南极适冷菌Psychrobacter sp.G 普遍胁迫蛋白(USP)基因的克隆及其胁迫条件下的应答特征分析

根据南极适冷菌Psychrobacter sp.G基因组测序草图发现并克隆到了1个普遍胁迫蛋白(Universal StressProtein,USP)基因Usp1141。该基因ORF长462bp,编码153个氨基酸,蛋白理论分子量为17kDa,pI为5.23。通过多序列比对发现该USP含有交替排列的4个α螺旋和5个β折叠,以及与ATP结合相关的氨基酸残基,表明该蛋白可能通过与ATP结合而发挥作用。利用实时定量PCR技术对该蛋白在不同胁迫条件下的表达情况进行分析发现,温度变化对Usp1141基因的表达没有显著影响;低盐度(0,15)胁迫会显著抑制其表达,而高盐度(90,120)胁迫则会显著促进其表达;在温度与盐度协同胁迫条件下,当培养基盐度低于菌株生长的最适盐度时,无论温度高低,Usp1141基因的表达均受到抑制,而当培养基盐度高于最适盐度时,无论温度高低,Usp1141基因的表达均会提高。实时定量PCR研究表明,Usp1141基因可能在南极适冷菌Psychrobactersp.G对环境渗透压变化的适应性中发挥作用。     

Combined Effects of Temperature and Salinity on the Zoeal Development of the Crab Liocarcinus puber (Decapoda: Portunidae)

Abstract. Larval development of the crab Liocarcinus puber was studied from hatching to the megalopa stage under 12 different temperature and salinity combinations, 10°C, 15°C, and 20°C (T): 20, 25, 30, and 35% (S), with a view to establishing optimum conditions for zoeal development. High larval mortalities were recorded at 10°C in all the experimental salinities, indicating the sensitivity of the larvae to low temperatures. Highest zoeal survival was obtained in the higher salinity and temperature combinations. First zoeae completed their development in all T-S combinations tested, while successive stages showed increased sensitivity and suffered high mortalities in lower salinities at 15° and 20°C and in all salinities at 10°C. The duration of zoeal development decreased with increasing temperature, whilst the lower salinities delayed development. Q₁₀ values did not differ much at higher salinities, with values ranging from 1.5 to 2.6.     

五种抗真菌药物对远海梭子蟹离体卵孵化的影响

研究了不同浓度的孔雀石绿、亚甲基蓝、制霉菌素、达克宁、里劳素等抗真菌药,对远海梭子蟹离体卵孵化及幼体培育的影响.结果发现,孔雀石绿和里劳素的抗真菌效果较好,它们的最适浓度分别为0.2×10-6,0.1×10-6.     

壳聚糖酶基因在解脂耶氏酵母中的表达及重组酶性质研究

为构建高效表达壳聚糖酶工程菌株,以Microbacteriumsp.基因组为模板扩增壳聚糖酶基因,目的基因与表达载体pINA1317连接构建重组质粒,重组质粒NotⅠ酶切线性化后转化解脂耶氏酵母Y.liPolytica Po1h。所得结果为PCR扩增得到约1000bp的特异性片段,序列分析表明含有壳聚糖酶全长基因,开放阅读框801bp,编码266个氨基酸残基。转化子酶活力为10.4U/mL,是原菌株酶活力的3.15倍。重组蛋白经DEAE-Sepharose FF分离纯化,达到电泳纯,SDS-PAGE显示单一条带,分子量约41kDa。重组酶反应最适温度为45℃,最适pH为5.6,Km和Vmax分别是0.926mg/mL和6.15U/mL。质谱分析酶解产物主要为三糖和五糖。实现了壳聚糖酶在解脂耶氏酵母中的分泌表达,为壳聚糖酶的工业化生产奠定了理论基础。     

