双壳类软体动物贝壳DNA高效提取及方法学评价

通过对虾夷扇贝(Patinopecten yessoensis)、栉孔扇贝(Chlamys farreri)、牡蛎(Crassostrea gigas)和侏儒蛤(Mulimia lateralis)4种双壳贝类进行壳DNA提取实验,本研究提出一种适用于双壳类软体动物贝壳DNA提取方法。首先用EDTA(15%)脱钙液对贝壳进行预处理去除表面的杂质,经PBS缓冲液(4×)清洗后放入37℃烘箱30 min,称取重量为1 g的壳放入液氮中冷却后使用中通量组织研磨仪研磨成粉末,分装到2 mL离心管中,加入裂解液后在56℃、200 r/min条件下裂解3 h后,常温下12 000 r/min离心10 min得到上清液,加入无水乙醇和醋酸钠沉淀DNA。经微量紫外分光光度计检测、琼脂糖凝胶电泳分析和全基因组重测序比对分析来评价提取到的基因组DNA质量,结果显示该方法可获得完整性好的高质量DNA。本研究提出的贝壳DNA提取方法操作简单、快速,且成本低,为双壳贝类的遗传分析提供了一种方法,本方法在贝类遗传学和分子育种研究中具有广阔的应用前景。     

6种海洋双壳类贝壳中碳酸钙、碳酸镁含量的测定

为了了解海洋双壳类的固碳能力,以紫脲酸铵-萘酚绿B、铬黑T为指示剂,采用络合滴定法对泥蚶(Tegillarca granosa)、四角蛤蜊(Mactra veneriformis)、文蛤(Meretrix meretrix)、缢蛏(Sinonovacula constricta)、虾夷扇贝(Patinopecten yessoensis)和紫贻贝(Mytilus galloprovincialis)等6种海产双壳贝类贝壳中的碳酸钙含量、碳酸钙和碳酸镁总量进行了测定。6种贝类贝壳中的碳酸钙含量占干重比例介于93.99%~97.35%之间,其中虾夷扇贝的含量最多,紫贻贝的含量最少;碳酸镁含量介于0.61%~1.79%之间,其中紫贻贝的含量最多,虾夷扇贝的含量最少。目前的结果表明,六种贝类均具有较好的固碳能力,基于碳酸钙和碳酸镁总量估算,6种贝类单位湿重固碳力介于0.023 2~0.074之间。     

甲硫氨酸脑啡肽对栉孔扇贝血淋巴中2种自由基的影响

用分光光度技术对甲硫氨酸脑啡肽(met-Enk)对栉孔扇贝(Chlamysfarreri)血淋巴中自由基及其相关酶的影响进行了研究。结果表明,除了脑啡肽浓度为1μg/mL时血清和血细胞中过氧化氢酶(CAT)活力高于对照外,脑啡肽浓度分别为10和50μg/mL,作用时间分别为1,3和5h时,血清和血细胞中一氧化氮(NO)、一氧化氮合酶(NOS)、过氧化氢(H2O2)和CAT的含量或活力均低于对照;随着脑啡肽浓度的升高,其含量或活力均呈下降趋势。说明脑啡肽对栉孔扇贝血淋巴中的自由基NO和H2O2及其相关酶的含量或活力起副调节(抑制)作用。相同的脑啡肽浓度下,随着作用时间的延长,其含量或活力又有所回升。说明栉孔扇贝血淋巴中可能存在脑啡肽的拮抗物质。脑啡肽可能在栉孔扇贝的免疫调节中发挥重要的作用。     

