红白锦鲤人工雌核发育纯系的微卫星标记分析

MICROSATELLITE MARKER ANALYSIS ON ARTIFICIALLY GYNOGENETIC PURE LINE OF RED-WHITE ORNAMENTAL CARP

  • 摘要: 利用连续两代人工雌核发育操作,构建了极具观赏价值的红白锦鲤人工雌核发育系。应用对微卫星引物进行遗传标记研究,检测分析了该雌核发育系内个体间的遗传同质性及其基因座位的纯合度。实验结果显示,11尾经人工挑选具有相同表形、极具观赏价值的F1代性成熟个体的微卫星扩增图谱呈现出高度的一致性。同时,随机挑选的人工雌核发育F2代个体在所检测的基因座位均呈现纯合,扩增图谱呈现了高度的一致,从分子水平上证实了本实验所获得的红白人工雌核发育F2代是一个纯系。

     

    Abstract: One pure line of red white ornamental carp( Cyprinus carpio L.) was produced by two successive artificial gynogenesis with the combination treatment of UV irradiation sperm and cold shock (3-5℃) to block second polar body extrusion.The used heterozygous sperm came from bluntnose black bream( Megalobrama amblycephala Yih).Six microsatellite markers of the poly (CA) type isolated from common carp were used to assess genetic homogeneity among individuals of the artificially gynogenetic Red Write ornamental carp pure line.Electrophoretic patterns showed that the six pairs of microsatellite primers all produced well identifiable DNA fragments,and their size ranged from 100bp to 300bp.In this study,11 individuals similar to their mother in the color phenotype were selected from the artificially gynogenetic F1 progeny of Red-Write ornamental carps.The DNA fragment patterns of 11 individuals amplified by six primers were highly identical.One of them was successfully induced to spawn,and the ovulated eggs were used to produce artificial gynogenetic F2 progeny.Six individuals were randomly sampled from the F2 gynogenetic ornamental carps to analyze their genetic homogeneity.As a control,8 individuals reproduced from bisexual mating were also analyzed by the six microsatellite markers.The results indicated that the 6F2 gynogenetic individuals showed highly identical DNA patterns to their mother,and Mendel's genetic law of the six microsatellite markers were demonstrated by the control group.The current study provides a new technique to produce pure line in ornamental carp.Moreover,the artificially gynogenetic pure ling of red white ornamental carp will be useful for further studying on genetic mechanism of body color types in ornamental carp,and the molecular markers will be able to provide some valuable tools for marker-assisted selection breeding or gene mapping in the ornamental carp.

     

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