产脲酶细菌矿化修复Cd和Pb污染土壤效应和机制

为了修复重金属污染土壤,保障蔬菜品质安全,利用产脲酶筛选培养基从重金属污染生菜根际土壤中筛选产脲酶细菌.通过溶液吸附和土壤静置培养试验研究功能菌株对Cd和Pb的固定钝化机制;利用水培试验研究功能菌株对不同品种蔬菜生长及吸收Cd和Pb的影响.结果显示,从生菜根际土壤中共分离到10株产脲酶细菌,其中菌株Microbacterium foliorum CH6和Bacillus thuringiensis N3的产脲酶能力（分别为62.6,59.6mS/（cm·min））和吸附钝化Cd和Pb （84.6%~91.3%）的能力最高.扫描电镜-能谱、傅里叶红外光谱和X衍射分析表明,菌株CH6和N3能够通过细胞壁吸附和诱导重金属成矿反应,降低溶液中的Cd和Pb含量.土壤静置试验表明,与不接种菌株对照相比,菌株CH6和N3均能降低土壤中可交态Cd和Pb （46.3%~66.7%）的含量,增加碳酸盐结合态及残渣态Cd和Pb （40%~95.4%）的含量.砂培试验表明,菌株CH6和N3均能显著降低生菜、上海青和空心菜可食用部分中Cd （39.2%~81.4%）和Pb （27.4%~82.3%）的含量.产脲酶细菌CH6和N3能够矿化土壤中的Cd和Pb,降低土壤中重金属有效态含量,阻控生菜、上海青和空心菜对Cd和Pb的吸收.     

Effects of disinfection efficiency on microbial communities and corrosion processes in drinking water distribution systems simulated with actual running conditions

The effects of disinfection efficiency on microbial communities and the corrosion of cast iron pipes in drinking water distribution systems (DWDSs) were studied. Two annular reactors (ARs) that simulated actual running conditions with UV/Cl₂ disinfection and chlorination alone were used. High chlorine consumption and corrosion rate were found in the AR with UV/Cl₂. According to functional genes and pyrosequencing tests, a high percentage of iron recycling bacteria was detected within the biofilm of the AR with Cl₂ at early running stage, whereas siderophore-producing bacteria were dominant in the biofilm of the AR with UV/Cl_2. At the early running stage, the sequential use of UV light and an initial high chlorine dosage suppressed the biomass and iron-recycling bacteria in both bulk water and biofilms, thereby forming less protective scales against further corrosion, which enhanced chlorine consumption. Non-metric multidimensional scaling analysis showed that the bacterial communities in the ARs shaped from within rather than being imported by influents. These results indicate that the initial high disinfection efficiency within the distribution system had not contributed to the accumulation of iron-recycling bacteria at the early running stages. This study offer certain implications for controlling corrosion and water quality in DWDSs.     

Effects of EDTA and plant growth-promoting rhizobacteria on plant growth and heavy metal uptake of hyperaccumulator Sedum alfredii Hance

Phytoremediation is a cost-effective and environment-friendly strategy for decontaminating heavy-metal-contaminated soil. However, the practical use of phytoremediation is constrained by the low biomass of plants and low bioavailability of heavy metals in soil. A pot experiment was conducted to investigate the effects of the metal chelator ethylenediaminetetraacetic acid (EDTA) and EDTA in combination with plant growth-promoting rhizobacteria (Burkholderia sp. D54 or Burkholderia sp. D416) on the growth and metal uptake of the hyperaccumulator Sedum alfredii Hance. According to the results, EDTA application decreased shoot and root biomass by 50% and 43%, respectively. The soil respiration and Cd, Pb, Zn uptake were depressed, while the photosynthetic rate, glutathione and phytochelatin (PC) contents were increased by EDTA application. Interestingly, Burkholderia sp. D54 and Burkholderia sp. D416 inoculation significantly relieved the inhibitory effects of EDTA on plant growth and soil respiration. Compared with the control, EDTA + D416 treatment increased the Cd concentration in shoots and decreased the Pb concentration in shoots and roots, but did not change the Zn concentration in S. alfredii plants. Furthermore, EDTA, EDTA + D54 and EDTA + D416 application increased the cysteine and PC contents in S. alfredii (p < 0.05); among all tested PCs, the most abundant species was PC2, and compared with the control, the PC2 content was increased by 371.0%, 1158.6% and 815.6%, respectively. These results will provide some insights into the practical use of EDTA and PGPR in the phytoremediation of heavy-metal-contaminated soil by S. alfredii.     

