Molecular identification of Prorocentrum （Dinophyceae） species     

张诚韩笑天  邹景忠俞志明  宋秀贤ChristianSchuett 《中国海洋湖沼学报》2005,23(3):317-322

A fragment of a large sub-unit ribosomal DNA (LrDNA) of 12 strains of Prorocentrum species was amplified by polymerase chain reaction (PCR). The PCR products were digested by 3 restriction endonucleases (Cfo Ⅰ, Hae Ⅲ, and RSA Ⅰ) and then resolved in agarose gels. Results show that different species had different RFLP patterns, except for P arcuatum (ME 131), which had the same pattern to P. micans (ME 160 and 04).The same fragment of 19 strains of the genus was also amplified and subjected to denaturing gradient gel electrophoresis (DGGE). 11 different patterns were resolved. Different cultures of a same species had the same pattern. The results of RFLP and DGGE analyses showed that eight newly isolated epibenthic Prorocentrum species were different from each other, and also from other cultured ones examined in this study. P arcuatum(MEI32) could not be differentiated from P. micans (MEI60 and 04), it was probably mis-identified, since they are quite different morphologically. P. redfieldii (MEI38) could also not be distinguished form P. triestinium(MEI32), it should be regarded as a synonym ofP. triestinium. Unexpectedly, a restriction site was found in P.micans, compared with previous sequence data.  相似文献   

THE EFFECTS OF NITROGEN,PHOSPHORUS,VITAMINS AND TRACE METALS ON THE GROWTH OF THE RED TIDE ORGANISM PROROCENTRUM MICANS   总被引：3，自引：0，他引：3  

王正方  张庆  龚敏 《中国海洋湖沼学报》1995,(4)

Laboratory culture experiments showed that < 100μmol/L nitrate, amonium or mixture of aminoacids promote the growth of the red tide organism Prorocentrum micans Ehrenb, but that >100μmol/Lof ammonium, or mixture of glycine and glutamate was harmful to growth, and that orthophosphatewas P. micrns’main phosphorous soruce in the ocean. Presence of 80μmol/L EDTA, 0.5 to 1μmol/LFe~(3+), 1.0 to 20.0 μmol/L Mn~(2+) 0.1 to 0.4 μmol/L Co~(2+) in the culture medium could improvethe growth of P. micans. Vitamin B_1 promoted growth, but vitamin B_(12) and biotin did not. Theestimated minimum cell quotaS (q_ο) for nitrogen and phosphorus being 0.74 pmole/cell and 0.045pmole/cell show that phosphorus (more than nitrogen) limits the growth of P. micans in the study area.  相似文献   

聚苯乙烯微塑料对东海原甲藻生长的影响          下载免费PDF全文

孙炎  刘千龙  罗肇河  康建华  孙秀武  张元标  林辉  潘钟 《应用海洋学学报》2021,40(4):636-642

微塑料是海洋中的一种污染物,近年来已成为全球关注的热点环境问题。东海原甲藻（Prorocentrum donghaiense）是我国东海海域常见的有害赤潮藻,目前尚未有关于微塑料污染对东海原甲藻影响研究的报道。为了探讨聚苯乙烯（PS）微塑料对东海原甲藻生长的影响机制,阐明海洋微塑料对东海原甲藻生长的影响,本研究开展了东海原甲藻在聚苯乙烯微塑料差异化暴露条件下的生长动力学室内模拟实验。结果显示：①5 mg/L溶解性有机质（DOM）可促进东海原甲藻的生长;②10 mg/L（0.1 μm粒径）的聚苯乙烯微塑料对东海原甲藻前期生长的影响,表现为先抑制、后促进;③聚苯乙烯微塑料会抑制处于稳定期和衰退期东海原甲藻的生长;微塑料粒径越小、浓度越高抑制作用越强;浓度的抑制效应强于粒径的。我们研究发现聚苯乙烯微塑料会影响东海原甲藻的生长,这将为微塑料和赤潮藻复合污染生态效应以及海洋生态系统保护提供新的科学依据。  相似文献   

改性粘土对球形棕囊藻(Phaeocystis globosa)和东海原甲藻(Prorocentrum donghaiense)的去除作用     

邱丽霞  俞志明  曹西华  宋秀贤  刘扬  钟怡 《海洋与湖沼》2017,48(5):982-989

利用改性粘土去除藻华生物是目前有害藻华应急处置最常用的方法。本文研究了六种铝盐改性粘土(PAC-MC、PAFC-MC、PAFCs-MC、AC-MC、AS-MC、PAS-MC)对藻华生物球形棕囊藻(Phaeocystis globosa)、东海原甲藻(Prorocentrum donghaiense)的去除效率,考察了悬浮液pH、改性粘土颗粒表面电位及粒径分布等因子对去除效率的影响。结果显示:不同赤潮生物由于生物特征不同,其去除效率存在较大差异,PAC-MC、PAFC-MC、PAFCs-MC对东海原甲藻有较高的去除效率,但对于球形棕囊藻去除能力较差;AC-MC、AS-MC、PAS-MC对两种藻华生物均有较好的去除效果,但对水体酸碱扰动较大,在pH敏感水域应注意用量;对于同一种改性粘土,提高铝离子含量、增加改性粘土浓度有利于除藻效率的提升;自絮凝程度越低、表面正电性越强(或负电性越弱)、悬浮液pH值越低的改性粘土,除藻能力越强。本研究进一步为改性粘土应急处置有害藻华提供了参考。  相似文献   

