赤潮甲藻锥状斯氏藻rDNAITS区的序列测定和分析

锥状斯氏藻是赤潮甲藻的主要类群之一，其孢囊在我国南海大鹏湾底泥中占明显优势，是我国沿海增养殖区的潜在危害．采用ＰＣＲ及序列测定的方法，对其ｒＤＮＡＩｎｔｅｒｎａｌＴｒａｎ－ｓｃｒｉｂｅｄＳｅｑｕｅｎｃｅ（ＩＴＳ）区进行了序列测定。

一种快速制备甲藻细胞PCR扩增DNA模板的方法

报道了一种快速制备PCR扩增甲藻细胞模板DNA的方法。以赤潮甲藻塔玛亚历山大藻(Alexandriumtamarense)和链状亚历山大藻(A.catenella)为目标藻,对藻液预处理后直接用于PCR扩增,获得5.8S核糖体RNA基因及其两侧的核糖体RNA基因转录单元内基因间隔区(internaltranscribedspacer,ITS)序列片段,成功测序。此方法避免了复杂的DNA提取过程。

塔玛亚历山大藻对黑褐新糠虾存活、生长以及种群繁殖的影响

通过塔玛亚历山大藻 ( Alexandrium tamarense)对黑褐新糠虾 ( N eomysis awatschensis)的急性和慢性毒性作用研究 ,发现塔玛亚历山大藻对黑褐新糠虾的存活、生殖、生长等有不利影响 ,影响程度随塔玛亚历山大藻藻细胞密度的增加而增加。在 96 h急性毒性实验中 ,塔玛亚历山大藻对黑褐新糠虾的半致死密度为 70 0 0 cells/ml,去藻过滤液中糠虾的死亡率为 2 5 %。在 6 2 d的慢性毒性实验中 ,密度为 90 0 cells/ml的塔玛亚历山大藻对黑褐新糠虾的繁殖有严重影响 ,在此影响下的实验组亲虾产幼虾总数只有 2 7尾 ,仅为对照组产幼虾数目的 1 6 .4 % ;其总产幼虾天数、日最高产幼数分别只有对照的 32 %、4 1 % ,其初次产虾日期也推迟了 3d,并出现了 3次生殖中断。塔玛亚历山大藻对黑褐新糠虾亲虾的存活、生长也有一定的影响 ,处在密度为 90 0 cells/ml塔玛亚历山大藻中的黑褐新糠虾亲虾的存活率只有对照的 6 3% ,糠虾亲虾的体长和体重分别为对照组亲虾的 95 .6 %和 81 .9% ,但差异尚不显著 ( P>0 .0 5 )

Effects of Lugol's iodine solution and formalin on cell volume of three bloom-forming dinoflagellates

Fixatives are traditionally used in marine ecosystem research. The bias introduced by fixatives on.the dimensions of plankton cells may lead to an overestimation or underestimation of the carbon biomass. To determine the impact of traditional fixatives on dinoflagellates during short- and long-term fixation, we analyzed the degree of change in three bloom-forming dinoflagellates （Prorocentrum rnicans, Scrippsiella trochoidea and Nocfiluca scintillans） brought about by Lugol＇s iodine solution （hereafter Lugol＇s） and formalin. The fixation effects were species-specific. P. micans cell volume showed no significant change following long-term preservation, and S. trochoidea swelled by approximately 8.06% in Lugol＇s and by 20.97% in formalin as a percentage of the live cell volume, respectively. N. scintillans shrank significantly in both fixatives. The volume change due to formalin in N. scintillans was not concentration-dependent, whereas the volume shrinkage ofN. scintillans cells fixed with Lugol＇s at a concentration of 2% was nearly six-fold that in cells fixed with Lugol＇s at a concentration of 0.6%-0.8%. To better estimate the volume of N. scintillans fixed in formalin at a concentration of 5%, we suggest that the conversion relationship was as follows： volume of live cell=volume of intact fixed cell/0.61. Apart from size change, damage induced by fixatives on N. scintillans was obvious. Lugol＇s is not a suitable fixative for N. scintillans due to high frequency of broken cells. Accurate carbon biomass estimate ofN. scintillans should be performed on live samples. These findings help to improve the estimate of phytoplankton cell volume and carbon biomass in marine ecosystem.

