PCR-DGGE analysis of intestinal bacteria and effect of Bacillus spp. on intestinal microbial diversity in kuruma shrimp (Marsupenaeus japonicus)

In this study, the intestinal microbiota of kuruma shrimp (Marsupenaeus japonicus) was examined by molecular analysis of the 16S rDNA to identify the dominant intestinal bacteria and to investigate the effects of Bacillus spp. on intestinal microbial diversity. Samples of the intestines of kuruma shrimp fed normal feed and Bacillus spp. amended feed. PCR and denaturing gradient gel electrophoresis (DGGE) analyses were then performed on DNA extracted directly from the guts. Population fingerprints of the predominant organisms were generated by DGGE analysis of the universal V3 16S rDNA amplicons, and distinct bands in the gels were sequenced. The results suggested that the gut of kuruma shrimp was dominated by Vibrio sp. and uncultured gamma proteobacterium. Overall, the results of this study suggest that PCR-DGGE is a possible method of studying the intestinal microbial diversity of shrimp.

内蒙古多伦草原土壤微生物多样性对全球变化的响应

气候变暖和氮沉降是全球变化现象中最为重要和广为关注的两个方面，它们给陆地生态系统带来了深刻的影响。目前，已有众多研究就二者对地上植被的影响展开了大量工作;而作为陆地生态系统的另一重要生命组分，土壤微生物群落对二者的响应与适应机制还不明了，还需要相关工作的不断补充和完善。本文正是在温带典型草原通过设置增温、施氮等模拟控制实验，探讨增温、施氮及其交互效应对草原生态系统土壤微生物群落多样性和功能多样性的影响，使我们对该地区土壤微生物群落的响应与适应有了一个初步的了解，有助于全面分析和评估气候变化对温带典型草原生态系统的影响。 目前，全球变暖表现出时间上的不均衡性，即夜间增温的幅度要大于白天增温的幅度，而且这种现象在欧亚大陆尤为普遍。这种气候变暖的不均衡性是否会给草原生态系统土壤微生物群落带来显著的影响是我们所关注的问题之一。在第二章中，于多伦典型草原设置了对照、白天增温、夜间增温和全天增温处理，处理时间起始于2006 年5 月，并通过磷脂脂肪酸和BIOLOG 分析方法分别对土壤微生物的群落多样性和碳源利用能力（即功能多样性）进行分析。结果表明，不同增温处理对土壤微生物群落组成的影响均不显著，而夜间增温对土壤细菌群落的碳源利用能力具有显著的促进作用。这说明土壤微生物群落结构对短期增温处理的响应要滞后于功能的响应，而功能上的响应又首先出现在细菌群落，并且这种响应表现为白天增温和夜间增温的非加和效应。 气候变暖往往与降雨的变化相伴而生，而水分又是半干旱典型草原重要限制因子，因此探讨降雨的变化对土壤微生物的影响有助于更深入的了解土壤微生物对气候变暖的响应。在第三章中，分别设置了对照、增温、夏季增雨及增温×夏季增雨处理，处理时间起始于2005 年4 月，并同样分析了土壤微生物的群落组成和功能多样性。结果表明，增温和夏季增雨对土壤微生物群落结构和功能的影响具有明显的一致性，二者均降低了革兰氏阴性菌的相对含量，并且促进了细菌群落的碳源利用能力。增温和增雨在影响细菌群落结构（即，革兰氏阴性菌与阳性菌之比）上具有一定程度的交互效应。 陆地生态系统生产力普遍受到土壤氮素的限制，由于人类活动的干扰，尤其是农业生产和化石燃料燃烧，使土壤氮素含量增加，改变土壤理化环境和地上植物群落，这将直接或间接地影响土壤微生物群落结构和功能。就已有的研究结果来看施氮究竟如何影响土壤微生物群落尚无定论。第四章主要是针对相关研究结果上的分歧，在2003 年设置的0-64 g N m-2 yr-1 的指数施氮梯度实验上进行土壤理化因子、微生物量、净氮矿化和微生物碳源利用能力的分析。结果表明，土壤微生物群落对施氮的响应存在一个介于16 和32 g N m-2y-1 最优施氮量，即低于最优施氮量施氮处理表现为对土壤微生物的促进作用，而过高的施氮量则表现为抑制作用。这一结果说明施氮量是影响土壤微生物对施氮处理做出响应的重要方面之一，有助于解释目前相关研究中出现的结果分歧。 第五章的研究则是考虑了地形和火烧这两个草原生态系统普遍存在的干扰因素，比较分析二者与施氮处理对土壤微生物群落结构的影响。实验设置在多伦典型草原一个坡度为15﹪的南向阳坡，坡上坡下分别设置对照、施氮和火烧处理，处理的起始时间为2005 年4 月。结果表明，土壤微生物群落结构存在地形上的显著差异，表现为坡下以细菌类群占优势，而坡上真菌类群的相对优势较大;火烧对土壤微生物群落结构的影响显著，并表现出明显的地形上的差异，即坡下微生物对火烧的响应敏感性要高于坡上;而短期的施氮处理对土壤微生物群落结构的影响不显著。第六章的研究主要是针对增温、施氮和增温×施氮处理对土壤微生物群落结构和功能的影响，处理时间起始于2006 年5 月。结果表明，土壤微生物群落结构和功能年际变化显著;增温和施氮对土壤微生物的影响均不显著，但增温复合施氮处理中的土壤微生物群落结构表现出一定的响应趋势。