单环刺螠中肠和后肠交替氧化酶对硫化物的应激反应

外源性硫化物可通过抑制或阻断线粒体电子传递经典途径而对生物体产生损伤甚至致死,交替氧化酶(Alternative Oxidase,AOX)是线粒体硫化物氧化电子传递分支途径中的1个关键酶。为了研究硫化物环境中单环刺螠的生存对策,本文利用间接竞争性ELISA和酶活性分析等技术检测了单环刺螠在硫化物应激前后中肠和后肠中AOX的蛋白含量以及活性的变化。结果显示:在未处理的单环刺螠中,后肠的AOX蛋白含量和酶活性均高于中肠。当单环刺螠暴露在硫化物(50和150μmol·L-1)环境中时,2个器官的AOX含量和酶活性水平均随着硫化物浓度的提高和暴露时间的延长而提高,并且150μmol·L-1硫化物处理组的单环刺螠2个器官AOX蛋白含量和酶活性普遍高于相同处理时间下的50μmol·L-1组。结合已报道的单环刺螠硫化物应激下细胞色素c氧化酶活性的变化,提出单环刺螠体内存在线粒体电子传递分支途径;提高组织线粒体中AOX活性是其应对环境硫化物毒性的对策之一。     

Biomarkers of Exposure and Effects for Assessing Toxicological Risk of Endocrine Disrupters in Top Predators of the Mediterranean Sea

Abstract. Endocrine Disrupting Chemicals (EDCs) have the potential to alter hormone pathways that regulate reproductive processes in wildlife and fisheries. In this research the unexplored hypothesis that Mediterranean top predator species (such as large pelagic fish and marine mammals) are potentially at risk due to EDCs is investigated. In the Mediterranean environment, top predators accumulate high concentrations of polyhalogenated aromatic hydrocarbons (PHAHs) and toxic metals, incurring high toxicological risk. Here we illustrate the need to develop sensitive biomarkers for evaluation of toxicological risk in top marine predators (Xiphias gladius, Thunnus thynnus thynnus) and non-lethal techniques, such as non-destructive biomarkers, for the hazard assessment of threatened species exposed to EDCs, such as marine mammals ( Stenella coeruleoalba, Tursiops truncatus, Delphinus delphis and Balaenoptera physalus ).     

急性高温胁迫对虹鳟二倍体和三倍体幼鱼hsps基因表达的影响

本文以二倍体和三倍体虹鳟(Oncorhynchus mykiss)幼鱼为实验对象,探究急性高温胁迫下,不同倍性虹鳟热休克蛋白家族(hsps)基因表达水平。选取二倍体和三倍体虹鳟幼鱼,分别进行常温处理(14℃)和高温胁迫(22℃),并在高温胁迫48h后,将虹鳟置于常温(14℃)恢复48h。分别在实验开始的0,12,24,48,72和96h进行取样,测定实验虹鳟肝脏和肌肉组织中的hsp70s(hsp70a,hsp70b),hsp90s (hsp90a1a,hsp90a2a,hsp90a1b,hsp90a2a,hsp90b1,hsp90b2)mRNA水平。结果表明:hsp70s和hsp90s的各个亚型均能对热胁迫做出反应,并具有应答幅度的组织特异性和时间特异性。在胁迫6和24h后,上述基因mRNA水平达到最高,随后降低。三倍体幼鱼肝脏和肌肉hsps mRNA表达水平的峰值显著高于二倍体幼鱼。研究结果显示,三倍体虹鳟对高温环境更为敏感,需要消耗更多的能量用于适应高温环境。     

中华豆蟹对青岛海区贻贝的感染及危害初探

20 0 0年 10月～ 2 0 0 1年 12月 ,对青岛太平角养殖海区的贻贝寄生中华豆蟹的感染情况进行了调查。以肥满指数为指标 ,通过对感染和未感染贻贝的比较 ,分析了中华豆蟹对贻贝的危害程度。结果表明 ,中华豆蟹对贻贝的感染率为 2 .0 %～ 2 0 .4 %。2 0 0 0年 11月和 2 0 0 1年 11月出现 2次感染高峰 ,分别为 9.7%和 2 0 .4 % ;最低感染率出现于 2 0 0 1年 1月和 3月 (均为 2 .0 % )。感染强度为 1～ 5 ,多数为 1(占感染个体的 83.2 % )。贻贝所感染的豆蟹的性别及感染强度与宿主的大小有关。小个体贻贝仅见雄豆蟹感染 ,绝大多数雌豆蟹见于壳长大于 2 cm的贻贝。未发现同一宿主体内同时感染 2只或以上的雌豆蟹。在调查的 15个月中 ,有 11个月被感染贻贝的肥满度显著低于未感染的贻贝 ,且雌豆蟹对宿主肥满度的影响大于雄豆蟹     

