云斑尖塘鳢微卫星标记在两种虾虎鱼亚目鱼类群体间的适用性研究

Applicability of microsatellite markers developed from Oxyeleotris marmoratato Oxyeleotris lineolatus and Odontobutis potamophila

利用微卫星DNA-聚合酶链式反应(STR-PCR)技术对已开发的14对云斑尖塘鳢(Oxyeleotrismarmorata)微卫星标记在线纹尖塘鳢(Oxyeleotris lineolatus)和河川沙塘鳢(Odontobutis potamophila)群体间进行通用性分析。结果显示,14对云斑尖塘鳢微卫星标记在线纹尖塘鳢群体均能扩增出特异性条带,在河川沙塘鳢群体中也有10对引物可稳定扩增,其中9对在线纹尖塘鳢群体中具有高度多态性,6对在河川沙塘鳢群体中具有高度多态性。通过这10对通用微卫星标记检测3个种群的平均观测杂合度(Ho)为0.344 9,平均期望杂合度(He)为0.704 3,平均多态信息(PIC)含量为0.536 9。同时获得3个有着较高的通用性微卫星位点(H27、H142和H97),平均PIC值均大于0.5,具有高度多态性,在3种虾虎鱼亚目鱼类中具有较高的通用性,这3个位点在3种虾虎鱼亚目鱼类之间共检测得到6～22个不等的等位基因,并获得1个可以用于鉴别沙塘鳢科与塘鳢科特异基因型的等位基因位点(H97)。因此,通过已开发的云斑尖塘鳢微卫星标记来获得适用相近物种的微卫星标记是可行的。

Using microsatellite DNA-polymerase chain reaction（STR-PCR）,14 microsatellite primers developed in our laboratory from Oxyeleotris marmorata were tested for cross-amplification in Oxyeleotris lineolatus and Odontobutis potamophila.14 loci were amplified successfully in Oxyeleotris lineolatus and 10 in Odontobutis potamophila.The polymorphism of these scorable loci was assessed using corresponding population individuals,and the results showed that 9 loci were polymorphic in Oxyeleotris lineolatus and 6 loci were polymorphic in Odontobutis potamophila.The average observed heterozygosity of three populations was 0.344 9,the average of expected homozygosty was 0.704 3,and the average of PIC was 0.536 9.3 loci（H27,H142 and H97） revealed highly polymorphic（PIC〉0.5） in the three populations.The number of alleles per polymorphic locus ranged from 6 to 22 in these three loci.In addition,we found a genotype-specific locus（H97） which could differentiate average allelic length between family Odontobutidae and Eleotridae.Therefore,to apply developed microsatellite markers of Oxyeleotris marmorata to cross-amplification in closely related species was feasible and valid.