青蛤(Cyclina sinensis)组织蛋白酶L基因的表达与重组蛋白活性分析

利用荧光定量PCR方法检测了青蛤(Cyclina sinensis)在鳗弧菌(Vibrio anguillarum)胁迫下组织蛋白酶L基因(CsCPL)在各组织的表达差别及其在血淋巴中的时序表达关系。结果表明,CsCPL基因在青蛤血淋巴、肝脏、外套膜、鳃和闭壳肌等组织中普遍表达,其中以血淋巴表达水平最高。在鳗弧菌刺激后3—96h青蛤的血淋巴中CsCPL的表达量均出现明显上调,其中6h达到最大值并且与对照组有极其显著性差异(P0.01),说明该基因在青蛤免疫反应中具有重要作用。文章还对CsCPL基因进行了原核表达,其产物蛋白分子量约35kDa,经纯化及复性后具有明显的抑菌作用。     

双壳贝类CK1基因家族的鉴定及表达分析

酪蛋白激酶1 (CK1)是一种重要的蛋白激酶家族,其在DNA损伤应答和修复、细胞增殖和凋亡、胚胎发育和稳态等重要的生物学过程中都具有复杂多样的调控作用。为了理解CK1基因家族在双壳贝类中的特征、进化及生物学功能,实验采用比较基因组学及生物信息学的方法对双壳贝类CK1基因家族进行了鉴定分析,通过虾夷扇贝高温应激实验,研究了CK1基因家族在高温应激时的表达规律。结果显示,在虾夷扇贝、栉孔扇贝、长牡蛎与侏儒蛤中均存在CK1α,CK1αlike,CK1δ,CK1γ3,并发现CK1ε,CK1γ1,CK1γ2在双壳贝类中发生了丢失。时空表达分析发现,双壳贝类CK1基因均在胚胎发育早期集中表达,并以多细胞/囊胚期为界呈现出母源/合子特异性表达模式。CK1为关键基因的基因共表达模块显著富集在错配修复、核苷酸剪切修复等相关通路上,暗示了CK1基因在双壳贝类胚胎发育过程中参与调控DNA损伤修复过程,从而维持早期胚胎发育时的基因组稳定性。双壳贝类CK1基因在成体中呈现组织特异的表达模式,主要在鳃和雄性性腺中具有相对较高的表达量。虾夷扇贝受到高温应激后,其鳃中PyCK1α, PyCK1αlike和PyCK1δ...     

青蛤(Cyclina sinensis) TRAF6基因克隆及其在Poly I:C胁迫下的免疫应答

对青蛤(Cyclina sinensis)转录组文库筛选并克隆得到青蛤TRAF6(Cs TRAF6)基因的c DNA序列,其开放阅读框为2337bp,编码778个氨基酸,分子量为86.59k Da。Cs TRAF6蛋白包含RING型锌指结构,两个TRAF型锌指结构,coiled-coil结构域以及MATH结构域。经生物信息学及分子系统学分析表明Cs TRAF6属于TRAF6家族的一员。利用实时荧光定量PCR方法检测了Cs TRAF6基因在青蛤各个组织以及在Poly I:C侵染下在血淋巴中的时序性表达情况。结果表明,Cs TRAF6基因在青蛤血淋巴、闭壳肌、鳃、性腺、外套膜和肝脏中普遍表达,其中在血淋巴中的表达水平最高。在Poly I:C刺激后的3h,青蛤血淋巴中Cs TRAF6的表达量迅速升高,在6h达到最大值,说明青蛤的Cs TRAF6基因参与了病毒类似物侵染下的免疫应答反应。     

青蛤(Cyclina sinensis)TLR2基因的克隆与表达分析

利用转录组文库分析得到青蛤(Cyclina sinensis)的Toll样受体2(Toll-like receptor 2,TLR2)基因序列,采用荧光定量PCR法分析该基因的表达过程。结果显示,青蛤TLR2基因的cDNA开放阅读框为2082bp,编码693个氨基酸,具典型的LRRs、单次跨膜区和TIR结构域。该基因在血淋巴中的表达量最高,与肝脏、鳃、外套膜、闭壳肌和性腺有显著性差异(P0.05)。在鳗弧菌和Poly I:C刺激下,青蛤血淋巴中TLR2基因3h开始升高,48h达到最大值,与对照组和空白组有极显著性差异(P0.01);藤黄微球菌刺激下,血淋巴中CsTLR2基因3h开始升高,48h亦达到最大值,实验组与对照组有显著性差异(P0.05)。     

