太湖冬季军团菌（Legionella spp．）的分布和多样性

军团菌（Legionellas pp．）广泛存在于各种水体中,对人类的健康造成了一定的安全隐患．为了解太湖在冬季是否存在军团菌,以及它的分布和多样性,本研究于2010年2月,在全太湖32个点位采样,使用巢式PCR对样品中军团菌进行检测,并用变性梯度凝胶电泳（denaturing gradient gel eletrop...     

稻田土壤固碳功能微生物群落结构和数量特征

研究不同类型稻田土壤自养微生物数量和多样性差异及其影响因子,对全面认识稻田生态系统的固碳潜力及其机制具有重要意义.鉴于此,本文选取4种典型稻田土壤,通过室内培养实验对具备卡尔文循环途径碳同化微生物进行了研究.利用荧光定量PCR(qPCR)、克隆文库以及末端限制性长度多态性分析(T-RFLP)技术,研究了卡尔文循环关键酶(1,5二磷酸核酮糖羧化酶/加氧酶Rubis CO)的2种编码基因(cbbL和cbbM)的丰度和多样性.结果表明,与培养前相比,培养45 d后碳同化自养微生物数量有所增加,cbbL基因丰度比cbbM基因高3个数量级.不同稻田土壤中碳同化功能微生物优势种群存在差异,且这些微生物大多不能归类到已知的细菌类群中,部分可归类的与变形菌和放线菌有较高相似度.RDA分析结果显示土壤有机碳(SOC)、阳离子交换量(CEC)、pH、黏粒、粉粒和砂粒含量对碳同化功能微生物群落结构有显著影响.本文的研究结果对于理解微生物在碳循环过程中的作用具有一定的科学意义,也可以为稻田土壤肥力科学化管理和构建低碳农业提供科学依据.     

外源菌剂联合柠檬酸强化龙葵修复土壤镉污染

设置对照（CK）、摩西球囊霉菌（GM）、摩西球囊霉菌+柠檬酸（GM+CA）、摩西球囊霉菌+巨大芽孢杆菌（GM+BM）和摩西球囊霉菌+巨大芽孢杆菌+柠檬酸（GM+BM+CA）这5个处理,通过测定土壤全Cd、有效Cd和植株Cd吸收量及微生物群落变化,分析了外源菌剂和柠檬酸添加对龙葵修复Cd污染效果的影响.结果表明,相对于CK处理,GM处理龙葵根、茎和叶生物量显著增加35.67%、41.35%和65.38%,GM+BM+CA处理龙葵根和茎生物量显著增加73.38%和75.38%.GM处理提高了龙葵各部位Cd含量但差异不显著,GM+BM+CA处理龙葵叶部的Cd含量显著提高79.34%.GM处理龙葵茎和叶Cd累积量显著提高47.51%和89.58%.GM+BM+CA处理龙葵叶部Cd累积量显著提高226.84%.GM+BM+CA处理显著增加了Cd由龙葵茎向叶的转运系数,增幅为52.47%.GM+BM+CA处理显著增加了叶的富集系数,增幅为120.53%.此外,联合修复对根际微生物群落结构也产生了影响,特别体现在诱导某些关键微生物类群,如变形菌门、放线菌门、球囊菌门和油壶菌门相对丰度增加2.00%~5.77%、0.76%~9.96%、2.11%~3.63%和0.54%~2.98%.RDA分析发现变形菌门和放线菌门与土壤全Cd呈显著负相关,球囊菌门和油壶菌门与土壤全Cd呈负相关.关键微生物的变化增强了龙葵吸收根际养分和抗Cd胁迫的能力,提高龙葵Cd累积能力,有效降低土壤全Cd含量.综上,摩西球囊霉菌、柠檬酸与巨大芽孢杆菌通过共接种的方式活化了土壤中的难溶态Cd,有助于龙葵富集更多的Cd,同时也与摩西球囊霉菌产生联合修复的效果.富集植物-微生物联合修复Cd污染土壤有较好的应用潜力.     