Effect of Alexandrium tamarense on three bloom-forming algae

We investigated the allelopathic properties of Alexandrium tamarense(Laboar) Balech on the growth of Prorocentrum donghaiense Lu,Chattonella marina(Subrahmanyan) Hara et Chihara and Heterosigma akashiwo(Hada) Hada in a laboratory experiment.We examined the growth of A.tamarense,C.marina,P.donghaiense and H.Akashiwo in co-cultures and the effect of filtrates from A.tamarense cultures in various growth phases,on the three harmful algal bloom(HAB)-forming algae.In co-cultures with A.tamarense,both C.marina and H.akashiwo were dramatically suppressed at high cell densities;in contrast,the growth of P.donghaiense varied in different inoculative ratios of A.tamarense and P.donghaiense.When the ratio was 1:1(P.donghaiense:A.tamarense),growth of P.donghaiense was inhibited considerably,while the growth of P.donghaiense was almost the same as that of the control when the ratio was 9:1.The growth difference of P.donghaiense,C.marina and H.akashiwo when co-cultured with A.tamarense indicated that the allelopathic effect may be one of the important factors in algal competition and phytoplankton succession involving A.tamarense.In addition,the filtrate from A.tamarense culture had negative impacts on these three HAB algae,and such inhibition varied with different growth phases of A.tamarense in parallel with reported values of PSP toxin content in Alexandrium cells.This implied that PSP toxin was possibly involved in allelopathy of A.tamarense.However,the rapid decomposition and inactivation of PSP toxin above pH7 weakened this possibility.Further studies on the allelochemicals responsible for the allelopathy of A.tamarense need to be carried out in future.

Inhibitory activity of an extract from a marine bacterium Halomonas sp.HSB07 against the red-tide microalga Gymnodinium sp.(Pyrrophyta)

In recent years,red tides occurred frequently in coastal areas worldwide.Various methods based on the use of clay,copper sulfate,and bacteria have been successful in controlling red tides to some extent.As a new defensive agent,marine microorganisms are important sources of compounds with potent inhibitory bioactivities against red-tide microalgae,such as Gymnodinium sp.(Pyrrophyta).In this study,we isolated a marine bacterium,HSB07,from seawater collected from Hongsha Bay,Sanya,South China Sea.Based on its 16S rRNA gene sequence and biochemical characteristics,the isolated strain HSB07 was identified as a member of the genus Halomonas.A crude ethyl acetate extract of strain HSB07 showed moderate inhibition activity against Gymnodinium sp.in a bioactive prescreening experiment.The extract was further separated into fractions A,B,and C by silica gel column chromatography.Fractions B and C showed strong inhibition activities against Gymnodinium.This is the first report of inhibitory activity of secondary metabolites of a Halomonas bacterium against a red-tide-causing microalga.

Experimental Study on the Interspecific Interactions Between the Two Bloom-Forming Algal Species and the Rotifer Brachionus plicatilis

The interspecific interactions between the rotifer Brachionus plicatilis and two harmful algal blooms(HAB) species were investigated experimentally by single culture method. B. plicatilis population and the growth of the two algae were compared at different algal cell densities. The results demonstrated that the B. plicatilis obtained sufficient nutrition from Prorocentrum donghaiense to support net population increase. With exposure to 2.5×104 cells mL-1 of P. donghaiense,the number of B. plicatilis increased faster than it did when exposed to other four algal densities(5,10,15 and 20 ×104 cells mL-1),and the increase rate of B. plicatilis population(r) at this algal density was 0.104 ± 0.015 r d-1. Cell densities of P. donghaiense decreased due to the grazing of B. plicatilis. In contrast,Heterosigma akashiwo had an adverse effect on B. plicatilis population and its growth was largely unaffected by rotifer grazing. In this case,B. plicatilis population decreased and H. akashiwo grew at a rate similar to that of the control.

大亚湾三门岛珊瑚礁海域锥形宽水蚤(Temora turbinata)现场食物组成的分子生物学检测

浮游桡足类是连接海洋初级生产者和较高营养级生物的关键类群,了解其食物组成是理解海洋生态系统中物质与能量流通途径的基础。本文通过分子生物学手段研究大亚湾三门岛海域桡足类优势种锥形宽水蚤(Temora turbinata)现场食物组成,共检测出6种食物类群(住囊虫、甲藻、绿藻、有孔虫类、棘皮类、苔藓类),住囊虫是主要被摄食的生物类群(45.94%),其次是青绿藻纲(Prasinophyceae)的微微型真核自养生物(29.73%),另外还检测出2种锥形宽水蚤可能摄食的赤潮甲藻种类(Takayama acrotrocha和Karlodinium veneficum)。研究结果揭示了在食物限制环境下锥形宽水蚤灵活的摄食策略,突出了浮游动物在摄食传递过程及生态系统中的关键地位,将有助于解释浮游动物摄食行为在维持近岸生态系统平衡与稳定性中的作用。