基于高通量测序的二长棘鲷微卫星标记开发与评价  

Development and evaluation of microsatellite markers in Parargyrops edita

作　　者：杨兵[1,2] 林琳[1] 李纯厚[1] 徐姗楠[1] 刘永[1] 肖雅元[1] 陈作志[1] 

机构地区：[1]中国水产科学研究院南海水产研究所农业部南海资源开发利用重点实验室,广东广州510300 [2]上海海洋大学水产与生命学院,上海201306

出　　处：《南方水产科学》2015年第4期116-120,共5页South China Fisheries Science

基　　金：公益性行业(农业)科研专项经费项目(201403008);广东省自然科学基金项目(S2013010013752)

摘　　要：采用高通量测序法对二长棘鲷（Parargyrops edita）基因组进行高通量随机测序,从测序结果中搜索微卫星位点,根据搜索结果设计聚合酶链式反应（PCR）引物,并根据PCR产物质量对引物进行筛选,使用一个二长棘鲷野生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。结果共获得二长棘鲷基因组拼接序列0.12Gbp,从中搜索得到侧翼长度≥250 bp的微卫星序列292条。根据微卫星序列搜索结果设计PCR引物40对,共有15对引物通过了筛选。标记种群遗传学评价的结果显示,表观等位基因数（A）分布范围为2~16,平均为6.93;表观杂合度（HO）分布范围为0.268~0.979,平均为0.568;期望杂合度（HE）分布范围为0.300~0.914,平均为0.640。经Bonferroni校正后,除3个标记外,其余标记等位基因频率均符合＂哈迪-温伯格＂（Hardy-Weinberg）平衡预期。连锁不平衡检测表明各位点间无连锁不平衡现象。We isolated and characterized 15 polymorphic microsatellite markers in crimson seabream （Parargyrops edita） using high- throughput sequencing and polyacrylamide gel electrophoresis （PAGE）, having obtained 0. 12 Gbp spliced sequences of P. edita and searched 292 microsatellite sequences with suitable flanking regions from the spliced sequences. Then we designed 40 primer pairs in the microsatellite sequences for amplification by polymerase chain reaction （PCR）, among which 15 primer pairs were clearly ampli- fied and shown to be polymorphic. We found 2 - 16 alleles per locus （ averagely 6. 93 ）. The observed and expected heterozygosities ranged from 0. 268 to 0. 979 and from 0. 300 to 0. 914, with an average of 0. 568 and 0. 640, respectively. No loci showed significant deviation from Hardy-Weinberg equilibrium after Bonferroni correction, except for three loci. These markers will be useful for analy- zing the population genetic diversity characteristics of P. edita.

关 键 词：二长棘鲷 微卫星 高通量测序 

分 类 号：S968.3[农业科学—水产养殖]