红鳍笛鲷微卫星DNA标记的开发与遗传多样性分析

以三亚外海的红鳍笛鲷(Lutjanus erythropterus)自然群体为研究对象,利用磁珠富集法开发19个适用于红鳍笛鲷的微卫星DNA标记,其中16个为多态性标记,等位基因数为2～11,期望杂合度为0.242～0.896,观测杂合度为0.156～1.000,Ler 14、Ler 29、Ler 49等3个位点偏离哈迪-温伯格平衡(P<0.0031),Ler9-Ler18、Ler12-Ler19、Ler46-Ler47、Ler46-Ler49、Ler18-Ler50、Ler14-Ler51等6对位点间检测到明显的连锁(P<0.05),Ler29、Ler13、Ler49、Ler19、Ler24、Ler25、Ler14、Ler51、Ler18、Ler9等10个位点属于高度多态位点[多态信息含量(PIC)>0.5],Ler47、Ler12、Ler15、Ler46等4个为中度多态位点(0.25>PIC>0.5)。     

浅析基于Lecai DNA003水准仪在等级水准测量中的应用

介绍Lecai DNA003型电子水准仪在西煤东运煤炭基地三等水准测量项目中的实际应用情况,结合实际作业经验对仪器的不足之处讨论了解决办法,以提高工作效益。     

Sample preparation methods for quantitative detection of DNA by molecular assays and marine biosensors

The need for quantitative molecular methods is growing in environmental, food, and medical fields but is hindered by low and variable DNA extraction and by co-extraction of PCR inhibitors. DNA extracts from Enterococcus faecium, seawater, and seawater spiked with E. faecium and Vibrio parahaemolyticus were tested by qPCR for target recovery and inhibition. Conventional and novel methods were tested, including Synchronous Coefficient of Drag Alteration (SCODA) and lysis and purification systems used on an automated genetic sensor (the Environmental Sample Processor, ESP). Variable qPCR target recovery and inhibition were measured, significantly affecting target quantification. An aggressive lysis method that utilized chemical, enzymatic, and mechanical disruption enhanced target recovery compared to commercial kit protocols. SCODA purification did not show marked improvement over commercial spin columns. Overall, data suggested a general need to improve sample preparation and to accurately assess and account for DNA recovery and inhibition in qPCR applications.     

Genetic Analysis of Invertebrates From Great Salt Lake,Utah

正Great Salt Lake(GSL),in northern Utah,is one of the largest lakes in the United States,with a total surface area of 4400 square kilometers.Arthropods constitute the most conspicuous and abundant animals inhabiting the waters     

海洋线虫Litoditis marina早期发育的转录组分析

全球气候和环境变化影响海洋动物的生长发育和繁殖等生命过程,探究环境因子调控发育的分子机制,需要深入了解海洋动物早期发育的生理和分子特征。以实验室驯化的潜在模式动物海洋线虫Litoditis marina为研究对象,对其胚胎期和孵化后发育早期2h、4h和6h的L1幼虫样品进行了转录组测序和分析。结果表明,2h、4h和6h的L1幼虫间差异较小,而三个L1幼虫样品与胚胎期相比,基因表达发生了显著变化。通过KEGG富集分析,发现与胚胎期相比,三个L1幼虫样品的多个通路如核糖体、核糖体生物发生、糖酵解/糖原异生、TCA循环和氧化磷酸化通路相关基因发生了显著上调。另外还发现多个神经递质和神经肽受体基因如dop-和npr-等在L1期转录水平显著上调。与胚胎期相比,L1幼虫的多个DNA复制和修复相关、Notch、Hippo和Hedgehog信号和剪切体等通路相关基因发生了显著下调。发现的L. marina早期发育转录组变化模式与已经发表的陆生模式生物秀丽线虫Caenorhabditis elegans从胚胎期到L1幼虫的转录组变化特征非常相似,但同一上调或下调通路中具体发生表达变化的基因有些不同。另外,L1幼虫显著上调的核糖体生物发生通路相关基因在秀丽线虫中发生了显著下调。进一步通过基因编辑等技术和方法深入研究发育调控关键基因的功能将为海陆近缘线虫间的发育进化机制、海洋线虫对潮间带环境适应以及全球气候变化应答的分子机制研究提供新认知。     