褐藻寡糖对刺参体腔液和体壁免疫相关酶活性变化的影响

研究刺参基础饲料中添加0.1%的褐藻寡糖对刺参体腔液和体壁中过氧化物酶(Peroxidase, POD)、酸性磷酸酶(Phosphatase, ACP)、碱性磷酸酶(Alkaline phosphatase, AKP)和溶菌酶(Lysozyme, LSZ)活性的影响.实验周期为40 d,每隔10 d取样1次,检测刺参体腔液和体壁中免疫相关酶活性的变化.结果显示:褐藻寡糖组刺参体腔液中POD、AKP和ACP活性均呈现先增高后降低的趋势,POD在第10 d时极显著提高(P<0.01),比对照组提高306%,AKP实验期间均与对照差异显著(P<0.05),活性最高时比对照组提高298%,ACP和LSZ分别在第20和40天与对照组差异显著(P<0.05),分别比对照组提高66%和26%;褐藻寡糖组刺参体壁中POD、AKP、ACP和LSZ活性与体腔液相比提高幅度较小,POD、AKP和ACP活性分别在第20,20和30天与对照组差异显著(P<0.05),分别比对照组提高了24%、20%和35%,LSZ活性略有提高但无显著性差异(P>0.05).研究结果表明,褐藻寡糖均能提高刺参体腔液和体壁中POD、ACP、AKP和LSZ活性,显著增强了刺参非特异性免疫水平,将其应用为刺参免疫增强剂具有广阔的前景.     

不同类型附着基对刺参生长和存活的影响

附着基是刺参(Apostichopus japonicus, Selenka)池塘或围堰养殖水体中设置的专用养殖设施,其功能主要是为刺参提供避光场所和培养刺参的天然饵料.本实验比较研究了石头、瓦片、塑料管、空心砖和水泥管5种不同附着基处理对刺参(规格为5.0 g±2.0 g)生长和存活的影响.结果表明,不同附着基上异养菌含量差异显著,5月附着基上异养菌含量显著的低于11月;石头附着基刺参的成活率最高,但其生长速度较慢;空心砖刺参的生长速度最快,成活率略低于石头;空心砖和石头附着基处理刺参产量显著高于其他附着基,瓦片、塑料管和水泥管上海参产量之间差异不显著.仅从养殖效果上看,目前较为流行的5种刺参附着基以空心砖较好.     

盐度骤变对仿刺参hsp70及hsp90基因表达的影响

为研究夏季暴雨导致的养殖池塘盐度变化对仿刺参(Apostichopus japonicus)生理的影响,在实验室内模拟野外盐度变化,采用半定量RT-PCR的方法研究了其hsp70、hsp90a及hsp90b基因的表达。实验中,盐度先以每6h降2.5的速度由30降至20;保持96h后,再以同样的速度由20升至30,随后在盐度为30条件下保持96h。在盐度变化过程中随机取样。hsp70和hsp90b基因在盐度降至20后表达量开始显著升高(P<0.05),hsp90a基因在盐度降至22.5时表达量开始显著升高(P<0.05);但保持在盐度为20条件下,3个基因的表达量均逐渐降低至初始值。在之后的盐度升高及恢复阶段,3个基因表达量均有先升高后降低的趋势,表明hsp70、hsp90a和hsp90b基因是仿刺参在盐度胁迫下的重要响应因子。     

