Divergence of Parartemia and Artemia haemoglobin genes

Parartemia is a genus of brine shrimp endemic to Australia which is related to the more widespread and economically importantArtemia sp. The expression of a multimeric haemoglobin molecule inArtemia is well documented but inParartemia only trace levels of a possible haemoglobin have been observed. In this paper we describe the DNA sequence of a domain of a haemoglobin molecule inParartemia. The derived amino acid sequence suggests that the possible date of divergence about 85 million years ago of the two genera predates the divergence of the C and T polymers ofArtemia haemoglobin. This date would correlate with the physical and temporal isolation of Australia in the late Mesozoic.     

无机离子（Ni，Ge）和烷化剂对DNA Tm值的影响

在实验室条件下，选择几种有代表性的无机离子（包括宏观生命元素、微量生命元素、副生命元素），并保持一定的浓度和ＰＨ值（６．１８±０．２），研究了各无机离子和烷化剂对ＤＮＡＴｍ值的影响。结果表明，有益元素与有害元素对ＤＮＡＴｍ值高低的影响明显不同。前者使Ｔｍ升高，抑制ＤＮＡ热变性，后者使Ｔｍ值降低，促进ＤＮＡ热变性；这并非阴阳离子的离子强度造成的，而是由于加入的无机离子对ＤＮＡ分子中磷酸基因和碱基的作用不同所致。而且它们对ＤＮＡＴｍ值的影响幅度和方向与烷化剂的存在有很大关系。烷化剂可以大大降低Ｔｍ值，加入各种无机离子对烷化作用产生不同的影响。这表明，作为致癌的主要机理—烷化作用可能与各种离子的存在和在生物体内的平衡状态有关。所测结果还表明，烷化剂的作用比无机离子大得多。这些结果有助于区分有害与有益元素，对癌症的早期发现及阐明致癌、防癌机理有重要意义。     

两种海蜇毒素的分子标记研究

采用聚丙烯酰胺凝胶电泳技术研究了8种同工酶在海蜇的刺胞和中胶层的表达特异性,利用RAPD技术对海蜇刺胞组织的DNA标记进行研究。结果表明,作为生物体防御清除自由基的SOD,在刺胞和中胶层均有表达。而与酯类化合物代谢相关的EST、维持细胞正常的能量代谢的ATPase,能在海蜇和口冠海蜇的刺胞中表达,而不能在中胶层中表达,所以,EST和ATPase可作为刺胞毒素的分子标记。碳水化合物代谢中重要的酶类MDH和ADH、清除细胞内H2O2的POD、催化磷酸单酯水解的重要酶类(与磷脂的转移、消化、吸收等活动密切相关的)ACP、在体外碱性环境下能水解有机磷脂键而产生一个有机基团和无机磷酸根的ALP,这5种酶仅在毒性较强的口冠海蜇刺胞中表达,MDH和ACP活性很高,在毒性相对较弱的海蜇刺胞中不表达。所以,这5种酶可作为海蜇毒性强弱的标记。以两种海蜇刺胞DNA为模板,S11、S32、S38、S95等4个随机引物的RAPD谱图差异明显,亦可作为毒素强弱的间接分子标记。     

两种提取枝角类休眠卵基因组DNA方法的比较

休眠卵是枝角类在环境胁迫条件下通过有性生殖形成的,休眠卵的孵化对种群补充与恢复有着重要作用.沉积于底泥中的休眠卵保存了不同阶段的种群遗传信息,有效地提取枝角类休眠卵基因组DNA是进一步研究水体枝角类遗传多样性的关键环节.采用目前国际上使用的氯仿异戊醇提取法和玻璃乳提取法分别对流溪河水库底泥中表层和底层的盔型溞(Daphnia galeata)休眠卵进行基因组DNA提取的比较分析.结果显示,氯仿异戊醇提取法成功率为27.5%,平均浓度为14.25±1.84 ng/μl;玻璃乳提取法成功率为65.0%,平均浓度为28.37±2.56 ng/μl.无论是提取成功率还是提取浓度玻璃乳提取法都显著高于氯仿异戊醇提取法,且玻璃乳提取法所用试剂少,不涉及有毒试剂,操作步骤简单,整个提取过程所用时间短,玻璃乳提取法提取枝角类休眠卵基因组DNA是一种快捷实用的方法.     