接种Penicilliumsp.和根际分泌物对土壤甲磺隆的协同降解研究

通过接种Penicillium sp.和模拟小麦根际环境的方法,研究了甲磺隆在Penicillium sp.和小麦根际分泌物协同作用下的生物降解特性.结果表明,根系分泌物丰富了土著微生物和外源微生物,对甲磺隆的降解具有显著的促进作用.接种Penicillium sp.的根际土壤中甲磺隆降解半衰期为8.6 d,其降解速率是接种Penicillium sp.的非根际土壤的1.8倍,是普通根际土壤的2.7倍.继续追加甲磺隆的试验表明,接种菌株Penicillium sp.对甲磺隆的降解具有可持续性.     

Shinella zoogloeoides BC026对吡啶的降解特性研究

从首钢焦化厂的污水处理系统中分离1株能以吡啶为唯一碳、氮源的细菌BC026,它具有自絮凝特性,对卡那霉素、氨苄青霉素和壮观霉素具有抗性,可在阿须贝无氮培养基中良好生长.通过16S rRNA序列分析和Biolog微生物鉴定系统鉴定,确定该菌为Shinella zoogloeoides.纯菌对单基质的降解实验表明,在30℃、180 r/min和pH为7的条件下,当投菌量为0.1 g/L时,BC026可在17 h内将400 mg/L吡啶完全降解;在吡啶初始浓度为99～1 806 mg/L的无机盐培养基中,BC026均能保持降解活性,较高初始浓度的吡啶对BC026的生长产生一定抑制,但BC026在适应后对吡啶的降解速率较快;降解最适温度为30～35℃,最适pH为8.BC026对吡啶的代谢途径研究表明：降解的第一步是断开吡啶的2条C—N链,生成氨氮和戊二醛,随后戊二醛被氧化为戊二酸,并最终转化为二氧化碳和水;吡啶中的氮有59.5%转化成氨氮.     

武昌湖典型退化湿地菰草根际微生物群落结构

为研究淡水湖泊湿地微生物群落结构与退化湿地环境之间的关系,选取武昌湖湿地菰草根际水体、根际土壤、根下湖泊沉积物3组环境样本,提取样本微生物基因组DNA,并利用16S r DNA基因进行454高通量测序,共得到优质序列17 051条,产生2 062条OUTs。结果表明:武昌湖退化湿地菰草根际具有较为复杂的微生物群落结构,涉及15门48属,其中以变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、酸杆菌门(Acidobacteria)和绿弯菌门(Chloroflexi)为主要门类;菰草根际不同微环境微生物群落结构存在明显差异,生物多样性指数Shannon、Chao、ACE指数及OTUs均呈现根际土壤湖泊沉积物根际水体的变化特征。探究湿地退化过程中植被菰草根际微生物群落结构特征,对揭示长江中下游浅水湖泊湿地退化机理具有重要意义。     

珠江水体中细菌碱性磷酸酶基因phoD的多样性

碱性磷酸酶(APase)是最主要的有机磷水解酶,在无机磷匮乏的情况下,APase释放的无机磷供藻类利用,致使藻类大量繁殖,因此,对APase基因多样性的研究有利于揭示对水体富营养化有重要影响的微生物类群.基于此,本文以珠江水体为研究对象,构建了珠江广州段穗石、坦尾和南沙3个点细菌的pho D基因克隆文库.限制性酶切片段多态性(RFLP)分析结果表明,上述3个点的pho D基因分别含有15、17、13个操作分类单元(Operational Taxa Unit,OTU),穗石点的多样性(Shannon指数和Simpson指数)均高于其他两个点;pho D基因序列系统发育分析表明,珠江水体中pho D基因主要来源于Plesiocystis、Limnohabitans、Pirellula等属的细菌,而Limnohabitans属的pho D基因在文库中含量最高,占文库的44%~50%.研究表明,珠江水体中细菌的pho D基因具有较高的多样性,这些基因编码的碱性磷酸酶可能在珠江水体的富营养化过程中有重要的作用.     

