一株甲烷利用菌的分离、鉴定和性能研究

目的：研究土壤中能利用甲烷的细菌类型及特性。方法：运用传统的富集、分离、纯化方法得到1株甲烷利用菌,结果：该菌株在显微镜下呈球形,在液体培养基中生长72h后OD值可达到0.8左右,其最适生长温度30℃,最佳pH值7左右。对该菌株进行了16SrDNA扩增,测序结果表明其与Pseudoxanthomonas菌属的序列相似性为93%,其生理生化特征和分子鉴定结果表明,该菌与假黄单胞菌为不同的属。甲烷消耗气相色谱分析结论显示,120h后培养瓶中的甲烷浓度降低60%左右,说明该菌株具有较好的利用甲烷的性能。     

大蒜内生细菌的分离及拮抗菌筛选与鉴定

利用常规分离方法对大蒜鳞茎进行内生细菌的分离,采用对峙法和平板涂布法对分离的内生菌进行拮抗试验研究,并对菌株DSP6进行16S rDNA全序列鉴定。结果表明:分离得到19株内生细菌,其中10株菌对2种以上植物病原真菌有不同程度的抑制作用,占分离菌总数的52.6%,DSN7对番茄早疫病的抑菌圈半径最大,为13mm;17株菌对5种病原细菌中至少1种有抑制作用,占分离菌总数的89.5%,其中菌株DSP3对大肠杆菌的抑菌圈半径最大,达到10 mm;菌株DSP6对供试的9种病原菌有较强的抑菌作用,且抑菌圈平均半径最大,为6.88mm;16S rDNA全序列鉴定显示,菌株DSP6与芽孢杆菌属Bacillus axarquiensis相似性为100%,表明菌株DSP6为Bacillus axarquiensis。     

北极海泥菌群的分离鉴定及生物学特性的初步研究

对从北极冰川海面下1500-4000m处的海泥样品中分离的8株冷适应细菌进行了生理特征和分子生物学研究。其中5株嗜冷菌、3株耐冷菌,利用16SrDNA通用引物对5株嗜冷菌基因组DNA进行扩增,测序得到其部分16SrDNA序列。经Blast调出与菌株16SrDNA同源的序列,按照Neighbor-Joining方法构建16SrDNA系统发育树。对8株细菌进行酶检测试验,结果表明其中有部分细菌产低温酶:N014产淀粉酶,R151产明胶酶,P371产纤维素酶。研究结果为进一步开发利用冷适应微生物产物提供参考依据。     

大莲湖池杉林湿地土壤可培养细菌的分离与鉴定

采用牛肉膏蛋白胨培养基培养,从大莲湖池杉林土壤中共分离得到20个菌落形态不同的菌株。通过对这些菌株的形态、培养特征、生理生化特征的研究以及16S rDNA序列分析,初步确定这些菌株分别属于假单胞菌属(Pseudomonas)、芽胞杆菌属(Bacillus)、红球菌属(Rhodococcus)、北里孢菌属(Kitasatosporia)、金黄杆菌属(Chryseobacterium)、不动杆菌属(Acinetobacter)、黄杆菌属(Flavobacterium)、鞘氨醇杆菌属(Sphingobacte-rium)和丛毛单胞菌属(Comamonas)等9个属细菌。其中芽胞杆菌属和不动杆菌属细菌是优势菌,分离到的红球菌属、北里孢菌属、鞘氨醇杆菌属和丛毛单胞菌属细菌在国内湿地土壤中报道较少。     

一株抗玉米纹枯病内生细菌的分离鉴定及其抗病促生作用

玉米纹枯病是玉米主要的真菌病害,从优良玉米品种川单13号中分离到一株对玉米纹枯病菌具有明显抑制作用并能够促进玉米生长的内生细菌.通过形态、生理生化特性分析以及16SrDNA序列同源性比较,鉴定该菌株为枯草芽孢杆菌.盆栽试验表明,菌株能够抑制玉米纹枯病的发生,抑菌率为 43.01%,促生试验表明,该菌能够显著促进玉米苗的生长,促生作用与植物生长刺激素(IAA)和效果相似,显示出良好的开发前景.     