南海北部区域表层水微生物多样性及其分布特点研究

海洋中的微生物多样性是十分丰富的。南海北部区域表层水的微生物群落结构及物种多样性情况仍不十分清楚。本研究，采用构建基因克隆文库的方法，对该区域内表层水中的微生物多样性及分布特点进行研究。获得了8000多个细菌16S rDNA基因、真核微生物ITS 区基因及光合微型生物 psbA 基因单克隆。本研究结果表明：在南海北部区域表层水中存在两种不同微生物类群，即近海岸带海洋微生物类群和开阔海域海洋微生物类群。 16S rDNA基因克隆文库中，确定了507个OTUs。93.7% 的16S rDNA 序列定义在同一种水平上，1.4 %的16S rDNA序列定义在同一属的水平上，2.7%的16S rDNA序列定义在同一纲的水平上，1.2%的16S rDNA序列定义在同一门的水平上。值得一提的是有0.7%的南海表层水样品的16S rDNA 序列，属于目前数据库中的未知序列。系统育树分析表明这类序列可归属于4个不同的分枝群。与Venter’s Sorcerer II 海洋科考（马尾岛海域）的结果不同，南海北部区域表层水中，并没有发现SAR11分支细菌、丝状杆菌（Fibrobacter）和Rheinheimera细菌序列，但南海北部区域却发现了马尾岛海域未检测到的物种，如酸杆菌门、恐球菌-栖热菌门、厚壁菌门，硝化螺旋菌门，浮霉菌门以及疣微菌类细菌。除疣微菌外，其他5种细菌都是海洋环境样品中较为常见的细菌。变形菌门、蓝细菌及厚壁菌门细菌序列是南海北部表层样品16S rDNA基因克隆文库中的主要类群。 真核生物如浮游植物和海洋真菌是海洋表面生物质的主要组成部分之一。现有的研究多集中在环境样品的原核微生物的群落结构研究上，很少关注海洋微型真核生物的多样性及群落结构分布。本研究通过构建ITS基因克隆文库的方法，得到了3044条ITS序列，最终定义了1288个OTUs。其中，329个OTUs序列定义在同一种水平上，310个OTUs序列定义在同一属或纲的水平上，123个OTUs序列定义在同一门的水平上。值得注意的是有339个OTUs的序列，属于目前数据库中的未知序列。系统发育树分析表明它们分别归属于4个不同的分枝群。这表明以往对海洋真核微型生物的多样性仍知之甚少。盘菌亚门、体腔动物门和担子菌纲是南海北部表层样品ITS基因克隆文库中的主要类群。此外，在南海北部区域还发现了少数归属于绿藻、链形植物、定鞭金藻类、放射虫类、Stramenopiles、Typhlocoela、壶菌类、多孢囊霉目、子囊菌门、地位未定的物种、 酵母、领鞭毛虫门、不可培养的后生动物和海绵动物的ITS序列。 海洋初级生产力主要是依靠光合微型浮游生物进行光合作用完成的。利用新设计的psbA通用引物，对南海北部33个表层水样滤膜进行基因克隆文库建库分析，最终获得了南海北部区域表层水微生物多样性及其分布特点研究3062条部分psbA基因序列，并将其划分为957个 OTUs。其中蓝细菌和未培养的病毒序列在psbA基因库中的数量最多。本研究还发现了南海北部区域存在11个独立分支的新型psbA类群。研究证实psbA基因可以作为一种研究海洋光合微型浮游生物群落结构的指示基因。 克隆文库相似性分析发现，在所有的16S rDNA克隆文库中没有任意两个站点的克隆文库相似性超过50%。虽然N401和N420站点的16S rDNA克隆文库相似性最大，但它们在地理位置上并不接近。一些地理位置接近的站点，其16S rDNA克隆文库之间相似性比较接近。比如，海南岛区域的克隆文库之间就比较相似，且在同一分支。大多数地理环境相似的站点的16S rDNA克隆文库都聚在同一大分支上。例如，来自于珠江口区域站点的克隆文库之间的相似性比较接近，而且分布在一个大分支中；开阔海洋区域的16S rDNA克隆文库，也大多聚类在同一分支中。但也有例外的情况：比如 N107 和N400 站点的16S rDNA克隆文库，就聚类到一起，分析发现这两个文库中所处的环境都是甲烷产生区，其中都含有相似的与甲烷代谢相关的菌群。不过从整体来看，整个南海北部的细菌群落，大致分为两大类：中国大陆近海岸微生物群落和开阔海域微生物群落。33个ITS克隆文库的相似性分析发现：相似性在10%以下的类群，可以分成两大分支，而且该分类，比细菌群落的分布情况更接近南海北部的地理环境特征。对psbA基因克隆文库的相似性分析也验证了在南海北部区域表层水中存在两种不同微生物生态系统。 此外，本研究针对分子生态专业软件DOTUR程序在处理大量克隆文库数据时所遇到的

