黄河下游营养盐浓度季节变化及其入海通量研究

2001年3月～2002年2月期间在黄河下游采集溶解及颗粒态营养盐样品,分析了黄河径流中各形态的营养盐的浓度及其月动态,估算此时段内黄河的营养盐入海通量。研究发现,溶解无机氮是溶解总氮的主要存在形式,硝酸盐是黄河中氮的最主要存在形态,其季节变化与水量变化趋势相反,年平均含量为(260.6±84.0)μmol/L,显著高于世界其它河流,为世界背景值(7.14μmol/L)的20多倍;黄河中磷主要以颗粒态存在,颗粒态磷含量变化规律与SPM的分布一致,其年平均含量为(16.2±22.9)μmol/L,磷酸盐含量较低,年平均含量为(0.42±0.20)μmol/L,与世界河流的平均水平相当。硅酸盐年平均含量为(122.0±18.2)μmol/L。每年约有17 200 t的总氮和1 600 t的总磷输入渤海,氮通量表现出在春季3月较高;磷通量在9月出现最高值,春季3,4月也较高;硅酸盐通量在3月出现最高值。     

东海原甲藻线粒体细胞色素b(Cytb)基因的定量检测

建立了检测东海原甲藻线粒体细胞色素b(Cytb)基因mRNA含量的实时荧光定量PCR方法.以甲藻Cytb基因的简并引物扩增得到984 bp长的东海原甲藻Cytb基因片段,经克隆、测序,制备并纯化质粒.以光度法测定质粒浓度并作为标准品.对上述基因片段,利用PRIMER EXPRESS 3.0 设计引物,以梯度稀释的质粒标准品进行定量PCR反应,以SYBR Green I为荧光染料,制作标准曲线,得到基因拷贝数X与Ct值的关系为:Ct=-3.50 lg X+39.35(相关系数r为0.999).通过对不同生长时期的东海原甲藻样品的Cytb基因mRNA含量检测,发现该基因的表达量较稳定,平均值为(45.4±4.7)拷贝/细胞,受生长状态影响不大,可作为实时荧光定量PCR的内参基因.     

中国东部边缘海冬季硅酸盐的分布特征及主要来源

利用2007年1～2月的调查资料,分析讨论了中国东部陆架边缘海(南黄海、东海)冬季硅酸盐的分布特征及其主要影响因素。结果表明:近岸海域硅酸盐的高值区位于受长江冲淡水影响的区域;东海的硅酸盐浓度高于南黄海。长江冲淡水和黑潮水是影响东海和南黄海硅酸盐分布的主要因素。黑潮中层水是东海陆架区硅酸盐的主要来源。     

渤海和南黄海沉积物中氨氧化微生物对硝化潜势的相对贡献

硝化作用是海洋氮循环的核心过程。作为硝化过程关键步骤的氨氧化过程的主要参与者,氨氧化古菌和氨氧化细菌对硝化作用的相对贡献是海洋氮循环关注的热点问题之一。本文选取渤海和南黄海20个站位的表层沉积物,通过微宇宙培养实验研究了沉积物中氨氧化古菌和氨氧化细菌对硝化潜势的相对贡献。结果表明,渤海和南黄海海域表层沉积物中潜在硝化速率(以氮计,下同)为0.004 6～0.283 1μmol/(g·d),其中氨氧化古菌潜在硝化速率为0.004 3～0.274 3μmol/(g·d),氨氧化细菌潜在硝化速率为0.000 4～0.056 0μmol/(g·d)。氨氧化古菌是硝化潜势的主要贡献者,在渤海海域的贡献率为59.79%～97.95%,在南黄海海域的贡献率为18.47%～94.26%。渤海海域潜在硝化速率显著高于南黄海海域。此外,本研究海域中盐度是影响潜在硝化速率的关键环境因子,对渤海海域的分析则表明越高的NO₃^-浓度可能指示着越高的硝化潜势。在河口及近海沉积物中,氨氧化古菌在硝化过程中起着更加重要的作用;河口和近岸沉积物硝化潜势总体高于远海。本研究为进一...     

运用PCR-DGGE技术对3种原甲藻18S rDNA部分序列的差异性分析

采用3对特异性引物,对中国近海常见3种原甲藻的18S rDNA的部分序列进行PCR扩增,运用变性梯度凝胶电泳技术(Denaturing Gradient Gel Electrophoresis,DGGE)对PCR产物进行了差异性分析。结果表明,各对引物具有不同的区分效果,选择包含较多差异碱基的短片段进行扩增,可以对藻种进行更好的区分;PCR-DGGE技术具有较高的灵敏性,不同藻种间即使1个碱基的差异也可以区分开;PCR-DGGE技术可以对3种原甲藻进行区分。     

