深海沉积物中产淀粉酶细菌的rep—PCR基因指纹分析

利用淀粉酶筛选培养基平板从27个深海沉积物样品中筛选得到30个具有胞外淀粉酶活性的菌株．其中革兰氏阴性细菌有22株．阳性细菌有8株．对这些细菌的基因组DNA进行ERIC—PCR和BOX—PCR扩增．指纹图谱显示大部分菌株均存在数目不等的各自独特的带型．各特异性扩增的主带型能重复稳定出现．比较两种引物的指纹图谱．显示ERIC—PCR比BOX—PCR具有较丰富的图谱多态性．对产生的指纹图谱进行聚类学分析．30株细菌可分为10组．其中5株细菌分别独立成组．最多的第Ⅱ组包含有8株细菌．而在同一聚类组内的细菌可能是处于进化上亲缘关系较近的位置．研究显示，30株菌具有丰富的种属多样性．揭示了深海微生物资源的丰富和潜力．ERIC—PCR和BOX—PCR技术可用于对深海微生物群落组成和多样性的研究．     

热带盐场域近海及潮间带沉积物可培养细菌遗传多样性

为了研究热带盐场域近海及潮间带沉积物可培养细菌遗传多样性,从海南莺歌海盐场域近海及潮间带沉积物中分离可培养细菌,通过16S rDNA序列比对,分析菌株遗传多样性.结果表明,9株分离自近海沉积物的细菌菌株均属于Bacil-lus;8株分离自潮间带沉积物细菌菌株分为5个属,分别为Acinetobacter、Massilia、Pseudomonas、Pantoea、Paenibacillus;系统发育树结果表明,分离自近海沉积物的菌株YGHWSB2、YGHWSB3、YGHWSB4、YGHWSB5、YGHWSB6、YGHWSB10、YGHWSB21与B.amyloliquefaciens聚为一支,而菌株YGHWSB13、YGHWSB30与B.altitudinis、B.aerophilus、B.stratosphericus、B.xiamenensis聚为一支;分离自潮间带沉积物的菌株聚类较分散,YGHISB1、YGHISB2、YGHISB3、YGHISB4与A.johnsonii聚类;而其他4株菌株YGHISB5、YGHISB6、YGHISB7、YGHISB8分别与4个属的菌株M.haematophila、P.psychrotolerans、P.eucrina、P.peoriae亲缘关系较近.这些结果展现了分离自潮间带沉积物可培养细菌存在丰富的遗传多样性.可见莺歌海近海沉积物中可培养细菌资源分类地位单一,而潮间带沉积物由于所处环境变化多样,可培养细菌资源多样性丰富.     

安徽省西瓜枯萎病菌遗传多样性ISSR-PCR分析

以42株来自安徽省不同地区的西瓜枯萎病菌为试材,采用Inter-simple Sequence Repeats(ISSR)技术对其群体的遗传多样性进行分析,探究病菌的遗传分化与地理来源之间的相关性.结果表明,用筛选出的11个随机引物对西瓜枯萎病菌菌株进行ISSR-PCR扩增,共获得129个ISSR条带,其中多态性条带为94个,占72.9%,表明安徽省西瓜枯萎病菌存在较丰富的遗传多样性,菌株间的遗传相似系数为0.71~0.97.UPGMA聚类分析显示,供试菌株可区分为8个聚类组,菌株的遗传谱系与地理来源无相关性.     

