几株原甲藻核糖体大亚基RNA基因的部分克隆及序列分析

对7株赤潮原甲藻28S rDNA 5'端部分序列进行扩增、克隆和序列测定,并从GeneBank上获取14个原甲藻28S rDNA序列,用NJ法和ME法构建了原甲藻属的系统树,并对序列进行分析.结果表明,7株原甲藻28S rDNA扩增序列长度为950~958 bp,通过NJ法和ME法构建的系统树完全一致.大部分分离自不同海域的同种原甲藻的序列高度保守,而不同种间在序列高变区却有较大的差异.但来自南海海域的海洋原甲藻(Prorocentrum micans)与分离自其他海域的株系序列差异较大,甚至超出了有些种间的差异.由28S rDNA高变区获得的序列,有望成为浮游植物特异性分子探针设计的良好靶区域.     

利玛原甲藻腹泻性毒素的生物学测定

１９９４年和１９９５年分季度从海南省三亚海区采集附生于褐藻上的利玛原甲藻Ｐｒｏｒｏｃｅｎｔｒｕｍｌｉｍａ（Ｅｈｒｅｎｂｅｒｇ）Ｄｏｄｇｅ，经丙酮提取。提取液用ｐＨ＝３的水除去麻痹性贝毒（ＰＳＰ），取残留液注入小白鼠（ＢａＢ／Ｃ纯系），有腹泻性贝毒（ＤｉａｒｒｈｅｔｉｃＳｈｅｌｌｆｉｓｈＰｏｉｓｏｎｉｎｇ即ＤＳＰ）中毒的症状出现。初步确定此种藻中提取的毒素为ＤＳＰ。     

温度、盐度、光照强度和pH对海洋原甲藻增长的效应

采用微量移液稀释法对海洋原甲藻进行无菌培养，以研究温度、盐度、光照强度和pH对海洋原甲藻增长的效应。结果表明，海洋原甲藻最大增殖速率发生在温度为(25±1)℃、盐度为31、光照强度为3000lx、pH为8.0的环境条件下。海洋原甲藻在温度18—28℃、盐度为25—34、光照强度为1000—6500lx、pH为7.5—8.3范围内适宜生长。这一结果与观测到的海洋原甲藻赤潮发生时环境条件相近，其中温度的效应最为明显。     

东海原甲藻的分子鉴定

采用nrDNAITS序列及18S序列两种分子标记,对东海原甲藻和美国国家海洋藻种保藏中心(CCMP)的具齿原甲藻进行分子鉴定,结果表明,两者之间序列非常相似,两者的nrDNAITS序列碱基差异值仅为0.002,nrDNA18S序列的碱基差异值为0.000,根据分子数据,东海原甲藻和美国国家海洋藻种保藏中心的具齿原甲藻应为同一个种.从基因库获取原甲藻的另外3个种nrD-NAITS序列和8个种的18S序列,用计算机软件进行分析和构建分子系统树,结果显示18S序列用于原甲藻的分子鉴定过于保守,而nrDNAITS更适合于该种属界定的分子标准.     

赤潮发生机理研究──海洋原甲藻的氮营养生理特征

于１９９１年１月－１９９１年１２月，以不同浓度的硝态氮（２０，６０，２００，６００μｍｏｌ／Ｌ）研究对赤潮生物海洋原甲藻的生长和生化组成的影响，及其营养生理特性，结果表明，在氮浓度为２００和６００μｍｏｌ／Ｌ时，表现光合作用强，细胞生长快，指数生长期长，叶绿素含量高，提示了由原甲藻爆发引起的赤潮与海洋富营养化有直接关系，结果还表明，海洋原甲藻可以累积蛋白质和碳水化合物，当外界氮源不能满足生长需要时     

海水养殖池海洋原甲藻水华前后水化条件的变化

本文研究了海水养殖池塘实验围隔中海洋原甲藻水华暴发前、后水化条件的变化,得出了如下结果：海洋原甲藻水华一般多发生在连续晴朗的天气,而常在阴雨天气崩溃;从水化条件方面来说,该藻水华也常发生于ＮＯ３－Ｎ浓度以及Ｎ／Ｐ比较大时。在扁灌等绿藻类与海洋原甲藻交替成为优势种时,可以利用水体中Ｃh１。总量及Ｃｈ１．ｂ、Ｃｈ１．ｃ之间 的波动情况来反映该水华的发生与崩溃。     

