【摘要】 通过FIASCO法(Fast Isolation by AFLP Sequences Containing Repeats)筛选宽口光唇鱼Acrossocheilus monticola(Günter)基因组微卫星位点,利用生物素标记的寡核苷酸探针(AC)8、(CT)8、(GGT)8、(GATA)8和(GATT)7首次成功构建宽口光唇鱼基因组微卫星富集文库。从文库中共筛选495个克隆测序,成功设计163对微卫星引物,经PCR扩增检测,获得了18个多态性微卫星标记。利用这18对多态性引物,分析了赤水河赤水市宽口光唇鱼种群的遗传多样性,数据显示:18对多态性微卫星引物的等位基因数为8～31个,平均等位基因数为19.6个,平均期望杂合度为0.8701,平均观测杂合度为0.8312,其中引物Am07、Am35、Am47、Am69、Am78和Am127显著偏离哈迪温伯格平衡(P<0.05),以上数据表明赤水河赤水市宽口光唇鱼种群的遗传多样性水平较高。筛选的微卫星标记对于宽口光唇鱼的遗传背景分析和生物种质资源的保护有重要作用。更多还原

【Abstract】 Acrossocheilus monticola is the endemic fish in the upper reach of the Yangtze River.For a long time,little attention has been paid to this population because of its small size and low economic value.In recent years,the construction of hydroelectric projects and some anthropic factors changed the physical environment and hydrological conditions in the Yangtze River,which lead to some impacts on the rare and endemic fishes in the Yangtze River including A.monticola.In this study,we isolated microsatellite loci from A.monticola genome by FIASCO(Fast Isolation by AFLP Sequences Containing Repeats) procedure.Taking advantages of biotinylated oligo probes(AC)8,(CT)8,(GGT)8,(GATA)8 and(GATT)7,we successfully constructed a microsatellite-enriched genomic library,and 495 clones were sequenced.163 pairs of SSR-primers were designed afterwards,and from which 18 polymorphic microsatellite markers were screened out by PCR amplification.Using these polymorphic microsatellite markers,we analyzed the genetic diversity of the population of A.monticola collected from the Chishui city.The results showed that the average allele number of microsatellite loci was 19.6,the average observed heterozygosity(Ho) was 0.8312,and the average expected heterzygosity(He) was 0.8701.Six markers,Am07,Am35,Am47,Am69,Am78 and Am127,were deviated from Hardy-Weinberg equilibrium significantly(P<0.05).These results indicated that the genetic diversity of A.monticola was diversiform,and these polymorphic microsatellite primers could be used in A.monticola population genetic analysis as well as biological resource conservation.更多还原