鲫鱼遗传多样性的微卫星研究
摘要
本研究以鲫鱼的血液、肌肉、肝、肾为材料，采用苯酚-氯仿法、试剂盒法提取鲫鱼基因组DNA，结果表明不同方法提取的鲫鱼基因组DNA浓度在385.8-3167.5ng/μL之间，A_(260)/A_(280)比值处于1.66-1.87之间，所提取的DNA适合微卫星PCR扩增，但试剂盒法更适用于本研究。
用Crooijmans et al从鲤鱼中分离出来的17对微卫星引物(MFW14-MFW19，MFW21-MFW31)和依据鲫鱼的微卫星序列自行设计一对微卫星引物(MFW32)，共计18对引物，对四种群体的鲫鱼基因组DNA进行扩增，筛选扩增带型清晰、特异性强、重复性好的引物MFW14、MFW16、MFW18、MFW24、MFW25、MFW26、MFW28、MFW29、MFW32用作结果分析，其中引物MFW18、MFW24、MFW26、MFW29可作为区分四种鲫鱼群体的微卫星标记的有效引物。
从群体内来看，9个微卫星位点中共检测到46个等位基因，其中有18个等位基因为4种鲫鱼群体所共有，8个等位基因具有种群特异性；4种鲫鱼的平均遗传杂合度在0.607-0.720之间，白鲫为0.720、红鲫为0.652、彭泽鲫为0.629、鲫鱼为0.607；平均多态信息含量在0.530-0.650之间，白鲫为0.650、红鲫为0.587、彭泽鲫为0.549、鲫鱼为0.530，由此，白鲫的遗传多样性最高，鲫鱼的遗传多样性最低。
从群体间来看，四种群体中，鲫鱼和白鲫间的遗传相似性指数最小，为0.7143，其遗传距离为0.2857，说明这两种群体亲缘关系较远；白鲫与红鲫的遗传相似性指数最大，为0.8116，其遗传距离为0.1884，这可推断白鲫与红鲫亲缘关系较近。聚类分析结果表明，4种鲫鱼分为两组，白鲫和红鲫为1个组，而鲫鱼和彭泽鲫处于另1个组。
Using blood,muscle,liver and kidney from Auratus as the material, two different methods were compared in extraction of Auratus genomic DNA ,which belonged to Phenol-chloroform and Molecular biology Kits. The results show that the production concentration was 385.8-3167.5ng/uL and A260/A28o was 1.66-1.87. they can both be amplified by the the polymerase chain reaction(PCR) of Microsatellite DNA ,but Molecular biology Kits is feasible for the research.
This research adopts 18 primers of Microsatellite DNA that Crooijmans et al separated out 17 primers from the carp (MFW14-MFW19, MFW21-MFW31), and designing 1 primer (MFW32 ) according to microsatellite DNA sequence of Auratus by myself. The clear, peculiar and repetitive trips of the the polymerase chain reaction(PCR) products of the MFW14 , MFW16 , MFW18 , MFW24 , MFW25 , MFW26 , MFW28 , MFW29 , MFW32 among four populations of Auratus genomic DNA from different individuals using 18 primers are analyzed by the research.The prmers of MFW18, MFW24, MFW26, MFW29 can be differentiated from four populations of Auratus.
From four intrapopulations of Auratus, 9 Microsatellite DNA location measures 46 alleles, among them 18 alleles are in all by four populations of Auratus, 8 alleles are peculiar in a population among them. The average heterozygosity of four populations of Auratus is 0.607-0.720, C.auratus cuvieri is 0.720, C.auratus red va is 0.652, C.auratus auratus var. Pengze is 0.629, C.auratus auratu is 0.607; The average polymorphism information content is 0.530-0.650, C.auratus cuvieri is 0.650, C.auratus red var is 0.587, C.auratus auratus var. Pengze is 0.549, C.auratus auratus is 0.530, so the genetic diversity of C.auratus cuvieri is the highest and the genetic diversity of C.auratus auratus is the lowest.
From four populations of Auratus, it is 0.7143 that genetic similarity indices between C.auratus auratus and C.auratus cuvieri is minimum , its genetic distances is 0.2857, proving the phylogenetic relationships of two populations are relatively far. C.auratus cuvieri genetic similarity indices to C.auratus red var is most greatly 0.8116, its genetic distances is 0.1884, which infers that the phylogenetic relationships of C.auratus cuvieri and C.auratus red var are nearer. Cluster's analysis indicates four
populations of Auratus are divided into two groups , C.auratus cuvieri and C.auratus red var are a group, and C.auratus auratus and C.auratus auratus var.Pengze are in another 1 group.
