高通量测序技术研究典型湖泊岸边陆向深层土壤中厌氧氨氧化细菌的群落结构

为研究具有富营养化特征的白洋淀湖泊岸边陆向深层土壤中厌氧氨氧化细菌的群落结构组成,针对厌氧氨氧化的功能基因hzs B(联氨合成酶关键基因)进行高通量测序.结果显示,白洋淀湖泊岸边陆向深层土壤共8个样品处理后共得到77082条序列,在相似度90%下聚类得到40个OTUs.多样性分析(OTUs水平)表明,在白洋淀湖泊岸边近湖岸交界处,上层土壤中厌氧氨氧化细菌的群落多样性要高于下层,而在远湖岸交界处则相反,下层土壤中厌氧氨氧化细菌的群落多样性要高于上层土壤.PCo A分析(OTUs水平)表明,近湖岸交界处采样点各不同深度土壤层之间的厌氧氨氧化细菌的群落结构更相似,而远湖岸界面处采样点各不同深度之间的厌氧氨氧化细菌的群落结构差异较大.群落结构分析表明,在白洋淀湖泊岸边陆向深层土壤中,序列隶属于Candidatus‘Brocadia anammoxidans’(34.07%)、Candidatus‘Brocadia fulgida’(24.66%)、Candidatus‘Jettenia sp.’(3.00%)、Candidatus‘Kuenenia sp.’(0.22%)、Candidatus‘Scalindua sp.’(0.08%)及与Genbank中已探明的厌氧氨氧化细菌亲缘关系较远的Anammox-like cluster(37.98%).相关性分析(p0.10)表明,在白洋淀湖泊岸边陆向深层土壤中,厌氧氨氧化细菌α-多样性指数中丰富度Chao指数与含水率(MC)呈负相关,多样性Shannon指数与氨氮(NH+4)呈负相关,优势度Simpson指数与NH+4呈正相关;但厌氧氨氧化细菌群落组成中Candidatus‘Brocadia anammoxidans’的富集与总碳(TC)及总硫(TS)含量呈正相关.综上,本文针对厌氧氨氧化功能基因hzs B扩增进行高通量测序,一定程度上揭示了具有富营养化特征的白洋淀湖泊岸边陆向深层土壤中厌氧氨氧化细菌的多样性、群落组成及其与土壤环境因子的关系.     

岩溶地区不同土地利用方式土壤固碳细菌群落结构特征

固碳细菌是土壤碳循环重要的微生物群落,研究其群落结构特征对认识土壤生态系统的固碳机制具有重要意义.以桂林毛村岩溶实验场的岩溶区、混合区与非岩溶区为研究样区,采集稻田、玉米和柑橘园表层土壤,以cbb LR为固碳细菌的指示基因,采用高通量测序方法,对比在三类区域土壤中固碳细菌的群落丰度、组成及多样性特征的异同.结果表明,三类区域土壤中固碳细菌属于变形菌门和放线菌门.其中,变形菌门的α-变形菌纲(α-Proteobacteria,24.6%)为三类区域土壤中的优势纲,以根瘤菌为主的兼性自养菌是主要的固碳细菌.在岩溶区,伯克氏菌目(Burkholderiales)、红假单胞菌属(Rhodopseudomonas)、固氮螺菌属(Azospirillum)、费氏根瘤菌(Sinorhizobium fredii HH103)、豌豆根瘤菌(Rhizobium leguminosarum bv.trifolii)等微生物的丰度均高于混合区和非岩溶区;而慢生根瘤菌属(Bradyrhizobium)为混合区与非岩溶区土壤中的优势种群.冗余分析(redundancy analysis)表明,pH、土壤有机碳(SOC)、可溶性有机碳(DOC)、总氮(TN)和阳离子交换量(CEC)等土壤因子是影响固碳细菌群落结构差异的主要生态因子.以上结果表明,岩溶区的土壤特性对固碳细菌的群落结构有显著影响.     

碱性蚀刻废液的综合利用

介绍了利用含铜蚀刻废液回收铜的生产工艺、技术特点、工艺流程和产品质量. 研究了沉淀母液中残余铜的除去方法,使之再生,可回用于碱性蚀刻液的生产.碱性蚀刻废液的综合利用对于资源节约与环境保护都具有重要意义.     

