基于RAD-seq技术的长体圆鲹二、三核苷酸重复微卫星标记开发与评价  

Development and evaluation of di-/tri-nucleotide-repeated microsatellites by RAD-seq in Decapterus macrosoma

作　　者：孔啸兰[1] 李敏[1] 陈作志[1] 龚玉艳[1] 张俊[1] 张鹏[1] KONG Xiaolan;LI Min;CHEN Zuozhi;GONG Yuyan;ZHANG Jun;ZHANG Peng(South China Sea Fisheries ResearchInstitute,Chinese Academy of Fishery Sciences,Guangzhou 510300,China)

机构地区：[1]中国水产科学研究院南海水产研究所农业农村部外海渔业开发重点实验室

出　　处：《南方水产科学》2019年第3期97-103,共7页South China Fisheries Science

基　　金：国家重点研发计划项目(2018YFC1406502);农业农村部财政专项(NFZX2018);中国水产科学研究院基本科研业务费专项(2017YB23);广东省促进经济发展专项(GDME-2018E004);中国水产科学研究院南海水产研究所中央级公益性科研院所基本科研业务费专项资金资助(2015TS04);农业农村部东海与远洋渔业资源开发利用重点实验室开放课题(201720761)

摘　　要：通过对长体圆鲹(Decapterus macrosoma)基因组进行RAD-Seq高通量测序,共获得58 180条微卫星序列,选取112条二、三核苷酸重复的微卫星序列设计引物,经筛选后,共获得27个具有多态性的微卫星标记。利用一个长体圆鲹群体对通过筛选的微卫星标记的种群遗传学特征进行评价。结果显示,27对引物扩增的序列中18个位点为二核苷酸重复,重复次数为9~14次,9个位点为三核苷酸重复,重复次数为6~10次,等位基因数(Na)为5~17 (平均10.6),表观杂合度(Ho)为0.342 9~0.857 1 (平均0.631 7),期望杂合度(He)为0.538 3~0.911 8(平均0.796 8),多肽信息含量(PIC)为0.497~0.886 (平均0.780 9),除1个位点外,其他位点PIC值均大于0.500,表明开发的微卫星位点具有较高的多态性。"哈迪-温伯格"平衡(HWE)检测结果显示,19个标记等位基因频率符合HWE。连锁不平衡检测表明各位点间无连锁不平衡现象。该研究开发的27个微卫星标记可为长体圆鲹种群遗传学研究提供基础。A total of 58 180 microsatellites were developed by RAD-Seq high-throughput sequencing technology in Decapterus macrosoma. One hundred and twelve microsatellite loci were randomly selected from di-/tri-nucleotide repeat microsatellite loci. Finally, twenty-seven highly polymorphic markers were developed. The population genetic analysis finds 18 dinucleotide-repeated microsatellite loci(9-14 repeated) and 9 trinucleotide-repeated microsatellite loci(6-10 repeated). The number of alleles(Na) ranged from 5 to 17(mean 10.6). The observed and expected heterozygosities ranged from 0.342 9 to 0.857 1(mean 0.631 7) and 0.538 3 to0.911 8(mean 0.796 8), respectively. The polymorphism information content(PIC) ranged from 0.497 to 0.886(mean 0.780 9), and the PIC at all loci except for one was geater than 0.500,indicating that the microsatellites were highly polymorphic. The HardyWeinberg equilibrium(HWE) test shows that 19 microsatellite markers conformed to HWE. There is no linkage imbalance among the 27 loci. The 27 microsatellites developed by this study will be useful for further population genetic analysis.

关 键 词：长体圆鲹 高通量测序 微卫星标记 

分 类 号：S931[农业科学—渔业资源] Q347[农业科学—水产科学]