假单胞杆菌抗镉基因的克隆

以pBR322为载体,将抗镉的假单胞杆菌R4染色体的抗镉基因克隆至大肠杆菌HB101,从5000个重组体中筛选出一株抗镉基因克隆株,命名为大肠杆菌HB101(pBC311)。重组质粒pBC311经PstⅠ酶解和琼脂糖凝胶电泳证实抗镉基因位于3.6kb长的PstⅠ片段上。克隆菌株可在含100μg/ml CdCl_2的L-肉汤中生长,其生长停滞期比受体大肠杆菌HB101短得多,说明克隆菌株含有抗镉基因。     

假单胞菌的氯儿茶酚1,2-双加氧酶基因的克隆和表达

 从土壤中分离得到1株降解2,4-二氯酚(2,4-DCP)能力较强的细菌菌株GT241-1,克隆了该菌株的3,5-二氯儿茶酚1,2-双加氧酶基因(dcpB),采用的克隆策略为:用Southern杂交对dcpB进行定位后,构建重组质粒,再用斑点杂交从重组质粒中筛选目的转化子.经序列测定得知dcpB亚克隆片段全长4303bp,其中dcpB基因编码区765bp.核苷酸和推测的氨基酸序列分析表明,dcpB与已在GenBank登记的相关基因有一定的差异.dcpB基因能够在大肠杆菌转化子中成功地表达有生物活性的酶.     

镉铅对小麦醇脱氢酶(ADH)基因表达影响的初步研究

经过镉，铅４５ｈ污染处理，运用Ｎｏｒｔｈｅｉｎ斑点杂分析和酶活性测定的方法探测萌发期小麦ＡＤＨ酶基因的表达情况。实验结果表明，ＡＤＨ基因表达在ＲＮＡ和酶活性两个层次均有变化，并且呈现相似的变化趋势，反映得金属的影响是来源于基因转录水平。     

一株高效苯酚降解菌的分离、鉴定及降解特性的研究

从污泥中分离到1株高效苯酚降解细菌JF-10.在以1000mg·L-1苯酚作为唯一碳源并加有0.05%酵母粉的无机盐培养基中,JF-10能在24h内将苯酚完全降解.该菌被鉴定为假单胞菌,与P.citronellolis、P.nitroreducens、P.aeruginosa亲缘关系较近,但生理生化特征又有差异.JF-10与P.citronelloli gyrB基因的序列相似性只有75%,与P.aeruginosa也只有80%.该菌在pH值4.0～9.0时对苯酚的降解达80%以上,加入一定浓度Mn2 、Cu2 、M2 、Pb2 对苯酚降解有促进作用;200mg·L-1的α-萘酚、苯胺能显著抑制苯酚降解,但甲苯对苯酚的降解无影响,苯酚可以促进邻甲酚的降解.     

学习借鉴,走跨越式发展之路

论述了为实现全面建设小康社会的发展目标，必须走跨越式发展之路。     

Implications of Macroalgal Isolation by Distance for Networks of Marine Protected Areas

The global extent of macroalgal forests is declining, greatly affecting marine biodiversity at broad scales through the effects macroalgae have on ecosystem processes, habitat provision, and food web support. Networks of marine protected areas comprise one potential tool that may safeguard gene flow among macroalgal populations in the face of increasing population fragmentation caused by pollution, habitat modification, climate change, algal harvesting, trophic cascades, and other anthropogenic stressors. Optimal design of protected area networks requires knowledge of effective dispersal distances for a range of macroalgae. We conducted a global meta‐analysis based on data in the published literature to determine the generality of relation between genetic differentiation and geographic distance among macroalgal populations. We also examined whether spatial genetic variation differed significantly with respect to higher taxon, life history, and habitat characteristics. We found clear evidence of population isolation by distance across a multitude of macroalgal species. Genetic and geographic distance were positively correlated across 49 studies; a modal distance of 50–100 km maintained F_ST < 0.2. This relation was consistent for all algal divisions, life cycles, habitats, and molecular marker classes investigated. Incorporating knowledge of the spatial scales of gene flow into the design of marine protected area networks will help moderate anthropogenic increases in population isolation and inbreeding and contribute to the resilience of macroalgal forests. Implicaciones del Aislamiento por Distancia de Macroalgas para Redes de Áreas Marinas Protegidas     