家蚕肠道产淀粉酶细菌的分离鉴定及其酶基因的克隆与表达

从家蚕(Bombyx mori L.)5龄幼虫肠道分离鉴定产淀粉酶细菌菌株以用作微生态制剂的研究,并对该菌α-淀粉酶基因进行克隆、序列分析及在大肠杆菌中原核表达.通过含淀粉NA培养基筛选分离得到产淀粉酶菌,通过形态学观察及16S rDNA序列分析鉴定其种属,DNS法测定酶活,并设计了淀粉酶基因引物进行克隆,构建了DZ-...     

一株中度嗜盐菌的分离鉴定及特性研究

目的：从盘锦红海滩泥样中分离中度嗜盐菌,对其进行鉴定及特性研究。方法：CZ培养基分离纯化,通过形态学、生理生化实验、16SrDNA序列比对分析进行鉴定,吸光光度法测定生长特性。结果：分离得到一株中度嗜盐菌CNY0802,该菌革兰氏阳性反应,菌体杆状,宽度0.7μm～0.9μm,长度2μm～3μm,产芽孢,接触酶、酯酶、明胶液化和硝酸盐还原反应阳性,氧化酶、淀粉酶、MR和VP反应均为阴性。生长的盐度范围0.5%～25%（NaCl,W/V）,最适盐度5%;温度范围10℃～45℃,最适温度30℃;pH范围4.0～12.0,最适pH8.0～10.0;不同的阴阳离子对CNY0802生长影响显著。经16S rDNA序列比对及系统发育树分析,与Halobacillus salinus亲缘关系最近。结论：该菌的分离鉴定对我国辽宁沿海地区极端环境微生物资源的开发研究有一定意义。     

一个甲烷氧化菌株的分离、鉴定及其特性研究

实验室自行设计方案分离多株以甲烷为唯一碳源的菌株, 对其中的1株QJ16进行了研究, 根据该菌株形态特征与16S rDNA序列的同源性分析, 证实该菌株是一个与其最近的甲基单胞菌属各成员都不相同的菌株。对该菌株的培养条件和利用甲烷的特性进行研究结果表明, 氮源以氯化铵和硝酸钾共同作用最好, 碳源以甲烷最佳, 最佳生长温度为30℃, 最佳生长pH为6~7; 在批式实验时菌株利用甲烷的最适pH为6.5左右, 微量元素Cu2+的浓度为15 mmol/ L。     

一个甲烷氧化菌株的分离、鉴定及其特性研究

实验室自行设计方案分离多株以甲烷为唯一碳源的菌株,对其中的1株QJ16进行了研究,根据该菌株形态特征与16S rDNA序列的同源性分析,证实该菌株是一个与其最近的甲基单胞菌属各成员都不相同的菌株.对该菌株的培养条件和利用甲烷的特性进行研究结果表明,氮源以氯化铵和硝酸钾共同作用最好,碳源以甲烷最佳,最佳生长温度为30℃,最佳生长pH为6～7;在批式实验时菌株利用甲烷的最适pH为6.5左右,微量元素Cu2 的浓度为15 μmol/L.     

Isolation of Arsenic-Resistant Bacteria from Bengal Delta Sediments and their Efficacy in Arsenic Removal from Soil in Association with Pteris vittata

The potential of arsenic-resistant bacteria in association with Pteris vittata to reduce the level of arsenic from soil was studied. The physicochemical characteristics of contaminated paddy soil were analyzed, and 3 bacterial isolates amongst 11 were screened and were selected for further study. These three isolates were characterized by 16S rDNA sequencing and identified as Bacillus altitudinis Strain SS8 (KJ432582), Bacillus megaterium Strain SS9 (KJ432583) and Lysinibacillus sp. Strain SS11 (KJ432584). Of these, Lysinibacillus sp. Strain SS11 displayed arsenic tolerance of 3256 mg L^?1 for arsenate and 1136 mg L^?1 for arsenite. Additionally, it showed bioaccumulation capacity of 23.43 mg L^?1 for arsenate and 5.65 mg L^?1 for arsenite. It also showed resistance to other heavy metals, especially towards iron, copper and chromium. It was also observed that Pteris vittata was able to take up more arsenic and iron from soil in the presence of these bacterial strains than in their absence, leading to contaminant-free soil. Thus, this system appears to be an effective bioremediating process to remove arsenic from contaminated soil.     

