生物强化膜生物反应器(MBR)处理邻苯二甲酸二乙酯(DEP)效果及微生物群落结构分析

为强化邻苯二甲酸二乙酯(DEP)降解,将从活性污泥中分离的高效DEP降解菌Arthrobacter sp.LMS13运用到MBR反应器,考察强化系统对DEP去除效果,并通过实时荧光定量(q-PCR)和Illumina Mi Seq技术分别对系统运行期间邻苯二甲酸双加氧酶降解基因(phtA)数量以及微生物群落结构特征进行了分析.结果表明,强化系统能加快反应器启动,对进水浓度为800 mg·L-1的DEP,强化系统和未经强化系统在运行后期(61 d)对DEP平均去除率分别为81%和19%.q-PCR结果显示,在驯化阶段,phtA基因拷贝数上升,但当DEP浓度大于400 mg·L-1时,phtA基因数量下降;phtA基因拷贝数与DEP降解率呈正相关.Mi Seq测序结果进一步表明,高浓度DEP导致系统中群落多样性指数下降,但对强化系统多样性影响相对较小.反应器运行过程中,原活性污泥中占有较高比例的拟杆菌门(Bateroidetes)和厚壁菌门(Firmicutes)丰度降低,ε-变形纲(ε-Proteobacteria)、绿弯菌门(Chloroflexi)在反应系统中逐渐被淘汰,而β-变形纲(β-Proteobacteria)和放线菌门(Actinobacteria)成为系统中的优势菌门,其中红环菌属(Rhodocyclus)、博得氏杆菌属(Bordetella)、节杆菌属(Arthrobacter)为优势菌属,在DEP降解系统中起着主要作用,是保证DEP生物处理效果和维持反应器稳定运行的重要菌属.     

Responses of activities, abundances and community structures of soil denitrifiers to short-term mercury stress

The responses of activities, abundances and community structures of soil denitrifiers to mercury (Hg) stress were investigated through a short-term incubation experiment. Four soil treatments with different concentrations of Hg (CK, Hg25, Hg50, and Hg100, denoted as 0, 25, 50, and 100 mg Hg/kg dry soil, respectively) were incubated for 28 days. Soil denitrification enzyme activity (DEA) was measured at day 3, 7 and 28. The abundances and community structures of two denitrification concerning genes, nirS (cd₁-nitrite reductase gene) and nosZ (nitrous oxide reductase gene), were analyzed using real-time PCR and denaturing gradient gel electrophoresis (DGGE). Results showed that soil DEA was significantly stimulated in the treatments of Hg25 and Hg50 compared with others at day 7. Meanwhile, no difference in the abundances of soil nirS and nosZ was found between Hg spiked treatments and CK, except the lower abundance of nirS (P<0.05) in the Hg added treatments compared with that in the CK at day 28. The community structures of denitrifiers based on nirS gene presented obvious change at day 7 along with the Hg additions, however, no variation was found in all treatments based on the nosZ gene. The results indicated that Hg (Hg25 and Hg50) had a strongly short-term stimulation on soil DEA, and nirS gene is more sensitive than nosZ gene to Hg stress.     

Quantification of microcystin-producing and non-microcystin producing Microcystis populations during the 2009 and 2010 blooms in Lake Taihu using quantitative real-time PCR

Lake Taihu, a large, shallow hypertrophic freshwater lake in eastern China, has experienced lake-wide toxic cyanobacterial blooms annually during summer season in the past decades. Spatial changes in the abundance of hepatotoxin microcystin-producing and non-microcystin producing Microcystis populations were investigated in the lake in August of 2009 and 2010. To monitor the densities of the total Microcystis population and the potential microcystin-producing subpopulation, we used a quantitative real-time PCR assay targeting the phycocyanin intergenic spacer (PC-IGS) and the microcystin synthetase gene (mcyD), respectively. On the basis of quantification by real-time PCR analysis, the abundance of potential toxic Microcystis genotypes and the ratio of the mcyD subpopulation to the total Microcystis varied significantly, from 4.08× 10⁴ to 5.22× 10⁷ copies/mL, from 5.7% to 65.8%, respectively. Correlation analysis showed a strong positive relationship between chlorophyll-a, toxic Microcystis and total Microcystis; the abundance of toxic Microcystis correlated positively with total phosphorus and ortho-phosphate concentrations, but negatively with TN:TP ratio and nitrate concentrations. Meanwhile the proportion of potential toxic genotypes within Microcystis population showed positive correlation with total phosphorus and ortho-phosphate concentrations. Our data suggest that increased phosphorus loading may be a significant factor promoting the occurrence of toxic Microcystis bloom in Lake Taihu.     

