太平洋牡蛎微卫星引物对三角帆蚌的适用性研究(英文)

Preliminary study on applicability of microsatellite primers developed from Crassostrea gigas to genomic analysis of Hyriopsis cumingii

三角帆蚌(Hyriopsiscumingii)是我国特有种,它形成的珍珠具有珠质光滑细腻、色泽鲜艳等方面的优点,是淡水蚌中育珠质量最佳者,已成为最主要的淡水养殖珍珠蚌。本研究选取已发表的32对太平洋牡蛎的微卫星引物在三角帆蚌基因组DNA中进行PCR扩增,探讨太平洋牡蛎的引物用于三角帆蚌基因组微卫星分析的可能性。通过优化PCR反应条件,筛选13对引物可在三角帆蚌基因组中扩增出特异性条带(占总数的41%);其中10对引物(占总数的31%)在洞庭湖三角帆蚌小群体中即检测到了个体间等位基因的多态性,共出现40条多态性条带。10个位点杂合度大小在0.125到0.693之间,其中7个微卫星位点(Cgi10,Cgi22,Cgi24,Cgi26,Cgi29,Cgi30,Cgi32)为高度多态性位点,杂合度大于0.500,而其余3个微卫星位点(Cgi1,Cgi18andCgi25)杂合度小于0.500,为低度多态性位点。初步的结果表明,部分太平洋牡蛎的微卫星引物可以用于三角帆蚌基因组的分析,这为其它贝类微卫星标记的开发奠定了基础。

Hyriopsis cumingii, which produces pearl oftbe highest quality, is a kind of freshwater species unique in China. The development of microsateUite markers of H. cumingii has not been reported. However, many microsateUite markers have been developed from Crassostrea gigas. The conservativeness of the side sequences of the microsatellite in species of close genetic relationship has already been discovered. In order to determine the applicability of micmsatellite primers developed from C. gigas to genomic analysis in H. cumingii, 32 polymorphic micmsatellite primers identified in the Pacific oyster were employed to amplify in the genome of mussel. The conditions of polymerase chain reaction（PCR） were optimized for the fidelity of DNA synthesis during PCR amplification. It was found that 19 loci failed to be amplified and 13 loci （about 41% ） amplified specific products successfully. Among 13 loci, 3 （Cgi-6, Cgi-27and Cgi-28） were monomorphic and 10 （about 31% ） （Cgi-l,Cgi-10,Cgi-18,Cgi-22,Cgi-24,Cgi- 25, Cgi-26, Cgi-29, Cgi-30 and Cgi-32） were polymorphic. The analysis of the genetic diversity showed the average heterozygosity of l0 microsateUites loci of H. cumingii ranged betwen 0. 125 and 0.693 and 7 loci （Cgi-10, Cgi-22, Cgi-24, Cgi-26, Cgi-29, Cgi- 30, Cgi-32 ） are high polymorphic（He 〉 0.500）, whereas 3 loci （Cgi-1, Cgi-18 and Cgi-25 ） are low polymorphic（He 〈0.500）. This study confirmed that 10 Pacific oyster pimers could be used for the analysis of the genetic diversity in H. cumingii. This result showed some of the microsateUite primers can be used for genetic analysis of mussel without high coat costing and time consuming. And it suggested that this method could be useful in genetic analysis of other species of mussel.