大麻哈鱼SRAP体系建立及其遗传多样性研究
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摘要
本研究将正交设计与单因子试验相结合,建立了大麻哈鱼SRAP-PCR优化反应体系：在15μL反应液中,Taq DNA聚合酶0.75U,引物浓度O.3μmol/L, dNTPs浓度0.3mmol/L, Mg2+浓度2.5mmol/L, DNA模板100ng。
利用该体系对大麻哈鱼四个群体(绥芬河野生群体、绥芬河养殖群体、抚远野生群体、抚远养殖群体)进行遗传多样性分析,从100对引物组合中筛选出47组条带清晰、多态性丰富的组合,不同引物组合检测位点数5-14不等。在大麻哈鱼四个群体中共检测到381个位点,其中多态位点281个,多态座位比例(P)73.75%,显示了较高的多态性,表明大麻哈鱼群体遗传多样性丰富,种质资源好。四个群体的多态位点比例由高到低依次为：绥芬河野生群体(62.20%)>抚远野生群体(58.27%)>抚远养殖群体(52.76%)>绥芬河养殖群体(51.97%); Shannon's信息指数(I)水平由高到低依次为：绥芬河野生群体(0.3493)>抚远野生群体(0.3354)>抚远养殖群体(0.3080)>绥芬河养殖群体(0.2738)。这两方面数据均表明,大麻哈鱼野生群体的遗传多样性相对高于养殖群体,可见鱼类的生存环境与其遗传多样性是相关联的。
采用同工酶标记对大麻哈鱼群体遗传多样性进行研究,所得结果与SRAP标记结果一致,从蛋白质水平上验证了SRAP研究结果的科学性和准确性。两种方法在不同水平上进行遗传多样性分析,从不同角度对大麻哈鱼群体遗传多样性做出更为科学全面的评价。
This research unifies the orthogonal design and the single factor test, establishes SRAP-PCR optimization reacting system on Oncorhynchus keta. The optimal PCR system is obtained as Taq DNA polymerase 0.75U, primer 0.3μmol/L, dNTPs 0.3mmol/L, Mg2+ 2.5mmol/L, and DNA template 100ng in 15μL reaction system.
Applying this system to the genetic diversity analysis of four Oncorhynchus keta populations (Suifenhe wild population, Suifenhe cultured population, Fuyuan wild population and Fuyuan cultured population).47 primer combinations are selected from 100 which produced clear bands and polymorphic combinations. Different primer combination finds the different number loci from 5 to 14. From four Oncorhynchus keta populations 281 polymorphic loci are detected from 381 loci totally, and the percentage of polymorphic loci is 73.75%.The high polymorphism indicates that the genetic diversity of Oncorhynchus keta is rich and has an ideal germplasm. The percentage of polymorphic loci from four populations in order is:Suifenhe wild population (62.20%)>Fuyuan wild population (58.27%)>Fuyuan cultured population (52.76%)>Suifenhe cultured population (51.97%). The Shannon's Information index in order is:Suifenhe wild population (0.3493)>Fuyuan wild population (0.3354)>Fuyuan cultured population (0.3080)>Suifenhe cultured population (0.2738). Considering all the data of two respective aspects above, it's clear that the genetic diversity of Oncorhynchus keta wild population is relatively higher than Oncorhynchus keta cultured population. As a result, the living environment of fish and their genetic diversity is associated.
Using isozyme markers to study the genetic diversity of Oncorhynchus keta populations, the results are consistent with the SRAP marker, and also demonstrate that the results from SRAP marker are scientific and accurate. Using these two methods to analyze genetic diversity from different levels could make a more scientific and comprehensive evaluation on the genetic diversity of Oncorhynchus keta from different angles.
利用该体系对大麻哈鱼四个群体(绥芬河野生群体、绥芬河养殖群体、抚远野生群体、抚远养殖群体)进行遗传多样性分析,从100对引物组合中筛选出47组条带清晰、多态性丰富的组合,不同引物组合检测位点数5-14不等。在大麻哈鱼四个群体中共检测到381个位点,其中多态位点281个,多态座位比例(P)73.75%,显示了较高的多态性,表明大麻哈鱼群体遗传多样性丰富,种质资源好。四个群体的多态位点比例由高到低依次为：绥芬河野生群体(62.20%)>抚远野生群体(58.27%)>抚远养殖群体(52.76%)>绥芬河养殖群体(51.97%); Shannon's信息指数(I)水平由高到低依次为：绥芬河野生群体(0.3493)>抚远野生群体(0.3354)>抚远养殖群体(0.3080)>绥芬河养殖群体(0.2738)。这两方面数据均表明,大麻哈鱼野生群体的遗传多样性相对高于养殖群体,可见鱼类的生存环境与其遗传多样性是相关联的。
采用同工酶标记对大麻哈鱼群体遗传多样性进行研究,所得结果与SRAP标记结果一致,从蛋白质水平上验证了SRAP研究结果的科学性和准确性。两种方法在不同水平上进行遗传多样性分析,从不同角度对大麻哈鱼群体遗传多样性做出更为科学全面的评价。
This research unifies the orthogonal design and the single factor test, establishes SRAP-PCR optimization reacting system on Oncorhynchus keta. The optimal PCR system is obtained as Taq DNA polymerase 0.75U, primer 0.3μmol/L, dNTPs 0.3mmol/L, Mg2+ 2.5mmol/L, and DNA template 100ng in 15μL reaction system.
Applying this system to the genetic diversity analysis of four Oncorhynchus keta populations (Suifenhe wild population, Suifenhe cultured population, Fuyuan wild population and Fuyuan cultured population).47 primer combinations are selected from 100 which produced clear bands and polymorphic combinations. Different primer combination finds the different number loci from 5 to 14. From four Oncorhynchus keta populations 281 polymorphic loci are detected from 381 loci totally, and the percentage of polymorphic loci is 73.75%.The high polymorphism indicates that the genetic diversity of Oncorhynchus keta is rich and has an ideal germplasm. The percentage of polymorphic loci from four populations in order is:Suifenhe wild population (62.20%)>Fuyuan wild population (58.27%)>Fuyuan cultured population (52.76%)>Suifenhe cultured population (51.97%). The Shannon's Information index in order is:Suifenhe wild population (0.3493)>Fuyuan wild population (0.3354)>Fuyuan cultured population (0.3080)>Suifenhe cultured population (0.2738). Considering all the data of two respective aspects above, it's clear that the genetic diversity of Oncorhynchus keta wild population is relatively higher than Oncorhynchus keta cultured population. As a result, the living environment of fish and their genetic diversity is associated.
Using isozyme markers to study the genetic diversity of Oncorhynchus keta populations, the results are consistent with the SRAP marker, and also demonstrate that the results from SRAP marker are scientific and accurate. Using these two methods to analyze genetic diversity from different levels could make a more scientific and comprehensive evaluation on the genetic diversity of Oncorhynchus keta from different angles.
引文
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