Identification of Prorocentrum minimum and Takayama pulchella by fluorescence in situ hybridization through epifluorescence microscopy and flow cytometry

Partial rDNA sequences of Prorocentrum minimum and Takayama pulchella were amplified, cloned and sequenced. and these sequence data were deposited in the GenBank. Eight oligonucleotide probes (DNA probes) were designed based on the sequence analysis. The probes were employed to detect and identify P. minimum and T. pulchella in unialgal and mixed algal samples with a fluorescence in situ hybridization method using flow cytometry. Epifluorescence micrographs showed that these specific probes labeled with fluorescein isothiocyanate entered the algal cells and bound to target sequences, and the fluorescence signal resulting from whole-cell hybridization varied from probe to probe. These DNA probes and the hybridization protocol we developed were specific and effective for P. minimum and T. pulchella, without any specific binding to other algal species. The hybridization efficiency of different probes specific to P. minimum was in the order: PM18S02 > PM28S02 > PM28S01 > PM18S01, and that of the probes specific to T. pulchella was TP18S02 > TP28S01 > TP28S02 > TP18S01. The different hybridization efficiency of the DNA probes could also be shown in the fluorescent signals between the labeled and unlabeled cells demonstrated using flow cytometry. The DNA probes PM18S02, PM28S02; TP18S02 and TP28S01, and the protocol, were also useful for the detection of algae in natural samples.

环境因子对有毒赤潮生物微型原甲藻（Prorocentrum minimum）生长增殖的影响

本文研究了温度、盐度、PH、营养盐（N、P）；维生素（B_(12)、B_1）、微量元素（Fe、Mn）、底泥抽提液和赤潮生物骨条藻与微型原甲藻生长的关系。结果表明，微型原甲藻增殖生长的适温范围为20－30 ℃，28 ℃时有最大生长率；最适盐度为10－35‰，25‰盐度生长率最高；PH 7.18-8.72范围内生长良好，影响不明显。N、P营养盐能促进微型原甲藻增殖生长，N盐以硝态氮效果最好，容易被吸收利用；氨盐为唯一N源时效果较差，浓度超过100uM时则有抑制作用；亚硝酸盐作用不明显。微型原甲藻主要是利用无机磷；有机磷作唯一P源时不能有效地被吸收利用，对微型原甲藻生长不利；维生素B_(12)促进微型原甲藻增殖生长效果明显，B_(12)与B_1综合效果更好；底泥抽提液、微量元素Fe、Mn都能有效地促进微型原甲藻增殖生长；研究证实底泥中腐植酸（HFA）虽有促进生长的作用，但效果尚不及抽提液，推测可能还有其他有效成分。研究发现中肋骨条藻有刺激微型原甲藻生长的作用，但其具体作用成分还有待研究。所得这些结果对进一步研究阐明微型原甲藻赤潮成因和机理有重要科学意义。

Three-dimensional fluorescence characteristics of dissolved organic matter produced by Prorocentrum donghaiense Lu

Filtration and cross-flow ultrafiltration techniques were used to separate culture media of Prorocentrum donghaiense at the exponential growth, stationary and decline stages into < 0.45 mu m filtrate, 100 kDa-0.45 mu m, 10-100 kDa and 1-10 kDa retentate and < 1 kDa ultrafiltrate fractions. The fluorescence. properties of different molecular weights of dissolved organic matter (DOM) were measured by excitation-emission matrix spectra. Protein-like and humic-like fluorophores were observed in the DOM produced by P. donghaiense. The central positions of protein-like fluorophores showed a red shift with prolonged growth duration, shifting from tyrosine-like properties at the exponential growth stage to tryptophan-like properties at the stationary and decline stages. The excitation wavelengths of protein-like fluorophores exhibited some change in the exponential growth and stationary stages with increased molecular size, but showed little change in the decline stage. However, the emission wavelengths in the decline stage exhibited a blue shift. Very distinct C type and A type peaks in humic-like fluorophores were observed. With a prolonged culture time, the intensities of both of the peaks became strong and the excitation wavelengths of peak A showed a red shift, while the A:C ratios fell. More than 94% of fluorescent DOM was in the lower than 1 kDa molecular weight fraction.