梯度淹水胁迫下全日潮海区秋茄幼苗的生长和生理反应

以秋茄(Kandelia candel L.Druce)幼苗为材料,在全日潮海区广西英罗湾开展为期5个月的野外梯度淹水胁迫实验。3座平台作为重复,每座设置8个梯度,相邻高程组间的高度相差10cm。实验5个月后测定幼苗的生长状况和一些生理指标。结果表明:秋茄幼苗的生长高度、节数、叶数和大根数这4个外部形态特征指标的最大值均出现在360cm高程组。生长高度、节数、叶数的曲线较为一致,幼苗大根数的略有不同,但总体上可看出太低的高程均不利于这4个指标的增长。叶面积对淹水胁迫的反应与叶数一致,但高程组之间的差异更大。叶绿素含量则表现长时间淹水胁迫促进含量增高。在任一高程组,秋茄幼苗各新生器官生物量分配均表现为:茎>根>叶。各新生器官及全株生物量的最大值出现在360cm高程组,它们的干重曲线表现比较一致。幼苗根系中的活性氧清除酶类活性均高于叶片。根系中SOD酶和POD酶的分布规律较为一致,都以360cm高程组的活性最高。叶片中SOD酶和POD酶的变化则相反,叶片SOD酶随高程增大而降低,POD酶则随高程增大而增大。根系中活性氧清除酶类活性的水平与幼苗全株生物量的关系均呈显著相关,叶片中的则无此相关性。360cm及以上高程组幼苗全部存活,较低高程组有部分死亡。综合本文实验结果,可初步判断北部湾沿海秋茄造林的滩涂高程不宜低于当地平均海平面。     

四种单胞藻和海洋红酵母对刺参浮游幼虫生长与变态成活的影响

本实验研究4种单胞藻与海洋红酵母(Rhodomonas sp.)搭配以及单胞藻混合投喂对刺参浮游幼虫体长生长及变态存活率的影响。采用三角褐指藻(Phaeodactylum tricornutum)、小新月菱形藻(Nitzschia closteriumf.minutissima)、牟氏角毛藻(Chaetoceros muelleri)和等鞭金藻(Isochrysis galbana)与海洋红酵母分别以1∶0、0.8∶0.2、0.6∶0.4、0.4∶0.6、0.2∶0.8和0∶1的比例搭配投喂刺参浮游幼虫,观测浮游幼虫每天的体长变化以及变态成活率。结果表明,单胞藻与海洋红酵母以适当比例混合投喂能提高刺参幼虫的体长日增长率,单独投喂海洋红酵母的刺参变态成活率也显著高于单独投喂单胞藻组。采用2种单胞藻以0.5∶0.5的比例混合投喂浮游幼虫,结果表明牟氏角毛藻与小新月菱形藻混合投喂组刺参浮游幼虫的变态成活率高于牟氏角毛藻与等鞭金藻的混合组。     

Hatching of an Estuarine Crab, Sesarma Haematochier: Factors Affecting the Timing of Hatching in Detached Embryos, and Enhancement of Hatching Synchrony by the Female

The zoeas of the estuarine terrestrial crab Sesarma haematocheir hatch in synchrony with nocturnal high tides. Hatching is also highly synchronized among embryos: i.e., female-attached embryos would all hatch within 1 h, if the female is maintained at about 23°C. This study examined the factors affecting the timing of hatching in female-attached and detached embryos. The timing of hatching in the embryos detached from the female was somewhat affected by continuous light, salinity, and cluster size, but it was much more strongly affected by low temperature. In addition, a notable feature was that the influence of low temperature was markedly different between female-attached embryos and detached embryos. Hatching of this crab occurs after a special developmental process for hatching ("hatching program") that requires 48–49.5 h. This program would be mediated by a temperature-dependent process. Hatching of this crab cannot be fully explained in terms of osmotic effects. Since hatching of female-attached embryos was highly synchronized, the female should release an (unknown) stimulus enhancing hatching synchrony among embryos. Highly synchronized hatching among embryos would enable the female to release hatched zoeas into the water all at one time.     

