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微卫星的亲子鉴定技术既能减少分池饲养带来的遗传性状的差异又能减少其所占用饲养池的数量和管理强度,在水产养殖和品种选育中得到了广泛的应用。本研究应用21对微卫星标记对鲤(Cyprinus carpio)杂交家系(49-同胞家系:德国镜鲤(C.carpio var.mirror)♀×荷包红鲤(C.carpio var.wuynanensis)♂;24-同胞家系:荷包红鲤♀×德国镜鲤♂)进行遗传多样性及遗传特性的评价,并检测影响微卫星标记鉴定准确性的几个特性。研究结果标明,(1)筛选出的13对标记:在49-同胞家系中,平均有效等位基因数(K)、期望杂合度(He)、观测杂合度(Ho)和多态信息含量(PIC)分别为0.505、0.648、0.791和0.5889;在24-同胞家系中,K、He、Ho和PIC分别为0.548、0.670、0.819和0.6138。(2)筛选出的13对标记:在49-同胞家系和24-同胞家系中的联合排除率(CPE)分别达到0.999997978和0.999999583。(3)在两个杂交家系中,4个核心位点(MFW29,HLJ392,HLJ044,HLJ855)的联合排除率均高于99%。(4)三种情况下(一个亲本已知,NE-1P;另一个亲本已知,NE-2P;双亲都已知,NE-PP;)的联合非父排除率在49全胞家系中分别为1.34E-02、3.91E-04和2.02E-06;在24全胞家系中分别为1.34E-02、3.91E-04和2.02E-06。(5)模拟分析显示(置信度为99%),当使用11个或更多标记时,在49全胞家系中亲本与子代的配对率为100%;当使用9个或更多标记时,在24全胞家系中亲本与子代的配对率大于98%。(6)UPGMA聚类图表明:在混养条件下,筛选出的13对标记可以明显地将子代与无亲缘关系的家系区分开。本研究筛选出13对微卫星标记完全适用于荷包红鲤与德国镜鲤杂交家系的鉴定,为品种选育提供有效的鉴定工具。
Microsatellite paternity testing technology can not only reduce the genetic traits brought by sub-pooling, but also reduce the number and management intensity of the rearing ponds. It has been widely used in aquaculture and breeding. In this study, 21 pairs of microsatellite markers were used to breed Cyprinus carpio hybrids (49-sibling family: C.carpio var.mirror × C.carpio var. Woynanensis ♂; Sibling family: purse red carp × Germany mirror carp ♂) for genetic diversity and genetic characteristics of the evaluation, and test the impact of microsatellite markers identified several of the characteristics of accuracy. The results showed that: (1) 13 pairs of markers were screened: the average number of effective alleles (K), expected heterozygosity (He), observed heterozygosity (Ho) and polymorphism information content in 49- PIC) were 0.505, 0.648, 0.791 and 0.5889, respectively; in the 24-sib family, K, He, Ho and PIC were 0.548, 0.670, 0.819 and 0.6138, respectively. (2) 13 pairs of markers screened: The combined elimination rate (CPE) of 49- and 24-sib families reached 0.999997978 and 0.999999583, respectively. (3) The combined exclusion rates of the four core loci (MFW29, HLJ392, HLJ044 and HLJ855) were higher than 99% in the two crossbred families. (4) The combined nonparent exclusion rate of the 49 full-length sib families in three cases (one parent known to NE-1P; the other parent is known, NE-2P; both parents are known, NE-PP; 1.34E-02, 3.91E-04 and 2.02E-06, respectively; and 1.34E-02, 3.91E-04 and 2.02E-06 in the 24-cell family. (5) The simulation analysis showed that the pairing rate of parents and offspring was 100% in the 49 cell lineage when using 11 or more markers (with a confidence of 99%); when using 9 or more markers , The pairing rate of parents and offspring in 24-cell family was more than 98%. (6) The UPGMA clustering map shows that under the conditions of polyculture, 13 pairs of markers screened can clearly distinguish offspring from unrelated families. In this study, 13 pairs of microsatellite markers were screened for the identification of the hybrid pedigree of Purse red carp and German mirror carp, so as to provide an effective identification tool for breed breeding.