中华绒螯蟹37个多态性微卫星标记在亲子遗传中的分离方式分析
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上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室,上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室,上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室

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国家科技支撑计划(2012BAD26B04);上海高校水产学一流学科建设项目;江苏农牧科技职业学院科研项目(NSFYB1306);上海市河蟹产业技术体系建设项目(D8003100208)


The segregation patterns of 37 polymorphism tri-nucleotide microsatellite markers in an F1 family of the Chinese mitten crab(Eriocheir sinensis)
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上海海洋大学,,. Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, Shanghai Ocean University, Ministry of Education

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    摘要:

    为了研究中华绒螯蟹基因组特征及评估基因组扫描方法获得的中华绒螯蟹候选微卫星标记用于构建遗传图谱的可靠性,随机选择60个候选三核苷酸微卫星标记,首先利用中华绒螯蟹一个F1家系双亲及其6个F1子代共8个样品进行PCR扩增验证和多态性检测,随后利用该家系80个子代对多态性微卫星标记的亲子遗传分离类型及连锁关系进行分析。结果显示,42个(70.00%)微卫星位点得到清晰扩增产物,家系中每个检测位点最多检测到4个等位基因,每个检测个体微卫星位点上有1或2个等位基因。42个位点中有5个单态微卫星位点和37个多态微卫星位点,多态位点中有23个位点的子代基因型比为1:1:1:1,11个位点基因型比为1:1,余下3个位点基因型比为1:2:1。在37个多态位点中有35个位点(94.59%)的分离比符合孟德尔分离规律,连锁关系分析scaffold240262_150253、scaffold216209_138892、scaffold293154_172768等3个标记之间发生连锁关系,scaffold285640_169721和scaffold427534_212914发生连锁关系,scaffold507500_231891和scaffold92860_68250标记发生连锁关系。研究表明,中华绒螯蟹是二倍体生物,利用F1家系结合开发的候选微卫星标记可构建中华绒螯蟹遗传图谱。

    Abstract:

    The aim is to study the genomic characteristics and evaluate the candidate microsatellites(or simple sequence repeat, SSR)isolated via a genomic survey for genetic linkage mapping of Chinese mitten crab Eriocheir sinensis.In this study, a set of 60 candidate tri-nucleotide microsatellite markers were randomly selected.Their validation and polymorphism were investigated in an F1 family of mitten crab using the genomic DNA of the parents and 6 offspring, and the segregation patterns and linkage relationship of polymorphic loci were subsequently investigated in 80 F1 individuals.The results are as follows:Forty-two primer sets(70.00%)successfully yielded amplicons.The number of alleles per locus of the family varied from 1 to 4, and the number of alleles for each individual per locus was 1 or 2.In the 42 SSRs, 5 SSRs were mono-morphism and 37 SSRs were polymorphism.In the 37 polymorphic loci analysed using the F1 family, twenty-three loci exhibited a segregation ratio of 1:1:1:1, eleven loci exhibited a segregation ratio of 1:1, and three loci exhibited a segregation of 1:2:1.Polymorphism analysis showed that 35(94.59%)SSRs consisted with Mendelian segregation ratio, while the other 2(5.41%)SSRs(scaffold430598_213690 and scaffold21303_16865)deviated from the expected Mendelian segregation pattern.Linkage analysis showed that scaffold240262_150253, scaffold216209_138892 and scaffold293154_172768 had linkage relationship, scaffold285640_16972 and scaffold427534_212914 had linkage relationship, and so had scaffold507500_231891 and scaffold92860_68250.The results suggest that the Chinese mitten crab were diploid and its genetic linkage map could be constructed using the candidate SSRs and an F1 family.

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熊良伟,刘志强,严银龙,邱高峰.中华绒螯蟹37个多态性微卫星标记在亲子遗传中的分离方式分析[J].水产学报,2015,39(2):174~181

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  • 收稿日期:2014-09-18
  • 最后修改日期:2014-11-03
  • 录用日期:2015-01-20
  • 在线发布日期: 2015-02-09
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