Gene discovery in the tropical scallop Nodipecten nodosus: Construction and sequencing of a normalized cDNA library

We report the construction and characterization of a normalized cDNA library from the digestive gland of the marine bivalve Nodipecten nodosus, a commercially valuable tropical scallop. A total of 288 clones were sequenced, and 250 unique sequences were obtained. The cDNA library showed a small sequence redundancy (2.3%) and high numbers of recombinant (99.9%) and independent clones (2.0 × 10⁶ cfu), indicating that the cDNA library generated in this study is a profitable resource for efficient gene discovery for N. nodosus. EST functional annotation by Gene Ontology term assignment revealed the identification of sequences potentially involved in aquaculture and ecotoxicology relevant processes such as apoptosis, growth, lipid metabolism, reproduction, development, response to stress and immunity.     

Instability of the Kuroshio in Luzon Strait: Effects of ridge topography and stratification

The spectral analyses of moored current velocities in the central Luzon Strait reveal northward (i.e., downstream of the Kuroshio) propagation of a frontal wave with a five-day period, with wave amplitude increasing northward. Estimated from both curve fitting and frequency domain Empirical Orthogonal Function methods, the characteristics of five-day variations have wave speeds ranging from 32 to 40 cm s⁻¹, wavelengths ranging from 130 to 150 km, and e-folding time scales for growth ranging from 0.8 to 3 days. An analytical two-layer model used to explore linear stability characteristics indicates that bottom topography (two meridional ridges) is important for the Kuroshio stability characteristics in the Luzon Strait. In the two-layer model with the two ridges, the flow is stabilized for the long-wave mode but destabilized for the short-wave mode (due to increasing vertical shear in the horizontal velocity). The analytical model produces wavelengths and phase speeds for the most unstable mode which is similar to the observation, but the growth rate is underestimated. However, a spectral numerical model applied with a more realistic stratification and velocity structure does obtain faster growth rates comparable to the observations. Parameter sensitivity tests were conducted using the analytical model. The characteristics of the most unstable mode are most sensitive to the surface front location relative to the bottom topography but not sensitive to varying the density difference and thickness of the upper layer.     

Earthquake of February 3, 2016 in the Middle Baikal Region: Source Parameters and Macroseismic Effects

Izvestiya, Atmospheric and Oceanic Physics - We study in detail the source parameters, seismotectonic position, and macroseismic effects of the February 3, 2016, earthquake that occurred in the...     

Characterization of bacterial symbioses in Myrtea sp. (Bivalvia: Lucinidae) and Thyasira sp. (Bivalvia: Thyasiridae) from a cold seep in the Eastern Mediterranean

Cold seeps have recently been discovered in the Nile deep‐sea fan (Eastern Mediterranean), and data regarding associated fauna are still scarce. In this study, two bivalve species associated with carbonate crusts and reduced sediment are identified based on sequence analysis of their 18S and 28S rRNA‐encoding genes, and associated bacterial symbioses are investigated using 16S rRNA gene sequencing and microscopy‐based approaches. The specimens are closely related to Myrtea spinifera and Thyasira flexuosa, two species previously documented at various depths from other regions but not yet reported from the Eastern Mediterranean. Both species harbour abundant gammaproteobacterial endosymbionts in specialized gill epithelial cells. The Myrtea‐associated bacterium is closely related to lucinid symbionts from both deep‐sea and coastal species, whereas the Thyasira‐associated bacterium is closely related to the symbiont of a T. flexuosa from coastal waters off the U.K. An epsilonproteobacterial sequence has also been identified in Thyasira which could correspond to a helicoid‐shaped morphotype observed by electron microscopy, but this was not confirmed using fluorescent in situ hybridization. Virus‐like particles were observed within some symbionts in Thyasira, mostly in bacteriocytes localized close to the ciliated zone of the gill filament. Overall, results indicate that very close relatives of shallow species M. spinifera and T. flexuosa occur at cold seeps in the Eastern Mediterranean and harbour chemoautotrophic symbioses similar to those found in their coastal relatives.