Carbamoyl phosphate synthetase I deficiency: molecular genetic findings and prenatal diagnosis

We report a Japanese boy who died at Day 28 of life because of severe carbamoyl phosphate synthetase I (CPS1) deficiency that was proven by enzyme assay. By analysis of cDNA and genomic DNA, he was shown to be a compound heterozygote with two point mutations of the CPS1 gene, 840G>C leading to an aberrant splicing and 1123C>T (predicting Q375X). The 840G>C was a mutation described in another Japanese family. Since his parents carried each mutation heterozygously, we performed prenatal diagnosis at 16 weeks of his mother's next gestation by multiplex PCR and melting curve analysis in a single capillary containing two-color fluorescent (LC-Red 640 and LC-Red 705) probes on LightCycler. We analyzed genomic DNA extracted from amniotic cells and found that the fetus was homozygous for the wild-type alleles. At term a healthy girl was born without hyperammonemia. Copyright © 2001 John Wiley & Sons, Ltd.     

生物炭对非常规水源灌溉下土壤-作物病原菌的影响

采用根箱实验,基于定量PCR技术研究非常规水源（再生水和养殖废水）灌溉及生物炭添加后根际土、非根际土及玉米根部病原菌的赋存情况.结果发现,灌溉时间、土壤类型可显著影响土壤中病原菌的整体分布.相较于蒸馏水灌溉下根际土、非根际土的单个病原菌,再生水灌溉下病原菌变化为-0.74~0.17个数量级,养殖废水灌溉下为-0.14~0.60个数量级.生物炭对土壤中病原菌的影响因灌溉水源、灌溉时间而改变,可使再生水及养殖废水灌溉下病原菌检出量最高降低0.35、0.39个数量级.但再生水及养殖废水灌溉可使玉米根部病原菌检出量最高增加1.17、2.20个数量级,生物炭又使非常规水源灌溉下其检出量最高增加0.60、1.08个数量级,带来潜在的健康风险.     

温带亚高山针叶林土壤碳循环微生物群落的时空动态

选取参与碳固定的二磷酸羧化/加氧酶基因（cbbM）、有机碳降解的淀粉酶基因（amylase）和纤维素酶基因（cellulase）作为分子标记,用实时定量PCR方法对温带亚高山华北落叶松（Larix gmelinii var.principis-rupprechtii）林、白杄（Picea meyeri）林、青杄（P.wilsonii）林和油松（Pinus tabulaeformis）林土壤碳循环功能微生物类群丰度的时空动态开展研究.结果显示,总碳（TC）、总氮（TN）、总硫（TS）、有机质（OM）和有机碳（TOC）、pH值、铵态氮（NH₄⁺-N）、硝态氮（NO₃^--N）、过氧化氢酶、蔗糖酶和脲酶活性在4种森林土壤中都有不同程度的差异,且有显著的季节变化特征.高海拔华北落叶松林土壤TC、TN、TS、C/N、OM和TOC含量最高,而pH值最低.土壤TC、TN、亚硝态氮（NO₂^--N）含量、蔗糖酶和脲酶活性,与碳循环微生物类群的丰度呈极显著相关.土壤NO₃^--N含量与有机碳分解和固碳微生物类群的相对丰度显著相关;土壤C/N、NO₂^--N、pH值、OM、TOC、过氧化氢酶及脲酶活性,与降解易分解碳（labile C）和难分解碳（recalcitrant C）的微生物类群的相对丰度呈极显著相关.植被类型和季节变化共同影响土壤碳循环微生物类群的丰度,而季节变化是主导因素.植被和土壤环境因子通过调控微生物群落碳代谢功能类群的结构,影响森林土壤碳源-汇的平衡.     

Normal pregnancy after preimplantation DNA diagnosis of a dystrophin gene deletion

To perform preimplantation DNA diagnosis for Duchenne muscular dystrophy (DMD) in a female carrier of a dystrophin gene deletion of exons 3–18, we developed a polymerase chain reaction (PCR)-based assay of exon 17 sequences. Exon 17 was efficiently amplified in all 50 single blastomeres of normal control embryos and in five blastomeres of one male embryo of the DMD carrier obtained after a first preimplantation diagnosis (PID) for gender determination. In ten blastomeres of another two male embryos of the DMD carrier, no PCR signals were observed, probably as a result of the deletion. After intracytoplasmic sperm injection, embryos were analysed for exon 17 and three of the four embryos showing normal PCR signals were replaced, resulting in a singleton pregnancy. Prenatal diagnosis showed a female karyotype and DNA analysis indicated that the fetus was not a DMD carrier.     

亚硝酸细菌研究进展

从硝化细菌的生长特性 ,分类 ,检测 ,亚硝酸细菌氨单加氧酶等方面概要的叙述了国外在硝化细菌方面研究的进展 ,并展望了以后的研究和应用