填埋场中亚硝酸还原酶测定条件的优化

填埋垃圾中的含氮化合物经一系列生物脱氮作用,最终使得填埋场中的氮素得以消减,在这一过程中亚硝酸还原酶起着十分重要的作用。以填埋场中的填埋垃圾为研究对象,在土壤亚硝酸还原酶测定方法的基础上,对亚硝酸还原酶活性测定条件进行了优化。结果表明,填埋垃圾中的亚硝酸还原酶活性测定的最优条件为:垃圾样品风干温度25℃,2mL的1%NaNO2溶液和2.5 mL的1%葡萄糖溶液,抽气5 min,置于25℃的培养箱中培养24 h。优化后的测定条件相对标准偏差(RSD)小于1.27%,表明该方法具有较高的灵敏度和精密度。     

Xα21基因导入水稻及转基因植株的鉴定

以水稻愈伤组织为受体、质粒pCXK1301为Xα21基因供体，用农杆菌EHA105和PDS－1000／He基因枪转化受体，经Hyg浓度梯度筛选，获得一些转基因水稻植株，GUS检测、PCR扩增和Southern杂交分析表明，报告基因（GUS）、选择标记基因（Hyg^r）和抗白叶枯病基因Xα21都已整合入转化水稻基因组中，Southern杂分析显示了Xα21基因在转化水稻基因组中整合的拷贝数。田间接菌鉴定表现抗白叶枯病。     

Use of reporter-gene based bacteria to quantify phenanthrene biodegradation and toxicity in soil

A phenanthrene-degrading bacterium, Sphingomonas paucimobilis EPA505 was used to construct two fluorescence-based reporter strains. Strain D harboring gfp gene was constructed to generate green fluorescence when the strain started to biodegrade phenanthrene. Strain S possessing gef gene was designed to die once phenanthrene biodegradation was initiated and thus to lose green fluorescence when visualized by a live/dead cell staining. Confocal laser scanning microscopic observation followed by image analysis demonstrates that the fluorescence intensity generated by strain D increased and the intensity by strain S decreased linearly at the phenanthrene concentration of up to 200 mg/L. Such quantitative increase and decrease of fluorescence intensity in strain D (i.e., from 1 to 11.90 ± 0.72) and strain S (from 1 to 0.40 ± 0.07) were also evident in the presence of Ottawa sand spiked with the phenanthrene up to 1000 mg/kg. The potential use of the reporter strains in quantitatively determining biodegradable or toxic phenanthrene was discussed.     

Hydroxytyrosol, the phenolic compound of olive oil protects human PBMC against oxidative stress and DNA damage mediated by 2,3,7,8-TCDD

The protective effect of hydroxytyrosol (HT), a strong antioxidant compound from extra virgin olive oil, against TCDD induced toxicity was investigated in human peripheral blood mononuclear cells (PBMC). PBMC (1 × 10⁶ cells mL⁻¹) were divided into four groups and were incubated in a CO₂ incubator (5% CO₂) for 12 h with vehicle, TCDD (10 nM), TCDD + HT (10 nM + 100 μM) and HT alone (100 μM) respectively. To clarify the role of HT against TCDD induced cytotoxicity, oxidative stress and the levels of antioxidant enzymes were assessed. Incubation of PBMC with TCDD significantly decreased cell viability, catalase (CAT) and glutathione peroxidase (GPx) and increased the levels of superoxide dismutase (SOD), glutathione reductase (GR) and oxidative stress markers such as lipid peroxidation products (LPO), protein carbonyl content (PCC) and reactive oxygen species (ROS). Whereas, HT had an effective antioxidant property as observed by the increased cell viability, normalization of antioxidant enzymes and decreased levels of LPO, PCC and ROS in PBMC co-treated with HT and TCDD. Apoptosis detection and comet assay results shows that HT, by acting as an antioxidant, prevents the damage to DNA induced by TCDD. In addition light microscopic and histopathological observations revealed that the cells are apoptotic and degenerated during TCDD treatment, whereas cells showed intact morphology during co-treatment with HT. On the whole, the results reveal that HT exerts a promising antioxidant potential in protecting the PBMC against TCDD induced oxidative stress, which might be due to the presence of catechol moiety in its structure.     

