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Identification of polymorphic microsatellite DNA loci and paternity testing of Amur tigers

ZHANG Yu-Guang ①② LI Di-Qiang① RAO Li-Qun② XIAO Qi-Ming② LIU Dan③ (① Institute of Forestry Ecology, Environment and Protection, Chinese Academy of Forestry, Beijing 100091, China) (② Hunan Agricultural University, Changsha 410128, China) (③ Ha'erbin Amur Tiger Park, Harerbin 150001, China)  
We identified the paternity of some Amur tigers (Panthera tigris altaica) in Ha'erbin Tiger Park and constructed clear pedigrees using microsatellite loci.Four blood (No.209, No.232, No.233, No.244) and twenty-three hair specimens from Amur tigers were examined. Eighteen polymorphic microsatellite DNA loci identified in the domestic cat (Felis catus) were amplified in the Amur tiger genome. It was found that four of the loci failed to be amplified, eight were monomorphic and six (Fca005, Fca075, Fca094, Fca152, Fca161, Fca294) were polymorphic. At the same time, we designed eight pairs of microsatellite primers based on Sumatran tiger (Panthera tigris sumatrae) gene sequences for amplification in the Amur tiger. The results show that four pairs of primers produced a monomorphic pattern and the other four (Pti002, Pti003, Pti007, Pti010) gave a polymorphic pattern. Altogether, 38.5% of the twenty six microsatellite markers were polymorphic, and inheritance patterns of alleles at these loci show close agreement with the well documented pedigrees.Seven tigers (No.067, No.130, No.132, No.209, No,232, No.233, No.244) had ambiguous paternity. We identified their biological fathers (No.048, No.048, No.057, No.048, No.019, No.081, No.075, respectively) according to the inheritance patterns of ten polymorphic microsatellite loci. This analysis revealed that male tiger No.048 had a relatively high propagation competence while some other males had fathered relatively few progeny. In this study, we obtained the genome DNA from hairs using the non-invasive DNA technology. We demonstrated that DNA derived from hair samples is as good as that obtained from blood for the analysis of microsatellite polymorphism. These results imply that microsatellite DNA markers and non-invasive DNA technology can help establish the detailed pedigrees required for the captive husbandry of the Amur tiger. We suggest that this method could be useful in the captive management of other endangered species[Acta Zoologica Sinica 49(1):118-123,2003].
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