环境因子对海蜇生长发育影响的研究进展

<正>海蜇(Rhopilema esculenta Kishinouye)隶属于刺胞动物门(Cnidaria),钵水母纲(Scyphozoa),根口水母目(Rhizostomeae),根口水母科(Hizostomadae),海蜇属(Rhopilema)[1]。海蜇为进化比较保守的两胚层动物[2]。丁耕芜等[3]首次报告了海蜇的整个生活史。海蜇生活史具有刺胞动物门典型的世代交替特征,分为两个世代:水螅体世代和水母体世代。在水螅体世代整个过程中,海蜇螅状体于海底基质上营固着生活,以无性生殖     

Phytoplankton diversity in the East China Sea and Yellow Sea measured by PCR-DGGE and its relationships with environmental factors

Relationships between phytoplankton community composition and environmental variables in the East China Sea (ECS) and Yellow Sea (YS) were investigated using geochemical and molecular microbiology methods. The diversity of phytoplankton was characterized using cultivation-independent PCR-based denaturing gradient gel electrophoresis (DGGE). Groups resulting from unweighted pair-group method with arithmetic averages clustering of the DGGE profiles showed good consistency with the eco-environmental characteristics of the sea area they belonged to. Additionally, the clustering results based on DGGE fingerprinting and those based on morphological compositions were practically identical. The relationship of phytoplankton diversity to environmental factors was statistically analyzed. Temperature, dissolved inorganic nitrogen (DIN), and silicate-Si were found significantly related to the phytoplankton community composition. Canonical correspondence analysis (CCA) was performed to reveal the relationship between community composition and these three environmental factors. Generally, values of the ECS are clearly separated from those of the YS in the CCA biplot, due to mainly the effect of temperature and DIN.     

Development of a real-time PCR assay (SYBR Green I) for rapid identification and quantification of scyphomedusae Aurelia sp.1 planulae

The complicated life cycle of A urelia spp., comprising benthic asexually-reproducing polyps and sexually-reproducing medusae, makes it hard for researchers to identify and track them, especially for early stage individuals, such as planulae. To solve this problem, we developed a real-time PCR assay(SYBR Green I) to identify planulae in both cultured and natural seawater samples. Species-specific primers targeting Aurelia sp.1 mitochondrial 16 SrDNA(mt 16 SrDNA) regions were designed. Using a calibration curve constructed with plasmids containing the A urelia sp.1 mt 16 SrDNA fragment and a standard curve for planulae, the absolute number of mt 16 SrDNA copies per planula was determined and from that the total number of planulae per sample was estimated. For the field samples, a 100-fold dilution of the sample DNA combined with a final concentration of 0.2 μg/μL BSA in the PCR reaction mixture was used to remove realtime PCR inhibitors. Samples collected in Jiaozhou Bay from July to September 2012 were subsequently analyzed using this assay. Peak A urelia sp.1 planula abundance occurred in July 2012 at stations near Hongdao Island and Qingdao offshore; abundances were very low in August and September. The real-time PCR assay(SYBR Green I) developed here negates the need for traditional microscopic identification, which is laborious and time-consuming, and can detect and quantify jellyfish planulae in field plankton samples rapidly and specifically.     

东海表层沉积物中氨氧化微生物群落的空间分布和季节性变化

Ammonia oxidation plays a significant role in the nitrogen cycle in marine sediments. Seasonal and spatial distribution of ammonia-oxidizing archaea(AOA) and betaproteobacteria(β-AOB) in surface sediments from the East China Sea(ECS) were investigated using ammonia monooxygenase α subunit(amo A) gene. In order to characterize the community of AOA and β-AOB, real-time quantitative polymerase chain reaction(q PCR) was carried out in this study, along with environmental parameters. The abundance of β-AOB amo A gene(2.17×106–4.54×107 copy numbers per gram wet weight sediment) was always greater than that of AOA amo A gene(2.18×105–9.89×106 copy numbers per gram wet weight sediment) in all sampling stations. The q PCR results were correlated with environmental parameters. AOA amo A gene copy numbers in April were positively related to temperature and nitrite concentration(p0.05). β-AOB amo A gene copy numbers in August correlated negatively with salinity(p0.01), and correlated positively with ammonium concentration(p0.05). With the increase of salinity, the amo A gene copy ratio of AOB to AOA had a tendency to decrease, which suggested β-AOB dominated in the area of high level ammonium and AOA preferred high salinity area.     

免疫学方法在有害赤潮藻细胞检测中的应用

赤潮可以严重危害海洋渔业、海水养殖业和旅游业,甚至危害到人类的生命健康[1,2],它的频繁发生已经引起各国政府的高度重视。虽然可以通过每日多次实时检测海水的水文、化学要素等来推测发生赤潮的可能性[3],但是这些方法都不能直接反映有害赤潮藻的种类和数量,从而不能为赤潮监测和预报提供最直接的参考数据。实际上,对海水中的藻类进行分类、鉴定和数量分析是监测和预报赤潮发生的最直接有效的方法,这包括对水样采集、藻种类鉴定、毒性鉴定和赤潮藻定量计数等。传统的形态分类技术可以解决赤潮监测中的多数问题,但需要有经过专门训练的分…