Demographic history,range size and habitat preferences of the groundwater amphipod Niphargus puteanus (C.L. Koch in Panzer, 1836)

Niphargus puteanus is the oldest described species of its genus and, in the past, was used as a taxonomic annotation for any subterranean amphipod record. For that reason, no clear knowledge exists about its actual range size and habitat preferences. We here applied a molecular taxonomic and phylogeographical approach based on mitochondrial and nuclear DNA to shed light on its distribution and to infer its demographic history. Furthermore, we analysed aquifer types and water flow regimes to provide a clearer picture of the species’ ecological requirements.Our results indicate that N. puteanus is widely distributed north of the Alps, having its core range in the geomorphological natural region of the ‘South German Scarplands’ (SGS). Additionally, isolated satellite populations exist in the Taunus and the Sauerland, and two single individuals were collected in Luxembourg and in Austria, respectively. The species’ maximal distribution range reaches 756 km between the two single-specimen records and 371 km within the SGS. A very high haplotype diversity was observed, revealing the presence of seven haplotype groups. All the haplogroups were present in the SGS and exhibited distinct spatial-genetic patterns. We thus inferred a high degree of population isolation, with the SGS being a potential long-term refugium for N. puteanus. Historical drift of specimens along major riverine networks (Rhine, Danube) may provide an evolutionary dispersal mechanism explaining the formation of satellite populations.From an ecological perspective, populations of N. puteanus were found to be closely associated with fast-flowing spring environments within fissured aquifers. This species should therefore be classified as both crenophile and stygophile, effectively exploiting resources of surface as well as subsurface spring waters.     

中国大陆近海菖鲉属鱼类新记录种——三色菖鲉(Sebastiscus tertius)的形态特征与DNA条形码研究

2018年4月在浙江省舟山近海海域的渔业资源调查中采集到19尾菖鲉属鱼类标本,经鉴定发现为三色菖鲉(Sebastiscus tertius),为中国大陆近海海域的新记录种。本研究对采集到的19尾三色菖鲉标本拍摄照片并对其进行形态特征以及DNA条形码研究。三色菖鲉具有以下主要形态特征:背鳍数ⅩⅡ-12,胸鳍数18—19,腹鳍数Ⅰ-5,臀鳍数Ⅲ-5;第一鳃弓总鳃耙数6—7+13—17;前鳃盖骨有5枚硬棘,鳃盖骨后沿有2枚硬棘;眶前骨下后角有1枚硬棘;胸鳍处有一明显黑斑。测定了19尾标本的12S rRNA和COI序列,结合GenBank中所有学名为Sebastiscus tertius的同源序列进行分析,发现12S rRNA序列完全一致,可作为该物种的微型DNA条形码,而COI序列中所有个体明显分为两个分支,两者遗传距离达到0.044,表明两分支可能为不同的有效种。COI同源序列分析结果显示, GenBank中学名为Sebastiscus tertius的鱼种与褐菖鲉(Sebastiscus marmoratus)聚为一支,表明GenBank中的Sebastiscus tertius存在错误鉴定的情况。     

Bacterial communities fluctuate in abundance and diversity under simulated oil-contaminated seawater conditions

Marine bacteria have recently been identified as a potent solution for petroleum hydrocarbon degradation in response to hazardous oceanic oil spills. In this study, a mesocosm experiment simulating a petroleum spill event was performed to investigate changes in the abundance, structure, and productivity of bacterial communities in response to oil pollution. Cultured heterotrophic bacteria and total bacteria showed a consistent trend involving an immediate decrease in abundance, followed by a slight increase, and a steady low-level thereafter. However, the changing trend of bacterial productivity based on bacterial biomass and bacterial volume showed the opposite trend. In addition, the density of oil-degrading bacteria increased initially, then subsequently declined. The change in the bacterial community structure at day 0 and day 28 were also analyzed by amplified ribosomal DNA restriction analysis (ARDRA), which indicated that the species diversity of the bacterial community changed greatly after oil pollution. Alphaproteobacteria (40.98%) replaced Epsilonproteobacteria (51.10%) as the most abundant class, and Gammaproteobacteria (38.80%) became the second most dominant class in the whole bacterial community. The bacterial communities in oil-contaminated seawater (32 genera) became much more complex than those found in the natural seawater sample (16 genera). The proportion of petroleum-degrading bacteria in the oil-contaminated seawater also increased. In this study, culture-dependent and culture-independent approaches were combined to elucidate changes in both bacterial productivity and community structure. These findings will contribute to a better understanding of the role that bacteria play in material cycling and degradation in response to oil pollution.     