栉孔扇贝血淋巴中谷胱甘肽还原酶的活性研究

用酶标仪采用DTNB比色法测定栉孔扇贝(Chlamys farreri)血淋巴中谷胱甘肽还原酶(GR)的活性,并研究pH值、温度、激活剂和抑制剂对血清中该酶活性的影响及酶的热稳定性。结果表明,栉孔扇贝血细胞中GR比活力明显高于血清。血清中GR的最适pH值因缓冲液的不同而稍有差异:用PBS为缓冲液时,最适pH值为7.0;Tris-HCl为缓冲液时,最适pH值为7.5。酶的最适温度为60℃。将血清在20℃,30℃,40℃,50℃,60℃,70℃和80℃温度的水浴中放置30min后于20℃,pH7.5条件下测定该酶的热稳定性,结果显示50℃时酶的比活力最高,低于或高于此温度的酶比活力均下降,80℃时酶活性完全丧失。栉孔扇贝血淋巴中GR属中温型中性酶类,低温下较稳定。EDTA对该酶活性有促进作用,硫酸铜有抑制作用。     

低溶解氧对凡纳滨对虾酚氧化酶原激活系统和相关免疫因子的影响

本文研究了凡纳滨对虾在低溶解氧条件下酚氧化酶原系统的激活机制。结果表明:低溶氧(1.5mg/L,3.0mg/L)胁迫对凡纳滨对虾血淋巴中多巴胺含量、血细胞数量、酚氧化酶原激活系统相关酶活力、血淋巴中类α2-巨球蛋白活力影响显著(P<0.05),且各指标表现出明显的时间效应性。在实验时间内,DO为3.0mg/L处理组,总血细胞、小颗粒细胞和透明细胞数量均在12h内降低而后趋于稳定,大颗粒细胞数量在24h内降低后保持稳定,DO为1.5mg/L处理组,总血细胞数量和三类血细胞数量均在24h内降低而后趋于稳定;DO为3.0mg/L和1.5mg/L处理组凡纳滨对虾血淋巴中多巴胺含量分别在24h、12h内呈峰值变化,至12h、6h时达到最大值,在24h、12h后恢复至对照组水平并趋于稳定;血细胞中酚氧化酶原活力分别在12h、6h时显著降低,且均在24h时趋于稳定;丝氨酸蛋白酶活力在6h时均显著降低,在24h时达到最低值,而后趋于稳定,蛋白酶抑制剂活力显著降低,但类α2-巨球蛋白活力48h内呈峰值变化,与对照组比较均显著升高,48h后趋于稳定。实验期内对照组DO为5.5mg/L各指标变化不显著。     

双壳贝类积累转化麻痹性贝毒的研究进展

麻痹性贝毒(Paralytic shellfish toxins, PST)是一类分布广、危害大的海洋毒素。滤食性双壳贝类在摄食、消化产毒单胞藻和细菌等过程中积累代谢PST,并通过食物链进行传递,给人类生命健康和水产业带来不利影响。随着贝类基础生物学和养殖产业的发展,以及基因组学和毒素检测技术的不断进步,近年来,各国学者对贝类吸收、转运和代谢PST的规律有了更深入的认识,为养殖贝类食品安全风险防控提供了理论参考。为更全面了解贝类积累和利用PST的研究进展,本文从PST在双壳贝类中的分布、积累转化特征与分子机制等方面进行了综述。     

Model of the post-Cenozoic evolution of the cryolithozone of the shelf of the western part of the Laptev Sea