水肥处理对黄瓜土壤养分、酶及微生物多样性的影响

以津优1号黄瓜为试材,设3个土壤相对含水量水平(50%～60%、70%～80%、90%～100%)和2个肥料追施量(600kgN·hm-2和420kgP2O5·hm-2,420kgN.hm-2和294kgP2O5·hm-2)处理,研究了不同水肥供应对日光温室黄瓜土壤养分、酶活性及微生物多样性的影响.结果表明:土壤中NH4+-N含量随施肥量的增加而提高,随土壤相对含水量的增加而降低;水肥供给的增加有利于提高土壤中速效磷含量和蔗糖酶活性;肥料增加使土壤中蛋白酶活性降低,而水分降低使土壤中脲酶活性提高.土壤中微生物多样性与土壤中养分含量无显著相关性,与土壤脲酶活性呈显著正相关,与蔗糖酶活性呈显著负相关.土壤相对含水量70%～80%、氮肥追施量600kgN·hm-2和420kgP2O5.hm-2处理的土壤养分含量、蔗糖酶、磷酸酶和脲酶活性较高,且土壤中微生物多样性和均匀度显著高于其他处理,土壤生产潜力最优.

氮肥缺失对旱地土壤细菌群落多样性的影响

本文旨在研究氮肥缺失对旱地土壤细菌群落多样性的影响。采用直接提取土壤微生物总DNA的方法,对不施肥(CK)、适量施肥(F1)、和缺氮施肥(F2)3种不同施肥水平土样DNA进行提取,扩增细菌16S rDNA基因片段,建立克隆文库。用限制性内切酶HhaI和RsaI进行PCR-RFLP分析,分别得到146、187、11个酶切类型。采用α多样性的测度对试验结果进行分析统计结果表明,不同处理间土壤细菌的多样性(H′、Ds和Dg)和物种丰富度(dMa、R2和E)均为F1>CK>F2;λ、dMa、E和H′指数在不同施肥处理间的变异系数达到56.96%~163.1%,尤其Simpson指数λ是非常敏感的指标,处理间的差异最大,表明氮肥缺失严重影响土壤细菌群落多样性,合理施肥有利于土壤细菌的多样性。

