几株原甲藻核糖体大亚基RNA基因的部分克隆及序列分析
Cloning and analysis of 28S rDNA partial sequences of several strains of Prorocentrum
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摘要: 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.Abstract: The 28S rDNA partial sequences of several strains of Prorocentrum were cloned and sequenced, and other 14 corresponding sequences of Prorocentrum were obtained from Genbank.Then the phylogenetic trees of Prorocentrum were constr ucted with the methods of NJ(neighbour-joining) and M E(minimume-volution).The results demonstrated that the sequenced partial 28S rDNA of 7 strains was 950~958 bp.The phylogenetictree based on the NJ method was similar with that based on the ME method.The sequences of the different strains within the same species are highly conservative;other wise, the diversity among the sequences from different species was obvious.P.micans separated from the South China Sea was very different from the P.micans strains separated from other area of the world, and the dist ances between them were even long erthan the distances between two species.The sequences from the 28S rDNA would be helpful to design species-specific probes of the phytoplankton.
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Key words:
- Prorocentrum /
- 28S rDNA /
- phylogenetic analysis
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齐雨藻.中国沿海赤潮[M].北京:科学出版社,2003.19-21. 王金辉,黄秀清.具齿原甲藻的生态特征及赤潮成因浅析[J].应用生态学报,2003,14(7):1 065-1 069. MORTONSL.Maximizing okakaic acid content from Prorocentrum hoffmannianum Faust[J].J Applic Phycol,1994,6(1):41-44. GRZEBYKD,DENARDOUA,BER Land B,et al.Evidence of a newt oxin in the redtide dinofl agellate Prorocentrum minimum[J].J Plankton Res,1997,19(8):1111-1124. LEVASSEURM,COUTUREJY,SAUVEG,et al.Contamination of molluscs in Quebec by diarrhetic(DSP)and amnesic(ASP)phycotoxins and research into possible sources for DSP phycotoxins[J].Can Tech Rep Fish Aquat Sci,2001,2 350:51. STOCKDW,WHITTGS.Evidence from 18S riboromal RNA sequences that lampreys and hagfishes form a natural group[J].Science,1992,257:787-789. 陈月琴,王宁,周惠,等.棕囊藻赤潮原因种的分子鉴定和起源分析[J].海洋学报,2002,24(6):99-103. 李文红,胡自民,覃志彪,等.细基江蓠及其繁枝变种的RAPD 和ITS分析[J].海洋学报,2004,26(6):89-95. 张纹,王军,苏永全,等.两种新贝尼登虫28SrRNA序列分析及其亲缘关系探讨[J].海洋学报,2004,26(5):116-122. ELLISJ T,AMOYALG,RYCEC,et al.Comparison of the large subun it ribosomal DNA of Neos pora and Toxoplasma and devel opment of a new genetic marker for their differentiation based on the D2 domain[J].Molecular and Cellular Probes,1998,12:1-13. SHOKOHOSO -ITANABE,SAKOYOSHIHIKO.Rapid detection of natural cells of Alexandriumt amarense and A.catenella(Dinophyceae)by fluores cence in situ hybridization[J].Harmful Algae,2005,4(2):319-328. MIKULSKICM,MORTONSL,DOUCETTEGJ.Devel opment and application of LSU rRN Aprobes for Karenia brevis in the Gulf of Mexico,USA[J].Harmful Algae,2005,4(1),49-60. 吕颂辉,张玉宇,陈菊芳.东海具齿原甲藻的扫描电子显微结构[J].应用生态学报,2003,l 4(7):1070-1072. 辜小莲,齐雨藻.大亚湾水域原甲藻调查与鉴定[J].热带亚热带植物学报,2002,10(4):335-339. 胡晗华,石岩峻,丛威,等.微小原甲藻的生长及其对锌限制的响应[J].应用生态学报,2003,14(7):1140-1142. 谷颖,项有堂.象山港海域富营养化与赤潮的关系[J].海洋环境科学,2002,21(3):67-69. 黄贯虹,黄伟健,方刚,等.大鹏湾优势藻引发赤潮的灰分析[J].生态学报,2002,22(6):822-827. DANIELG,YOSHIHIKOS.Phylogenetic analysis of nine species of Prorocentrum(dinoph yceae)inferred from 18S ribosomal DNA sequence,Morphological comparisons,and description of Prorocentrum panamensis,sp.nov.[J].J Phycol,1999,34:1055-1068. ZARDOYAR,COSTASE,LOPEZ-RODAV,et al.Revised dinoflagel late phylogeny in ferred from molecular analysis of large-subunitribosomal RNA gene sequence[J].J Mol Evol,1995,41:637-645. 苟万里,孙军,何闪英,等.几种角毛藻亲缘关系的分子生物学分析[J].高技术通讯,2003,13:68-73. THOMPSONJD,GIBSONTJ,PLEWNIAKF,et al.The Clustal Xwindows interface:flexible strategies for multiple sequence alignmentaided by quality analysis tools[J].Nucleic Acids Res,1997,24:4876-4882. KIMURAM.Asimple method for estimating evolution ary of base substation through comparative studies of nucleotide sequences[J].J Mol Evol,1980,16:111-120. SAITOU N,NEIM.The neighbour-joining method:A new method for reconstructing phylogenetic trees[J].Mol Bio Evol,1987,4:406-425. FELSENSTEINJ.Confidence limits on phylogenies:An approach using the bootstrap[J].Evolution,1985,39:783-791. 王波,米铁柱,吕颂辉,等.五种/株原甲藻核糖体小亚基(18S rRNA)基因克隆及序列分析[J].海洋与湖沼,2006,37(5):450-456. CORTES-ALTAM IRANOR,SIERRA-BELTRANAP.Morphology and taxonomy of Prorocentrum mexicanum and reinstatement of Prorocentrum rhathymum(dinophy ceae)[J].Journal of Phycology,2003,39(1),221-230. 陆斗定,齐雨藻,Jeanette Goebel,等.东海原甲藻修订及相关原甲藻的分类学比较[J].应用生态学报,2003,14(7):1060-1064. TYRRELLJV,CONNELLLB,SCHOLINCA.Monitoring for Heterosig maaka shiwo using a sandwich hybridization assay[J].Humful Algae,2002,1(2):205-214. ANDERSONDM,KULISDM,KEAFERBA.Detection of the toxic Din of lagellate A lex and rium Fundyende(Dinophyceae)with oligonu cleotide and antibody probes:variablith in labeling intensity with physiological condition[J].J Phycol,1999,35:70-883. MILLERPE,S CHOLINCA.Ident ification of cultured Pseudo-nitzschia(Bacillariophyceae)using species-specific LSU rRNA-targeted fluorescent probes[J].J Phycol,1996,32:644-655.
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