Non-specific association between filamentous bacteria and fungus-growing ants

Fungus-growing ants and their fungal cultivar form a highly evolved mutualism that is negatively affected by the specialized parasitic fungus Escovopsis. Filamentous Pseudonocardia bacteria occurring on the cuticle of attine ants have been proposed to form a mutualistic interaction with these ants in which they are vertically transmitted (i.e. from parent to offspring colonies). Given a strictly vertical transmission of Pseudonocardia, the evolutionary theory predicts a reduced genetic variability of symbionts among ant lineages. The aim of this study was to verify whether actinomycetes, which occur on Acromyrmex octospinosus leaf-cutting ants, meet this expectation by comparing their genotypic variability with restriction fragment length polymorphisms. Multiple actinomycete strains could be isolated from both individual ant workers and colonies (one to seven strains per colony). The colony specificity of actinomycete communities was high: Only 15% of all strains were isolated from more than one colony, and just 5% were present in both populations investigated. Partial sequencing of 16S ribosomal deoxyribonucleic acid of two of the isolated strains assigned both of them to the genus Streptomyces. Actinomycetes could also be isolated from workers of the two non-attine ant species Myrmica rugulosa and Lasius flavus. Sixty-two percent of the strains derived from attine ants and 80% of the strains isolated from non-attine ants inhibited the growth of Escovopsis. Our data suggest that the association between attine ants and their actinomycete symbionts is less specific then previously thought. Soil-dwelling actinomycetes may have been dynamically recruited from the environment (horizontal transmission), probably reflecting an adaptation to a diverse community of microbial pathogens.     

三角帆蚌对白洋淀底泥氮磷释放及微生物的影响探究

为了解三角帆蚌对白洋淀底泥的影响,以白洋淀淀区水和底泥为试验材料,通过室内模拟试验并结合16S rRNA高通量测序技术,考察了三角帆蚌的活动对底泥氮、磷释放以及微生物丰度和多样性的影响.结果表明:①三角帆蚌的生物扰动作用显著提高了底泥氮、磷的释放,上覆水中TN、NH₃-N和TP浓度分别提高了0.18、1.1和1.2倍,颗粒态NH₃-N和可溶性磷浓度的增加是上覆水中NH₃-N、TP浓度增加的主要来源.②三角帆蚌的生物扰动作用轻微提高了上覆水中COD_Cr的浓度（P < 0.05）,并降低了底泥中全氮、全磷的含量,将底泥中全氮、全磷的削减率分别提高了26.0%和10.9%,底泥氮、磷释放规律的不同是导致全氮削减率高于全磷的主要原因.③三角帆蚌的活动可显著降低水中蓝细菌的数量,为其他微生物提供了更多生态位,从而提高了脱氮除磷功能菌丰度和微生物多样性,但是三角帆蚌对底泥中微生物丰度和多样性影响较小.研究显示,三角帆蚌能极大地促进白洋淀底泥氮、磷释放,同时降低内源污染风险,并提高上覆水脱氮除磷功能菌丰度和微生物多样性,可依据实际情况用于白洋淀湿地修复中.      

简青霉Penicillium simplicissimum木质素降解能力

从土壤中分离得到一株真菌经鉴定为简青霉Penicillium simplicissimum,将该菌用于木质素类化合物利用、染料脱色、天然木素降解及产酶特性等研究,充分证实该菌具有木质素降解能力.简青霉降解木质素是木质素过氧化物酶(LiP)、漆酶(Lac)和木聚糖酶共同作用的结果,降解主要发生在LiP与木聚糖酶活高产的初级代谢阶段,与白腐真菌表现出不同的作用机制.25d培养可使稻草木质素绝对量损失0.23g,降解率达14.94%,同时纤维素、半纤维素也有较高程度的降解.而pH值、Cu²⁺和Mn²⁺浓度对木质素的降解有较大影响.     

芦苇抑藻化感物质的分离及其抑制蛋白核小球藻效果研究

采用乙醇提取、溶剂萃取法从芦苇中分离出2组能抑制蛋白核小球藻生长的化感物质. 该2组化感物质对蛋白核小球藻的半效应浓度(EC₅₀)分别为0.68mg/L和2.65mg/L. 将前者用碱性氧化铝层析柱分离得到一组抑藻活性更强的化感物质和一组能够促进蛋白核小球藻生长的化感物质: 前者对蛋白核小球藻的EC₅₀为0.50mg/L, 是目前报道的抑藻能力最强的化感物质; 后者在浓度为0.63mg/L时能显著促进蛋白核小球藻的生长. 芦苇化感物质可以用来控制淡水中有害藻类.     