Morphological and genetic study of Prorocentrum donghaiense Lu from the East China Sea, and comparison with some related Prorocentrum species

In this paper, the detailed morphology of Prorocentrum donghaiense Lu from both field samples and cultures was examined, and a taxonomic comparison was made between P donghaiense and some related Prorocentrum spp. using morphological and molecular data and other published information. There were distinct differences among these species in morphological characteristics that historically have been presented as conservative features. The discrepancies extended beyond that of individual variations within the same species due to environmental factors. Therefore, these morphological features may not be conservative but, rather, polymorphic depending on environmental conditions. Based on this analysis, we suggest that the high-biomass bloom-forming species in the East China Sea, previously reported as Prorocentrum dentatum Stein, is P donghaiense Lu. The species reported from the East China Sea and Japanese and Korean waters appear to be the same species. Molecular data also suggest that P. dentatum (CCMP1517) and P. donghaiense are genetically identical. Therefore, the geographic distribution of P. donghaiense may be much wider than expected. (C) 2004 Elsevier B.V. All rights reserved.

A comparison of future increased CO2 and temperature effects on sympatric Heterosigma akashiwo and Prorocentrum minimum

Very little is known about how global anthropogenic changes will affect major harmful algal bloom groups. Shifts in the growth and physiology of HAB species like the raphidophyte Heterosigma akashiwo and the dinoflagellate Prorocentrum minimum due to rising CO2 and temperature could alter their relative abundance and environmental impacts in estuaries where both form blooms, such as the Delaware Inland Bays (DIB). We grew semi-continuous cultures of sympatric DIB isolates of these two species under four conditions: (1) 20 degrees C and 375 ppm CO2 (ambient control), (2)20 degrees C and 750 ppm CO2 (high CO2),(3) 24 degrees C and 375 ppm CO2 (high temperature), and (4) 24 degrees C and 750 ppm CO2 (combined). Elevated CO2 alone or in concert with temperature stimulated Heterosigma growth, but had no significant effect on Prorocentrum growth. P-Bmax (the maximum biomass-normalized light-saturated carbon fixation rate) in Heterosigma was increased only by simultaneous CO2 and temperature increases, whereas P-Bmax in Prorocentrum responded significantly to CO2 enrichment, with or without increased temperature. CO2 and temperature affected photosynthetic parameters alpha, Phi(max), E-k, and Delta F/F'(m) in both species. Increased temperature decreased and increased the Chl a content of Heterosigma and M Prorocentrum, respectively. CO2 availability and temperature had pronounced effects on cellular quotas of C and N in Heterosigma, but not in Prorocentrum. Ratios of C:P and N:P increased with elevated carbon dioxide in Heterosigma but not in Prorocentrum. These changes in cellular nutrient quotas and ratios imply that Heterosigma could be more vulnerable to N limitation but less vulnerable to P-limitation than Prorocentrum under future environmental conditions. In general, Heterosigma growth and physiology showed a much greater positive response to elevated CO2 and temperature compared to Prorocentrum, consistent with what is known about their respective carbon acquisition mechanisms. Hence, rising temperature and CO2 either alone or in combination with other limiting factors could significantly alter the relative dominance of these two co-existing HAB species over the next century. (c) 2007 Elsevier B.V. All rights reserved.

