Mitochondrial DNA variation pattern and postglacial history of the Siberian larch (<Emphasis Type="Italic">Larix sibirica</Emphasis> Ledeb.)

Mitochondrial DNA (mtDNA) variation has been studied in populations of the Siberian larch with the use of five PCR-based markers characterizing 29 haplotypes. The studied populations from the main parts of the Siberian larch range belong to several geographically isolated groups with their specific sets of haplotypes, which have probably originated from separate refugia for forest vegetation of the last glacial maximum. Conclusions concerning the possible location of these refugia and the ways and pattern of the postglacial recolonization of the recent Siberian larch range are drawn. The spatial distribution of haplotypes within individual populations is nonrandom. The typical size of the area occupied by a group of plants with the same haplotype is 50–100 m across.     

久效磷农药对金鱼肝细胞DNA的损伤及其机制研究

以金鱼(Carassius auratus)为模式生物,研究了久效磷农药暴露导致的DNA损伤类型及其作用机制.结果表明:0.01,0.10,1.00mg/L的久效磷农药暴露24,48,96,168h均导致金鱼肝细胞DNA损伤程度显著升高,48h损伤最严重;暴露48h采用碱性、pH值12.1和中性彗星电泳发现DNA损伤类型主要为碱不稳定位点形成,其次为单/双链断裂;采用Endo Ⅲ酶和FPG酶处理的碱性彗星电泳,发现DNA出现氧化损伤;暴露24h肝脏谷胱甘肽过氧化物酶(GSH-Px)活性显著降低,丙二醛(MDA)含量显著升高达到峰值,96~168h超氧化物歧化酶(SOD)和GSH-Px活性显著升高,MDA含量与24h相比有所降低,表明活性氧自由基(ROS)含量在短期暴露升高(24h)、在96~168h暴露逐渐降低.影响抗氧化酶活性、干扰ROS清除过程是久效磷农药造成DNA损伤的主要机制.     

大亚湾浮游植物DNA指纹及其与群落结构的关系

于2012年5月、6月采集了大亚湾9个站点的表层水样,利用PCR-变性梯度凝胶电泳(PCR-DGGE)技术对浮游植物DNA指纹进行了分析,同时通过显微镜观察对浮游植物进行了定性定量分析.研究结果显示,大亚湾浮游植物种类丰富,共分析鉴定出浮游植物72种;浮游植物DNA指纹也较为丰富,2个月份指纹条带数分别为26条和28条.DGGE指纹条带数一般远远低于浮游植物种类数,在剔除了相对含量小于0.1%的非优势种后,DNA条带数与种类数变化趋势相近,说明DNA指纹图谱能较大程度地反映浮游植物优势种群的组成,而对于相对含量较少的物种,可能会由于优势种的屏蔽作用而被掩盖.DNA指纹图谱与浮游植物群落结构的聚类分析结果相近,富营养化的近岸站点聚在一起,而远岸站点聚为一类.虽然目前大亚湾浮游植物群落结构中以硅藻占据优势,但在某些站点大量出现的甲藻和蓝细菌值得进一步关注.本研究结果表明PCR-DGGE技术可在一定程度上运用于浮游植物群落结构分析.     

DNA甲基化结合蛋白

DNA甲基化是哺乳动物细胞中最重要的表观遗传学修饰之一,大约70%—80%的CpG发生这种甲基化修饰.异常的甲基化在许多癌症中频发,启动子CpG岛的高甲基化作为普遍的失活机制介导抑癌基因沉默.甲基化信号由甲基化结合蛋白来转译,它们能够特异性识别并结合至甲基化位点通过募集辅阻遏复合物例如组蛋白去乙酰化酶(Histone Deacetylase,HDAC)等建立沉默的染色质,从而在DNA甲基化和基因沉默中起桥梁作用.目前,哺乳动物中已鉴定出的甲基化结合蛋白有三类,分别是:MBD(Methyl-CpG-Binding Domain)、Kaiso以及SRA(Set and Ring finger-associated)家族.本文就这三大家族(以MBD为主)各自的结构、功能、结合甲基化DNA的特性以及它们在某些疾病发生中的作用做一综述.     