辽河干流底质微生物群落结构及芘降解菌研究

辽宁省是以石油化工、煤炭化工和钢铁工业为主的重工业基地。辽河流域近年来经济发展迅速，城市化水平不断提高，但由于产业结构的不合理和污染治理水平的相对滞后，致使辽河流域水体污染严重。对辽河流域水体污染状况、污染物化学与生物学的相互作用、微生物群落结构与功能的关系开展调查研究，对开展污染水体的生物生态修复具有重要的指导意义。 本论文选取辽河流域干流8个水文监测站点的不同时期（丰水期和平水期）底质为研究对象，调查了有机污染物（总油TPHs）和有毒污染物（多环芳烃PAHs）污染程度以及主要来源；采用变性梯度凝胶电泳（DGGE）和磷脂脂肪酸（PLFA）两种分析方法，对其微生物群落结构及多样性进行了分析；并以13C标记的菲和芘为代谢底物，以PLFA为生物标记物，采用气相色谱-稳定同位素比率质谱（GC-c-IRMS）分析技术，鉴定了底质样品中参与菲和芘代谢的主要微生物类群；并利用不同的吸附性载体进行了芘降解菌的富集和筛选。 研究结果表明： 1）平水期总石油烃污染比丰水期严重，TPH 含量分别在276.1～560.6mg/kg（平水期）和157.9～462.2mg/kg（丰水期）之间，辽河入海口TPH污染最重；PAHs含量分别在124.1～270.4ug/kg（平水期）和93.5～209.1ug/kg（丰水期）之间；主要来源于石油类污染物和化石燃料的热解，汽车尾气的污染等。 2) 采用DGGE和PLFA两种方法分析微生物群落结构得到基本一致的结果。微生物多样性与总石油烃含量、总多环芳烃含量无显著相关性，多样性指数是多种污染物和环境因子综合影响的结果。 3) 稳定同位素代谢示踪实验表明，底质中存在菲和芘的降解菌群；参与菲和芘降解的微生物均以G-细菌为主，真菌次之；G+细菌和放线菌也参与代谢；参与菲和芘代谢的菌群有一定的相似性。 4) 利用不同吸附性载体从污染底质样品中筛选到6株降解菌。