废蚀刻铜液制取氧化铜及废液的再生条件研究

采用酸性蚀刻废液与碱性蚀刻废液混合沉铜的方法制取氧化铜,讨论了酸度对沉淀的影响;同时,对沉铜后母液用于制取碱性蚀刻液的指标的调整、蚀刻速率进行了研究,达到了废液重复使用的目的,是一种较理想的再生利用电路板废液和回收铜的方法。     

不同养分对磷酸盐固定的污染土壤中铜锌生物有效性的影响

采用盆栽试验,在磷酸盐固定铜、锌单一及复合的三级污染(Cu 200 mg·kg^-1、Zn 400 mg·kg^-1)红壤和水稻土中,种植黑麦草40　d,测定不加养分、加入KCl和NH₄Cl、K₂SO₄和(NH₄)₂SO₄后土壤中有效态铜锌含量、黑麦草生物量及其铜锌吸收量,以讨论养分对污染土壤中磷酸盐固定态铜锌的活化作用.结果表明,磷酸盐固定的铜单一、锌单一和铜锌复合污染红壤中,增加养分,有效态铜锌的含量都较未加入养分的增高,并且养分KCl和NH₄Cl的增高幅度大于K₂SO₄和(NH₄)₂SO₄,其中在磷酸盐固定的锌单一污染红壤中,有效态锌含量较未加入养分时增幅高达133.4%.虽然加入K₂SO₄和(NH₄)₂SO₄,促进了红壤中黑麦草植株的生长,最大增幅达22.2%,但同时也增加了植株21.5%～112.6%的铜锌吸收量.调节养分对水稻土中铜锌的生物有效性没有显著影响.施入氮钾等养分能够改变磷酸盐固定的污染红壤中铜锌的生物有效性,对固定的铜锌都有活化作用.因此,在重金属污染土壤的修复中要充分考虑施肥等养分条件对固定钝化的重金属活性的影响.     

铜和锌离子对真鲷幼鱼组织酶活性的影响

研究水中Ｃｕ＾２＋、Ｚｎ＾２＋对真鲷幼鱼组织ＣＡＴ、ＧＰＴ、胃蛋白酶活性的影响。结果表明：真鲷幼鱼暴露于不同浓度Ｃｕ＾２＋（０．５、０．７５、１．５ｍｇ／Ｌ）、Ｚｎ＾２＋（１、２、４ｍｇ／Ｌ）海水中４ｄ后,Ｃｕ＾２＋、Ｚｎ＾２＋对鳃,肝脏ＣＡＴ,胃蛋白酶有抑制作用。     

1株反硝化除磷菌的鉴定及其反硝化功能基因研究

研究反硝化除磷菌(denitrifying polyphosphate-accumulating organism,DPAO)的筛选、鉴定及反硝化功能基因.利用反硝化培养基分离得到菌株ZQN2,经好氧吸磷试验,硝酸盐还原产气试验并辅助异染颗粒和PHB颗粒染色,确定其为反硝化除磷菌.厌氧释磷/缺氧吸磷试验结果表明,菌株ZQN2在厌氧段释磷并合成PHB(poly-β-hydroxybutyrate),在缺氧段以NO3-为电子受体氧化PHB并过量吸磷,进行了同步反硝化除磷.对菌株ZQN2进行16S rRNA基因序列分析,并构建系统进化树,结果表明该菌株与GenBank数据库中多株Bacillus cereus菌株16S rRNA基因的同源性在99%以上.结合生理生化检测,判断菌株ZQN2为蜡状芽胞杆菌(Bacillus cereus).对其反硝化功能基因的研究表明,菌株ZQN2为nirS+和nirK-型,从分子生物学角度确认其具有反硝化功能;菌株ZQN2的nirS序列的系统发育分析结果表明,其nirS序列与多株Pseadomonas aeruginosa的亲缘关系最接近,这与基于16S rRNA的系统发育分析的结果不一致.     