The analysis of the structure of the cryolithozone, facies, and thicknesses of the Quaternary sediments and the results of the physicochemical mathematical modeling of the modern shelf of the western part of the Laptev Sea support the influence of the Late Pleistocene glaciations on the heat conditions and the distribution of the permafrost in the area. A ～200-m thick glacier formed under aerial conditions from atmospheric precipitation represented the metamorphosed snow cover. According to the modeling, the long-living (from 60?50 to 10?4 ky) glacier reduced the thickness of the permafrost rocks in the reviewed shelf area for 280–360 m. The Holocene marine transgression additionally decreased the thickness from 50–140 m on the inner shelf to 220–350 m on the outer shelf. The modern submarine cryolithozone 450-0 m thick is wide-spread in the studied region from the coast to the shelf boundary (isobaths of 130–140 m), where it pinches out at a distance of ～380 km from the coast at a depth of ～250 m above the sea level.     

Investigation of the mechanism of the long-term evolution of the carbon cycle in the ecosystem of the Sevastopol bay

We analyze the components of the carbon system of the Sevastopol bay waters and the balance of main sediment-forming substances using the data of field investigations in 1998–2008. The interannual variations of total inorganic carbon and the equilibrium partial pressure of carbon dioxide in the bay water are noted. An increase in the flux of carbon dioxide into the bay and in the content of organic carbon in bottom sediments is revealed, and an explanation of this phenomenon is given. The priority accumulation of organic carbon in the sediments of the bay is established. We assess the interannual variation in the relative abundances of organic and inorganic carbon as an index of the carbon cycle stability.     

Contemporary state of the investigation of the influence of the discharge of rivers on the hydrologic structure of the Black Sea

We generalize and analyze the state of investigations of the influence of river waters on the hydrologic structure of the Black Sea. Specific features of the water, regime and hydrography of rivers flowing into the sea, data on the discharge of rivers, and intrayear and interyear variability of the discharge are presented. We discuss and analyze works studying the transformation of river waters, routes of desalinized sea waters, and their influence on the thermohaline structure and dynamic regime of the sea. Translated by Peter V. Malyshev and Dmitry V. Malyshev     

Modelling of the ecosystem of the Black-Sea shelf zone near the estuary of the Danube

We analyze the results of numerical calculations performed according to the three-dimensional interdisciplinary model of an ecological system of the Black-Sea shelf zone near the estuary of the Danube. The complete system of equations of hydrothermodynamics is solved together with transport equations of the advection-diffusion-reaction type used to describe the transformation of a substance (nitrogen) between the components of the characteristic vectors of the ecosystem: plankton, detritus, and biogenic elements (nitrates). We describe the distinctive features of the circumcontinental distribution of components obtained as a result of numerical experiments and present arguments for the conclusion that the ecosystem of the Danube estuary water area plays the role of a buffer zone between the press of the Danubian biogenic pollutions and the neighbouring areas of the shelf zone and open sea. Translated by Peter V. Malyshev and Dmitry V. Malyshev     

黄河入海径流对河口海域盐度影响研究

为研究黄河入海径流变化条件下河口附近海域盐度扩散特征,以更好地保护河口海域生物资源多样性,本文以黄河下游利津水文站的长序列实测径流数据资料为基础,利用近海水动力模型FVCOM,分析径流变化对黄河口海域盐度的影响规律。结果表明：黄河口与莱州湾之间存在顺时针的环流系统,在余流作用以及涨落潮方向的影响下,黄河冲淡水长期向莱州湾扩散;丰水期黄河冲淡水几乎影响了整个莱州湾,27盐度锋可以到达莱州湾中部,27等盐线的表层包络面积为2 665.61 km²,占莱州湾的1/4左右,枯水期低盐度水只有向南扩散的趋势,27以下的低盐度水集中分布在黄河口门附近,27等盐线的表层包络面积只有199.65 km²;5月份,随着入海径流量增加,27等盐线扩散的范围、距离、方向都会发生明显变化。在对近海生物资源有迫切保护需求的情景下,适当减少其他用水户供水量以增加入海生态径流量,可以有效改善黄河口海域附近的盐度情况,为生物资源的生长繁殖创造良好条件。     

