基于转录组测序的高原鼢鼠多态性微卫星标记筛选  

Screening of polymorphic microsatellite markers in the plateau zokor based on transcriptome sequencing

作　　者：刘倩倩 谭宇尘 姚宝辉 康宇坤 苏军虎[1,2,3] LIU Qianqian;TAN Yuchen;YAO Baohui;KANG Yukun;SU Junhu(Pratacultural College/Key Laboratory of Grassland Ecosystem(Ministry of Education)/Pratacultural Engineering Laboratory of Gansu Province,Sino–U.S.Centers for Grazing land Ecosystem Sustainability,Gansu Agricultural University,Lanzhou 730070,Gansu,China;Gansu Agricultural University–Massey University Research Centre for Grassland Biodiversity,Gansu Agricultural University,Lanzhou 730070,Gansu,China;Gansu Qilianshan Grassland Ecosystem Observation and Research Station,Lanzhou 730070,Gansu,China)

机构地区：[1]甘肃农业大学草业学院/草业生态系统教育部重点实验室/中–美草地畜牧业可持续发展研究中心,甘肃兰州730070 [2]甘肃农业大学–新西兰梅西大学草地生物多样性研究中心,甘肃兰州730070 [3]甘肃省祁连山草原生态系统野外科学观测研究站,甘肃兰州730070

出　　处：《草业科学》2021年第12期2481-2489,共9页Pratacultural Science

基　　金：国家自然科学基金项目(31760706);甘肃省草原生态修复治理科技支撑项目(LCJ2021020)。

摘　　要：利用第二代高通量测序技术对高原鼢鼠(Eospalax baileyi)大脑组织进行转录组测序,筛选微卫星标记,分析其组成与特征,并检验其多态性,为高原鼢鼠微卫星标记的开发及后续研究等奠定基础。结果发现:在获得的29090个微卫星标记中单核苷酸重复A/T是重复次数最多的类型(54.31%),其次为二碱基重复CA/TG(10.46%),其他碱基重复类型仅占16.79%。随机选取104个微卫星位点进行引物设计,发现有43个位点扩增成功。12个多态性位点经毛细管电泳检测发现,等位基因数为3~12个,观测杂合度(Ho)为0.263~0.937,平均值为0.651,期望杂合度(He)为0.285~0.844,平均值为0.600。使用Bonferroni校正后,有2个微卫星位点显著偏离哈迪–温伯格平衡(P<0.01),10个微卫星位点未观察到显著的连锁不平衡(P>0.01)。多态信息含量(PIC)在0.251~0.816,全部为中度多态性(0.250<PIC<0.500)以上的位点,且有7个微卫星位点表现为高度多态性(PIC>0.500)。这些微卫星标记为高原鼢鼠的亲权关系鉴定、种群遗传学等研究提供了基础数据。The second generation of high-throughput sequencing technology was used for transcriptional sequencing of the brain tissue of the plateau zokor(Eospalax baileyi).Microsatellite markers were screened,their composition and characteristics were analyzed,and their polymorphisms were examined,which laid a foundation for the development of microsatellite markers in the plateau zokor.The results showed that among the 29090 microsatellite markers obtained,the main repeat types were mononucleotide repeats A/T(54.31%),followed by the dinucleotide repeats CA/TG(10.46%),and other base repeat types only accounted for 16.79%.A total of 104 microsatellite loci were randomly selected for primer design,and 43 loci were successfully amplified.After 12 polymorphic loci were detected by capillary electrophoresis,we found that the number of alleles was 3~12,the observed heterozygosity(Ho)was 0.263~0.937,with an average value of 0.651,and the expected heterozygosity(He)was 0.285~0.844,with an average value of 0.600.After Bonferroni correction,two microsatellite loci deviated significantly from Hardy–Weinberg equilibrium(P<0.01),and no significant linkage disequilibrium was observed in the 10 microsatellite loci(P>0.01).The polymorphism information content(PIC)ranged from 0.251 to 0.816,all loci were moderately polymorphic(0.250<PIC<0.500),and seven microsatellite loci were highly polymorphic(PIC>0.500).These microsatellite markers provided basic data for the identification of parental relationships and population genetics of the plateau zokor.

关 键 词：高原鼢鼠 啮齿动物 转录组 微卫星 多态性 分子标记 筛选 

分 类 号：S812.6[农业科学—草业科学]