三种鲍线粒体基因测序及系统发育分析

发布时间：2018-03-04 11:23

本文选题：皱纹盘鲍　切入点：黑唇鲍　出处：《山东大学》2017年硕士论文　论文类型：学位论文

【摘要】：线粒体16SrRNA、CO Ⅰ、COⅡ基因是物种线粒体基因研究中比较普遍的基因序列,在群体遗传结构,种系发生及亲缘关系研究等方面应用广泛,是比较忠实的遗传分子标记。本文选取中国的皱纹盘鲍(H.discus hannai),澳大利亚的黑唇鲍(H.rubra)和新西兰的黑足鲍(H.iris)3种典型经济鲍作为研究对象,通过测定其线粒体16SrRNA、COⅠ、COⅡ基因序列片段,来分析其单核苷酸多态性及种内种间一致性和系统发育关系,为鲍的新品种引进、杂交育苗提供理论支持。本研究主要获得以下结果:(1)三种鲍线粒体16S rRNA基因片段测序及系统发育分析采用基因测序方法,对皱纹盘鲍(H.hannai)、黑足鲍(H.iris)和黑唇鲍(H.rubra)线粒体16S rRNA基因序列片段进行PCR扩增和测序。试验得到的序列长度分别为皱纹盘鲍544bp,黑唇鲍550bp,黑足鲍546bp;A+T碱基含量比例分别为皱纹盘鲍58.1%,黑唇鲍56.2%,黑足鲍56.4%;种内一致性分别为皱纹盘鲍96.37%,黑唇鲍97.51%,黑足鲍99.06%。三种鲍的种间一致性为93.94%。皱纹盘鲍的碱基突变率为0.37%,黑足鲍的碱基突变率为0.55%,黑唇鲍的碱基突变率为0.36%。黑足鲍与黑唇鲍间的遗传距离为0.148,皱纹盘鲍与黑唇鲍间的遗传距离为0.116,皱纹盘鲍与黑足鲍间的遗传距离为0.077。(2)三种鲍线粒体CO Ⅰ基因片段测序及系统发育研究分析采用基因测序方法,对皱纹盘鲍(H.hannai)黑足鲍(H.iris)和黑唇鲍(H.rubra)线粒体COⅠ基因序列片段进行PCR扩增和测序。实验得到的序列长度分别为皱纹盘鲍554bp,黑唇鲍573bp,黑足鲍552bp;A+T碱基含量比例分别为皱纹盘鲍55.4%,黑唇鲍52.3%,黑足鲍53.5%;种内一致性分别为皱纹盘鲍97.66%,黑唇鲍98.49%,黑足鲍97.42%。三种鲍的种间一致性为89.12%。皱纹盘鲍的碱基突变率为0.36%,黑足鲍的碱基突变率为0.36%,黑唇鲍的碱基突变率为2.09%。黑足鲍与黑唇鲍间的遗传距离为0.204,皱纹盘鲍与黑唇鲍间的遗传距离为0.200,皱纹盘鲍与黑足鲍间的遗传距离为0.186。(3)种鲍线粒体COⅡ基因片段测序及系统发育研究分析采用基因测序方法,对皱纹盘鲍(H.hannai)、黑足鲍(H.iris)和黑唇鲍(H.rubra)线粒体COⅡ基因序列片段进行PCR扩增和测序。实验得到的序列长度分别为皱纹盘鲍151bp,黑唇鲍150bp,黑足鲍153bp;A+T碱基含量比例分别为皱纹盘鲍58.3%,黑唇鲍53.7%,黑足鲍56.5%;种内一致性分别为皱纹盘鲍98.23%,黑唇鲍99.33%,黑足鲍91.83%。三种鲍的种间一致性为92.51%。皱纹盘鲍的碱基突变率为3.31%,黑足鲍的碱基突变率为19.61%,黑唇鲍的碱基突变率为2.00%。黑足鲍与黑唇鲍间的遗传距离为0.124,皱纹盘鲍与黑唇鲍间的遗传距离为0.192,皱纹盘鲍与黑足鲍间的遗传距离为0.071。以上结果表明:线粒体16SrRNA、COⅠ、COⅡ基因序列片段适用于鲍种间遗传分析、系统发生及亲缘关系研究。数据表明线粒体16SrRNA序列最为保守,COⅡ基因序列碱基突变率最高。皱纹盘鲍与黑足鲍间的亲缘关系最近。属于同一地理种群的鲍亲缘关系较近。
[Abstract]:Mitochondrial 16SrRNA, CO I and CO II gene is the gene sequence comparison research of mitochondrial genes in common species, in terms of population genetic structure, phylogeny and genetic relationship of genetic molecular markers is widely used, more loyal. This paper selects the China Abalone (H.discus hannai), Australia (black lip abalone H.rubra) and new Zealand h.iris (H.iris) of 3 typical economic abalone as the research object, through the determination of the mitochondrial 16SrRNA, CO 1, CO 2 gene fragment, to analyze the single nucleotide polymorphism and intraspecific and interspecific relationship and a system development, the introduction of new varieties of abalone, provide hybrid breeding the theory of support. The main results are as follows: (1) analyzed by gene sequencing method of three kinds of abalone mitochondrial 16S rRNA gene fragment sequencing and phylogenetic of Abalone (H.hannai), h.iris (H.iris) and black lip Abalone (H.rub RA) of mitochondrial 16S rRNA gene fragment was amplified by PCR and sequenced. Sequence length tests were Haliotis 544bp, black lip abalone 550bp, h.iris 546bp; A+T base content ratio was 58.1% black abalone, abalone lip 56.2%, h.iris 56.4%; internal consistency were wrinkles 96.37% black