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鲴亚科和相关类群线粒体DNA的分子进化及鲴亚科与寄生六鞭毛虫协同进化的研究
肖武汉
Subtype博士
Thesis Advisor潘金培 ; 王伟俊 ; 张亚平
1997
Degree Grantor中国科学院水生生物研究所
Place of Conferral中国科学院水生生物研究所
Degree Discipline水生生物学
Keyword鲴亚科 鲤科 六鞭毛虫科 线粒体dna 限制性片段长度多态性 Cytb基因序列 Nd4l-nd4基因序列 系统发育 协同进化
Abstract鲴亚科(Xenocyprinae)属鲤形目(Cypriniformes)、鲤科(Cyprinidae),是一类主要分布于东亚地区的淡水鱼类,在属级和种级水平,其分类都存在许多争议,有关其系统发育的研究也很贫乏。鲴亚科与寄生六鞭毛虫相互间的关系,也令人绕有兴趣。本研究中:I(1)建立了制备鱼类线粒体DNA地高辛标记探针的简易方法;(2)选择21种限制性内切酶,利用Southern blot方法,分析了鲴亚科4属7种12个群体103个个体线粒体DNA的限制性片段长度多态性,共检测到32种单倍型。利用UPGMA和NJ法构建了32种单倍型的分子系统树。(3)构建了17种酶、27种单倍型的限制性内切酶图谱。运用简约法、距离法,进行了系统发育分析,结果表明:银鲴与黄尾鲴亲缘关系最近。湖北圆吻鲴和鲴属的亲缘关系较近,而与圆吻鲴的关系较远。鲴亚科中,似鳊的分化较早。(4)银鲴四个自然群体的遗体结构研究表明:湖北和湖南群体的遗传多样性较为丰富,福建与四川群体的遗传多样性则相对较贫乏。湖南和湖北群体间的关系较近;湖南、湖北群体与福建群体的亲缘关系近,而与四川群体的亲缘关系较远。湖南和湖北两地银鲴和似鳊群体mtDNA基因型间存在较大差异,可能是由于这两种鱼不同的生态习性决定的。II(1)建立了从Formalin固定标本中提取大片段DNA(1.2kb),并进PCR扩增和测序的方法。(2)测定了鲴亚科4属10种30个个体的确Cytb基因1140bp和17个体的ND4L,ND4,tRNA~(His)和tRNA~(ser)四个基因约1812bp的DNA序列。运用简约法、距离法和最大似然法对鲴亚科进行了系统发育分析,结果表明:鱼尾鲴和银鲴的亲缘关系最近,它们单倍型间的交叉可能是由于存在杂交渐渗。湖北圆吻鲴与鲴属的亲缘亲系较近,而与圆吻鲴的亲缘关系较远细鳞鲴与方氏鲴组成的一支与鲴属其它种的分化明显。似鳊和圆吻鲴与鲴属的分化较早。在鲴亚科鱼类,mtDNA的进化速度较快,接近哺乳类的进化速率。(3)测定了鲤科11亚科(不含鲴亚科)22个属,29个个体的Cytb基因1140bp的序列,并利用从Genebank中查得的鱼类Cytb基因序列,进行了系统发育分析,结果表明:软骨鱼类与硬骨鱼类的分化较早,多鳍鱼可能与硬骨鱼的亲缘关系较近;鲤亚目、鲤科可能都为单系群;鲤科鱼类可分为鱼巴系和雅罗鱼系。鲴亚科可能不是一个单系群。(4) 测定了鲤科8个亚科(不含鲴亚科)15个种的ND4L,ND4,tRNA~(His),tRNA~(ser)四个基因约1814bp的DNA序列,系统发育分析也表明:鲤科鱼类可能为一单系群,可分为鱼巴系和雅罗鱼系;鲴亚科不是一个单系群;ND4基因提供的系统发育信息比Cytb基因好。III(1)选择24个显微和超微结构特征,利用支序系统学原理和方法对鲴亚科寄生六鞭毛虫17个种进行了系统发育分析。NJ树和简约树的拓扑结构有很大差异;简约树各分枝的支持率都太低,可能表明:所选用特征含信息量太小。(2)根据鲴亚科寄生六鞭毛虫的区系组成,利用PAUP3.1.1构建了宿主的系统进化树,发现:各分枝的支持率都在50%以上,其拓扑结构与依据mtDNA序列数据构建的分子系统树高度一致,该结果表明:寄生虫的区系组成,能够反映宿主间的亲缘关系。
Other AbstractXenocyprinae belonging to Cyprinidae, Teleostei, is a group of freshwater fishes mainly distributed in East-Asia. Not only are there many contradictions about its taxonomy at both genus and species level, but also little is know about its phylogeny. Furthermore, the relationship between Xenocyprininae and parasitic Hexamita is very special and interesting. In this study, the results have been obtained as the following: (1)A simple method for getting digoxigenin labelling probe of fish mtDNA has been established. (2)The RFLP analysis of mtDNA of 103 individuals belonging to Xenocyprinae being detected and the phylogram of the haplotypes was constructed by UPGMA and NJ. (3)The restrction maps of 27 haplotypes were constructed by double digested. The phylogenetic analysis by parsimony and neighbor-joining methods suggests that Xenocypris argentea and X. dividi are closely related and the relationship between Distoechodon hupeiensis and the genus Xenocypris is closer than that of between D. hupeiensis and D. tumirostris. The