生姜化感作用机理及其栽培模式研究

近年来，随着对作物重茬（连年种植）障碍原因的深入研究，植物的化感作用越来越受到国内外众多学者的重视。而作为重要调料和药用植物的生姜，其连作障碍也备受关注，系统地研究生姜化感作用将有助于理解和最终解决生姜连作障碍问题。本文通过研究生姜不同部位、不同浓度的水浸液对与其间作的两个物种（大豆和四季葱）种子的萌发及幼苗生长的影响，从而证明生姜化感作用的存在；并通过温室盆栽实验研究了生姜的自毒作用（即研究生姜不同部位、不同浓度的水浸液对其幼苗的形态、生理生化、光合作用、土壤酶、土壤微生物多样性及土壤养分的影响），从而揭示生姜退化和衰老的机制，并为生姜筛选出合适的间作物种提供科学依据，对生姜连作障碍提出科学的解决方法。主要研究结果如下： 1. 与对照相比，生姜所有部位（根茎、茎、叶）、所有浓度（10、20、40、 80 g l-1）的水浸液均抑制了大豆种子和葱籽的萌发率、幼苗生长、水分吸收和脂肪酶活性，并且其抑制程度随着水浸液浓度的增加而增强，其生姜各部位水浸液抑制效应的强弱顺序为茎＞叶＞根茎。这一结果表明生姜根茎、茎、叶含有能够抑制大豆种子和葱籽种子萌发和幼苗生长的水溶性化感物质。根茎是生姜的主要收获部位，而生姜的残株（主要是茎和叶）应该从大田中处理掉以减轻其抑制效应。生姜水浸液中主要化感成分包括：根茎水浸液中主要是丁香酸和伞花内脂；茎水浸液中主要是阿魏酸，且其含量最高为73.4 ug/g；叶水浸液中除了阿魏酸，其他六种物质均检测出来，但含量较高的主要有丁香酸、伞花内脂和香豆酸。 2. 生姜茎和叶不同浓度的水浸液均显著抑制了生姜幼苗的株高、每株叶片数和叶面积，其抑制程度随着水浸液浓度的增加而有所增强，而生姜幼苗每株分枝数差异不显著；同时生姜水浸液也极大程度地影响了生姜幼苗的生物量（包括地下生物量、地上生物量和总生物量，均为鲜重）。在同一浓度下，茎水浸液对生姜幼苗形态指标及生物量指标均显示出最强的抑制作用，叶水浸液次之，根茎水浸液最弱。与对照相比，低浓度的生姜根茎水浸液提高了生姜幼苗叶片内四种抗氧化酶（SOD、POD、CAT、APX）活性，高浓度的根茎水浸液抑制了四种抗氧化酶活性，而茎和叶水浸液均随着浓度的增加而抑制了四种抗氧化酶活性，三种水浸液均随着浓度的增加降低了生姜幼苗叶片内叶绿素的含量，而增加了生姜幼苗叶片的相对电导率和丙二醛含量。同时，三种水浸液均随着浓度的增加降低了生姜幼苗的光合参数（包括胞间CO2浓度、气孔导度、蒸腾速率及净光合速率）。 3. 三种生姜水浸液对所测六种土壤酶活性均产生了不同程度的影响，其中影响最大的是酸性磷酸酶和蔗糖酶，在10 g l-1 时就达到了显著水平，并且所有酶均有随着水浸液浓度增加而增大的趋势；相同部位的水浸液随着浓度的增加，细菌和真菌的数量呈增加趋势，而放线菌的数量呈减少趋势；三种生姜水浸液均随着浓度的增加降低了土壤中有机质的含量，加剧了土壤中硝态氮含量的积累，根茎水浸液对土壤有效磷、速效钾和铵态氮均显示出低浓度提高其含量而高浓度降低其含量的趋势，而茎和叶水浸液则随着浓度的增加均降低了其含量。 4. 与生姜单作相比，所有间作系统均在旺盛生长期和收获期不同程度地提高了土壤酶活性，同时也增加了土壤细菌数量及土壤微生物总数但不显著；所有间作系统在旺盛生长期和收获期均不同程度地影响了土壤真菌及放线菌数量（增加或减少），所有间作系统间的多样性指数差异不显著，除了旺盛生长期四种作物（生姜-大豆-四季葱-大蒜）的间作模式显著降低了多样性指数，其值仅为生姜单作的33.18%；生姜与大豆间作不仅提高了19.6%的生姜产量而且获得了较好的经济效益，并且，所有间作系统均显著抑制了生姜姜瘟病的发生。 5. 不同栽培模式不同程度地影响了收获期生姜的株高、分枝数、根茎产量及内在品质。其中处理2显著地促进了生姜的分枝（10.5%），同时处理2、3和4也促进了生姜的生长（株高分别增加了15.0%、11.4%和14.0%），并且这三个处理提高了生姜的产量；处理2和3能有效提高生姜块茎中维生素C（分别较单作生姜显著提高了3.29%和4.05%）、处理3显著提高了可溶性糖（8.2%）、姜辣素（4.6%）和蛋白质等有益物质的含量，降低硝酸盐有害物质的含量（处理2显著降低了14.0%），改善了姜块的外观和内在品质。并且，生姜与大豆间作具有最高的纯收入和产投比，分别较生姜单作提高了24.80%和8.8%。Recently, allelopathy has been more and more paid attentions by national and foreign scholars with profound research on reasons of crop replanted (continuous planted) obstacle. Ginger rhizome is valuable all over the world either as a spice or herbal medicine and ginger replanted obstacle is also paid attentions. Systematic research on ginger allelopathy will contribute to understanding and ultimate solving problem of ginger replanted obstacle. The effects of ginger aqueous extracts with different parts and concentrations on seed germination and early seedling growth of soybean and chive were studied in this article to testify that ginger existed allelopathy. Furthermore, ginger autotoxicity was also studied by pot experiment in greenhouse (namely research on effects of ginger aqueous extracts with different parts and concentrations on morphological indexes, physiological and biochemical indexes, photosynthesis, soil enzymes, soil microbial diversity and soil nutrients) to reveal mechanism of ginger degeneration and senescence, provide scientific basis for selecting appropriate intercropping species and put forward scientific resolvent for ginger replanted obstacle. The main results were as follows: 1. All aqueous extracts at all concentrations inhibited seed germination, seedling growth, water uptake and lipase activity of soybean and chive compared with the control, and the degree of inhibition increased with the incremental extracts concentration. The degree of toxicity of different ginger plant parts can be classified in order of decreasing inhibition as stem＞leaf＞rhizome. The results of this study suggested that rhizome, stem and leaf of ginger contained water soluble allelochemicals which could inhibit seed germination and seedling growth of soybean and chive. The rhizome is the main harvested part of ginger. The residue (mainly stems and leaves) of the ginger plant should be removed from the field so as to diminish its inhibitory effect. The main allelopathic components of three kind of aqueous extracts were as follows: Rhizome extract chiefly contained syringic acid and vmbelliferone and stem extract mainly contained frulic acid whose content was the highest (73.4 ug/g). The other six substances were detected except of frulic acid, but only contents of syringic acid, vmbelliferone and p-coumaric acid were higher. 