褐牙鲆耐热性状相关的微卫星分子标记筛选

The Screening of Microsatellites Molecular Markers with Correlationship to Heat-resistance of Paralichthys olivaceus

采用微卫星分子标记技术分析褐牙鲆耐热相关特性,为耐高温褐牙鲆的分子辅助育种提供合适的分子标记.褐牙鲆经过热处理,将其区分为耐热组与不耐热组,用于实验分析.采用酚-氯仿抽提法抽提褐牙鲆肌肉组织的DNA,进行微卫星引物PCR(SSR-PCR)扩增,所用引物为已知的20个褐牙鲆微卫星位点的侧翼保守序列.对PCR扩增出的差异条带进行个体统计,最后进行微卫星位点与耐热性状的相关性分析.结果表明,有6个微卫星位点的某等位基因片段与褐牙鲆耐热性状存在一定的正相关性,其中位点Po13(AB046746)跟耐热性有极显著的正相关性,相关系数为0.466;有3个微卫星位点的某等位基因片段与耐热性存在负相关性,其中位点Po42(AB046754)与其有极显著的负相关性,相关系数为-0.408.微卫星位点Po13与Po42所扩增出的等位基因片段可作为分子标记指导耐热褐牙鲆的辅助育种.

The present work refers to the correlative analysis between microsatellites and the heat-resistance of Paralichthys olivaceus. From this project,we screened out some suitable microsatellite molecular markers for MAS（Molecular Assistant Selection）. The studied flounders were divided into two categories： heatresistant group and thermolabile group by heating the water they existed in. Then DNAs were extracted out from these individuals and PCR-amplified with 20 microsatellite primers which were already registered in GeneBank. The PC.R mixture were then electrophoresed through the PAGE gel and siliver stained. After that,the differential bands were analysed and figured out. Finally, we analysed the correlationship between the upper thermal tolerance and the microsatel- lite loci. The results show that 3 loci are significantly negative-correlated with upper thermal tolerance（UTT） ,of which Po42 has extreme significance of - 0. 408^＊＊. Otherwise, 6 loci is positive-correlated with UTT, of which Po13 has extreme significance of 0. 466^＊＊. These microsatellites and their differential bands could be used for molecular assistant selection of heat-resistant flounders.