东海三叶原甲藻的形态特征及其ITS序列分析

报道了首次分离于东海海域的三叶原甲藻(Prorocentrum triestinum Schiller)藻株号(LAMB100721),通过利用光学显微镜、荧光显微镜、扫描电镜及分子生物学方法,对其形态特征、显微结构和分子系统进化进行了详尽描述和鉴定。细胞长卵形或披针形,后端细长且尖,前端圆,最宽部位于细胞中央。顶刺长而显,三角状。壳面光滑,无刺或突起物。刺丝胞孔稀疏而不规则地分布于壳面边缘。叶绿体无具体形状,分布于整个细胞中,细胞核球形,位于中下部。老化的细胞可见细胞边缘的间接带,且间接带表面光滑。细胞长为19~25 μm,平均值为(22.6±0.9)μm;宽为11~16 μm,平均值为(13.2±1.1) μm。所测目标藻株的rDNA ITS序列长度为569 bp,其中GC含量为47.6%。三叶原甲藻系赤潮种,为东海原甲藻春季大规模赤潮的伴随种。加强有害赤潮的预防和监测工作是减少危害的有效途径,而对赤潮原因种的准确识别和鉴定则是基础和关键。     

FIVE RED TIDE SPECIES IN GENUS PROROCENTRUM INCLUDING THE DESCRIPTION OF PROROCENTRUM DONGHAIENSE LU SP. NOV. FROM THE EAST CHINA SEA

A new planktonic dinoflagellate, Prorocentrum donghaiense I_a sp. nov., is described in the present paper. The water sample was collected from the Changjiang Estuary, the East China Sea. The species identification is based on shape, size, surface micro-morphology, ornamentation of thecal plates and the architecture of the pel‘iflagellar area and the intercalary bands as seen by fight and scanning electron microscope. Prorocentrum donghaiense Lu sp. nov. is compared with other prorocentrum species with respect to morphological characteristics and bloom behavior. It is not known whether Prorocentrum donghaiense Lu sp. nov produces phycotoxins like some other Prorocentrum species. Four other red tide species in the family Prorocentraceae (Dinophyceae), namely P. balticum, P. minimum, P.micans, P. triestinum, were examined and identified by light and scanning electron microscope. They have been recorded as bloom-forming species. Some aggregates of Prorocentrum are observed at the end of blooms. An event of strong discoloration caused by P. donghaiense could be detected by satellite sensor in the East China Sea in the late spring, of 1995.     

东海原甲藻抗体的制备及酶联免疫检测方法的建立

通过制备针对东海原甲藻细胞破碎物的多克隆和单克隆抗体,建立了基于双抗体酶联免疫分析定量检测东海原甲藻(Prorocentrum donghaiense)的检测方法。利用该方法对单一藻种、混合藻种和现场样品中的东海原甲藻进行检测的结果与镜检结果相一致,最低检测限度为1×103 cells/m L。该方法的建立对中国近海赤潮暴发的实时监控具有重要意义。     

Optical detection of Prorocentrum donghaiense blooms based on multispectral reflectance

Prorocentrum donghaiense is one of the most common red tide causative dinoflagellates in the Changjiang （Yangtze） River Estuary and the adjacent area of the East China Sea. It causes large-scale blooms in late spring and early summer that lead to widespread ecologic and economic damage. A means for distinguish- ing dinoflagellate blooms from diatom （Skeletonema costatum） blooms is desired. On the basis of measure- ments of remote sensing refectance [Rrs（λ）] and inherent optical parameters, the potential of using a mul- tispectral approach is assessed for discriminating the algal blooms due to P. donghaiense from those due to S. costatum. The behavior of two reflectance ratios [R1 = Rrs（560）/Rrs（532） and Re = Rrs（708）/Rrs（665）], suggests that differentiation of P. donghaiense blooms from diatom bloom types is possible from the current band setup of ocean color sensors. It is found that there are two reflectance ratio regimes that indicate a bloom is dominated by P. donghaiense; （1） R1 〉 1.55 and R2 〈 1.0 or （2） R1 〉 1.75 and R2 ≥ 1.0. Various sensitivity analyses are conducted to investigate the effects of the variation in varying levels of chlorophyll concentration and colored dissolved organic matter （CDOM） as well as changes in the backscattering ratio （bbp/bp） on the efficacy of this muitispectral approach. Results indicate that the intensity and inherent op- tical properties of the algal species explain much of the behavior of the two ratios. Although backscattering influences the amplitude of Rrs（λ）, especially in the 530 and 560 nm bands, the discrimination between P. donghaiense and diatoms is not significantly affected by the variation of bbp/bp. Since aCDOM （440） in coastal areas of the ECS is typically lower than 1.0 m-1 in most situations, the presence of CDOM does not interfere with this discrimination, even as SCDOM varies from 0.01 to 0.026 nm-1. Despite all of these effects, the dis- crimination of P. donghaiense blooms from diatom blooms based on multispectral measurements of Rrs（λ） is feasible.