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11.
东海原甲藻线粒体细胞色素b(Cytb)基因的定量检测 总被引：3，自引：0，他引：3
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. 相似文献
12.
用双特异探针技术定性定量分析微型原甲藻 总被引：5，自引：0，他引：5
针对微型原甲藻核糖体大亚基和小亚基RNA分别设计了大亚基探针(LSU probe)和小亚基探针(SSU probe),发展了对微型原甲藻进行定性和定量分析的双特异探针技术.分析数据表明针对微型原甲藻核糖体大亚基RNA的LSU probe能够特异地将微型原甲藻和其他7种微藻分开,而且检测信号的强弱在一定的范围内与细胞数呈线性关系;由于针对微型原甲藻核糖体小亚基的SSU probe探针由于与具齿原甲藻存在核酸序列一致性,该探针与具齿原甲藻有严重的交叉反应,但未发现与海洋原甲藻和其他藻的交叉反应.另外,研究还优化了破碎微型原甲藻细胞所需的超声时间以及获得探针的特异性所需的S1酶反应温度.本研究为实现对微型原甲藻海水样品的快速准确监测奠定了基础. 相似文献
13.
中国东部边缘海冬季硅酸盐的分布特征及主要来源 总被引：1，自引：0，他引：1
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。 相似文献
14.
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. 相似文献
15.
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。 相似文献
16.
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… 相似文献
17.
环境因子对海蜇生长发育影响的研究进展 总被引：1，自引：0，他引：1
<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 相似文献
18.
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 总被引：2，自引：0，他引：2
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. 相似文献
19.
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. 相似文献
20.
悬浮颗粒物中生物硅测定方法的改进与应用 总被引：1，自引：3，他引：1
硅是河流和海洋中非常重要的营养盐,硅藻、放射虫、硅质海绵、硅鞭毛虫的生长和骨骼形成都离不开硅[1].硅在河流和沿岸海域生态系统中扮演着非常重要的角色,在浮游植物大量繁殖季节,由于硅藻的大量繁殖,使活性硅酸盐含量急剧下降,甚至使硅藻的生长繁殖受到限制.在其他环境条件适宜的情况下,充足的硅又将成为发生硅藻赤潮的物质基础[2].生物硅是用化学方法测定的无定形硅的含量[3],称为生物蛋白石或简称蛋白石,是地球化学、古海洋学研究中非常重要的参数[4].根据世界部分河流生物硅和溶解硅的平均值计算,河流输送入海的硅中有16%的是生物硅,因此河流输送的生物硅的量是全球海洋硅收支中不可忽略的部分[5]. 相似文献