2. Stem and leaf aqueous extracts of ginger with different concentrations significantly inhibited plant height, leaf numbers per plant and leaf area, and the degree of inhibition increased with the incremental extracts concentration. However, tiller number per plant of ginger seedling showed no significant difference. At the same time, ginger aqueous extracts also influenced biomass including under-ground biomass, above-ground biomass and total biomass (fresh weight) to a large extent. Under the same concentration, stem aqueous extract showed the mostly inhibitory effect on morphological indexes and biomass indexes of ginger seedling. Rhizome aqueous extract showed the leastly inhibitory effect and leaf aqueous extract was intervenient. Enhanced concentration of ginger aqueous extracts significantly reduced total chlorophyll content, accompanying with increases in memberane permeability (REL) and lipid peroxidation (MDA). Compared with the control, rhizome ginger aqueous extract of lower concentration (10 g l-1) increased the activities of major antioxidant enzymes (superoxide dismutase, SOD; peroxidase, POD; catalase, CAT; ascorbate peroxidase, APX) of ginger leaf tissue and higher concentration inhibited the activities of four antioxidant enzymes. However, stem and leaf aqueous extract inhibited the activities of four antioxidant enzymes with increase in concentration. Meanwhile, enhanced concentration of ginger aqueous extracts significantly reduced photo-parameters of ginger seedling (including CO2 concentration, stoma conductivity, net photosynthesis rate and transpiration rate). 3. Rhizome, stem and leaf ginger aqueous extract showed different effect on six soil enzyme activities, and acid phosphatase and invertase showed significant effect when aqueous extract concentration got 10 g l-1. Furthermore, six soil enzyme activities increased with increase in aqueous extract concentration. Bcterial and fungi number tended to increase while antinomyces tented to decrease with the increase in aqueous extract concentration of identical part. Ginger aqueous extracts reduced soil organic matter content with increased concentration, accompanying with NO3-—N accumulation in soil. Rhizome aqueous extract showed the same tendency for available P, available K and NH4+—N, namely lower concentration increased their contents in soil and higher concentration reduced their contents. While stem and leaf aqueous extracts reduced their contents with the increamental concentration. 4. All intercropping systems increased soil enzyme activities to different extent both at VGS and at HS compared to solo ginger. All intercropping systems increased the colony numbers of soil bacteria and total of soil microbe but not significantly either at VGS or at HS. All intercropping systems increased the colony numbers of soil fungi and actinomytes to a different extent (increase or decrease) both at VGS and at HS. For DI, difference between all cultivation patterns and S-G was not significant either at VGS or at HS except that G-S-C-G whose value was only 33.18% of S-G at VGS significantly decreased. G-S not only increased ginger yield by 19.6% but also obtained better economic benefit. Furthermore, all intercropping systems significantly inhibited occurrence of bacterial wilt of ginger. 5. Different cultivated pattern influenced plant height, tiller numbers, rhizome yields and intrinsic quality of ginger. Treatment 2 significantly facilitated tiller occurring (10.5%). Treatment 2, 3 and 4 promoted ginger growth (plant height respectively increased 15.0%、11.4% and 14.0%) and enhanced rhizome yields. Treatment 2 and 3 effectively increased vitamin C content (significantly increased 3.29% and 4.05% compared to solo ginger). Treatment 3 significantly increased contents of beneficial substances such as soluble sugar (8.2%), gingerols (4.6%) and protein. Treatment 2 significantly decreased contents of deleterious substance namely nitrate (14.0%) and improved appearance and intrinsic quality of ginger rhizome. Furthermore, treatment 2 (ginger/soybean intercropping) could obtain better economic benefit and showed the highest net income and ratio of benefit and cost whose values respectively increased by 24.80% and 8.8% compared to solo ginger.

