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采用微卫星DNA指纹图谱技术对奥利亚罗非鱼、尼罗罗非鱼和橙色莫桑比克罗非鱼进行品种鉴定和遗传多样性分析。从103对罗非鱼微卫星引物中筛选出82对多态性高、带型清晰稳定的引物,用这82对引物检测3种罗非鱼的遗传结构,共检测到605个等位基因,平均等位基因数7.37个,数据经Popgen32计算和EXCEL工具作图,构建3种罗非鱼的DNA指纹数据库,并绘制了DNA指纹图谱模式图。结果显示,奥利亚罗非鱼、尼罗罗非鱼和橙色莫桑比克罗非鱼的平均观测杂合度分别为0.179 6、0.530 1和0.312 2,平均期望杂合度分别为0.203 2、0.678 6和0.291 3,平均多态信息含量分别为0.075 4、0.608 3和0.152 8,由此可见,尼罗罗非鱼群体的遗传多样性水平最高,奥利亚罗非鱼群体遗传多样性最低。同时筛选得到16对具特异性条带的核心引物组合可将3种罗非鱼完全区分,每个位点可在其中一种罗非鱼中扩增出独有的条带,从而将这种罗非鱼与其它两种区分开来,将16对特异引物的图谱数据转化成计算机可以识别的数码指纹,可以方便地应用于罗非鱼及其杂交种的鉴定研究。为解决罗非鱼品种混杂遗传渐渗问题和保护罗非鱼种质资源提供技术支撑。
Microsatellite DNA fingerprinting was used to characterize and analyze the genetic diversity of tilapia (Oreochromis niloticus), Nile tilapia (Oreochromis niloticus) and orange (Mozambique) tilapia. A total of 82 pairs of tilapia microsatellite primers were screened from 103 pairs of high polymorphic bands with clear and stable bands. The 82 pairs of primers were used to detect the genetic structure of three species of tilapia, 605 alleles were detected, The average number of alleles was 7.37. The data were calculated by Popgen32 and EXCEL tools to construct DNA fingerprinting database of three kinds of tilapia, and the DNA fingerprinting patterns were drawn. The results showed that the average observed heterozygosities of O. tilapia, O. niloticus and O. mozambique were 0.179 6, 0.530 1 and 0.312 2, respectively. The average expected heterozygosities were 0.203, 2.0678 6 and 0.291 3, and the average polymorphic information content was 0.075 4,0.608 3 and 0.152 8, respectively. Therefore, the genetic diversity of Nile tilapia population was the highest, and the genetic diversity of tilapia population was the lowest. At the same time, we obtained 16 pairs of core primers with specific bands to completely distinguish the three kinds of tilapia, and each site could amplify the unique bands in one of the tilapias, Tilapias are distinguished from the other two by converting 16 pairs of specific primers into computer-readable digital fingerprints, which can be easily applied to the identification of tilapia and its hybrids. To provide technical support to solve the problem of mixed genetic introgression of tilapia varieties and to protect germplasm resources of tilapia.