Interactions between the bloom-forming dinoflagellates Prorocentrum donghaiense and Alexandrium tamarense in laboratory cultures

Interactions between Prorocentrum donghaiense and Alexandrium tamarens, two bloom-forming dinoflagellates, were investigated using bi-algal cultures. All R donghaiense died, but A. tamarense was hardly affected by the end of the experiment when the initial cell density was set at 1.0 X 10(4) cells mL(-1) for P. donghaiense and 0.28 x 10(4) cells mL(-1) for A. tamarense. However, significant growth suppression occurred in either species when the initial cell density of P donghaiense increased to I. 0 X 105 Cells mL(-1) in the bi-algal culture, but no out-competement was observed. The simultaneous assay on the culture filtrates showed that P donghaiense filtrate prepared at a lower initial density (1.0 X 10(4) cells mL(-1)) stimulated growth of the co-cultured A. tanzarense (0.28 x 10(4) cells mL(-1)), but filtrate at a higher initial density (1.0 x 10(5) cells mL(-1)) depressed its growth. The filtrate of A. tamarense at a density of 0.28 x 10(4) cells mL(-1) killed all R donghaiense at a lower density (1.0 x 10(4) cells mL(-1)), but only exhibited an inhibitory effect on it at a higher density (1.0 x 10(5) cells mL(-1)). It is likely that these two species of microalgae interfere with each other mainly by releasing allelochemical substance(s) into the culture medium, and a direct cell-to-cell contact was not necessary for their mutual interaction. The allelopathic test further proved that A. tamarense could affect the growth of co-cultured P. donghaiense by producing allelochemical(s); moreover, A. tamarense culture filtrate at the stationary growth phase (SP) had a strongly inhibitory effect on P donghaiense compared to that at the exponential phase (EP). Results also demonstrated a dose-dependent relationship between the microalgal initial cell density and the degree of the allelopathic effect. The growth of R donghaiense and A. tamarense in the bi-algal cultures was simulated using a mathematical model to quantify the interaction. The estimated parameters from the model showed that the inhibition exerted by A. tamarense on P. donghaiense was about 17 and 8 times stronger than the inhibition P. donghaiense exerted on A. tamarense, when the initial cell density was set at 1.0 X 10(4) and 1.0 X 10(5) cells mL(-1) for P donghaiense, respectively. and 0.28 x 10(4) cells mL(-1) for A. tamarense in the bi-algal cultures. A. tamarense seems to have a survival strategy that is superior to that of P. donghaiense in bi-algal cultures under controlled laboratory conditions. (c) 2006 Elsevier B.V. All rights reserved.

泉州湾赤潮藻类优势种演替影响因子探讨

于2006年5—11月对东海泉州湾赤潮监控区设定4个采样站位,进行赤潮常规监测。结果表明,泉州湾藻类优势种由中肋骨条藻(Skeletonema costatum)、太平洋海链藻(Thalassiosira pacifica)、微小原甲藻(Prorocentrum minimum)、尖刺拟菱形藻(Pseudo-nitzschia pungens)、丹麦细柱藻(Leptocylindrus danicus)及旋链角毛藻(Chaetoceros curvisetus)等赤潮生物组成,虽然中肋骨条藻为最主要的优势

尖尾藻（Oxyrrhis marina）染色体固定特性的比较研究

本工作采用单纯戊二醛、单纯四氧化锇、戊二醛加四氧化锇双固定及Ryter和Kellenberger的细菌类核体“标准固定方法”等四种固定方法，在尖尾藻与典型涡鞭毛虫双甲藻(Prorocentrum micans)、典型原核生物肠道细菌、眼虫(Euglean sp.)、典型真核细胞小鼠胸腺细胞之间比较了电镜下所观察到的染色体和类核体的固定特性。实验结果表明尖尾藻染色体的固定特性异于典型涡鞭毛虫染色体的固定特性。而与眼虫染色体的相同，与典型真核细胞异染色质的相似。这些结果支持关于重新确定尖尾藻分类地位的设想；实验结果还指出尖尾藻染色体和眼虫染色体在碱性蛋白被抽提尽后，它们的固定特性均与细菌类核体的相似，这提示除了染色体碱性蛋白或组蛋白的含量是染色体固定特性的一个决定因素外，染色体碱性蛋白与DNA纤维的结合方式也可能影响染色体的固定特性。

