应用微卫星DNA标记进行蓝狐（Alopex lagopus）亲权鉴定和分子遗传多样性研究
摘要
蓝狐(Alopex lagopus)是我国重要的毛皮经济动物,养殖规模逐年扩大。为进一步了解中国蓝狐养殖群体的遗传多样性,并推动蓝狐养殖遗传管理工作的开展,本研究利用15个微卫星标记对5个蓝狐养殖群体的288个个体进行DNA多态性测定和统计分析。计算了等位基因频率、遗传杂合度、多态信息含量和奈氏标准遗传距离,采用NJ法构建了系统发生树。并对大兴安岭地区养殖的6窝蓝狐进行了单亲亲权鉴定。结果表明:
1.本文从20个微卫星位点中筛选出的15个微卫星位点多态信息含量较高,杂合性好,均呈现高度多态性和稳定的扩增效果,可以作为蓝狐遗传学研究的可靠的遗传标记。
2.5个地区蓝狐养殖群体在15个微卫星位点上共检测到100个等位基因,每个微卫星位点的等位基因数量为5～11个,平均每个位点6.67个,基因频率分布在0～1之间。说明蓝狐养殖群体的遗传多样性还比较丰富。
3.5个蓝狐养殖群体中锦州地区蓝狐养殖群体遗传多样性最为丰富,群体多态信息含量、平均杂合度和有效等位基因数亦为最高,其次依此为伊春地区、大兴安岭地区、大连地区和哈尔滨地区。但是多态信息含量、群体杂合度及有效等位基因数反映多态性并不完全一致。
4.由Nei氏(1972)标准遗传距离绘制出5个地区蓝狐养殖群体NJ聚类关系图。NJ聚类关系图可以看出5个地区的群体全部聚为一支,大连地区和大兴安岭地区先聚在一起,再和哈尔滨地区聚为一支,进而和锦州地区聚为一支,最终和伊春地区的养殖群体合为一支。
5.本文用15个微卫星标记进行了单亲亲权鉴定,找出了4个系谱吊牌丢失后裔的母本,对其余有明确系谱记录的6窝蓝狐的子代中有2只子代记录错误。亲子鉴定的排除概率和亲权指数较高,累积排除概率达0.999996,结果可靠。
Blue fox(Alopex lagopus)was for important for fur farming,creating lots of economy benefits every year in China.Blue fox farming had a long history in China,and the population was increasing each year.In order to further investigate the genetic diversity of Chinese blue fox farming,and improve the genetic management,this study screened total of 5 populations (n=288)using microsatellite markers to analyze statistically the DNA diversity of these colonies respectively.Allele frequency,heterozygosity value(H),polymorphism information content(PIC),Nei's standard distance was calculated.Dendrogram was constructed using the Neighbor-Joining methods.Father-son Parentage of 6 families in Greater Hinggan Mountains was identified.The results as following:
1.By using fluorescent-labeling-based automated DNA sequencing and microsatellite polymorphism analyzing techniques,of 20 microsatellite loci,15 were proved polymorphic,reproducible and reliable in amplifying blue fox genomic DNA.They were credible genetic markers in blue fox genetic study.
2.Five populations were observed 100 alleles in 15 microsatellite loci.Each microsatellite loci had 5～11 alleles,average number of allele was 6.67,allele frequency was 0～1.These results showed there were richer genetic diversity in blue fox farming.
3.Among five populations Jinzhou's population had the richest genetic diversity, polymorphism information content(PIC),heterozygosity and effective allele were the highest.The following were Yichun,Greater Hinggan Mountains,Dlian and Harbin.But polymorphism information content(PIC),mean heterozygosity(H),effective number of alleles(Ne)did not have good consistency to affect genetic diversity.
4.According to Nei's standard distance(1972),dendrogram was constructed using the Neighbor-Joining methods.Based on NJ dendrogram,5 Blue fox populations could be clustered into one group.The first group included Dalian and Greater Hinggan Mountains, the second clustered with Harbin,the third clustered with Jinzhou,finally clustered with Yichun.