饲料乳酸菌的分离鉴定及优良菌株筛选

对从饲料玉米、高粱、麦秆及棉花中筛选出的乳酸菌进行分类鉴定和综合性分析。用MRS+CaCO3固体培养基从棉花中分离出乳酸菌18株、高粱中30株、饲料玉米中18株、麦秆中18株。经形态学、生理生化试验进行初步鉴定并按产酸试验,耐盐及耐酸试验挑选出32株产酸率强的乳酸菌对其进行16S rDNA分子鉴定。结果显示,32株菌都具有良好的耐盐、耐酸能力;经生理生化和16S rDNA基因序列鉴定可知32株乳酸菌分属于两个属,即乳杆菌属、肠球菌属,4个种,即干酪乳杆菌(Lactobacilluscasei)、肠道球菌(Entercoccus faecium)、植物乳杆菌(Lactobacillus plantarum)、海氏肠球菌(Entercoccus hirae)。4种饲料原料中肠道球菌普遍存在。除了这种乳酸菌以外,棉花有干酪乳杆菌、植物乳杆菌、海氏肠球菌,玉米和麦秆内有植物乳杆菌。从饲料中筛选出4株具有较强产酸能力的乳酸菌,可进一步研发成青贮饲料添加剂。     

滑桃树不同器官伴生细菌多样性的分子特征

应用不依赖于培养的16S rRNA基因技术,揭示滑桃树根皮、茎皮以及种子伴生细菌的种类多样性,为建立伴生细菌的宏基因组文库奠定基础。基于我们新近发表的植物伴生菌富集方法,建立富集和未富集样品的全长16S rRNA基因文库,随机挑取至少100个克隆进行酶切分型并测序,根据16S rDNA的部分序列推测其所属伴生菌的种类。结果表明,所检测的细菌克隆大部分属于γ-Proteobacteria,有少量来源于α-Proteobacteria或放线菌（Actinobacteria）,也包括不可培养或分类地位不明确的细菌。经过富集,16S rRNA基因文库中细菌克隆的比例和序列多样性显著增加,根皮的伴生细菌种类最为丰富,其次是成熟种子和茎皮;幼嫩种子16S rDNA文库的细菌克隆比例较小（1．18％）,说明幼嫩种子的伴生菌最少。     

Isolation of haemolytic bacilli from field‐collected Culicoides oxystoma and Culicoides peregrinus: potential vectors of bluetongue virus in West Bengal,India

Two haemolytic bacterial strains of Bacillus pumilus (CU1A, CU1B) and one blood‐utilizing strain of Bacillus licheniformis (CU2B) were isolated from relatively low numbers of field‐collected females of Culicoides oxystoma and Culicoides peregrinus (Diptera: Ceratopogonidae). A total of 36 females, including 18 of each of C. oxystoma and C. peregrinus (consisting of one and a pool of eight blood‐engorged specimens, and one and a pool of eight non‐engorged specimens for each species), were tested. In C. oxystoma, all three strains of bacteria were isolated from the one non‐engorged, the pool of non‐engorged and the pool of blood‐engorged females tested, but CU1A and CU2B were not found in the one blood‐engorged female tested. In C. peregrinus, all three strains were present in the pool of blood‐engorged females. However, the strain CU2B was not found in the pool of non‐engorged females. In the one blood‐engorged and one non‐engorged female tested, CU1A and CU2B were detected. The bacterial strains were identified based on Gram staining, enzyme activity (amylase and protease) and alignment of the 16S rRNA partial gene sequence to that available in the National Center for Biotechnology Information (NCBI) database GenBank. The functional role and significance of these haemolytic and blood‐digesting bacteria within the genus Culicoides remain to be determined.     

1株虎源致病性肠球菌的分离鉴定及序列分析

从病死老虎肺脏中分离到1株肠球菌,并对该菌做了生理生化鉴定、药敏试验,致病性试验。本菌对多种抗生素高度耐药,对小白鼠有强致病性,其LD50为2.7×109.2cfu。并用PCR方法扩增分离菌株16S rDNA基因,获得1 415 bp片段,该片段核苷酸序列提交GenBank,登陆号为HM346186,将分离株的16S rDNA核苷酸序列与GenBank上其他肠球菌进行同源性分析。结果表明,分离株的16S rDNA核苷酸序列与肠球菌(EU285587)的同源性为100%,因此该分离菌株被鉴定为致病性肠球菌,命名为YN-1株(云南-1株)。     

一株絮凝菌的鉴定、絮凝基因定位及其絮凝剂成分分析

从武汉市综合废水处理的活性污泥中筛选得到一株絮凝剂产生菌株MBF03,其絮凝率达到90％以上,絮凝效果稳定.通过16S rDNA鉴定,该菌为泛菌属,扫描电镜结果显示为杆状.通过质粒转化与质粒消除试验,初步证实了MBF03菌的絮凝基因位于染色体上.提取该菌所产的絮凝剂,进行紫外、红外分析,结果表明,该絮凝剂主要成分是胞外多糖类物质,不含蛋白质和核酸.     