八个灵芝属菌株的分子标记与亲缘关系鉴定

采用RAPD技术对八个不同的灵芝株系进行遗传多样性分析。经筛选得到四个随机引物,共扩增出52条DNA片断,其中多态性条带为43条,多态性位点频率为82.7%,说明灵芝间遗传多样性较为丰富。利用POPGENE软件计算遗传距离并建立UPGMA聚类图,可将八种灵芝株系分为两个组群。通过分析DNA指纹图谱,找到了有代表性的特异条带,为进一步开发稳定的、优势菌株特异的分子标记奠定了基础。     

海绵Pseudoceratina sp.共栖细菌多样性及其抑制甘蔗鞭黑粉菌活性研究

【目的】探讨广西斜阳岛附近海域海绵共附生细菌的多样性,并筛选对甘蔗鞭黑粉菌有抑制作用的活性物质,为发现潜在的细菌新物种及开发农用生防菌肥奠定基础。【方法】采用稀释涂布法,通过6种复合营养分离培养基,从海绵Pseudoceratina sp.中分离可培养的共附生细菌。采取细菌形态学观察去除冗余,通过PCR扩增菌株的16SrRNA基因序列,并通过构建系统发育树分析物种多样性。采用牛津杯法进行抑制甘蔗鞭黑粉菌活性筛选实验。【结果】从海绵Pseudoceratina sp.中共分离到可培养细菌54株,经16SrRNA基因序列对比后,获得24株细菌,隶属于13科14属。其中,菌株BGMRC 2118与已发表有效菌株的16SrRNA基因序列的最高相似率为96.46%,可能为潜在新种。抑菌筛选实验结果显示,有8株细菌的发酵粗提物对甘蔗鞭黑粉菌具有很强的抑菌活性。【结论】广西斜阳岛附近海域中海绵共附生细菌具有较高的物种多样性,蕴藏着丰富的新物种资源,富含生物活性菌株。     

863课题“深海微生物活性物质的挖掘及其利用技术”2013年度进展

2013年为863课题"深海微生物活性物质的挖掘及其利用技术"实施的第二年。该年度该课题进展顺利,完成了预期的研究计划。取得的研究进展主要分为以下5个方面:(1)在深海沉积环境样品中分离、纯化、保藏869株菌株,确定了729个菌株的系统进化地位,在深海海水样品中分离了海洋微生物237株,完成了其中168株的细菌测序工作;其中发现一个潜在的新属级类群。(2)对21株深海微生物进行次生代谢产物的分离和纯化,从中分离鉴定代谢产物186个,其中新结构化合物71个;其中1个对称的环四肽类新骨架化合物;发现具有较好药用活性的化合物25个;对Staurosporine的衍生物HDZ-115和Xanthocillin X进行深入的药理药效研究,确定为下一步成药性评价的先导化合物。(3)从深海来源微生物中发现对植物病原真菌具有良好抗性的脂肽类活性物质;筛选得到一系列抗污损活性的活性物质,其中1个单体化合物在海洋挂板实验中较好抗海洋污损生物活性,具有进一步开发价值。(4)完成了1株深海海洋放线菌SCSIO ZH66的全基因组扫描测序,克隆鉴定了Grincamycin,Lobophorin的生物合成基因簇,完成了Grincamycin,marinacarboline,lobophorin的生物合成途径。(5)从深海微生物中分离纯化两种具有重要经济价值的新蛋白酶,分离得到到新型高温α淀粉酶和生淀粉酶基因,成功实现异源表达,并对酶学性质进行了研究。     

不同16SrDNA靶序列PCR-DGGE分析油页岩细菌多样性

为全面了解油页岩细菌群落组成结构,同时筛选出适于油页岩PCR-DGGE的最佳引物,采用改进SDS高盐提取法,提取抚顺西露天矿油页岩微生物总DNA,并用968F/1401R,338F/518R,341F/907R和1055F/1406R,对16SrDNAV6-V8,V3,V8和V9区进行PCR-DGGE分析.结果表明,4组引物均能较好扩增出目的基因,但不同靶序列对多样性检出有显著影响(P<0001);与其他引物相比,968F/1401R和338F/518R获得的指纹图谱物种丰富度更高,检出细菌类群更丰富,较适合油页岩样品.油页岩细菌群落组成较简单,优势细菌包括不动杆菌属、假单胞菌属和埃希氏菌属,同时还有假单胞菌目和肠杆菌目尚未被鉴定的一些种类.     

