三角帆蚌四个群体遗传多样性的SSR分析
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摘要
三角帆蚌是我国主要的淡水育珠蚌，在我国东部、南部及中部有着广泛的分布，关于它的研究涉及了形态学、生态学、生物化学、细胞生物学等领域。随着淡水珍珠养殖业的发展，三角帆蚌在我国的养殖范围与规模越来越大，所产生的经济效益越发显著。但随着蚌类栖息地的破坏和环境污染加剧，贝类捕捞强度加大，以及蚌类人工繁殖和集约化养殖规模的扩大，三角帆蚌的遗传多样性受到不同程度的破坏，自然资源不断减少，开展三角帆蚌遗传多样性研究，有利于保护其种质资源，为进行大规模养殖和获得高产提供相应理论依据。
本研究从Epioblasma capsaeformis和Margaritifera margaritifera的20个微卫星DNA标记中筛选出了在三角帆蚌(Hyriopsis cumingii)中扩增产物多态性较好的8个微卫星标记，利用这8个微卫星标记对三角帆蚌四个群体进行了遗传检测。这四个三角帆蚌群体分别为湖南养殖群体(HY)、洞庭湖野生群体(DY)、鄱阳湖野生群体(PY)、太湖野生群体(TY)。用非变性聚丙烯酰胺电泳检测微卫星的PCR扩增产物，计算了三角帆蚌四个群体的个体样本在8个微卫星座位上的等位基因频率、有效等位基因数、微卫星的多态信息含量(PIC)、各群体的遗传杂合度(H)、群体间的遗传距离，并根据遗传距离对这几个群体进行了聚类分析。
结果表明，所选择的8个微卫星位点在所检测群体中都表现出了较好的多态性，平均每个微卫星位点检测到5.0个等位基因和3.1个有效等位基因，8个微卫星标记的平均多态信息含量为0.5824，这表明所选择的微卫星标记适合用于三角帆蚌的遗传多样性分析。四个三角帆蚌群体的遗传杂合度均较高，其中太湖野生群体的群体杂合度最高(0.7500)；其次为洞庭湖野生群体(0.7083)和鄱阳湖野生群体(0.6500)；湖南养殖群体的群体杂合度最低(0.6167)。结果表明我国的三角帆蚌野生群体的遗传多样性比较丰富，可以进一步进行合理的选择和利用。对遗传距离计算的结果表明各群体间的遗传距离较小，彼此之间有较近的亲缘关系，聚类分析的结果表明洞庭湖野生群体和湖南养殖群体的亲缘关系最近。
Hyriopsis cumingii is the major freshwater pearl mussel, spread widely in the eastern, southern and center of China. The reseach of Hyriopsis cumingii constitutes of morphology, ecology, biochemistry, cell biology and so on. As the result of pearl production increased, the cultured scales of Hyriopsis cumingii expanded. With the large quantity catching and high-density culturing, the genetic diversity of Hyriopsis cumingii was destroyed in different extent, the resources deduce year by year. Working about the genetic diversity of Hyriopsis cumingii will benefit protecting the resources of Hyriopsis cumingii, and provide the scientific evidence for breeding and output in great large scale.Genetic diversity of four Hyriopsis cumingii groups was evaluated on the basis of eight microsatellite markers with polymorphism, selected from twenty microsatellite markers which belong to Epioblasma capsaeformis and Margaritifera margaritifera. The samples of four geographic populations of Hyriopsis cumingii were collected from Dongting Lake (DY), Poyang Lake (PY), Tai Lake (TY) and Hunan cultured population (HY). The PCR amplified products of microsatellites were detected by polyacrylamide gel electrophoresis (PAGE). Allele frequency, effective numbers of alleles (n_e), polymorphism information content (PIC), gene heterozygosity (H), and genetic distances (D_s) between these groups were estimated. And then, cluster analysis based on those calculated genetic distances was performed for the four Hyriopsis cumingii populations.The results showed that every microsatellite locus analyzed were polymorphic in those Hyriopsis cumingii populations with an average number of alleles per locus and the polymorphism information content (PIC) equal to 5 and 0.5824 respectively. The results suggested that microsatellite markers can be used to evaluated genetic diversity of Hyriopsis cumingii. Rates of hetrozygosity in four groups are all high, and that is the highest in TY (0.7500), the second is DY (0.7083), the third is PY (0.6500), and the last is HY (0.6500), which suggested that those Hyriopsis cumingii groups had abundant genetic diversity and potential of selection. The result of genetic distance indicated that there were close genetic relationship among those Hyriopsis cumingii groups. The results of cluster analysis suggest that the relationship between DY and HY is the nearest.