窖水中微生物降解污染物的关键细菌

为探究以氮、磷及有机物污染为主要特征的、窖水中参与污染物降解的关键细菌及它们之间潜在的相关关系,基于16S rRNA的微生物组学截面数据,分析了窖水中细菌群落结构与功能及其与水质因子间的相关性,并通过微生物物种的同现或相关种间作用推断模型,构建了7个微生物菌属间的共现性关联网络.结果表明,窖水中存在具有相对特定生态功能的细菌,进行着诸多活跃的与新陈代谢功能基因相关的代谢活动;窖水微生物类群共现性关联网络中大部分节点的菌属营互利共生类型的生态关系;Lacibacter、Arthrobacter、Candidatus Protochlamydia、Methylocaldum、Sulfuritalea、Mycobacterium、Aquirestis、Rhodobacter、Methylotenera等菌属拥有较高的点度中心度;较强的互作关系发生在Sulfuritalea-Rhodobacter、Azospirillum-Rhodobacter、Methylocaldum-Rhodobacter、Arthrobacter-Rhodobacter、Rhodoplanes-Rhodobacter、Candidatus Protochlamydia-Rhodobacter、Methylotenera-Rhodobacter、Rhodobacter-Aquirestis、Mycobacterium-Rhodobacter、PlanctomycesCandidatus Solibacter、Planctomyces-Legionella、Hymenobacter-Adhaeribacter、Luteolibacter-Crenothrix之间.综合分析节点微生物相关性、点度中心度及菌属间的互作强度,认为Rhodobacter、Methylocaldum、Methylotenera、Acinetobacter、Novosphingobium、Planctomyces、Hymenobacter、Luteolibacter为参与窖水污染物微生物降解的关键细菌,Rhodobacter为关键细菌的代表属.研究结果加深了对窖水中污染物微生物降解机制的认识.     

人类活动对三峡消落带土壤亚硝酸盐型甲烷厌氧氧化菌群落的影响

由"Candidatus Methylomirabilis oxyfera(M.oxyfera)"主导的亚硝酸盐型甲烷厌氧氧化过程(nitrite-dependent anaerobic methane-oxidizing,ndamo)是碳循环中新发现的一个过程.已有研究发现三峡库区消落带有n-damo菌的存在.为了解人类活动对n-damo菌群落结构的影响,对三峡库区受人类活动干扰程度不同的消落带n-damo菌群落进行了研究.结果表明,人为干扰水平高的小河消落带n-damo菌基因丰度(8.98×102~3.31×105copies·g~(-1))整体上要高于白家溪(4.72×102~5.32×104copies·g~(-1)).此外,淹水都会导致两个地点n-damo菌基因丰度的增加.白家溪的n-damo菌丰度与累积淹水时间呈线性正相关,而小河的n-damo菌丰度与土壤总氮显著负相关.系统发育树和多样性分析表明,淹水前白家溪和小河相近高程土壤n-damo菌群落结构比较类似,但淹水后出现较大差异;较低高程土壤的群落多样性要明显高于较高高程土壤.消落带土壤中的n-damo菌群落具有明显的时空异质性,且淹水会导致这种异质性增加.淹水和人类活动对于n-damo菌的多样性具有同等的重要性.这些结果表明,淹水和人类活动都会影响消落带土壤中n-damo菌群落结构,而人类活动可能进一步增加n-damo菌的丰度和多样性.     