Helical single-lamellar crystals thermotropically formed in a synthetic nonracemic chiral main-chain polyester

Phase structures and transformation mechanisms of nonracemic chiral biological and synthetic polymers are fundamentally important topics in understanding their macroscopic responses in different environments. It has been known for many years that helical structures and morphologies can exist in low-ordered chiral liquid crystalline (LC) phases. However, when the chiral liquid crystals form highly ordered smectic liquid crystal phases, the helical morphology is suppressed due to the crystallization process. A double-twisted morphology has been observed in many liquid crystalline biopolymers such as dinoflaggellate chromosomes (in Prorocentrum micans) in an in vivo arrangement. Helical crystals grown from solution have been reported in the case of Bombyx mori silk fibroin crystals having the beta modification. This study describes a synthetic nonracemic chiral main-chain LC polyester that is able to thermotropically form helical single lamellar crystals. Flat single lamellar crystals can also be observed under the same crystallization condition. Moreover, flat and helical lamellae can coexist in one single lamellar crystal, within which one form can smoothly transform to the other. Both of these crystals possess the same structure, although translational symmetry is broken in the helical crystals. The polymer chain folding direction in both flat and helical lamellar crystals is determined to be identical, and it is always along the long axis of the lamellae. This finding provides an opportunity to study the chirality effect on phase structure, morphology, and transformation in condensed states of chiral materials. [S0163-1829(99)01042-5].

Development of rRNA and rDNA-targeted probes for fluorescence in situ hybridization to detect Heterosigma akashiwo (Raphidophyceae)

Heterosigma akashiwo (Hada) is a fragile, fish-killing alga. Efforts to understand and prevent blooms due to this harmful species to mitigate the impact on aquaculture require the development of methods for rapid and precise identification and quantification, so that adequate warning of a harmful algal bloom may be given. Here, we report the development and application of rRNA and rDNA-targeted oligonucleotide probes for fluorescence in situ hybridization (FISH) to aid in the detection and enumeration of H. akashiwo. The designed probes were species specific, showing no cross-reactivity with four common HAB causative species: Prorocentrum micans Ehrenberg, P. minimum (Pavillard) Schiller, Alexandrium tarmarense (Lebour) Balech, and Skeletonema costatum (Greville) Cleve, or with four other microalgae, including Gymnodinium sp. Stein, Platy-monas cordiformis (Karter) Korsch, Skeletonema sp.1 Greville and Skeletonema sp.2. The rRNA-targeted probe hybridized to cytoplasmic rRNA, showing strong green fluorescence throughout the whole cell, while cells labeled by rDNA-targeted probe exhibited exclusively fluorescent nucleus. The detection protocols were optimized and could be completed within an hour. For rRNA and rDNA probes, about a corresponding 80% and 70% of targeted cells could be identified and quantified during the whole growth circle, despite the inapparent variability in the average probe reactivity. The established FISH was proved promising for specific, rapid, precise, and quantitative detection of H. akashiwo. (C) 2008 Elsevier B.V. All rights reserved.