Dynamics of the ice cover and peculiarities of the ice transportation of the sediments at the tidal flats of the Kandalaksha Gulf of the White Sea

Ten years of semistationary studies of the interaction of the ice cover and the coastal relief of the Kandalakhsa Gulf in the White Sea allowed us to reveal the peculiarities of these processes in the warm and cold years and the mechanisms of the origination of ice forms, including ice tents, which govern the character of the tidal flats. Three dynamic fast ice zones are distinguished by their geomorphological location, morphology, and ice dynamics. The dependence of the mechanisms and volumes of the enrichment of the fast ice in the sediments on the coastal zone??s relief was established. At the coast of the Kindo Peninsula (Velikaya Salma Strait), the ice mostly contains fine-sandy sediments. Coarser sediments, including boulders, are occasional and frozen into the ice at the contact of the fast ice and the bottom. At the expense of the ice??s spreading by 4?C6 times in the warm years and 2?C4 times in the cold years, several thousand tons of sediments mostly from the tidal flat??s surface (100?C700 m³ from 1 km of coast in the wide tidal flats and <50 m³ on the steep coasts) are delivered to the water area.     

The influence of the bottom contour on the deformed state of the ice cover due to the motion of the submarine

The authors have previously determined that the effectiveness and failure pattern of the ice cover caused by flexural-gravity waves generated by a submerged body motion near the bottom ice can greatly depend on the depth of the water area. In its turn, the presence of a ledge on the ice surface may affect a wave propagation pattern. This paper presents an experimental study of the bottom contour influence on the deflection and length of flexural-gravity waves. The authors describe a numerical model for the analysis of the deformed state of ice caused by hydrodynamic loads due to a submarine motion, taking into account the bottom contour. The experiments are carried out in the ice tank. The results of calculations and experiments are compared.     

国产XBT海上比测方法研究及结果分析

随着国产投弃式温深仪(XBT)性能的提高,其使用范围和规模也逐步扩大,受某专项办的委托,厦门大学在南海东北部开展了"海洋监测设备适用性检验规范及海上试验研究(国家863计划)"项目,针对海洋技术领域研发的技术较为成熟、具备产品化条件的XBT开展检验与验证,对国产XBT的性能和实际使用情况进行分析和总结。试验中,在不同站点进行温盐深仪(CTD)现场测量,以此数据为真值对国产和进口XBT精度做评估。在较高速度走航测量时,则以进口XBT数据为参照,对国产XBT进行评估。试验结果表明,国产XBT整体上表现良好,成功率大概为80%,有些型号的国产XBT精度与进口XBT精度接近,但国产XBT无效剖面较多。同时也发现,进口XBT的精度无法满足预期比测的要求,不能准确计算出国产XBT精度,但结果仍然具有参考价值。本次试验可以为国产XBT的规模化应用和推广提供数据支持,也可为其他型号XBT/XCTD/CTD等仪器的比测提供借鉴。     

Modelling of the ecosystem on the shelf of the Republic of Guinea

Three mathematical models are considered: a two-dimensional model for calculating the concentration fields of the ecosystem components on the shelf of the Republic of Guinea, a pointwise model for the annual variation of the components, and a two predators-prey model. These models are used to study the effect of hydrodynamic conditions on the state of the shelf ecosystem and for research into the causes which distort the balance between marketable and unmarketable fish.Translated by Mikhail M. Trufanov.     

Evolution of the slope of the buried depression of the Black Sea

The first data on the temporal evolution of the slope of the buried depression of the Black Sea are obtained. The sharp change in the rate of the decrease in the slope steepness that occurred in the Late Miocene-Pliocene time reflects a reorientation of the vector of mutual movement and a related intensification of the collision between the Arabian and Eurasian lithospheric plates.