divergent time of Pseudobrama simoni was early in the subfamily. (4)The population genetic structure among four natural populations of X. argentea was analysed. The populations of Hubei and Hunan is rich in genetic diversity, but the populations of Fujian and Sichuan is poor. The relationship between Hubei-Hunan population and Fujian population is closer than that of between Hubei-Hunan and Sichuan. (5)A method of extracting large fragment DNA(1.2kb) from formalin-fixed specimens was developed. (6)The mitochondrial Cytb, ND4L, ND4, tRNA~(His), and tRNA~(ser) genes for 10 species from 4 genera of Xenocyprinae have been sequenced. A total of 2952 characters for each individuals were obtained. Our interpretation of the results from parsimony, distance and maximum likelihood based phylogenetic analysis suggests : X. argentea and X. dividi are closely related, and the relationship between D. hupeiensis and Xenocypris is closer than that of between D. hupeiensis and D. tumirostris. X. microlepis and X. fangi are sister groups., the ratio of mtDNA divergence in Xenocyprinae is seemly faster than that of other fishes having been reported and is nearly equal to that of mammals. (7)The mitochondrial Cytb genes for 22 genera from 11 subfamilies of Cyprinidae have been sequenced and the Cytb gene sequences of fishes in 9 orders have also been copied from Genebank. The results of phylogenetic analysis suggest : both the order Cypriniformes and the family Cypriniae could be regarded as monophyletic group. The family Cypriniae could be divided into two series, Leuciscini and Barbini. The subfamily Xenocyprinae may not be regarded as a monophyletic group. (8)The mitochondrial ND4L, ND4, tRNA~(His), and tRNA~(ser) genes for 15 species from 8 subfamilies have been sequenced. The results obtained in phylogenetic analysing of ND4L-ND4 gene sequences were the same as analysing Cytb gene, but the boostrap is higher at each branch. The results also show that as a phylogenetic maker ND4 gene may be better than Cytb gene. (9)The cladictic analysis using 24 characters of 17 species belonging to Hexamita parasitic in Xenocyprinae could not illustrated their phylogenetic relationships. (10)The phylogenetic tree of Xenocyprinae constructed by the fauna of parasitic Hexamita is the same as that of constructed by the method of molecular systematics. The results suggests that the fauna of parasite could reflect the relationship of host.
Pages166
Language中文
Document Type学位论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/12514
Collection学位论文
Recommended Citation
GB/T 7714
肖武汉. 鲴亚科和相关类群线粒体DNA的分子进化及鲴亚科与寄生六鞭毛虫协同进化的研究[D]. 中国科学院水生生物研究所. 中国科学院水生生物研究所,1997.
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