改性粘土治理有害赤潮对生态环境的影响初探

本文以室内实验为基础，研究了改性粘土治理有害赤潮的方法对生态环境的影响。研究结果表明：有机和无机改性粘土对溶解氧（DO）、化学耗氧量（COD）、pH等主要水质因子均有改善作用；对营养盐，尤其是磷酸盐有一定的吸附作用，在外加磷酸盐为0-0.3μmol/ml范围内，吸附量随水体中磷酸盐浓度的增加而增大，有机改性粘土对海水中磷酸盐的吸附能力为：有机改性粘土Ⅰ>有机改性粘土Ⅱ>有机改性粘土Ⅲ。通过有机改性粘土对磷酸盐的吸附－再释放情况，进一步探讨了其磷酸盐的释放对赤潮异弯藻（Heterosigma akashiwo）、东海原甲藻（Prorocentrum donghaiense）等赤潮生物生长的影响。实验结果显示，经过有机改性的粘土有利于提高其对磷酸盐的吸附能力，降低对磷酸盐的解吸率，利用有机改性粘土治理赤潮可以缓解海水富营养化程度，即使被吸附的部分磷酸盐能缓慢释放，但仍不足以维持赤潮生物的正常生长。无机改性粘土对磷酸盐和硝酸盐与有机改性粘土有相似的吸附性能。其添加剂PAC的重金属含量符合沉积物排放要求，不会对环境造成压力。 其次，以太平洋牡蛎(Crassostrea gigas)幼贝为对象，研究了有机改性粘土Ⅱ和无机改性粘土在对海洋底栖生物的影响。急性毒性试验中二者对牡蛎幼贝的半致死浓度（LC50）分别为4.62 g/L和2.67 g/L，均远远大于改性粘土去除赤潮藻的浓度。在能够有效去除赤潮微藻的粘土浓度条件下，经慢性毒性试验发现改性粘土对牡蛎幼贝成活率、生长和摄食略有降低，但无明显影响，在牡蛎幼贝鳃组织和消化道组织的超微结构中未发现机械损伤。 最后，研究了改性粘土在对去除有毒赤潮藻过程中对底栖生物的影响。以能产生麻痹性贝毒（PSP）的塔玛亚历山大藻（A. tamarense，ATHK）为例，研究了PSP在水体中和粘土沉积层中的分布，发现改性粘土去除塔玛亚历山大藻能有效降低水体中藻细胞浓度及其所携带的PSP毒素。由于藻细胞破裂, 虽然沉积层中藻细胞体内部分PSP毒素溢出，但通过塔玛亚历山大藻细胞不同组分对牡蛎的影响发现，细胞内容物和细胞碎片对牡蛎没有明显毒副作用，由此推断改性粘土对藻细胞絮凝沉降后,能降低对生物的不良影响。此外，室内实验模拟了去除赤潮生物塔玛亚历山大藻（A. tamarense，ATHK）、东海原甲藻 (P. donghaiense )、赤潮异弯藻 (H. akashiwo)过程中，比较改性粘土和赤潮微藻对生物体的影响发现改性粘土在去除赤潮藻过程中能提高牡蛎的存活率，进一步证明改性粘土在赤潮生物防控中，是一种有效的应急措施。

