几种典型藻华生物的分子分类学分析  

MOLECULAR TAXONOMY ANALYSES OF THE RELATIONSHIPS IN SEVERAL REPRESENTATIVE HARMFUL ALGAL BLOOM SPECIES

作　　者：侯建军[1,2,3,4,5] 赖红艳[2] 黄邦钦[3] 刘绍平[1] 

机构地区：[1]中国水产科学研究院长江水产研究所,农业部淡水鱼类种质资源与生物技术重点开放实验室,荆州434000 [2]湖北师范学院生命科学学院,黄石435002 [3]厦门大学近海海洋环境科学国家重点实验室,厦门361005 [4]中国科学院水生生物研究所,淡水生态与生物技术国家重点实验室,武汉430072 [5]华中农业大学水产学院,武汉430070

出　　处：《水生生物学报》2008年第5期711-719,共9页Acta Hydrobiologica Sinica

基　　金：国家重点基础研究发展规划973项目(编号:CEOHAB2001CB409704);中国博士后基金项目(编号:20060400854);教育部博士点基金项目(编号:20070504076);淡水生态与生物技术国家重点实验室开放课题(编号:2008FB005);农业部淡水鱼类种质资源与生物技术重点实验室开放课题(编号:LFB20070611)资助

摘　　要：通过PCR克隆测序、rDNA序列分析、随机PCR引物扩增结合DGGE技术等三个层次的分子分类水平对厦门海域的典型藻华生物进行分子生物学分析。结果表明,基于DGGE检测的18S rDNA序列过于保守,分类的精确度不高;AP-PCR则是基于基因组的差异进行分析,结果较为精确和全面;而rDNA序列分析相对可靠,尤其是针对ITS的长度和全序列分析以及28S rDNA的D1、D2区的序列分析,可以提供更为准确的分类信息,并可为设计检测探针提供基础。据此对分离自厦门海域的3种典型甲藻及其相关藻株建立了系统进化树。针对23株甲藻ITS序列建立的系统进化树显示Takayama pulchella(AY764179)和Karlodinium micrum的距离最近,并能够把Akashiwo属、Karenia属、Gyrodinium属、Karlodinium属、Takayama属与Gymnodinium属等区分开。用28S rDNA序列建立的系统发育树只能把Karlodinium属、Karenia属和Takayama属区分开,但不能很好地把无纹环沟藻与Akashiwo属和Gymnodinium等的藻区分开。With rapid economic development and increasing pollution,some Chinese coastal waters have become eutrophic,which has resulted in a high frequency of harmful algal blooms.Rapid and unequivocal identification of harmful species has become a focal point of recent toxic algal research.At present,morphological criteria are the primary means to identify and classify harmful algae,but it is often difficult to distinguish morphologically-similar species and differentiate non-toxic from toxic algae.Some new molecular taxonomy methods and related technologies must be developed and applied to differentiate non-toxic from toxic algae and to monitor the development of algal blooms in coastal waters.Application of the ribosomal DNA（rDNA） sequences to identification and classification harmful algal bloom species are important complements for the traditionally morphological identification,and can give us more exact taxonomic informations and valuable results sometimes.Moreover,analysis of phylogenies and relationships among harmful algal bloom species using sequences of the rDNA can provide a basis on molecular probes design and can detect target algae quantitatively and rapidly.Molecular taxonomy has proved particularly useful to separate closely related species and has being the most common focuses on phytoplanktonic ecology.In the present study,the gene fragments of 18S,28S rDNA and ITS（Internal transcribed spacer） of several representative harmful algal bloom species from Xiamen harbor were amplified by polymerase chain reaction and cloned,and then their sequences were mensurated and analyzed by software.The variable regions of 18S rDNA were amplified by specific primer PCR and then analyzed by denaturing gradient gel electrophoresis.The genomic differences of these harmful algal bloom species were analyzed by AP-PCR.The different levels of molecular taxonomy were described by the above-mentioned methods,which were used to analyze the molecular taxonomy relationships in these harmful algal bloom species from Xiamen h

关 键 词：藻华生物 分子分类学 核糖体DNA 变形梯度凝胶电泳 随机引物PCR 

分 类 号：Q344.1[生物学—遗传学]