土地利用变化后不同种植年限香榧土壤微生物群落的组成及多样性

为探究毛竹林改种香榧后以及香榧种植年限的增加对土壤微生物群落结构特征的影响,并研究其与土壤肥力因子的关系,运用高通量测序技术,研究毛竹林、香榧5a、香榧+山稻5a、香榧10a和香榧30a 土地利用方式下土壤微生物群落组成及多样性的变化.结果表明土地利用改变后,土壤中细菌群落的Shannon指数和Chao l指数显著增加...     

凤眼莲对富营养化水体中氨氧化和反硝化微生物的影响

凤眼莲近年来广泛应用于富营养化淡水湖泊的生态修复中,但其对微生物的相互作用和对水体中氮素的去除鲜有报道.本研究在氮素去除过程中对比凤眼莲和细菌的相对重要性,并且检测浮游植物对硝化细菌和反硝化细菌相对丰度及多样性的影响.水体中氮素的去除率以及硝化和反硝化作用的潜在能力使用定量聚合酶链式反应(qPCRs)对硝化作用基因amoA和反硝化作用基因nirS/K进行检测,从微观角度研究富营养化水体中是否会受到凤眼莲存在的影响.结果表明,TN的减少在70d的实验周期中所有处理组表现较为一致,但凤眼莲存在的实验组在24h内TN和NH_4~+-N的去除上有显著的降低,并且amoA的丰度有所增加,nirS/K的丰度有所降低.T-RFLP结果表明亚硝化单胞菌在氨氧化微生物中占优势.凤眼莲的种植可以实现富营养化水体中NH_4~+-N的快速有效减少,且微生物的相互作用可以充分利用到淡水生态系统的修复中.     

Degradation of pyrene by immobilized microorganisms in saline-alkaline soil

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) is very difficult in saline-alkaline soil due to the inhibition of microbial growth under saline-alkaline stress. The microorganisms that can most effectively degrade PAHs were screened by introducing microorganisms immobilized on farm byproducts and assessing the validity of the immobilizing technique for PAHs degradation in pyrene-contaminated saline-alkaline soil. Among the microorganisms examined, it was found that Mycobacterium sp. B2 is the best, and can degrade 82.2% and 83.2% of pyrene for free and immobilized cells after 30 days of incubation. The immobilization technique could increase the degradation of pyrene significantly, especially for fungi. The degradation of pyrene by the immobilized microorganisms Mucor sp. F2, fungal consortium MF and co-cultures of MB+MF was increased by 161.7% (P < 0.05), 60.1% (P < 0.05) and 59.6% (P < 0.05) after 30 days, respectively, when compared with free F2, MF and MB+MF. Scanning electron micrographs of the immobilized microstructure proved the positive effects of the immobilized microbial technique on pyrene remediation in saline-alkaline soil, as the interspace of the carrier material structure was relatively large, providing enough space for cell growth. Co-cultures of different bacterial and fungal species showed different abilities to degrade PAHs. The present study suggests that Mycobacterium sp. B2 can be employed for in situ bioremediation of PAHs in saline-alkaline soil, and immobilization of fungi on farm byproducts and nutrients as carriers will enhance fungus PAH-degradation ability in saline-alkaline soil.     

