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| 三倍体牙鲆(Paralichthys olivaceus)群体遗传多样性的微卫星分析 |
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刘 慧1, 吴志昊2, 李 桢1, 朱香萍1, 尤 锋2, 宋宗诚3, 李 军4, 胡金伟2
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1.青岛农业大学 青岛 266109;2.中国科学院实验海洋生物学重点实验室 中国科学院海洋研究所 青岛 266071;3.威海圣航水产科技有限公司 威海 264200;4.海洋生物工程技术研究发展中心 中国科学院海洋研究所 青岛 266071
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| 摘要: |
| 采用微卫星遗传标记技术对三倍体牙鲆群体及二倍体对照群体进行比较分析。取2012年冷休克诱导和培育至11月龄的三倍体及同期二倍体对照牙鲆个体各32尾，通过高盐法提取肌肉组织总DNA；利用筛选得到的21对微卫星引物进行PCR扩增，并经14%聚丙烯酰胺凝胶电泳分型后，进行人工判读和分析。结果显示，21个微卫星位点均为多态，二倍体和三倍体群体的平均等位基因数(A)、基因型总数、平均有效等位基因数(ae)、平均观测杂合度(HO)和平均期望杂合度(HE)分别为5.6和5.0、190和142、3.7和2.8、0.613和0.868、0.681和0.629。二倍体和三倍体群体的多态信息含量(PIC)分别为0.642和0.589，个体识别率(DP)为0.660和0.609，非父排除率(PPE)为0.482和0.399，其累积个体识别率和非父排除率均达到0.999，表明所选座位均属中高识别力的遗传标记，可以将它们应用于今后牙鲆雌核发育群体的遗传变异分析以及进一步的遗传育种的研究中。二倍体和三倍体群体间的遗传距离(D)为0.312，遗传相似系数(I)为0.732，基因分化系数(GST)为0.971。以上结果说明，三倍体诱导对牙鲆的遗传结构造成了一定影响，导致三倍体遗传多样性水平相对二倍体有所下降，二倍体和三倍体牙鲆的基因型也存在一定差异。 |
| 关键词: 牙鲆 三倍体 人工诱导 微卫星 遗传多样性 |
| DOI：10.11758/yykxjz.20150105 |
| 分类号: |
| 基金项目:国家863计划资助项目(2012AA10A402)、山东省自然科学基金项目(ZR2011CM035)、山东省科技发展计划项目(2013GHY11526)和鲆鲽类产业技术体系和苗种繁育项目科学家课题(NYCYTX-50)共同资助 |
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| Genetic Diversity Analysis on Artificial Triploid Stock of Paralichthys olivaceus using Microsatellite DNA Marker |
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| Abstract: |
| In this study, we compared the extent of genetic diversity of the triploid and diploid flounders using the microsatellite method. The triploid were induced using cold shock method in 2012, and then they were cultured under the same conditions as their diploid control. Genomic DNAs were isolated using the high salt-extraction method from the muscle tissues of 32 triploid and 32 diploid flounders of 11-month old. The genetic variations of the triploid and diploid flounders were assessed based on the selected 21 microsatellite loci. The PCR products were isolated using 14% PAGE electrophoresis, and the microsatellite alleles were artificially identified and analyzed. The results showed that in the triploid and diploid stocks, the average numbers of alleles per locus (A) were 5.6 and 5.0; the total numbers of genotype were 190 and 142; the average numbers of effective alleles per locus (ae) were 3.7 and 2.8; the values of average observed heterozygosity (HO) were 0.613 and 0.868; and the unbiased expected heterozygosity (HE) were 0.681 and 0.629, respectively. The values of the polymorphism information content (PIC), the power discrimination (DP) and the probability of paternity exclusion (PPE) in the triploid and diploid stocks were 0.642 and 0.589, 0.660 and 0.609, 0.482 and 0.399, respectively. The accumulation of PIC and PPE both achieved 0.999, which indicated that these microsatellite loci were very sensitive and could be used in the parentage and kinship determination in future genetic breeding studies. The genetic distance (D), the genetic similarity (I) and the coefficient of gene differentiation (GST) between the triploid and the diploid stocks were 0.312, 0.732 and 0.971 respectively. The genetic diversity of the triploid stock was moderately reduced compared to the diploid stock. There were also some differences in the genotypes between the triploid and diploid flounders. |
| Key words: Paralichthys olivaceus Triploid Artificial induction Microsatellite DNA marker Genetic variation |