基质浓度对ABR厌氧氨氧化反硝化脱氮除碳效能影响及动力学特征

采用厌氧折流板反应器(ABR)为研究对象,以一定COD、NH+4-N和NO-2-N比例增加进水基质浓度,以明确基质负荷提高对ABR厌氧氨氧化和反硝化协同体系脱氮除碳的影响,并通过基质去除模型获得反应器对基质的耐受程度.研究表明,ABR反应器能够实现厌氧氨氧化反硝化耦合脱氮除碳,当进水基质COD、NO-2-N和NH+4-N浓度从220、168和60 mg·L~(-1)提高至420、270和110 mg·L~(-1)时,反应器脱氮效能下降,COD、NO-2-N、NH+4-N和TN去除率分别为97%、94%、30%和78%,厌氧氨氧化对TN去除的贡献率从43.08%骤降至16.49%,反硝化脱氮贡献率从53.81%增至82.07%.动力学模型拟合发现,Stover-Kincannon模型(R2=0.937,TN;R2=0.975,COD)较一级基质去除模型(R2=0.314,TN;R2=0.016,COD)更适合评价反应器对基质的承受力;Stover-Kincannon模型表明,反应器对TN和COD的最大基质利用率分别为1.43 g·L-1·d-1和3.33 g·L-1·d-1,饱和常数(KB)分别为1.2和3.79,研究认为ABR协同脱氮除碳体系理论上还有继续提升基质负荷的潜力.     

温度对沉水植物腐解释放DOM及微生物群落多样性的影响

在5,10,20,35℃下研究了黑藻和马来眼子菜腐解过程中DOM的特性、细菌和真菌群落结构多样性的变化.结果表明,腐解结束时5,10,20,35℃下黑藻的干物质剩余量分别为初始干物质质量的59.13%、43.91%、32.61%和29.57%,马来眼子菜的干物质剩余量分别为初始干物质质量的69.13%、51.3%、30.87%和29.57%.升高温度一定程度上促进了植物有机碳和全氮释放,对全磷无明显影响（P>0.05）.采用平行因子分析法得到黑藻中含有2种类腐殖酸组分C1、C2和1种类蛋白质组分C3,马来眼子菜中含有3种类腐殖酸组分C1、C2和C3.温度升高使得腐解水体中DO和电导率的变化程度加剧,总体上4种温度下2种植物腐解水体中DO均呈现先下降后上升的趋势.通过高通量测序得到腐解前期（0~16d）参与黑藻和马来眼子菜腐解的主要细菌分别为变形菌门（Proteobacteria）和拟杆菌门（Bacteroidetes）;腐解中后期（16~68d）参与黑藻和马来眼子菜腐解的细菌分别以厚壁菌门（Phylum Firmicutes）和变形菌门（Proteobacteria）为主,温度对细菌群落结构无明显影响,而参与2种植物腐解的真菌均以子囊菌（Ascomycetes）最为活跃.     

包埋厌氧氨氧化的脱氮特性及其微生物群落结构

采用聚乙二醇二丙烯酸酯作为载体,分别以N,N,N,‘N’-四甲基乙二胺（TEMED）,过硫酸钾（KPS）作为促进剂和引发剂,对厌氧氨氧化菌进行包埋固定化.采用正交试验优化包埋条件,得到的最佳条件为：10% PEGDA单体,0.25% KPS,0.5% TEMED,最佳的操作条件为：聚合温度20℃,聚合时间控制在5min左右,菌胶比选取1：1.包埋颗粒的连续流实验表明,颗粒经过短暂的活性恢复后,脱氮效果不断提升,并且对水力负荷的提高有一定的抗冲击能力.扫描电镜（SEM）表明本实验包埋材料具有很好的生物相容性,且具有良好的传质性能.高通量测序显示,稳定运行一段时间后,颗粒内微生物多样性略有减小,厌氧氨氧化功能菌Candidatus Kuenenia占整个微生物体系比例由6.58%上升至9.8%,微生物种群的变化说明了包埋后厌氧氨氧化性能能够得到更大的提升.     

蜡样芽孢杆菌中甲酸脱氢酶基因产氢研究

通过基因筛选,成功分离并克隆到蜡样芽胞杆菌XN12（Bacillus cereus XN12）的甲酸脱氢酶基因fdhF（formate dehydrogenase）,该基因全长2937bp,GC含量39.3%,编码978个氨基酸,与已报道的蜡样芽孢杆菌Q1的fdhF基因（GenBank No.CP000227.1）同源性达到100%.将其连接在表达载体pET32a上并融合His标签,构建了重组质粒pET32a-FDHF-His,转入大肠杆菌BL21（Escherichia coli BL21）后获得了高效表达.重组菌株经IPTG诱导后经WesternBlot分析表明,重组蛋白分子量约为108kDa.通过对重组菌株产氢性能试验表明,重组菌对提高产氢率具有一定促进作用,产氢量为每消耗1mol的葡萄糖和甲酸盐分别能产生0.73mol和0.20mol的氢气.