用Crooijmans et al从鲤鱼中分离出来的17对微卫星引物(MFW14-MFW19，MFW21-MFW31)和依据鲫鱼的微卫星序列自行设计一对微卫星引物(MFW32)，共计18对引物，对四种群体的鲫鱼基因组DNA进行扩增，筛选扩增带型清晰、特异性强、重复性好的引物MFW14、MFW16、MFW18、MFW24、MFW25、MFW26、MFW28、MFW29、MFW32用作结果分析，其中引物MFW18、MFW24、MFW26、MFW29可作为区分四种鲫鱼群体的微卫星标记的有效引物。
从群体内来看，9个微卫星位点中共检测到46个等位基因，其中有18个等位基因为4种鲫鱼群体所共有，8个等位基因具有种群特异性；4种鲫鱼的平均遗传杂合度在0.607-0.720之间，白鲫为0.720、红鲫为0.652、彭泽鲫为0.629、鲫鱼为0.607；平均多态信息含量在0.530-0.650之间，白鲫为0.650、红鲫为0.587、彭泽鲫为0.549、鲫鱼为0.530，由此，白鲫的遗传多样性最高，鲫鱼的遗传多样性最低。
从群体间来看，四种群体中，鲫鱼和白鲫间的遗传相似性指数最小，为0.7143，其遗传距离为0.2857，说明这两种群体亲缘关系较远；白鲫与红鲫的遗传相似性指数最大，为0.8116，其遗传距离为0.1884，这可推断白鲫与红鲫亲缘关系较近。聚类分析结果表明，4种鲫鱼分为两组，白鲫和红鲫为1个组，而鲫鱼和彭泽鲫处于另1个组。
Using blood,muscle,liver and kidney from Auratus as the material, two different methods were compared in extraction of Auratus genomic DNA ,which belonged to Phenol-chloroform and Molecular biology Kits. The results show that the production concentration was 385.8-3167.5ng/uL and A260/A28o was 1.66-1.87. they can both be amplified by the the polymerase chain reaction(PCR) of Microsatellite DNA ,but Molecular biology Kits is feasible for the research.
This research adopts 18 primers of Microsatellite DNA that Crooijmans et al separated out 17 primers from the carp (MFW14-MFW19, MFW21-MFW31), and designing 1 primer (MFW32 ) according to microsatellite DNA sequence of Auratus by myself. The clear, peculiar and repetitive trips of the the polymerase chain reaction(PCR) products of the MFW14 , MFW16 , MFW18 , MFW24 , MFW25 , MFW26 , MFW28 , MFW29 , MFW32 among four populations of Auratus genomic DNA from different individuals using 18 primers are analyzed by the research.The prmers of MFW18, MFW24, MFW26, MFW29 can be differentiated from four populations of Auratus.
From four intrapopulations of Auratus, 9 Microsatellite DNA location measures 46 alleles, among them 18 alleles are in all by four populations of Auratus, 8 alleles are peculiar in a population among them. The average heterozygosity of four populations of Auratus is 0.607-0.720, C.auratus cuvieri is 0.720, C.auratus red va is 0.652, C.auratus auratus var. Pengze is 0.629, C.auratus auratu is 0.607; The average polymorphism information content is 0.530-0.650, C.auratus cuvieri is 0.650, C.auratus red var is 0.587, C.auratus auratus var. Pengze is 0.549, C.auratus auratus is 0.530, so the genetic diversity of C.auratus cuvieri is the highest and the genetic diversity of C.auratus auratus is the lowest.
From four populations of Auratus, it is 0.7143 that genetic similarity indices between C.auratus auratus and C.auratus cuvieri is minimum , its genetic distances is 0.2857, proving the phylogenetic relationships of two populations are relatively far. C.auratus cuvieri genetic similarity indices to C.auratus red var is most greatly 0.8116, its genetic distances is 0.1884, which infers that the phylogenetic relationships of C.auratus cuvieri and C.auratus red var are nearer. Cluster's analysis indicates four
populations of Auratus are divided into two groups , C.auratus cuvieri and C.auratus red var are a group, and C.auratus auratus and C.auratus auratus var.Pengze are in another 1 group.
引文
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