Development of a real-time PCR method (Taqman) for rapid identification and quantification of Prorocentrum donghaiense

Prorocentrum donghaiense is a dinoflagellate that is widely distributed in the East China Sea and has become increasingly involved in Harmful Algal Blooms (HABs). Therefore, it is necessary to study this dinoflagellate to monitor HABs. In this study, 13 pairs of primers specific to P. donghaiense (within its internal transcribed spacer (ITS) regions) were designed for SYBR Green I real-time PCR. As the SYBR Green I real-time PCR could not identify P. donghaiense in a specific manner, a Taqman real-time PCR method was developed by designing a set of specific primers and a Taqman probe. A 10-fold serial dilution of recombinant plasmid containing ITS regions of P. donghaiense was prepared as standard samples and the standard curve was established. Additionally, we quantified the genomic DNA in P. donghaiense cells and utilized this DNA to prepare another 10-fold serial dilution of standard sample and accordingly set up the standard curve. The mathematic correlation between the cell number and its corresponding plasmid copy number was also established. In order to test the efficiency of the real-time PCR method, laboratory samples and P. donghaiense HAB field samples were employed for identification and quantitative analysis. As to laboratory samples, as few as 102 cells of P. donghaiense could be quantified precisely utilizing both centrifugation and filtration techniques. The quantification results from field samples by real-time PCR were highly similar to those by light microscopy. In conclusion, the real-time PCR could be applied to identify and quantify P. donghaiense in HABs.     

夏季台湾暖流的水文化学特性及其对东海赤潮高发区影响的初步探讨

依据2006年夏季对东海区水文化学数据的现场调查, 对台湾暖流的水文化学特性进行了初步分析, 并对台湾暖流自身的水文化学特性对东海赤潮高发区的影响进行了初步探讨。结果表明, 夏季, 台湾暖流水具有台湾海峡水和黑潮次表层涌升水两个来源, 分别构成台湾暖流的表层水和深层水。通过亚硝酸盐含量的多少能够对表层水和深层水进行明显的区分。此外, 通过对台湾暖流水文特征的分析, 发现夏季台湾暖流在浙江沿海出现的上升流给赤潮生物提供了适宜的温度。并通过对台湾暖流的营养盐含量进行分析, 发现相对于硝酸盐含量, 台湾暖流含有较高的磷酸盐浓度, 能够缓解海区“过剩氮”导致的磷限制。     

Occurrence of Pyrodinium bahamense var. compressum along the southern coast of the Baja California Peninsula

As part of a continuing toxic microalgae monitoring program, 22 phytoplankton samples were collected from July to November 2010 at several sampling stations along the southern coast of the Baja California Peninsula. For the first time, the toxic dinoflagellate Pyrodinium bahamense var. compressum was found along the southeastern and southwestsern coasts of the peninsula. P. bahamense var. bahamense was first observed off San José del Cabo, which is an extension of the range of this variety. Both varieties occur as solitary cells. P. bahamense var. compressum occurred at temperatures ranging between 24.5 °C and 31 °C, whereas var. P.bahamense occurred at 28.5 °C to 29 °C, indicating its tropical and subtropical nature. Occurrence of P. bahamense var. compressum along this coastline may be related to El Niño 2009-2010.     

东海甲藻硅藻现场生长营养盐相关参数的计算

应用生态动力学模型简化法,建立了现场船基培养实验计算营养盐半饱和常数和浮游植物细胞内营养盐比例的方法,并获得了东海甲藻和硅藻的营养盐吸收半饱和常数、营养盐氮磷比和硅氮比的取值范围及建议取值。甲藻:氮半饱和常数2.50μmol/L、磷半饱和常数0.16μmol/L、营养盐氮磷比建议取值为15.80;硅藻:氮半饱和常数3.99μmol/L、磷半饱和常数0.38μmol/L、硅半饱和常数3.35μmol/L、营养盐氮磷比20.45、营养盐硅氮比建议取值为0.49。     

Study on impact of dinoflagellate Alexandrium tamarense on life activities of marine bivalves

The effects of a PSP producing dinoflagellate Alexandrium tamarense on marine bivalvesat their several important life stages: egg, D- shape larva, eyespot larva, juvenile and adult, were studied. The results show that the hatching, survival, activity, filtration and growth were adversely affected by the alga and the impact was significantly increased with the increase of algal density. The inhibitory effect on egg hatching was most significant, which the hatching rate was only 30% of the control when exposed to the alga at 100 cell/cm3 after 36 h. Further experiments show that the algal culture, re-suspended cells and cell fragments had the inhibitory effect, while no such effect was from the cell-free medium, cell contents and standard STX. The results indicate that the alga could produce unknown toxins, rather than PSP, associated with the cell surface.     

长江口邻近海域春季藻华期间浮游植物生物量与环境要素的拟合分析

基于2005年长江口邻近海域春季现场调查资料,本文采用非线性拟合方法,构建了该海域春季硅藻藻华和甲藻藻华期间浮游植物生物量与无机营养盐、温度和盐度之间的定量关系方程,进而分析了春季硅、甲藻藻华发生的最适环境条件。结果表明,硅藻藻华出现的最适环境条件为:溶解态无机氮浓度10.1μmol/L、磷酸盐浓度0.21μmol/L、硅酸盐浓度6.39μmol/L、温度11.9℃、盐度30.5;甲藻藻华暴发的最适环境条件为:溶解态无机氮浓度3.98μmol/L、磷酸盐浓度0.18μmol/L、温度22.0℃、盐度27.2。上述硅、甲藻藻华发生的最适环境条件可作为硅、甲藻藻华形成的关键环境参数,为开展长江口邻近海域藻华形成和演替过程的数值模拟和参数优化提供参考。