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Alternative TitleIsolation of microsatellite markers and population genetic analysis
Thesis Advisor桂建芳
Degree Grantor中国科学院水生生物研究所
Place of Conferral水生生物研究所
Keyword大黄鱼 银鲫 微卫星 遗传分析 克隆鉴定
Abstract大黄鱼与银鲫分别是我国海水与淡水重要的经济鱼类。本研究依据研究对象不同分为两大部分。在大黄鱼的研究中,我们首先利用FIASCO 方法分离出微卫星标记,找到15个高度多态的特异性引物。选其中12个微卫星标记,以及4对AFLP选择性引物用于官井洋大黄鱼的一个自然群体和三个养殖群体的研究。所有结果经分析说明养殖行为已经造成了野生以及养殖大黄鱼多态性的下降以及一定程度的遗传分化。 在银鲫方面,我们使用数据挖掘和FIASCO法从银鲫中开发微卫星标记。利用这些引物对本实验室所保种的(A,D,L和F)四个雌核发育克隆系进行鉴别研究并在二倍体鲫鱼中进行扩增。找到12对可以区分全部品系的微卫星标记,分析品系间亲缘关系发现,D 与 L遗传距离最小,而与 F距离最大。在鲫鱼中扩增,除YJ0022失败外,其余18对引物均获得稳定扩增结果,并且发现14个多态性标记。利用12个微卫星标记对银鲫六个地理群体以及2个鲫鱼地理群体进行分析,认为目前长江流域存在天然银鲫,同时也有相当比例养殖银鲫渗入天然群体中,并且银鲫在天然水体中可能主要行两性生殖。研究结果支持淇河银鲫来自中国江汉平原区系的二倍种的假说,并进一步认为新疆银鲫也发源于长江流域,但不同银鲫地理群体起源不同。
Other AbstractThe large yellow croaker (Pseudosciaena crocea Richardson) and the gibel carp (Carassius auratus gibelio) are both commercially important fish species in China. This dissertation consists of two sections. One is the study of the large yellow croaker as following: Firstly Some polymorphic microsatellites from enriched-genomic libraries of the large yellow croaker were isolated using the Fast Isolation by AFLP of Sequences COntaining repeats (FIASCO) protocol and fifteen of them were designed primers and characterized. All of the fifteen microsatellite markers were polymorphism enough for populational genetic research of the large yellow croaker. Secondly, twelve polymorphic microsatellite loci and four pairs of AFLP selective primers were used to evaluate the genetic variation of one wild-captured and three cultured populations of the large yellow croaker. The results indicate the hatchery operations have lessened the genetic diversity of the large yellow croaker populations in guanjingyang and led to genetic divergence among natural population and cultured populations of the large yellow croaker. The other part is about the gibel carp. One hundred and nine microsatellites were isolated from gibel carp (Carassius auratus gibelio) by screening through genes found in GenBank, Gibel Carp Gene Database (GCGD) and a genomic DNA library enriched for AG and GATA contructed by using FIASCO protocol. Nineteen microsatellites were usedto identify four clones (A, D, L, F)of the gibel carp and cross-amplification in nineteen crucian carp individuals (Carassius auratus auratus Linnaeus). Twelve microsatellite markers sucessfully identified all the four clones, two are able to identify some clones but disable to distinguish others and five fail to identify four clones. Intraclonal homogeneity and interclonal heterogeneity were observed in these clones. A phylogenetic tree was constructed using UPGMA cluster analysis based on genetic distance matrix of the four clones (A, D, L, F). The genetic trees indicated clones D and L converge firstly, and then merge with A, finally with F. Except YJ0022, eighteen microsatellites successfully cross-amplified in crucian carp. Fourteen of them were polymorphic (average PIC=0.779, average HE=0.827). In this study, we identified 108 clones from total 292 gibel carp individuals from six geographical populations by using 12 microsatellite markers. And after comparing the microsatellite patterns among six gibel carp populations and two crucian carp populations, we confined the existance of the natural gibel carp clones and observed the releasing of cultured gibel carp individuals. We suggest that the gibel carp prefer to produce by gonochoristic mechanisms in nature. Additionally, our resultes support the hypothesis that the gibel carps of Heilongjiang river originate from the diploid crucian carp of Jianghan plain area, and we further propose that the gibel carps of Xingjiang also originate from Changjiang water system and different geographical populations of the gibel carps originate from different diploid crucian carp populations.
Document Type学位论文
Recommended Citation
GB/T 7714
郭葳. 大黄鱼与银鲫微卫星标记的分离及其群体遗传分析[D]. 水生生物研究所. 中国科学院水生生物研究所,2007.
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