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基于粪便DNA的小麂亲权鉴定和婚配制研究
引用本文:任鹏,龚堃,鲍毅新,黄相相,周晓,韩金巧.基于粪便DNA的小麂亲权鉴定和婚配制研究[J].生态学报,2017,37(20):6933-6944.
作者姓名:任鹏  龚堃  鲍毅新  黄相相  周晓  韩金巧
作者单位:浙江师范大学生态研究所, 金华 321004,浙江师范大学生态研究所, 金华 321004,浙江师范大学生态研究所, 金华 321004,浙江师范大学生态研究所, 金华 321004,浙江师范大学生态研究所, 金华 321004,浙江师范大学生态研究所, 金华 321004
基金项目:古田山国家级自然保护区资助项目
摘    要:2014年4月至2015年1月,在古田山国家级自然保护区内共收集634份粪便样本,2份肌肉样本。通过严格筛选,最终获得390份可用于PCR扩增的样本。用多态性较高的8个微卫星位点进行基因型分型,共识别出177个小麂个体。SRY基因性别鉴定显示研究样本中雄性84只,雌性93只。所使用的8个微卫星位点在177个样本中,平均等位基因数(A)为11,平均观测杂合度(Ho)在0.960—1.000之间,平均值为0.9685,平均期望杂合度(He)在0.799—0.887之间,平均值为0.8429,多态信息含量(PIC)在0.766—0.872之间,平均多态信息含量为0.8214,基因杂合度水平较高,为遗传多样性丰富的种群。采用Cervus3.0进行亲权分析,当置信度为95%和80%时,8个微卫星位点的鉴定率均达到100%。共鉴定出父-母-子24对,母-子23对,父-子19对,涉及到104只个体。根据亲缘关系分析小麂的婚配制,结果发现小麂的婚配制属于1雄多雌,但并不是目前所知的亚型,而可能是一种被称作"检查策略"的一雄多雌制。

关 键 词:小麂  亲权鉴定  遗传多样性  婚配制
收稿时间:2016/8/2 0:00:00

Parentage verification and mating system of Chinese muntjac (Muntiacus reevesi) based on fecal DNA
REN Peng,GONG Kun,BAO Yixin,HUANG Xiangxiang,ZHOU Xiao and HAN Jinqiao.Parentage verification and mating system of Chinese muntjac (Muntiacus reevesi) based on fecal DNA[J].Acta Ecologica Sinica,2017,37(20):6933-6944.
Authors:REN Peng  GONG Kun  BAO Yixin  HUANG Xiangxiang  ZHOU Xiao and HAN Jinqiao
Institution:Institute of Ecology, Zhejiang Normal University, Jinhua 321004, China,Institute of Ecology, Zhejiang Normal University, Jinhua 321004, China,Institute of Ecology, Zhejiang Normal University, Jinhua 321004, China,Institute of Ecology, Zhejiang Normal University, Jinhua 321004, China,Institute of Ecology, Zhejiang Normal University, Jinhua 321004, China and Institute of Ecology, Zhejiang Normal University, Jinhua 321004, China
Abstract:Microsatellite technology was utilized in the present study to investigate paternity testing, mating system, genetic diversity, and population quantity of Chinese muntjac (Muntiacus reevesi) to increase the knowledge of relationships between members of the species, reproductive strategies, and evolution potential. Therefore, the present study aims to assist the facilitation of in-depth research and protection of the species. We collected 634 fecal samples and 2 muscle samples from Chinese muntjac (M. reevesi) in Gutianshan National Nature Reserve in January 2013 and April 2014. After repeated experiments, 8 microsatellite loci were selected, which had stable amplification and high polymorphism information, and 442 samples were utilized for polymerase chain reaction (PCR) amplification. Based on the preliminary identification in terms of the fecal morphology, a verification test was conducted with the mitochondrial cytochrome b gene (Cyt b), which was regarded as a supplement to the species identification. Individual identification for Chinese muntjac was conducted by contrasting the fingerprints synthesized by the eight microsatellite marker genotypes. The SRY gene was amplified at three times to reduce any false negative influence on the fecal samples, where the target band was identified as male when it appeared more than two times. The SRY gene and 8 microsatellite loci were utilized for sexual and individual identification, respectively, and 177 individuals, including 94 females and 83 males, were identified. The results showed that the total number of microsatellite alleles were 88, and the mean number of alleles was 11, ranging from 8 to 16, and the mean observed and expected heterozygosities were 0.9685 (0.960-1.000) and 0.8429 (0.799-0.887), respectively. The polymorphism information content ranged from 0.766 to 0.872, with an average of 0.821. Based on the above data, the Chinese muntjac population in Gutianshan National Nature Reserve has a high level of genetic diversity. The identification of 8 microsatellite loci identification rate from Cervus3.0 was 100%, when the confidence level was 95% and 80%. Among 24 parentage groups, 19 father-child relationships and 23 mother-child relationships were obtained among 104 individuals. The result of the relationship analysis showed that the mating system of Chinese muntjac belongs to a polygyny. However, it does not belong to any known subtypes, and this polygyny might be called a "check strategy". During the female reproductive period, once the female individual is pregnant she will not mate with any other males. However, the relationship is not fixed and the female might still mate with another male in the next breeding season. Therefore, in the present study, the mating phenomenon of a female and several males appeared in different breeding periods.
Keywords:Chinese muntjac (Muntiacus reevesi)  parentage identification  genetic diversity  mating system
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