abalone, abalone lip 97.51%, a consistency between h.iris 99.06%. three kinds of abalone Haliotis discus hannai 93.94%. mutation rate was 0.37%, h.iris mutation rate was 0.55%, the black lip abalone mutation rate of 0.36%. h.iris and genetic distance between the black lip abalone 0.148, the genetic distance of abalone Haliotis with black lip abalone between 0.116 abalone abalone and h.iris genetic distance between the 0.077. (2) and three sequencing of mitochondrial CO gene fragment of abalone system development research and analysis by gene sequencing method of Abalone (H.hannai) h.iris (H.iris) abalone and black lips (H.rubra) of mitochondrial CO gene fragment was amplified by PCR and sequenced. The sequence length obtained from the experiment were discus hannai Ino 573bp 554bp, black lips, h.iris 552bp; A+T base content ratio was 55.4% black abalone, abalone lip 52.3%, h.iris 53.5%; internal consistency. For the 97.66% black abalone, abalone lip 98.49%, a consistency between h.iris 97.42%. three kinds of abalone Haliotis discus hannai 89.12%. mutation rate was 0.36%, h.iris mutation rate was 0.36%, the black lip abalone base mutation rate of the genetic distance 2.09%. h.iris and black abalone between lips the genetic distance was 0.204, abalone abalone and black lip abalone between 0.200 abalone abalone and h.iris genetic distance between the 0.186. (3) of the mitochondrial CO fragment and sequencing system Bao II gene phylogenetic analysis by gene sequencing method of Abalone (H.hannai), h.iris (H.iris) and black lip abalone (H.rubra) CO II mitochondrial gene fragment was amplified by PCR and sequenced. The sequence length obtained from the experiment were discus hannai Ino 150bp 151bp, black lips, h.iris 153bp; A+T base content ratio was 58.3% black abalone, abalone lip 53.7%, h.iris 56.5%; internal consistency. For the 98.23% black abalone, abalone lip 99.33%, a consistency between h.iris 91.83%. three kinds of abalone Haliotis discus hannai 92.51%. mutation rate was 3.31%, h.iris mutation rate was 19.61%, the black lip abalone mutation rate of 2.00%. h.iris and Powell left among the black lip transmission distance is 0.124, the genetic distance of abalone Haliotis with black lip abalone between 0.192 abalone abalone and h.iris genetic distance of 0.071. between the above results show that mitochondrial 16SrRNA, CO 1, CO 2 gene fragment for abalone interspecific genetic analysis, occurrence and genetic relationship of system data table. Open wire The 16SrRNA sequence was the most conservative. CO II gene sequence had the highest mutation rate. The relationship between Haliotis discus and abalone Haliotis was the closest.

【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S917.4

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