用12SrRNA基因序列研究潮龟科（BATAGURIDAE）闭壳龟类的进化

测定了潮龟科（Ｂａｔａｇｕｒｉｄａｅ）具闭壳结构的齿缘摄龟属（Ｃｙｃｌｅｍｙｓ）、锯缘摄龟属（Ｐｙｘｉｄｅａ）、闭壳龟属（Ｃｕｏｒａ）共６种线粒体１２ＳｒＲＮＡ基因片段的序列，结合潮龟科其它一些属的序列数据，探讨了具闭壳结构龟类的起源与进化以及闭壳龟属的分类学问题．认为闭壳龟属与锯缘摄龟属为姐妹群，两者与齿缘摄龟属的亲缘关系较远，提示这些具闭壳结构的龟类可能不是由一个共同祖先分化而来．本文的结果不支持将闭壳龟属拆分为闭壳龟属和盒龟属两属     

家蚕斑纹限性品种SC_1的RAPD差异DNA片段的克隆及其酶切图谱分析

利用RAPD技术对家蚕限性普斑品种SC1的雌雄体分别进行PCR扩增通过120种引物的筛选，得到1个雌体特异的RAPDDNA片段将该片段克隆到pUC19质粒，井进行了限制性内切酶图谱分析.     

Monitoring aromatic hydrocarbon biodegradation by functional marker genes

The development of biological treatment technologies for contaminated environments requires tools for obtaining direct information about the biodegradation of specific contaminants. The potential of functional gene array analysis to monitor changes in the amount of functional marker genes as indicators of contaminant biodegradation was investigated. A prototype functional gene array was developed for targeting key functions in the biodegradation of naphthalene, toluene and xylenes. Internal standard probe based normalization was introduced to facilitate comparison across multiple samples. Coupled with one-colour hybridization, the signal normalization improved the consistency among replicate hybridizations resulting in better discrimination for the differences in the amount of target DNA. During the naphthalene biodegradation in a PAH-contaminated soil slurry microcosm, the normalized hybridization signals in naphthalene catabolic gene probes were in good agreement with the amount of naphthalene-degradation genes and the production of 14CO2. Gene arrays provide efficient means for monitoring of contaminant biodegradation in the environment.     

连续施用发酵猪粪对土壤中四环素抗性基因数量的影响

为了研究连续施用发酵猪粪之后,土壤中抗性基因的数量以及发酵猪粪的施用量对土壤抗性基因的影响,采用实时荧光定量PCR方法,对稻麦轮作模式下连续6年施用4.5t/hm2和9.0t/hm2发酵猪粪的土壤进行了四环素抗性基因(TRGs)的检测和定量分析.在施用发酵猪粪的土壤中能够检测到9种TRGs.施用发酵猪粪显著增加了土壤中tet G、tet L、tet B(P)、tet O、tet W的绝对数量,其中tet B(P)、tet W、tet O的绝对数量受到了施肥用量的影响;而tet Z、tet C和tet S的绝对数量则没有受到施肥的影响.上述8种TRGs在同一施肥处理的0~5cm、5~10cm以及10~20cm土壤中的相对丰度分布均无显著差异.发酵猪粪的施用也显著增加了tet G、tet L、tet B(P)和tet O的相对丰度,但是仅有tet O的相对丰度受到了施肥用量的显著影响.本研究表明,在稻麦轮作模式下,农田土壤中的TRGs数量会受到发酵猪粪中残留TRGs的影响,连续施用该粪肥会显著增加土壤中tet G、tet L、tet B(P)和tet O的绝对数量和相对丰度.因此,需要优化畜禽粪堆肥发酵工艺,以减少抗性基因在粪肥中的残留.     

微生物聚磷及其酶学调控

本文论述了微生物细胞内磷酸盐转运机制和与聚磷相关的酶—PPK、PPX及两者对微生物聚磷行为的调控,并对今后研究重点做出展望.论文认为对于微生物聚磷的研究应从纯种微生物转移到强化生物除磷系统的混合微生物菌群,运用分子生物学和生物信息学手段分析聚磷优势菌种的PPK和PPX酶的结构特征及其在活性污泥中的变化规律,进一步深入认识生物除磷调控机理,开辟寻求提高系统除磷效率的新途径.