东海大规模赤潮对微型浮游动物群落结构影响的研究

本文从赤潮发生前的现场模拟实验、赤潮发生时的现场培养实验和赤潮发生过程中的现场调查等方面，较为系统地研究了东海大规模赤潮对微型浮游动物群落结构的影响。 2005年长江口及邻近海区赤潮发生前，赤潮973MC2005－03航次在本海区进行了综合调查，期间分别于4月27日、5月4日和5月8日，在zzf1、zc18a和ra5三个站位利用现场船基培养的方法，研究了添加到赤潮密度106 cells L-1的东海原甲藻（Prorocentrum donghaiense）对微型浮游动物群落结构的影响。结果发现，赤潮密度的东海原甲藻对小型无壳纤毛虫的种群数量影响较大，而对中大型砂壳纤毛虫的影响较小，从而使得微型浮游动物群落有向中大型砂壳纤毛虫演替的趋势。在zzf1站位，小型无壳纤毛虫占绝对优势，添加东海原甲藻72 h后，优势种由管游虫（Cyrtostrombidium sp.）演替为另一种小型无壳纤毛虫急游虫4（Strombidium sp.4）；在以中型砂壳纤毛虫百乐拟铃虫（Tintinnospsis beroidea）为优势种的zc18a站位，仍然是以此为优势种，但其在群落中的优势度更加明显；ra5站位也是以小型无壳纤毛虫为主，添加东海原甲藻72 h后，其群落结构由急游虫2（Strombidium sp.2）向中大型砂壳纤毛虫纤毛虫百乐拟铃虫和亚速岛网纹虫（Favella azorica）演替。不同站位微型浮游动物群落结构变化的差异与其本身的群落结构组成有一定关系，也与浮游植物的群落组成有关。zzf1和ra5站位实验组中微型浮游动物的总丰度和总生物量都低于对照组，而zc18a站位变化不明显，这是由于前两个站位实验组中的优势种管游虫（zzf1站位）和急游虫2（ra5站位）的丰度和生物量迅速下降，而zc18a站位实验组的优势种百乐拟铃虫的丰度和生物量比较稳定造成的。 2005年长江口及邻近海区赤潮发生时，赤潮973 MC2005－04航次在本海区进行了综合调查，期间分别于5月26日、5月28日、5月28日和5月29日，在xzm1、srb、sra1和hb8a四个站位利用现场培养的方法，研究了东海大规模赤潮对微型浮游动物群落结构的影响。结果发现，东海大规模赤潮对微型浮游动物群落结构的影响与赤潮藻的密度密切相关：xzm1、srb和sra1站位是赤潮区，两种赤潮藻的总密度分别为，1.3×106 cells L-1、1.8×106 cells L-1和5.6×106 cells L-1，而hb8a站位位于非赤潮区，两种赤潮藻的总密度仅为5×105 cells L-1；实验进行72 h后，在以大型砂壳纤毛虫网纹虫和筒壳虫（Tintinnidium sp.）为主的xzm1、srb和sra1站位，仍然是以这些大型砂壳纤毛虫为主，且其在群落中的百分比显著增大，尤其在赤潮藻密度最高的的sra1站位，这种演替趋势表现的最明显；而在以小型无壳纤毛虫急游虫2为主的hb8a站位，微型浮游动物群落向中型砂壳纤毛虫真丁丁虫（Eutintinnus sp.）演替。培养过程中微型浮游动物总丰度的变化以及总生物量的变化都与赤潮藻的密度密切相关：赤潮藻密度较低的xzm1站位和srb站位变化不大；赤潮藻密度较高的sra1站位则快速下降；而非赤潮区的hb8a站位呈快速上升的趋势。以上结果进一步表明东海大规模赤潮能使微型浮游动物群落向中大型砂壳纤毛虫演替。 在2006年4月18日－5月30日，赤潮973项目MC2006航次在长江口及其邻近海区围绕东海原甲藻和米氏凯伦藻赤潮进行了综合调查。为了研究该过程中微型浮游动物群落结构的变化，我们在南北5个断面共计21个站位进行了取样，样品涉及赤潮发生前期、发生期和消退期。本次调查共发现纤毛虫66种，其中砂壳纤毛虫有8属37种，寡毛类无壳纤毛虫25种，另外还有前口类3属4种。在赤潮发生前之前，小型无壳纤毛虫的丰度普遍较高，平均值为1574 ind L-1，而中大型砂壳纤毛虫的丰度普遍偏低，平均值仅为14 ind L-1；赤潮发生以后，在大多数站位小型无壳纤毛虫的丰度出现不同程度的下降，平均值降为171 ind L-1，而中大型砂壳纤毛虫的丰度出现一定程度的上升，平均值增至216 ind L-1。赤潮消退时，小型无壳纤毛虫丰度的平均值降为109 ind L-1，而中大型砂壳纤毛虫的丰度继续增长，变为401 ind L-1。不同微型浮游动物百分比的统计结果也表明：微型浮游动物群落由小型无壳纤毛虫向中大型砂壳纤毛虫演替的趋势：小型无壳纤毛虫在群落中百分比的平均值由赤潮发生前的62 %变为赤潮发生时的32 %，至赤潮消退时又降为15 %；而中大型砂壳纤毛虫百分比的平均值从赤潮发生前的10 %变为赤潮发生时的24 %，至赤潮消退时增至50 %。典型站位za3、za5、zb7和zb9的结果再次表明了赤潮发生过程中微型浮游动物群落由小型无壳纤毛虫向中大型砂壳纤毛虫演替的趋势。 由以上赤潮发生前的现场模拟实验、赤潮发生时的现场培养实验和赤潮发生过程中的现场调查的结果均可见，东海大规模赤潮可以改变微型浮游动物的群落结构，呈现由小型无壳纤毛虫向中大型砂壳纤毛虫演替的趋势，进而有可能影响中大型浮游动物等摄食者的种群数量和群落结构，最终可能会影响整个海洋生态系统的结构和功能。