浅层地下水升降对菜地土壤剖面硝化/反硝化微生物丰度的影响

为揭示湖泊近岸浅层地下水升降对菜地土壤剖面硝化与反硝化功能微生物基因丰度的影响,以洱海湖滨带菜地土壤剖面为研究对象,通过模拟地下水升降过程,分析了水位升高(S1)、水位降低(S2)及落干(S3)过程中土壤剖面AOA-amoA、AOB-amoA、nirK、nirS、nosZ基因丰度的变化特征,探讨了功能基因与土壤环境因子的耦合关系.结果表明:S3阶段的土壤剖面AOA-amoA和AOB-amoA基因丰度显著高于S1和S2;S1阶段的土壤剖面nirK、nirS、nosZ基因丰度均显著高于S2和S3.AOA-amoA基因丰度显著高于AOB-amoA基因丰度,nirS基因丰度显著高于nirK、nosZ基因丰度;不同取样时期的土壤剖面AOA-amoA、AOB-amoA、nirK、nirS、nosZ基因丰度均表现为A层B层C层D层.水位升降对土壤剖面AOA-amoA、AOB-amoA、nirK、nirS、nosZ基因丰度有显著影响,且AOA-amoA和nirS基因对水位升降更敏感,分别在硝化与反硝化作用中占主导地位;pH、有机碳(SOC)、全氮(TN)为功能基因AOA-amoA、AOB-amoA的环境驱动因子,而功能基因nirK、nirS、nosZ的环境驱动因子为土壤含水量(W)、铵态氮(NH~+_4-N)、硝态氮(NO~-_3-N)、TN、SOC、pH.该研究结果可为揭示浅层地下水升降过程中菜地土壤剖面氮素循环的微生物学机制提供科学依据.     

两种沉水植物附着生物种群特征对水深的响应研究

选取浅水湖泊中两种常见沉水植物上的附着生物作为研究对象,通过显微观测和16S rRNA高通量测序的方法,探究原位实验条件下水深(分别以T1、T2、T3、T4代表0.6、1.2、1.8和2.4 m)对狐尾藻(Myriophyllum spicatum)和马来眼子菜(Potamogeton malainus)附着藻类和附着细菌种群组成及其多样性特征的影响.结果表明,沉水植物附着生物种群特征与水深和生物种类有关,试验组中所有水深梯度下的狐尾藻附着生物群落的自养指数(AI)均大于马来眼子菜,且试验组中较小水深0.6~1.2 m的AI值较高,较大水深1.8~2.4 m的AI值较低.两种沉水植物表面附着藻类组成随水深变化差异显著(p0.05),绿藻门和硅藻门仍占种群绝对优势;附着细菌种群随水深梯度的变化差异也显著(p0.05),但主要包括有Proteobacteria、Firmicutes和Bacteroidetes等优势菌群,同种沉水植物或相似水层深度间的附着细菌组成和丰度有较高的相似性,并且试验组中较小水深0.6~1.2 m中拥有更多特有的细菌种类.研究发现,相同水深时狐尾藻附着藻类的多样性大于马来眼子菜,而马来眼子菜附着细菌多样性大于狐尾藻,且位于中等水深1.2~1.8 m时两种沉水植物附着藻类和附着细菌的多样性往往最高.通过水深对沉水植物附着生物种群特征影响的研究,可为揭示沉水植物表面微生态作用规律提供参考.     

集约化柑橘种植抑制土壤磷循环微生物活性

微生物是调节土壤磷循环的关键驱动力.阐明土壤解磷菌的微生物矿化过程对于提高植物养分吸收率和作物产量具有重要意义.通过测定柑橘园与毗邻的自然林地土壤编码碱性磷酸酶基因（phoD）丰度、解磷细菌群落多样性和土壤无机磷组分,探究柑橘种植对土壤微生物获取磷策略的影响机制.结果表明,柑橘种植导致土壤pH下降,土壤有效磷累积,ω（有效磷）平均值高达112 mg·kg^-1,显著高于毗邻的自然林地（3.7 mg·kg^-1）.柑橘种植也会影响土壤磷素组成,柑橘土壤含有较高的可溶态磷（CaCl₂-P）、柠檬酸提取态磷（Citrate-P）和矿物结合态磷（HCl-P）.自然林地土壤各磷组分均显著低于柑橘土壤,而phoD基因丰度和碱性磷酸酶活性显著高于柑橘土壤.高通量测序结果表明,柑橘土壤解磷细菌Shannon指数（4.61）显著低于自然林地（5.35）,群落结构也有别于自然林地.柑橘种植改变了土壤解磷菌的群落组成,自然林地变形菌门的相对丰度显著低于柑橘土壤.土壤有效磷含量与碱性磷酸酶活性呈显著负相关,表明土壤高磷累积抑制土壤解磷细菌的活性.柑橘种植改变了土壤微生物对磷的获取策略,在柑橘园中,土壤微生物主要依赖外源磷,而自然林地土壤微生物主要以微生物分泌碱性磷酸酶矿化有机磷来获取磷的方式满足其生长需求.     