无介体双室微生物燃料电池产电性能初步研究

本文对无介体双室微生物燃料电池的产电性能进行了初步研究，并根据不同运行阶段产电性能的优劣，对其中微生物的差异性进行了比较分析。全文分为两个部分： 第一部分：以乙酸钠为阳极原料构建双室微生物燃料电池（MFC），研究不同阴极受体、外接电阻、乙酸钠浓度和pH等因素对电池产电性能的影响，研究结果表明：在500mL的阴阳极反应体系中，选用乙酸钠作为阳极底物，质量浓度为6.46 g/L， pH 7.0，接入500Ω外电阻，阴极电子受体选择高锰酸钾的情况下，微生物燃料电池产电性能最好，最大电功率密度达到294.72 mW/m2，库伦效率能达到25.87%。在确定最适外接电阻阻值的同时对MFC内阻进行测定，阻值为871.87Ω。 第二部分：微生物燃料电池运行中，比较以厌氧污泥作为接种源的第一阶段和只接入附着有大量微生物电极的第二阶段的产电性能，得出第二阶段产电性能优于第一阶段，最大电功率密度达到353.57mW/m2，比第一阶段提高58.85 mW/m2；库伦效率为39.35%，增幅达52％左右；针对微生物燃料电池运行过程中，底物CH3COONa可能存在其它的代谢途径，本实验进行了第二阶段产电性能与CH3COONa消耗率关系以及阳极液面上方气体成分和含量的研究，发现第二阶段50h前CH3COONa的大量消耗主要用于微生物的生长，在整个运行过程中，阳极液面上方含有CH4和CO2；对气体测定的同时还发现，振荡能增强电功率密度的输出；通过对电极上和污泥中微生物差异性分析得出，δ-变形菌纲、β-变形菌纲和拟杆菌门的菌种更适应微生物燃料电池的运行环境，能在电极上大量富集，提高电池的产电性能，只接入附着有大量微生物的电极能有效降低热袍菌纲的菌种数量，降低了CH3COONa的无为消耗，有效提高了电池的库伦效率。 Electricity production in the mediator-less two-chambered microbial fuel cell(MFC) was researched. Based on the result in the different operation phase in the MFC, the microbial diversity was analysed. The paper involved two parts: Part 1: A two-chambered microbial fuel cell (MFC) was constructed with high-concentration sodium acetate as fuel in the anode. The influence of different electron acceptors in the cathode, external resistance value, pH value and concentration of sodium acetate on electricity generation in MFC was investigated. The result showed that the maximum power density of 294.72 mW/m2 and the coulombic efficiency of 25.87% was achieved at sodium acetate concentration of 6.46 g/L, pH 7.0, external resistance 500Ωin the anode and when using potassium permanganate as electron acceptor in the cathode. While decided the value of resistor, we found that shaking has effect on electricity production in the MFC. Part 2: Comparing the electricity production in different operation phases when using anaerobic sludge as inoculum in the first phase and microbes in the anodic electrode as inoculum in the second phase, the result showed that electricity production in the second phase was more than that in the first phase, the maximum power density of 353.57 mW/m2 and the coulombic efficiency of 39.35% was achieved, 58.85 mW/m2 and 52％ more than that in the first phase, respectively. According to the fact that CH3COONa might be metabolized in other pathway in the running process in the MFC, we determining the relationship between electricity production and CH3COONa consumption, and the gas content in the anode, we found that CH3COONa was mainly used for microbe growth before 50h, and the anode contained CH4 and CO2. At the same time, we found that shaking could improve power density. The analysis on diversity of microbe in the anodic electrode and anaerobic sludge showed that δ-proteobacterium, β-proteobacterium and Bacteroidetes adapted themselves to the running environment of MFC. The anode could enrich them to improve the electricity production while reduced the quantity of Thermotogales, which were obligately anaerobic organotrophs with a fermentative metabolism, to increase the coulombic efficiency effectively.

