| 中华绒螯蟹微卫星多重PCR体系的建立及其亲子鉴定应用 |
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| 引用本文： | 肖起珍,刘青,李清清,郑海地,吴旭干,成永旭.中华绒螯蟹微卫星多重PCR体系的建立及其亲子鉴定应用[J].中国水产科学,2018,25(2):325-335. |
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| 作者姓名： | 肖起珍 刘青 李清清 郑海地 吴旭干 成永旭 |
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| 作者单位： | 上海海洋大学农业部淡水种质资源重点试验室;浙江澳凌水产种业科技有限公司;上海海洋大学上海市教委水产动物遗传育种协同创新中心 |
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| 基金项目： | 农业部现代农业产业技术体系项目[农科（产业）函[2017]第23号]；上海市科技兴农推广项目[沪农科推字（2015）第1-7号]；上海市科委工程技术中心能力提升项目（16DZ2281200）；上海高校水产学高峰学科建设项目（2015-62-0908）；深圳市澳华农牧有限公司横向项目（D-8006-15-0054）. |
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| 摘 要： | 利用自主开发的微卫星标记对中华绒螯蟹(Eriocheir sinensis)进行微卫星多重PCR体系构建,最终成功建立4组多重体系,每组体系包含4个微卫星位点,并成功应用于3个家系的亲子鉴定中。结果显示:(1)本研究筛选的16个微卫星标记,平均观测杂合度为0.8206,平均期望杂合度为0.8164,平均多态信息含量为0.7927,具有丰富的多态性;(2)运用Cervus 3.0软件对已知系谱信息的3个中华绒螯蟹家系共95个子代个体进行亲子鉴定分析,结果显示,选用任意两组微卫星多重PCR体系时,累积实际正确鉴定率均超过94.74%;使用任意三组微卫星多重PCR体系时,累积实际正确鉴定率均大于98.95%;使用四组微卫星多重PCR体系时,累积实际正确鉴定率达到100%。并且当选用组合1、2和组合3,或者选用组合1、3和组合4时,对3个家系的累积正确鉴定率达到了100%,因此选用这些组合不但可以获得准确的系谱信息,还能减少工作量,降低成本。本研究构建的微卫星多重PCR体系能为中华绒螯蟹的种群选育和家系管理提供便捷、高效的途径。
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| 关 键 词： | 中华绒螯蟹 微卫星 多重PCR 亲子鉴定 |
| 修稿时间： | 2018/4/24 0:00:00 |
Development and application of multiplex PCR panels of microsatellites in Chinese mitten crab (Eriocheir sinensis) |
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| XIAO Qizhen,LIU Qing,LI Qingqing,ZHENG Haidi,WU Xugan,CHENG Yongxu.Development and application of multiplex PCR panels of microsatellites in Chinese mitten crab (Eriocheir sinensis)[J].Journal of Fishery Sciences of China,2018,25(2):325-335. |
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| Authors: | XIAO Qizhen LIU Qing LI Qingqing ZHENG Haidi WU Xugan CHENG Yongxu |
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| Institution: | 1. Key Laboratory of Freshwater Aquaculture Genetic Resources, Ministry of Agriculture;Shanghai Ocean University, Shanghai 201306, China;2. Zhejiang Aoling Aquatic Seed Industry Technology Co., Changxing 313106, China;3. Collaborative Innovation Center of Aquatic Animal Breeding Center Certified by the Shanghai Municipal Education Commission;Shanghai Ocean University, Shanghai 201306, China |
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| Abstract: | In order to increase the analysis efficiency of microsatellites in Chinese mitten crab (Eriocheir sinensis), we report four highly informative, multiplex PCR panels developed from 16 microsatellite markers. The multiplex PCR panels were used to evaluate the genetic diversities and parentage assignment for 95 individuals in 3 families. The results showed that the 16 microsatellite markers screened in this study were rich in polymorphism, for example the average of observed heterozygosity (Ho) was 0.8206, the average of expected heterozygosity (He) was 0.8164, and the average of polymorphism information content (PIC) was 0.7927. Cervus 3.0 software was used to determine parentage assignment for 95 individuals in 3 families, and the results revealed that the accuracy of parentage assignment using any 2 groups of multiplex PCR panels was more than 94.74%. When using any 3 groups of multiplex PCR panels, the accuracy of parentage assignment was greater than 98.95%. The accuracy of parentage assignment was greater than 100% using all four multiplex assays. Using the combination of set 1, set 2, and set 3, or the combination of set 1, set 3, and set 4, the accuracy of parentage assignment also reached 100%, so the selection of these combinations of multiplex PCR panels can not only get accurate genealogy information, but also reduce workload and costs. Therefore, these multiplex PCR panels can provide a convenient and efficient way for breeding and family management of E. sinensi. |
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| Keywords: | Eriocheir sinensis microsatellites multiplex PCR parentage assignment |
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