几种赤潮微藻的形态和系统进化分析及荧光原位杂交检测

赤潮也称红潮，通常是指由于一些海洋浮游生物在水体中过度繁殖或聚集而使海水变色的现象。赤潮特别是有害赤潮造成了严重的生态环境问题，给水产养殖业和滨海旅游业造成了巨大损失，并可直接危害人类健康。研究赤潮，进而预防和控制赤潮，首先要对引发赤潮的生物种类进行准确鉴定并对自然水域的赤潮生物进行监控，并建立赤潮藻的快速鉴定与检测方法。本文分别对几株赤潮微藻进行了形态和系统进化分析，并探讨了荧光原位杂交在赤潮检测中的应用。 分别对5株分离自中国沿海不同水域的中肋骨条藻[Skeletonema costatum (Greville) Cleve]类似种 (SK-BH、SK-FQ、 SK-HH、SK-DH和SK-XM) 进行光镜和扫描电镜观察，并PCR扩增了转录内间隔区 (含5.8S rDNA)(ITS) 和核糖体大亚基 (D1-D2)区 (LSU)，获得的序列与其它已报道的骨条藻的同源序列进行了进化分析，以探讨５株骨条藻与已报道的骨条藻之间的进化关系。5株骨条藻在形态上各不相同，其中，只有1株 (SK-XM)被鉴定为中肋骨条藻，而其余4株皆与已报道的骨条藻的形态学特征不符。ITS树和LSU树具有不同的拓扑结构，并表明5株骨条藻至少分属3个不同的种。遗传距离分析提示了在地理距离上靠近的种，在进化上也可能靠近。此外，还可以观察到这5株藻之间的细微的形态学“进化”关系。所有结果表明了中国沿海骨条藻属种的多样性。 对1株分离自赤潮水域的裸甲藻 (Gymnodinium)类似种进行了形态学分析，并探讨了该藻与裸甲藻、凯伦藻(Karena)、旋沟藻(Gyrodinium)、下沟藻(Karlodinium)和共生甲藻(Symbiodinium)的进化关系。光镜观察表明该藻具有裸甲藻的一些典型的形态学特征，而我们没能获得细胞形态保存完好的电镜样品；进化分析初步鉴定该藻为一种共生甲藻。 获得了赤潮异湾藻[Heterosigma akashiwo (Hada) Hada]的LSU和ITS序列，设计了以胞质rRNA和胞核rDNA为靶序列的特异性探针，建立了赤潮异湾藻的全细胞和细胞核荧光原位杂交技术，对探针的特异性进行了验证，并考察了杂交信号和检测率在整个细胞周期的变化情况。探针能分别使整个细胞和细胞核呈现明亮的绿色荧光。探针是特异性的，不与其它受试藻进行交叉反应。杂交信号在整个细胞周期内变化不明显，且检测率为70%–80%。整个检测过程不到1 ｈ，能实现赤潮异湾藻的快速、准确、特异和半定量检测。 获得了海洋原甲藻(Prorocentrum micans Ehrenberg)的LSU和ITS序列，设计了以胞质rRNA为靶序列的特异性探针，建立了海洋原甲藻的全细胞荧光原位杂交技术，并对探针的特异性进行了验证。探针能使整个细胞呈现强烈的绿色荧光。探针不与其它受试藻种进行交叉反应，表明是特异性的。