本研究从Epioblasma capsaeformis和Margaritifera margaritifera的20个微卫星DNA标记中筛选出了在三角帆蚌(Hyriopsis cumingii)中扩增产物多态性较好的8个微卫星标记，利用这8个微卫星标记对三角帆蚌四个群体进行了遗传检测。这四个三角帆蚌群体分别为湖南养殖群体(HY)、洞庭湖野生群体(DY)、鄱阳湖野生群体(PY)、太湖野生群体(TY)。用非变性聚丙烯酰胺电泳检测微卫星的PCR扩增产物，计算了三角帆蚌四个群体的个体样本在8个微卫星座位上的等位基因频率、有效等位基因数、微卫星的多态信息含量(PIC)、各群体的遗传杂合度(H)、群体间的遗传距离，并根据遗传距离对这几个群体进行了聚类分析。
结果表明，所选择的8个微卫星位点在所检测群体中都表现出了较好的多态性，平均每个微卫星位点检测到5.0个等位基因和3.1个有效等位基因，8个微卫星标记的平均多态信息含量为0.5824，这表明所选择的微卫星标记适合用于三角帆蚌的遗传多样性分析。四个三角帆蚌群体的遗传杂合度均较高，其中太湖野生群体的群体杂合度最高(0.7500)；其次为洞庭湖野生群体(0.7083)和鄱阳湖野生群体(0.6500)；湖南养殖群体的群体杂合度最低(0.6167)。结果表明我国的三角帆蚌野生群体的遗传多样性比较丰富，可以进一步进行合理的选择和利用。对遗传距离计算的结果表明各群体间的遗传距离较小，彼此之间有较近的亲缘关系，聚类分析的结果表明洞庭湖野生群体和湖南养殖群体的亲缘关系最近。
Hyriopsis cumingii is the major freshwater pearl mussel, spread widely in the eastern, southern and center of China. The reseach of Hyriopsis cumingii constitutes of morphology, ecology, biochemistry, cell biology and so on. As the result of pearl production increased, the cultured scales of Hyriopsis cumingii expanded. With the large quantity catching and high-density culturing, the genetic diversity of Hyriopsis cumingii was destroyed in different extent, the resources deduce year by year. Working about the genetic diversity of Hyriopsis cumingii will benefit protecting the resources of Hyriopsis cumingii, and provide the scientific evidence for breeding and output in great large scale.Genetic diversity of four Hyriopsis cumingii groups was evaluated on the basis of eight microsatellite markers with polymorphism, selected from twenty microsatellite markers which belong to Epioblasma capsaeformis and Margaritifera margaritifera. The samples of four geographic populations of Hyriopsis cumingii were collected from Dongting Lake (DY), Poyang Lake (PY), Tai Lake (TY) and Hunan cultured population (HY). The PCR amplified products of microsatellites were detected by polyacrylamide gel electrophoresis (PAGE). Allele frequency, effective numbers of alleles (n_e), polymorphism information content (PIC), gene heterozygosity (H), and genetic distances (D_s) between these groups were estimated. And then, cluster analysis based on those calculated genetic distances was performed for the four Hyriopsis cumingii populations.The results showed that every microsatellite locus analyzed were polymorphic in those Hyriopsis cumingii populations with an average number of alleles per locus and the polymorphism information content (PIC) equal to 5 and 0.5824 respectively. The results suggested that microsatellite markers can be used to evaluated genetic diversity of Hyriopsis cumingii. Rates of hetrozygosity in four groups are all high, and that is the highest in TY (0.7500), the second is DY (0.7083), the third is PY (0.6500), and the last is HY (0.6500), which suggested that those Hyriopsis cumingii groups had abundant genetic diversity and potential of selection. The result of genetic distance indicated that there were close genetic relationship among those Hyriopsis cumingii groups. The results of cluster analysis suggest that the relationship between DY and HY is the nearest.
引文
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