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用核微卫星的长度多态性刻画物种的种群遗传结构,推测其进化历史是现时景观遗传学、亲缘地理学的重要研究手段之一。选择具有合适多态性且突变机制相对明确的核微卫星对研究结果的准确性至关重要。通过收集15个已被广泛使用的梣属(Fraxinus)核微卫星,研究其在水曲柳(Fraxinus mandshurica)中的扩增稳定性,同时应用FIASCO方法针对水曲柳新开发了9对微卫星引物,并用克隆方法抽样检测能成功扩增的核微卫星序列,分析其长度变异的突变机制。研究结果表明,有66.7%的梣属核微卫星引物可以在水曲柳中稳定扩增,但其中有70%的引物在微卫星翼区普遍存在插入缺失,从而导致长度同塑。在新开发的9对引物中,仅33.3%的引物在翼区存在插入缺失。研究结果显示微卫星的长度同塑现象较为普遍,尤其是根据近缘物种开发的引物,这种现象应在利用微卫星的研究中得到重视。
Using the length polymorphism of nuclear microsatellite to characterize the population genetic structure of species, we speculate that its evolutionary history is one of the important research methods of landscape genetics and kinship geography. The selection of a nuclear microsatellite with a suitable polymorphism and a relatively clear mutation mechanism is crucial to the accuracy of the results. By collecting 15 widely used nuclear microsatellites of Fraxinus to study their amplification stability in Fraxinus mandshurica, 9 pairs of microsatellites have been newly developed for Fraxinus mandshurica using FIASCO method The primers were designed and the cloning method was used to detect the nuclear microsatellite sequences that could be amplified successfully. The mutation mechanism of length variation was analyzed. The results showed that 66.7% of Prunus nuclear microsatellite primers could be stably amplified in Fraxinus mandshurica, however, 70% of primers were ubiquitously inserted in the microsatellite wing region, resulting in the same length. Among the newly developed 9 pairs of primers, only 33.3% of primers had insertion deletion in the wing region. The results show that the length of microsatellites with the plastic phenomenon is more common, especially in accordance with the development of primers of related species, this phenomenon should be used in the study of microsatellites get attention.