四川传统烟区烤烟叶面可培养细菌的遗传多样性分析

为认识烤烟(Flue-cured tobaccos)叶面可培养细菌的多样性,挖掘叶面可培养细菌资源.本研究以四川省各传统烟区旺长期烤烟叶片为材料,通过纯培养法及测定菌株的产吲哚乙酸(IAA)能力、溶磷溶钾特性、纤维素降解活性、拮抗病原菌等,并通过反转录重复因子扩增(BOXA1R-PCR)技术、16S rRNA基因测序和系统发育分析研究叶面可培养细菌的遗传多样性和功能特征.结果表明:分离得到的86株叶面细菌中有12株菌株(占总分离菌株的13.95%)具有产IAA的能力,4株产量较高(57μg/mL);有9株(占总分离菌株的10.47%)表现溶磷活性,8株(占总分离菌株的9.30%)表现溶钾活性.基于BOXA1R-PCR图谱选取12株代表菌株进行16S rRNA基因序列测定,系统发育分析显示,86株菌株分别属于节杆菌属(Arthrobacter)、短小芽孢杆菌属(Lysinibacillus)、寡养单胞菌属(Stenotrophomonas)、无色杆菌属(Achromobacter)和假单胞菌属(Pseudomona),其中以假单胞菌属(Pseudomonas)为优势菌属.表明四川传统烟区烤烟叶面存在着丰富的细菌资源.     

一株多氯联苯降解菌的分离鉴定及基因分型

从深海底泥中分离筛选出一株以多氯连苯（PCBs）为惟一碳源和能源生长的菌株, 命名为pdi. 该菌在含有PCBs 7.5 mg/L的150 mL液体培养基中培养7d, 用GC MS检测其对PCBs的降解率达95.38%. 以细菌16SrDNA通用引物对pdi基因组进行PCR扩增, 得到~1　500　bp的片段. 经纯化, 测序后在Genbank上进行同源性比较分析及系统发育树构建, 该菌与Pseudomonsa spTS1138同源性达100%, 初步鉴定该菌为假单胞菌属. 对pdi基因组进一步运用RAPD分子标记技术进行随机引物扩增基因分型, 获得了稳定的DNA 指纹图谱.     

啤酒废水处理系统的高效菌株与微生物多样性

从啤酒废水处理系统SBR池取得的水样中分离筛选得到3株能高效降解啤酒废水COD的细菌,分别为12#、15#、23#.对3株高效菌株两两进行交互试验,考察混合菌株对废水的处理效果,并以高效菌株与活性污泥混合作用,23#与活性污泥交互作用明显,16 h后COD降解率达到75.13%;应用PCR-DGGE技术对啤酒废水生物处理系统的微生物多样性进行分析,对取自水解酸化池与SBR池中不同深度以及不同处理时段的水样,预处理后抽提基因组DNA,用细菌通用引物进行PCR扩增,PCR产物用变性梯度凝胶电泳进行分离,分析活性污泥样品中的微生物多样性.     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that     

On the Thermodynamic Limit for Responses to Static Electromagnetic Fields

For molecular systems,the quantum-mechanical treatment of their responses to static electromagnetic fields usually employs a scalar-potential treatment of the electric field and a vector-potential treatment of the magnetic field.Although the potential for each field separately is associated with the choice of an(unphysical)origin,the precise choice of the origin for the electrostatic field has little consequences for the results.This is different for the     

Call for Papers

<正>"The Journal of Shanghai Normal University:Mathematics"is published by Shanghai Normal University as regular issues of The Journal of Shanghai Normal University each year from 2014 in English.The editors-in-chief of the issues are professors Yuhao Cong and Maoan Han.The Journal of Shanghai Normal University was started in 1958 with     

Journal of Donghua University ( English Edition) Contents of Volume 31

正~~