5.This study had identified Father-son Parentage of 6 families in Greater Hinggan Mountains by 15 microsatellite marks.The biological parents of 4 blue foxes without pedigree recording were found,and false recording of 2 individuals were modified.Cumulative chance of exclusion(CCE)and Paternity index were higher.In father-son set,fifteen markers will exclude the alleged parent with a probability of 0.9986.
1.本文从20个微卫星位点中筛选出的15个微卫星位点多态信息含量较高,杂合性好,均呈现高度多态性和稳定的扩增效果,可以作为蓝狐遗传学研究的可靠的遗传标记。
2.5个地区蓝狐养殖群体在15个微卫星位点上共检测到100个等位基因,每个微卫星位点的等位基因数量为5～11个,平均每个位点6.67个,基因频率分布在0～1之间。说明蓝狐养殖群体的遗传多样性还比较丰富。
3.5个蓝狐养殖群体中锦州地区蓝狐养殖群体遗传多样性最为丰富,群体多态信息含量、平均杂合度和有效等位基因数亦为最高,其次依此为伊春地区、大兴安岭地区、大连地区和哈尔滨地区。但是多态信息含量、群体杂合度及有效等位基因数反映多态性并不完全一致。
4.由Nei氏(1972)标准遗传距离绘制出5个地区蓝狐养殖群体NJ聚类关系图。NJ聚类关系图可以看出5个地区的群体全部聚为一支,大连地区和大兴安岭地区先聚在一起,再和哈尔滨地区聚为一支,进而和锦州地区聚为一支,最终和伊春地区的养殖群体合为一支。
5.本文用15个微卫星标记进行了单亲亲权鉴定,找出了4个系谱吊牌丢失后裔的母本,对其余有明确系谱记录的6窝蓝狐的子代中有2只子代记录错误。亲子鉴定的排除概率和亲权指数较高,累积排除概率达0.999996,结果可靠。
Blue fox(Alopex lagopus)was for important for fur farming,creating lots of economy benefits every year in China.Blue fox farming had a long history in China,and the population was increasing each year.In order to further investigate the genetic diversity of Chinese blue fox farming,and improve the genetic management,this study screened total of 5 populations (n=288)using microsatellite markers to analyze statistically the DNA diversity of these colonies respectively.Allele frequency,heterozygosity value(H),polymorphism information content(PIC),Nei's standard distance was calculated.Dendrogram was constructed using the Neighbor-Joining methods.Father-son Parentage of 6 families in Greater Hinggan Mountains was identified.The results as following:
1.By using fluorescent-labeling-based automated DNA sequencing and microsatellite polymorphism analyzing techniques,of 20 microsatellite loci,15 were proved polymorphic,reproducible and reliable in amplifying blue fox genomic DNA.They were credible genetic markers in blue fox genetic study.
2.Five populations were observed 100 alleles in 15 microsatellite loci.Each microsatellite loci had 5～11 alleles,average number of allele was 6.67,allele frequency was 0～1.These results showed there were richer genetic diversity in blue fox farming.
3.Among five populations Jinzhou's population had the richest genetic diversity, polymorphism information content(PIC),heterozygosity and effective allele were the highest.The following were Yichun,Greater Hinggan Mountains,Dlian and Harbin.But polymorphism information content(PIC),mean heterozygosity(H),effective number of alleles(Ne)did not have good consistency to affect genetic diversity.
4.According to Nei's standard distance(1972),dendrogram was constructed using the Neighbor-Joining methods.Based on NJ dendrogram,5 Blue fox populations could be clustered into one group.The first group included Dalian and Greater Hinggan Mountains, the second clustered with Harbin,the third clustered with Jinzhou,finally clustered with Yichun.
5.This study had identified Father-son Parentage of 6 families in Greater Hinggan Mountains by 15 microsatellite marks.The biological parents of 4 blue foxes without pedigree recording were found,and false recording of 2 individuals were modified.Cumulative chance of exclusion(CCE)and Paternity index were higher.In father-son set,fifteen markers will exclude the alleged parent with a probability of 0.9986.
引文
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