Biological treatment of chicken feather waste for improved biogas production

A two-stage system was developed which combines the biological degradation of keratin-rich waste with the production of biogas. Chicken feather waste was treated biologically with a recombinant Bacillus megaterium strain showing keratinase activity prior to biogas production. Chopped, autoclaved chicken feathers (4%, W/V) were completely degraded, resulting in a yellowish fermentation broth with a level of 0.51 mg/mL soluble proteins after 8 days of cultivation of the recombinant strain. During the subsequent anaerobic batch digestion experiments, methane production of 0.35 Nm³/kg dry feathers (i.e., 0.4 Nm³/kg volatile solids of feathers), corresponding to 80% of the theoretical value on proteins, was achieved from the feather hydrolyzates, independently of the pre-hydrolysis time period of 1, 2 or 8 days. Cultivation with a native keratinase producing strain, Bacillus licheniformis resulted in only 0.25 mg/mL soluble proteins in the feather hydrolyzate, which then was digested achieving a maximum accumulated methane production of 0.31 Nm³/kg dry feathers. Feather hydrolyzates treated with the wild type B. megaterium produced 0.21 Nm³ CH₄/kg dry feathers as maximum yield.     

Cu2+和Pb2+在BS-12两性修饰膨润土上的吸附及其交互作用

两性表面修饰可以同时提高黏土矿物对有机物、重金属的吸附性能,研究两性修饰土对多种重金属共存条件下的吸附及交互作用对于其应用具有实际意义.采用红外光谱(FTIR)对膨润土(CK)和150BS-12修饰土(150BS-12)吸附金属离子前后进行表征,通过批处理法研究不同比例BS-12修饰膨润土对Cu~(2+)和Pb~(2+)在单一及Cu-Pb复合体系中的平衡吸附特征和差异,并探讨金属离子在修饰土样上的交互作用机制.结果表明,供试土样在单一及复合体系中对Cu~(2+)和Pb~(2+)的吸附量均呈150BS-12 100BS-12 50BS-12 CK的顺序,吸附均符合Freundlich和Langmuir模型; BS-12修饰更有利于Cu~(2+)的吸附,但Pb~(2+)在BS-12修饰土样上的吸附量及选择性系数均大于Cu~(2+); Cu-Pb复合体系中,两种金属离子存在相互拮抗的作用,BS-12修饰在减弱Pb~(2+)对Cu~(2+)的拮抗作用的同时增强了Cu~(2+)对Pb~(2+)的拮抗作用,但Pb~(2+)对Cu~(2+)的拮抗作用仍大于Cu~(2+)对Pb~(2+)的拮抗作用;温度升高、p H升高和离子强度降低均能增加对Cu~(2+)和Pb~(2+)的吸附,但其对两种金属离子的影响程度不同,也导致对金属离子间的交互作用产生影响.     

Nitrous oxide reductase gene (nosZ) and N2O reduction along the littoral gradient of a eutrophic freshwater lake

Lake littoral zones are characterized by heterogeneity in the biogeochemistry of nutrient elements. This study aimed to explore the relationship between the nitrous oxide reductase gene (nosZ)-encoding denitrifier community composition/abundance and N2O reduction. Five samples (deep sediment, near-transition sediment, transition site, near-transition land and land soil) were collected along a littoral gradient of eutrophic Baiyangdian Lake, North China. To investigate the relationship between the nosZ-encoding denitrifier community structure and N2O reduction, the nosZ-encoding denitrifier community composition/abundance, potential denitrification rate (DNR) and potential N2O production rate (pN2O) were investigated using molecular biological technologies and laboratory incubation experiments. The results showed that the average DNR of sediments was about 25 times higher than that of land soils, reaching 282.5 nmol N/(g dry weight (dw)·hr) and that the average pN2O of sediments was about 3.5 times higher than that of land soils, reaching 15.7 nmol N/(g dw·hr). In the land area, the nosZ gene abundance showed a negative correlation with the N2O/(N2O+N2) ratio, indicating that nosZ gene abundance dominated N2O reduction both in the surface soils of the land area and in the soil core of the transition site.Phylogenetic analysis showed that all the nosZ sequences recovered from sediment clustered closely with the isolates Azospirillum largimobile and Azospirillum irakense affiliated to Rhodospirillaceae in alpha-Proteobacteria, while about 92.3% (12/13) of the nosZ sequences recovered from land soil affiliated to Rhizobiaceae and Bradyrhizobiaceae in α-Proteobacteria. The community composition of nosZ gene-encoding denitrifiers appeared to be coupled with N2O reduction along the littoral gradient.