DNA03电子水准仪GSI-8数据格式转换软件开发

对徕卡DNA03电子水准仪线路测量模式及GSI-8数据存储格式进行了研究,基于Visual C++6.0平台MFC对话框,编写GSI-8数据格式向Excel电子表格水准记录簿格式的转换软件,实现DNA系列电子水准仪内置的5种线路测量模式GSI-8数据自动向满足国家水准测量规范的Excel电子手簿记录格式转换,测站各项限差指标自动检核并定位,水准测量等级满足从一等到四等以及等外水准的要求,相同等级和同一类型的线路数据文件实现批量格式转换。     

许氏平鲉(Sebastes schlegelii)速生选育群体的生长性能与遗传特征分析

本研究目的为开展许氏平鲉(Sebastes schlegelii)遗传改良和优质新品系选育,选取荣成(R)和长岛(C)的野生许氏平鲉为基础群体,采用群体选育(群体内和群体间)和家系选育相结合的方法,获得R-F1、RC-F1两个群体选育群体和R-F2、RC-F2、R-F3、RC-F3四个家系选育群体。对4个选育家系的生长性能进行比较分析,并利用20对高多态性的微卫星标记对8个群体进行遗传多样性检测。结果表明:(1)群体内和群体间家系选育子3代比家系选育子2代生长速度分别提高21.59%和30.99%。4个选育家系后代的绝对增重率随着天数的增加而上升,群体内家系选育(R-F3)的生长速度、绝对增重率显著大于群体间的杂交选育系(RC-F3),杂交系未表现出生长的杂种优势。(2)20对微卫星位点在所有群体中均表现出较高多态性,8个群体的平均有效等位基因数(Νe)为2.9849—7.9598,平均观测杂合度(oH)和期望杂合度(He)分别为0.7230—0.8405和0.6315—0.8716,平均多态信息含量(PIC)为0.6472—0.8478。经两代家系选育的R-F3和RC-F3平均等位基因数显著降低,但观测杂合度(Ho)和期望杂合度(He)均值仍达到0.7230、0.7549和0.6527、0.6315,还有维持了较高的遗传多样性水平,遗传潜力较大。Hardy-Weinberg平衡检测结果和遗传偏离指数(d)表明各群体在大约10个位点上表现出一定程度的杂合子缺失现象。群体间平均遗传分化系数(Fst)为0.1279,各群体间存在中等遗传分化程度。R与R-F1分别与RC-F3遗传距离最远(0.9959、0.9848),预测利用两组群体中生长优良个体分别进行杂交育种有可能获得杂种优势。     

Abschätzung der Eignung einer molekularbiologischen Methode zur Charakterisierung der Grundwasserbiozönose

Assessing the Suitability of a Molecularbiological Method To Characterise the Microbial Populations in Groundwater A molecularbiological technique was used to characterise the bacterial community structure of groundwater habitats. This method consists of the isolation of bacterial DNA from the samples, amplification of 16S rDNA by PCR (polymerase chain reaction), and separation of the amplified DNA by DGGE (denaturing gradient gel electrophoresis). By using more specific primer combinations in the PCR instead of universal eubacterial primers, also groups of microorganisms (Proteobacteria, sulfate reducer, Archaea) were determined. The resulting DGGE patterns that reflect the microbial diversity are compared and differences or similarities evaluated. In the present studies, groundwater from different sites (bank filtrate, artificially recharged groundwater, and natural groundwater) and with changing redox milieus (aerobic, anaerobic) were investigated as well as the solid aquifer material. Besides, samples were taken from the different stages of artificial groundwater recharge, i.e., from surface water to the drain tile. Samples from groundwater derived from sites with different hydrogeochemical or hydrological conditions like bank filtrate and recharged groundwater revealed great differences in DGGE patterns indicating a characteristic species composition in these habitats, while samples taken at different times from the same groundwater showed only small seasonal variations. Clearly different patterns were also found for groundwater and the adjacent solid material as well as for anaerobic and aerobic groundwaters. Looking at artificial groundwater recharge, almost identical patterns were found in raw water and samples from gravel and sand filtration. DGGE patterns from the resulting groundwater indicated a total change in community structure during underground passage. By using group specific primers, Desulfovibrionaceae, Desulfobacteriaceae, and Archaea could be detected in anaerobic groundwaters.The molecularbiological approach described here gives an increasingly comprehensive and more precise picture of the microbial population of different environments. It is especially suitable to compare the community structure from different habitats or to analyse changes for example due to environmental stress at the same site.