青海湖沉积柱Dinoflagellate藻类基因丰度变化及其古气候环境的指示意义

近年来,有学者通过古DNA方法成功地重建了海洋沉积记录中古微生物群落变化,并借此反演了当地古环境-气候变化。然而,此种方法对于陆地湖泊沉积记录是否适用仍然有待研究。本文采用聚合酶链式反应（PCR, Polymerase Chain Reaction）、变性梯度凝胶电泳（DGGE,Denaturing Gradient Gel Electrophoresis）与实时荧光定量聚合酶链式反应（Q-PCR,Quantitative PCR）相结合的综合分析技术手段,系统研究青海湖5.8米（时间跨度为～18500年）沉积柱dinoflagellate真核藻类多样性和丰度变化。研究结果显示青海湖Dinoflagellate藻18S rRNA基因序列主要与海洋型藻类Woloszynskia halophila和Scrippsiella hangoei相近（～98%序列相似性）。定量Q-PCR结果显示,每克沉积物含有dinoflagellate藻类18S rRNA基因丰度范围为2.27×103～8.55×106拷贝。另外,Dinoflagellate藻类18S rRNA基因丰度与总有机碳含量成显著正相关（r = 0.408,p = 0.0001）。对比分析揭示较高的藻类丰度对应高总有机碳含量和较低的可溶解性盐电导率;反之,较低的藻类丰度对应较高的可溶解性盐电导率和较低的总有机碳含量。在青海湖区,总有机碳指示着季风降雨变化并间接地指示着外源输入和湖泊营养状况变化,然而可溶性盐电导率则指示着湖泊盐度变化。综上所诉,青海湖沉积柱Dinoflagellate藻类丰度可能响应着历史时期湖泊营养状况和盐度波动情况。     

Time-course evaluation of ROS-mediated toxicity in mussels, Mytilus galloprovincialis, during a field translocation experiment

Harbours can be considered as model environments for developing and validating field monitoring procedures and to investigate mechanistic relationships between different biological responses. In this study several biomarkers were investigated in marine mussels caged for 4 weeks (June–July 2001) into an industrialized harbour of NW Italy. Organisms were collected at different time-intervals to better characterize the sensitivity, temporal variations and interactions of analysed responses. Besides single antioxidants the total oxyradical scavenging capacity (TOSC) assay was used to analyse the capability of the whole antioxidant system to neutralize specific forms of radicals: these data were further integrated by measurement of DNA integrity, oxidized bases and the impairment of lysosomal membrane stability in haemocytes. Results showed a biphasic trend for single antioxidants and TOSC, with an increase during the first 2 weeks of exposure to the polluted site followed by a progressive decrease up to a severe depletion in the final part of the experiment.     

Molecular evidence for Myripristis jacobus and Scarus taeniopterus new to Bay of Bengal: Sporadic appearance or preliminary colonization?

Individual specimens of Myripristis jacobus Cuvier, 1829 and Scarus taeniopterus Lesson, 1829 were collected from Gopalpur‐on‐sea, Odisha coast, Bay of Bengal. Both the fish samples were assigned species level tags using the conventional taxonomic methods. The capture of both the fish species is of particular interest in that they were new to the Bay of Bengal. Molecular analysis using the mitochondrial cytochrome oxidase subunit I (COI) gene showed high confidence sequence similarity in species identification. The resultant phylogenetic relationships strongly support the monophyly of both the genus with congruent clustering of both the species according to their morphological identification. In addition, the time tree produced is in complete agreement with the recent studies. These studies conclusively prove that both the species might have been originated during the middle to the late Miocene period. Furthermore, both fish species are typical inhabitants of stigmatized coral reefs, confined to the reef regions. These fish species perhaps migrated to the Bengal Bay from the Andaman and Nicobar Islands reef regions because of climatic changes.     

马氏珠母贝(Pinctada martensii)四种壳色选育系F5 的生长及遗传多样性分析

采用跟踪测量和微卫星(SSR)技术研究了马氏珠母贝四种壳色选育系F5和对照组的存活率、生长情况及遗传多样性。结果表明,四种壳色选育系和对照组的生长性状之间均存在显著差异(P<0.05)。从30对微卫星引物中扩增筛选获得8个多态位点,多态位点比例为26.67%,它们在4个壳色选育系共120个个体中产生了42个等位基因,平均每个多态位点产生5.25个。4个选育系的平均期望杂合度范围为0.6622—0.6850,平均观察杂合度范围为0.2708—0.4667,平均多态信息含量PIC值范围为0.6025—0.6190,说明4个选育系的遗传多样性处于较高水平,具有育种潜力;平均遗传偏离指数均为负值,4个选育系均存在不同程度的杂合子缺失。遗传分化和遗传距离分析表明白壳色选育系与红壳色选育系之间的亲缘关系最近,黑壳色与白壳色之间的遗传距离最大。     

Applications of three DNA barcodes in assorting intertidal red macroalgal flora in Qingdao, China

This study is part of the endeavor to construct a comprehensive DNA barcoding database for common seaweeds in China. Identifications of red seaweeds, which have simple morphology and anatomy, are sometimes difficult solely depending on morphological characteristics. In recent years, DNA barcode technique has become a more and more effective tool to help solve some of the taxonomic difficulties. Some DNA markers such as COI (cytochrome oxidase subunitⅠ) are proposed as standardized DNA barcodes for all seaweed species. In this study, COI, UPA (universal plastid amplicon, domain V of 23S rRNA), and ITS (nuclear internal transcribed spacer) were employed to analyze common species of intertidal red seaweeds in Qingdao (119.3 -121 E, 35.35 -37.09 N). The applicability of using one or a few combined barcodes to identify red seaweed species was tested. The results indicated that COI is a sensitive marker at species level. However, not all the tested species gave PCR amplification products due to lack of the universal primers. The second barcode UPA had effective universal primers but needed to be tested for the effectiveness of resolving closely related species. More than one ITS sequence types were found in some species in this investigation, which might lead to confusion in further analysis. Therefore ITS sequence is not recommended as a universal barcode for seaweeds identification.