粪便中大肠埃希菌的分离鉴定

2004年4月采集大连金州湾沿岸畜禽养殖场猪、牛、鸡和某中学人粪便样品。经改良的生化鉴定方法鉴定,得大肠埃希菌(Escherichia coli)猪源75株、牛源78株、鸡源69株、人源68株,占粪大肠菌比率分别为85.23%、92.86%、80.23%、80.00%。为验证改良的生化鉴定方法准确性,从中随机选取8株E.coli,扩增16S rRNA序列并与Genebank中E.coli16S序列比对,确定这8株细菌与E.coli同源相似率达99%以上,进而验证改良的生化鉴定方法的可行性。     

养虾池好氧反硝化细菌新菌株的分离鉴定及特征

利用间歇曝气选择性富集并对所获菌株的好氧反硝化活性进行检测。筛选到一株亚硝酸盐去除活性较高的好氧反硝化细菌。在溶解氧（D0）为3.80-5．21mg／L的培养条件下。该菌株10h内将亚硝态氮由26．18mg／L降至0;在盐度为0—25之间20h内均可达到同样的去除效果。通过形态学特征、生理生化反应及部分长度16SrDNA序列分析对筛选菌株进行鉴定,初步判定它为嗜麦芽寡养单胞菌Stertotrophomonas maltophilia。亚硝酸盐还原酶基因分析结果表明。该菌株只含亚硝酸盐还原酶nitS基因,其序列与Alcaligenes faecalis A15（后来被重新鉴定为Pseudomonas stutzeri）的nirS基因序列相似。     

Molecular Phylogenetic Analysis of Archaea and Bacteria in Wind Cave,South Dakota

The diversity of bacteria and archaea was characterized from sediments collected from Wind Cave located in Wind Cave National Park in the Black Hills of South Dakota. Wind Cave is a limestone dissolution cave with strata that started forming over 300 million years ago, making it one of the oldest in the world. Previous work suggested that the cave was largely a detritus based system ultimately dependent upon allochthonous energy and carbon from photosynthesis of the overlying vegetation, and algae growing near lights along the tour routes. In this work, we used a molecular phylogenetic approach to characterize the microbial structure and infer a corresponding ecosystem function where appropriate. Four bacterial divisions and subdivisions were found in the culture collection, which represented 14 phylotypes, whereas 12 divisions and subdivisions were identified in the clonal analysis comprising 49 phylotypes. The predominant groups were the γ-Proteobacteria and Acidobacteria. Although a few of the clones resembled sequences from other cave and subterranean systems, no cave-specific bacterial community was evident in this work. Archaeal phylotypes (20 Crenarchaeota and 2 Euryarchaeota) were detected, with a large proportion of the Crenarchaeota resembling sequences from a South African gold mine. One archaeal cluster in particular appears to be specific to the subterranean environment. Most of the microbial sequences were not related to known chemolithoautotrophs, therefore we conclude that this particular community is likely detritus based where allochthonous energy and carbon are transported into the cave by infiltrating waters.     

PCR-DGGE法在慢性胃炎病人舌苔菌群结构研究中的应用

在早间未刷牙和进食的情况下,刮取胃炎病人与正常人舌苔,去除杂质,分离菌体,采用酚/氯仿法抽提细菌基因组DNA,并对其中16S rDNA V3可变区进行聚合酶链式反应(PCR)扩增和变性梯度凝胶电泳(DGGE)测定.用Bionumerics软件对DGGE分子指纹图谱进行舌苔菌群结构相似性分析.实验结果表明,采用该方法成功地扩增出16S rDNA V3区片段,为230 bp.DGGE分子指纹图谱结果表明,正常人的舌苔菌群最高相似性为0.74,胃炎病人的舌苔菌群的最高相似性为0.52,正常人的舌苔菌群与胃炎病人的舌苔菌群相似性最高为0.38,即胃炎病人舌苔菌群结构发生了变化.