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Alternative TitleIsolation and characterization of microsatellites in Chinese Soft-shelled Turtle,Pelodiscus Sinensis
Thesis Advisor常剑波
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中华鳖 Fiasco 种质资源 微卫星 多态性
Abstract中华鳖(Pelodiscus sinensis(Wiegmann),1834)在我国分布很广泛,具有很高的营养价值和药用价值。在过去的10多年中,中华鳖野生群体资源受到了严重破坏,数量急剧下降。为了保护这一具有重要经济和科研价值的陆生野生动物,全国各地开展了大规模的人工养殖,然而人工养殖中华鳖的快速无序发展却导致了中华鳖的种质资源受到冲击。因此,中华鳖优良种质资源的准确、快速识别与保护成为当前中华鳖养殖中存在的主要问题。 目前,中华鳖不同品种的识别主要依靠其外部形态特征、经济性状特征,同工酶电泳及限制性片段长度多态性等方法,但是由于这些方法操作步骤较繁琐、结果不够准确,可重复性较差,因此有必要发展更加精确、可靠、简便的方法用于中华鳖的品种鉴定和良种选育工作。微卫星不仅多态性较高,而且操作简单、快速经济。微卫星分子标记在我国的种猪和大豆良种选育种广泛应用,国外在虹鳟、罗非鱼、大西洋鳕鱼和扇贝等经济动物中利用微卫星DNA来指导养殖和良种选育。 本研究采用微卫星FIASCO(Fast Isolation of Amplified Sequences COntainning repeat)富集方法,分离并获得了52个微卫星序列;根据这些位点设计合成其特异的44对微卫星引物对。所有的44对微卫星引物经过一个含有40个个体的中华鳖群体的检验后发现:20对微卫星引物具有高度多态性,其等位基因数为2~7,平均为3.45,位点的观测杂合度为0.03~0.98,平均为0.38。所有位点都不存在紧密连锁。这些位点具有较高的多态性水平,将为种群动态、群体遗传学及辅助育种等研究提供帮助。 这是首次报道中华鳖的高度多态性微卫星位点,将为中华鳖遗传背景的构建和辅助育种提供有力的工具,研究结果不仅有利于促进中华鳖养殖业的健康发展,也会促进中华鳖的良种生产和标准化管理。
Other AbstractChinese Soft-shelled Turtle, Pelodiscus sinensis, a Chinese fresh-water turtle, is widely distributed in Japan, Korea, Vietnam and other Southeast Asian countries. P. sinensis is favored for its supposed medical value and has been consumed as food for a long period of time in China. However, during the past two decades, habitat destruction, overfishing, general pollution and fragmentation of large rivers by dams have led to dramatical decline of the wild populations of this species. Consequently, the Chinese Soft-shelled Turtle was listed as a vulnerable species in the China Red Data Book of Endangered Animals in 1998. Knowledge on the genetic structure of P. sinensis populations is essential for effective protection measures of germplasm resources and sustainable management programs. However, little is known about the genetic diversity and genetic structure of once abundant wild populations of this species. A partial genomic library enriched containing (GT) n microsatellite was developed and 44 pairs of primers were created using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol for P. sinensis. Of all primers, twenty primers produced unambiguous polymorphic PCR products. The polymorphism of all twenty loci ranged from two to seven alleles with observed heterozygosities ranging from 0.03 to 0.98 (mean 0.38) in 40 individuals of one population. No significant linkage disequilibrium was detected among the loci. These novel microsatellite loci showed moderate to high levels of polymorphism in P. sinensis. It is the first time to report these 20 polymorphic microsatellite loci for P. sinensis. These loci will be helpful for understanding population structure at genetic level and markers assisted breeding of this vulnerable species.
Document Type学位论文
Recommended Citation
GB/T 7714
阙延福. 中华鳖微卫星DNA的分离与鉴定[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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