金矿区中国林蛙体内DNA甲基化水平

采集金矿附近山区河流水质、沉积物和当地两栖类动物中国林蛙样品,应用高效液相色谱仪测定污染区和对照区林蛙体内各组织器官的胞嘧啶(c)和5-甲基胞嘧啶(5-mc)含量,探讨因金矿开采引起的汞污染以及对林蛙体内组织器官DNA甲基化水平的变化,研究汞胁迫下,两栖类动物体内分子水平的影响。结果表明:金矿开采区河流水质和沉积物已受到甲基汞污染,污染区林蛙体内甲基汞含量远高于对照区;林蛙体内各组织器官中DNA甲基化水平发生不同的变化:污染区林蛙肝脏和皮肤中DNA甲基化水平高于对照区,肌肉和脑干DNA甲基化水平低于对照区;雄性林蛙肝脏和脑干DNA甲基化水平高于雌性,肌肉和皮肤DNA甲基化水平却低于雌性。以上结果说明汞胁迫下,中国林蛙体内组织器官DNA甲基化水平可以发生一定的变化,环境中重金属汞离子进入林蛙体内含量的不同,可以促进或抑制其体内甲基化水平的变化,引起基因毒性作用。     

基于流式细胞特性的LNA和HNA细菌相关性研究

采用流式细胞技术分析了7种不同陆地环境介质中LNA和HNA细菌的浓度变化及流式细胞特性特征.结果显示,在水体和土壤等环境中都存在LNA和HNA细菌的分类,其中,土壤环境中LNA细菌浓度(10⁷~10⁸cells/g数量级)高于水环境中浓度(10⁵~10⁶cells/mL数量级),但其相对比重(29.80%~33.94%)低于水环境(除地下水外,21.60%)中LNA细菌(42.25%~65.92%),主成分分析(PCA)显示水体和土壤环境介质中LNA和HNA流式细胞参数具有明显差异;相关分析表明,LNA与HNA细菌的流式细胞参数(绿色荧光信号FL1和侧向散色信号SSC)之间具有显著相关性(FL1:R²=0.711, P<0.01, SSC:R²=0.762, P<0.01),在不同生态系统中SSC变异大于FL1变异.结果表明,LNA和HNA细菌之间既不是各自对应的某一特殊生理阶段细菌,也不是两个完全相互独立的细菌群体,而是具有特殊共变关系的细菌.     

Assessment of the effect of environmental pollution in a marina on caged mussels using chemical and genotoxic analysis

An integrated approach using the contamination levels and DNA damage in mussels (Mytilus galloprovincialis) was applied in order to assess the chemical contamination in a marina (Eastern coastline of Aegean Sea). Mussels, which were harvested from a reference site (Foca), were transplanted into a marina situated along the coast of Izmir Bay. The transplanted mussels were collected at the 14th, 30th and 60th day of the experimental period. Polycyclic aromatic hydrocarbon (PAH) levels (27–51?ng?g^?1?wet?weight) detected in the mussels were similar to the levels detected in other coastal areas of the Mediterranean Sea. The marina’s sediment was found to be contaminated with PAHs (∑PAH?=?25?µg?g^?1) of pyrolytic origin and may become a source of pollution and a threat to the marine environment. In order to assess the DNA damage, the haemolymph and gill cells of the mussels were used for the comet analysis and considered as an indicator of exposure to genotoxic chemicals including 16 PAH compounds and metals. The highest levels of DNA damage expressed as %Tail-DNA (%T-DNA) were observed at the end of the experiment (21.5% T-DNA). The correlation analyses conducted between 2-, 3-, 4-ring PAHs in mussels and %T-DNA in haemolymph and gill cells showed a significant positive correlation. This investigation confirmed that transplanted mussel can be a useful tool to determine PAH contamination in marinas.     

基于彗星实验的洛克沙胂对秀丽隐杆线虫胚胎细胞DNA损伤的研究

为评价洛克沙胂的遗传毒性,采用单细胞凝胶电泳(彗星实验),研究了不同浓度的洛克沙胂对秀丽隐杆线虫胚胎细胞脱氧核醣核酸(DNA)的损伤作用。提取秀丽隐杆线虫的胚胎细胞,分别暴露于0(空白对照)、50、250、500μg·L~(-1)含洛克沙胂的溶液染毒1 h。用彗尾DNA百分比含量(TDNA%)、彗星尾长(TL)和Olive尾矩(OTM)作为DNA损伤的指标。实验结果表明,与空白对照组比较,处理组中彗尾DNA百分比含量、彗星尾长以及Olive尾矩显著增加(P0.01)。随着洛克沙胂浓度的增加,彗尾DNA百分比含量、彗星尾长以及Olive尾矩逐渐增加,其相关系数r0.99,说明在实验浓度范围内,存在极显著的浓度-效应关系。洛克沙胂对秀丽隐杆线虫胚胎细胞DNA具有损伤作用,彗星实验操作简便、快速、灵敏度高,能够反映出洛克沙胂的遗传毒性。因此,通过彗星实验建立实验室检测洛克沙胂遗传毒性的方法具有可行性。