Determination of the minimum number of microsatellite markers for individual genotyping in wild boar (Sus scrofa) using a test with close relatives
Determination of the minimum number of microsatellite markers for individual genotyping in wild boar (Sus scrofa) using a test with close relatives
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DOI：
10.1007/s10344-011-0588-9
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发表时间：
2012-06-01
影响因子：
2
通讯作者：
Schulz, R.
中科院分区：
文献类型：
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作者：
Kolodziej, K.;Theissinger, K.;Schulz, R.
为了开发一种无创的、可行的野猪种群大小估计方法，我们提出了一种逐步减少个体基因分型所需微卫星标记数量的方法。Step1：对12个微卫星位点的初始标记集进行非靶DNA的物种特异性测试，结果排除了两个标记。步骤2：关于杂合度和偏离Hardy-Weinberg平衡的变异性测试导致两个进一步的标记被拒绝。步骤3：用三个单独的野猪样本集测试其余8个标记在种群间的可转移性。步骤4：根据同一性值的概率，可以将标记从8个减少到5个。第五步：一项使用雌性野猪及其胚胎组织样本的新测试提供了证据，证明四个可变微卫星标记和一个性别标记足以用于近亲的个体识别。步骤6：最后使用粪便样本估计PCR （PS）和基因分型成功率（GS）。综上所述，我们推荐一种特定的四标记组合，PS和GS的比例均为50%，用于可靠的野猪非侵入性种群大小估计的个体识别。
In the context of developing a noninvasive, practicable method for population size estimation in wild boar, we present a stepwise procedure to reduce the number of required microsatellite markers for individual genotyping. Step1: an initial marker set of 12 microsatellite loci was tested for species specificity with nontarget DNA and resulted in an exclusion of two markers. Step 2: a variability test regarding heterozygosity and deviations from Hardy-Weinberg equilibrium led to the rejection of two further markers. Step 3: the remaining eight markers were tested for transferability across populations with three separate wild boar sample sets. Step 4: on the basis of probability of identity values, a reduction from eight to five markers was possible. Step 5: a novel test using tissue samples from female wild boars and their embryos provided evidence that four variable microsatellite markers and one sex marker are sufficient for individual identification of close relatives. Step 6: feces samples were finally used to estimate PCR (PS) and genotyping success (GS). In conclusion, we recommend a specific four-marker combination with both PS and GS > 50% for a reliable individual identification in noninvasive population size estimation of wild boar.