北部湾三角棘原甲藻(甲藻门原甲藻目)的形态观察和18S rDNA序列分析          下载免费PDF全文

龙超  陈宪云  陈波  何碧娟  高程海  王一兵 《热带海洋学报》2014,33(2):66-71

利用光学显微镜、分子生物学方法,对分离自广西北部湾海域的三角棘原甲藻Prorocentrum triestinum BBW-02藻株的形态特征进行了描述,并探讨了其分子系统进化关系。三角棘原甲藻BBW-02细胞呈长卵形或披针形,细胞长20~26μm,宽10~14μm,顶刺形如矛尖,刺丝胞孔数量较少,排列不规则,主要分布于壳板边缘,距离细胞底端1/3处壳板两侧各有2个近于对称排列的刺丝胞孔。18S rDNA序列同源检索和系统进化分析表明,三角棘原甲藻BBW-02与其他地理株系的三角棘原甲藻聚为一支,亲缘关系密切,遗传差异仅为0.000~0.001。与其他种类相比,三角棘原甲藻BBW-02与同属中的P.gracile亲缘关系最近。  相似文献   

Experimental study on the interspecific interactions between the two bloom-forming algal species and the rotifer Brachionus plicatilis     

Zhihao Xie  Hui Xiao  Xuexi Tang  Hengjiang Cai 《中国海洋大学学报(英文版)》2009,8(2):203-208

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 rd⁻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.  相似文献   

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

HOU Jianjun  LAI Hongyan  HUANG Bangqin  CHEN Jixin 《海洋学报(英文版)》2009,28(2)

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 fuorescence 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 hyrbridization efficiency of difierent probes specific to P.minimum was in the order:PMl8S02＞PM28S02＞PM28S01＞PM18S01,and that of the probes specific to T. pulchella was TP18S02＞TP28S01＞TP28S02＞TP18S01.The djfferent hybridization efficiency of the DNA probes could also be shown in the fuorescent signals between the labeled and unlabeled cells demonstrated using flow cytometry.The DNA probes PM18S02,PM28S02,TPl8S02 and TP28S01,and the protocol,were also useful for the detection of algae in natural samples.  相似文献   

Detection of Prorocentrum donghaiense using sandwich hybridization integrated with nuclease protection assay   总被引：1，自引：0，他引：1  

CHEN Jie  ZHEN Yu  MI Tiezhu  YU Zhigang 《海洋学报(英文版)》2009,28(2):121-126

Prorocentrum donghaiense is an important harmful algae bloom (HAB) causing creature in China’s seas, and the conventional visual detection can not cope with long-term monitoring and high-throughput sampling projects. An assay for P. donghaiense with sandwich hybridization integrated with nuclease protection assay (NPA-SH) was established. Tests with mixed samples and spiked field ones confirmed its good specificity and sensitivity. The cell number of P. donghaiense correlated well with the optical density, ...  相似文献   

Development of taxonomic rRNA-targeted probes of two harmful algae: Prorocentrum minimum and Karenia mikimotoi by fluorescence in situ hybridization     

CHEN Guofu  WANG Quanfu  ZHANG Chunyun  ZHANG Baoyu  WANG Guangce  LU Douding  XU Zhong  YAN Peishen 《海洋学报(英文版)》2013,32(2):66-75

Harmful algal blooms recently have been under the spotlight throughout the world, because of their negative impact on the marine environment, aquaculture, fisheries as well as public health. The development of methods for rapid and precise identification and quantification of causative species is essential for the warning and monitoring of blooms, among which the techniques based on taxonomic probes are the most favored. In this study, two harmful algae, i.e., Prorocentrum minimum and Karenia mikimotoi were taken into consideration. The partial large subunit rDNA (D1-D2) of both species were firstly PCR-amplified, cloned and sequenced. The obtained sequences were then introduced to carry out alignment analysis for gene specific regions. Three respective candidate probes for each species were designed and used to screen the optimal probe by performing fluorescence in situ hybridization (FISH) tests. The results showed that the probes Pmin0443 and Kmik0602 displayed the best hybridization for P. minimum and K. mikimotoi, respectively. Both the specific (taxonomic) (Pmin0443 and Kmik0602) and the control probes (UniC0512 and UniR0499) were used for cross-reactivity tests with other microalgae in our laboratory. The probes Pmin0443 and Kmik0602 are specific and could be served as taxonomic probes introduced into the techniques targeting rRNA, such as FISH, sandwich hybridization, and DNA-microarray assay of P. minimum and K. mikimotoi in the future. Finally, FISH analyses with both probes were performed on the simulated field samples. The probes could hybridize exclusively with the target cells well, and no significant difference (p >0.05) was observed in the cell densities of the samples determined by FISH and light microscopy (LM). All suggest that the probes are specific and could be introduced into FISH for the monitoring of both harmful algae.  相似文献   

FITC‐conjugated lectins as a tool for differentiating between toxic and non‐toxic marine dinoflagellates     

Lesley L. Rhodes  Allison J. Haywood  David W. Fountain 《新西兰海洋与淡水研究杂志》2013,47(3):359-365

FITC‐conjugated lectins proved to be effective probes for differentiating between morphologically similar dinoflagellate species isolated from New Zealand coastal waters. In particular the binding (fluorescence) of peanut (PNA) lectin differentiated G. mikimotoi from Gymnodinium sp. (Waimangu) and G. pulchellum and the non‐binding of Helix pomatia (HPA) and wheat germ (WGA) lectins discriminated between G. mikimotoi and the other Gymnodinium species tested. G. breve (Florida) was differentiated from the New Zealand isolates by binding with soy bean (SBA) lectin. Ulex europeus (UEA) distinguished the toxic species Alexandrium minutum from the morphologically similar, but non‐toxic, Cachonina hallii. Two strains of Prorocentrum lima (Spain and Rangaunu) were not differentiated by the lectins, but P. lima was differentiated from P. compressum.  相似文献