宁波三江口水域原核生物群落结构分析

首次应用16S rRNA基因-Illumina Miseq高通量测序技术对甬江流域宁波三江口区域的表层水体原核生物群落进行了分析,共获得215 504条高质量序列.多样性指数分析表明,该流域的水体原核生物群落具有较高的遗传多样性和丰富度.菌群分类分析发现,β-变形菌纲(β-Proteobacterium)、放线菌门(Actinobacteria)和拟杆菌门(Bacteroidetes)是主要的优势菌群,其相对丰度占总群落的78.88%.对比不同站位的原核生物群落,发现水文环境可能是影响三江口水域原核生物群落组成和结构的重要因素之一.与以往报道对比分析表明,甬江流域分布有多种污水和粪便污染指示菌,其中余姚江污染指示菌的丰度最高,说明其受污染的风险可能较高.病原菌BLASTN分析表明,在种和亚种水平上,分别检出76和18种潜在病原菌,占序列总量的2.19%和0.40%.本研究为系统认识甬江流域原核生物群落结构及生态功能提供了重要基础数据.     

Effect of reclaimed water effluent on bacterial community structure in the Typha angustifolia L. rhizosphere soil of urbanized riverside wetland, China

In order to evaluate the impact of reclaimed water on the ecology of bacterial communities in the Typha angustifolia L. rhizosphere soil,bacterial community structure was investigated using a combination of terminal restriction fragment length polymorphism and 16S rRNA gene clone library. The results revealed significant spatial variation of bacterial communities along the river from upstream and downstream. For example,a higher relative abundance of γ-Proteobacteria,Firmicutes,Chloroflexi and a lower proportion ofβ-Proteobacteria and ε-Proteobacteria was detected at the downstream site compared to the upstream site. Additionally,with an increase of the reclaimed water interference intensity,the rhizosphere bacterial community showed a decrease in taxon richness,evenness and diversity. The relative abundance of bacteria closely related to the resistant of heavy-metal was markedly increased,while the bacteria related for carbon/nitrogen/phosphorus/sulfur cycling wasn't strikingly changed. Besides that,the pathogenic bacteria markedly increased in the downstream rhizosphere soil since reclaimed water supplement,while the possible plant growth-promoting rhizobacteria obviously reduced in the downstream sediment.Together these data suggest cause and effect between reclaimed water input into the wetland,shift in bacterial communities through habitat change,and alteration of capacity for biogeochemical cycling of contaminants.     

石油污染土壤生物修复过程中氮循环功能基因的动态检测

氮循环相关功能微生物在土壤发挥其生态功能中起着重要的作用.为定量分析其中的固氮细菌、反硝化细菌和硝化细菌在石油污染土壤生物修复过程中的演变情况,采用了实时定量PCR技术对其相关功能基因nifH、narG和amoA的拷贝数进行检测.结果表明,污染土壤中nifH、narG和amoA基因拷贝数及其占总16S rRNA基因拷贝数的比例远低于正常土壤,修复后土壤中的同类分析结果与正常土壤接近.表明石油污染物破坏了氮循环相关菌落结构,而生物修复则使其得以恢复.进一步分析了不同修复方式下氮循环功能基因的恢复情况以及土壤中石油烃降解率.也表明同时添加秸秆和菌剂具有最好的修复效果,处理40 d后其nifH、narG和amoA基因拷贝数（以干土计）分别恢复到2.68×106、1.71×106和8.54×104g-1,石油烃降解率达到48%.投加真菌-细菌复合菌剂的效果优于只投加细菌菌剂的效果.本研究结果表明氮循环功能基因的动态监测可以从基因和物种水平上反映土壤修复的效果,为土壤修复的监控和效果评估提供参考.     

反硝化脱硫工艺中微生物群落结构及动态分析

为了研究反硝化脱硫工艺(denitrifying sulfide removal,DSR)中微生物群落与工艺运行的相关性,实验提取了反应器运行不同阶段污泥样品中微生物的全基因组DNA,利用高通量宏基因组学技术———基因芯片来解析各阶段功能微生物群落的结构特征及其演替过程.通过对功能基因的Simpson、Shannon多样性指数和聚类分析表明,微生物群落结构会随着反应器运行的阶段进行相应调整,且变化较大.在运行的前两个阶段,由于不适应环境,微生物群落的多样性指数比起始时明显减少,随着反应器的运行污泥逐渐成熟,多样性指数迅速增加,也标志着反应器进入稳定运行阶段.通过对反硝化脱硫过程中关键基因相对丰度的分析发现,功能基因的丰度不仅能反映功能菌群的活性,而且与工艺处理效果密切相关.