北京市典型土著沉水植物对水体中硝酸盐和正磷酸盐的耐受性研究

以北京市3种典型土著沉水植物轮叶黑藻、狐尾藻和金鱼藻为研究对象,构建模拟水生生态系统,研究3种沉水植物对水体中NO₃^-和PO₄^3-的耐受性并确定其耐受范围.同时,对植物体内过氧化氢酶、丙二醛、叶绿素和蛋白质4种指标进行检测.结果表明,轮叶黑藻对NO₃^-具有较强的耐受性,耐受浓度可达8 mg·L^-1,金鱼藻和狐尾藻次之,耐受浓度为3~5 mg·L^-1;狐尾藻对PO₄^3-的耐受性最差,耐受浓度约为0.2 mg·L^-1,轮叶黑藻和金鱼藻的耐受性相当,当PO₄^3-浓度达到0.4 mg·L^-1时开始出现显著胁迫.因此,在本研究的实验条件下,当水体NO₃^-浓度<5 mg·L^-1、PO₄^3-浓度<0.2 mg·L^-1时,建议3种沉水植物同时种植;当NO₃^-浓度>5 mg·L^-1时,建议种植轮叶黑藻;当PO₄^3-浓度为0.2~0.4 mg·L^-1时,建议种植轮叶黑藻和金鱼藻.研究结果可为北京市再生水补给河湖水库的水生态修复及其沉水植物群落的构建提供一定的理论指导.     

Scale-dependent analysis of an otter–crustacean system in Argentinean Patagonia

The Southern river otter or ‘huillin’, Lontra provocax, is an endangered species endemic of the Andean Patagonian region of Argentina and Chile. It feeds almost exclusively on the genera of macro-crustacea: Aegla and Sammastacus. The aim of this study was to analyse the role of food availability on the huillin’s distribution using a scale-dependent analysis of crustacean and otter distributions. We compared the distributions of otters and macro-crustaceans along a north–south regional gradient, between river basins of northern Patagonia, in an altitudinal gradient within a river basin, and between habitat types within a lake. We investigated the distribution of otters by sign surveys along lake shores, river banks and marine coasts, and of crustaceans using surveys in the water, undigested remains in mink (Mustela vison) scats, presence of external skeletons at the waterside and through interviews with local people. Our results show that there were heterogeneities in the distributions of macro-crustaceans at four scales and these were generally reflected in the distributions of freshwater otters. We conclude that the main factor limiting the distributions of L. provocax in freshwater environments is the availability of macro-crustaceans. This paper shows how scale-dependent type analyses of population distribution serves as a method for identifying key environmental factors for species for which the use of long-term demographies is unfeasible.     

Efficacy of two chemical coagulants and three different filtration media on removal of Aspergillus flavus from surface water

Aquatic fungi are common in various aqueous environments and play potentially crucial roles in nutrient and carbon cycling as well as interacting with other organisms. Species of Aspergillus are the most common fungi that occur in water. The present study was undertaken to elucidate the efficacy of two coagulants, aluminum sulfate and ferric chloride, used at different concentrations to treat drinking water, in removing Aspergillusflavus, as well as testing three different filtration media： sand, activated carbon, and ceramic granules, for their removal of fungi from water. The results revealed that both coagulants were effective in removing fungi and decreasing the turbidity of drinking water, and turbidity decreased with increasing coagulant concentration. Also, at the highest concentration of the coagulants, A. flavus was decreased by 99.6% in the treated water. Among ceramic granules, activated carbon, and sand used as media for water filtration, the sand and activated carbon filters were more effective in removing A.flavus than ceramic granules while simultaneously decreasing the turbidity levels in the test water samples. Post-treatment total organic carbon （TOC） and total nitrogen （TN） concentrations in the experimental water did not decrease; on the contrary, TN concentrations increased with the increasing dosage of coagulants. The filtration process had no effect in reducing TOC and TN in tested water.