Vertical distribution of bacterial and archaeal communities along discrete layers of a deep-sea cold sediment sample at the East Pacific Rise (similar to 13 degrees N)

The community structure and vertical distribution of prokaryotes in a deep-sea (ca. 3,191 m) cold sediment sample (ca. 43 cm long) collected at the East Pacific Rise (EPR) similar to 13 degrees N were studied with 16SrDNA-based molecular analyses. Total community DNA was extracted from each of four discrete layers EPRDS-1, -2, -3 and -4 (from top to bottom) and 16S rDNA were amplified by PCR. Cluster analysis of DGGE profiles revealed that the bacterial communities shifted sharply between EPRDS-1 and EPRDS-2 in similarity coefficient at merely 49%. Twenty-three sequences retrieved from DGGE bands fell into 11 groups based on BLAST and bootstrap analysis. The dominant groups in the bacterial communities were Chloroflexi, Gamma proteobacteria, Actinobacterium and unidentified bacteria, with their corresponding percentages varying along discrete layers. Pairwise Fst (F-statistics) values between the archaeal clone libraries indicated that the archaeal communities changed distinctly between EPRDS-2 and EPRDS-3. Sequences from the archaeal libraries were divided to eight groups. Crenarchaea Marine Group I (MGI) was prevalent in EPRDS-1 at 83%, while Uncultured Crenarchaea group II B (UCII B) abounded in EPRDS-4 at 61%. Our results revealed that the vertically stratified distribution of prokaryotic communities might be in response to the geochemical settings and suggested that the sampling area was influenced by hydrothermalism. The copresence of members related to hydrothermalism and cold deep-sea environments in the microbial community indicated that the area might be a transitional region from hydrothermal vents to cold deep-sea sediments.

Characteristics of the microbial communities in the integrated vertical-flow constructed wetlands

Microorganisms play an important role in removing pollutants from constructed wetlands. We investigated the microbial characteristics in a novel integrated vertical-flow constructed wetland (IVCW), which has been in operation in Wuhan, China since 1998. We used phospholipid fatty acid (PLFA) and amoA gene to analyze the structure and diversity of the microbial community within the IVCW. PLFA results suggested that the amount of bacterial PLFA was significantly higher than that of fungal PLFA, but the total microbial biomass represented by PLFA index was low in the system. Microbial spatial distribution showed significantly higher bacterial (both G(+) and G(-)) and fungal biomass in the surface than in the subsurface layers. The ratios of monounsaturated to branched PLFA demonstrated that an anaerobic layer sandwiched by two aerobic layers existed in the IVCW, consistent with the redox potential results. Analysis of the amoA revealed the presence of Nitrosomonas-like sequences in the surface substrate of the downflow chamber and apparent diversities of ammonia-oxidizing bacteria in the system. These results suggest that microorganisms, despite their relatively low biomass, have inhabited the IVCW, and the results will offer some valuable information on microbe to system designers and managers.

Genetic diversity of plankton community as depicted by PCR-DGGE fingerprinting and its relation to morphological composition and environmental factors in lake donghu

To collect information about the genetic diversity of the plankton community and to study how plankton respond to environmental conditions, plankton samples were collected from five stations representing different trophic levels in a shallow, eutrophic lake (Lake Donghu), and investigated by PCR-DGGE fingerprinting. A total of 100 bands (61 of 16S rDNA bands and 39 of 18S rDNA bands) were detected. The DGGE bands unique to any single station accounted for 38% of the total bands, whereas common bands detected at all five stations accounted for only 11%. Using UPGMA clustering and MDS ordination of DGGE fingerprints, stations I and II were found to initially group together into one cluster, which was later joined by station V. Stations III and IV were isolated into two separate groups of one station each. Some differences in grouping relationships were found when analysis was completed on the basis of chemical characteristics and morphological composition, with zooplankton composition showing the greatest variability. However, the most similar stations (I and II) were always initially grouped into one cluster. Moreover, stations that exhibited the same or similar trophic level (stations III and IV), but different concentrations of heavy metals, were further differentiated by the DGGE method. Results of the present study indicated that PCR-DGGE fingerprinting was more sensitive than the traditional methods, as other studies suggested. Additionally, PCR-DGGE appears to be more appropriate for diversity characterization of the plankton community, as it is more canonical, systematic, and effective. Most importantly, fingerprinting results are more convenient for the comparative analyses between different studies. Therefore, the use of the described fingerprinting analysis may provide an operable and sensitive biomonitoring approach to identify critical, and potentially negative, stress within an aquatic ecosystem.