基于高原鼠兔简化基因组数据的微卫星引物开发  

Development of Microsatellite Primers of Ochotona curzoniae with RAD-seq Data

作　　者：贺玉姣 胡万军 都玉蓉 王婷 杜雷 胡书立 HE Yujiao;HU Wanjun;DU Yurong;WANG Ting;DU Lei;HU Shuli(Qinghai Provincial Key Laboratory of Medicinal Plant and Animal Resources of Qinghai-Tibet Plateau,College of Life Science,Qinghai Normal University,Xining,810008,China;College of Physical Education,Qinghai Normal University,Xining,810008,China)

机构地区：[1]青海省青藏高原药用动植物资源重点实验室,青海师范大学生命科学学院,西宁810008 [2]青海师范大学体育学院,西宁810008

出　　处：《野生动物学报》2021年第3期730-736,共7页CHINESE JOURNAL OF WILDLIFE

基　　金：青海省科技计划项目青年自然科学基金(2019-ZJ-986Q)。

摘　　要：高原鼠兔(Ochotona curzoniae)是青藏高原的关键种,对其分子标记的开发有助于高原鼠兔种群生态等问题的研究。基于简化基因组测序技术获得了高原鼠兔简化基因组序列信息,并开发了微卫星分子标记。结果表明:通过MISA软件分析共检测到7064个SSR位点,其中单核苷酸和二核苷酸重复基序数量最多,各占46.15%和38.37%。四核苷酸重复基序的类型最多,达83种;其次是三核苷酸重复基序的类型,为50种。不同类型SSR位点基序的重复次数也不同。通过取SSR与InDel的交集,获得182处符合条件的SSR区域,并最终设计出90对引物。随机选择70对引物在8个高原鼠兔样本中进行验证,有42对引物具有单一条带,且多态性较好,占总检测引物的60%。研究表明,本方法在筛选高原鼠兔SSR标记中是可行的,所得引物可以应用于高原鼠兔遗传多样性、种群遗传分析等问题的研究中。Plateau pika(Ochotona curzoniae)is the keystone species on the Qinghai-Tibet Plateau.The development of molecular markers for pika contributes to the study of the population ecology of the species.We used restriction-site associated DNA sequencing(RAD-seq)technology to analyze simple sequence repeats(SSR)of plateau pika.A total of 7,064 SSR sites were detected by MicroSAtellite identification tool(MISA)analysis,among which the single nucleotide and dinucleotide repeat motifs were most abundant,accounting for 46%and 38%,respectively.The number of types in tetra-nucleotide repeat motifs was 83,followed by 50 in tri-nucleotide repeats.The number of repeats varied by SSR site.By taking the intersection of SSR and InDel,182 SSR regions were identified,and 90 pairs of primers were finally designed.Seventy pairs of primers were randomly selected for verification from eight samples.Forty-two pairs of primers had a single band and good polymorphism,accounting for 60%of the total primers selected.The results show that this method is feasible in screening SSR markers of plateau pika.The identified primers can be applied to the study of genetic diversity and population genetic analysis of plateau pika.

关 键 词：高原鼠兔 简化基因组测序 微卫星引物 

分 类 号：Q951[生物学—动物学]