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基于SLAF-seq技术开发蓝圆鲹微卫星标记及跨物种扩增检测
翟云,吴仁协,牛素芳,周玲莉,邱琦,张浩冉
0
(广东海洋大学水产学院,广东 湛江 524088)
摘要:
采用基于高通量测序平台的SLAF-seq技术开发蓝圆鲹(Decapterus maruadsi)微卫星标记,从获得的1 450个四至六碱基重复微卫星位点中随机挑选93个位点进行引物合成和多态性检测,最终开发出31个多态性微卫星标记.群体遗传学检测结果表明,31个位点的等位基因数为4~29(均值13.48),观测杂合度(Ho)为0.281~0.875(均值0.650),期望杂合度(He)为0.557~0.958(均值0.838),多态信息含量值(PIC)为0.456~0.939(均值0.803),表明所开发的31个微卫星位点均具有较高的多态性.Hardy-Weinberg平衡检测表明,17个位点符合HWE(“哈迪-温伯格”平衡),且各位点间不存在连锁不平衡现象.跨物种扩增结果显示,分别有27、20、20、18、9个微卫星标记在颌圆鲹(Decapterus macarellus)、长身圆鲹(Decapterus macrosoma)、无斑圆鲹(Decapterus kurroides)、竹荚鱼(Trachurus japonicus)及金带细鲹(Selaroides leptolepis)中具有较好的通用性.本研究开发的多态性长碱基重复微卫星标记可为蓝圆鲹渔业资源的科学管理和保护提供有力的遗传学评估手段,亦可为圆鲹属和鲹科鱼类系统进化研究提供新的分析手段.
关键词:  海洋生物学  蓝圆鲹  微卫星标记  SLAF-seq  跨物种扩增
DOI:10.3969/J.ISSN.2095-4972.2018.03.016
基金项目:广东省自然科学基金-博士启动资助项目(2016A030310329);广东海洋大学优秀青年骨干教师培养计划资助项目(HDYQ2017002);广东海洋大学科研启动费资助项目(R17040);广东省科技计划资助项目(2017A030303077)
Microsatellite loci developed by SLAF-seq in blue scad, Decapterus maruadsi, and cross-species amplification test
ZHAI Yun,WU Ren-xie,NIU Su-fang,ZHOU Ling-li,QIU Qi,ZHANG Hao-ran
(College of Fisheries, Guangdong Ocean University, Zhanjiang 524088, China)
Abstract:
In this study SLAF-seq technique based on the next generation sequencing platform was used to develop Decapterus maruadsi microsatellite markers. Ninety-three microsatellite loci were randomly selected from 1 450 multiple nucleotides microsatellite loci with tetra-nucleotide to hexa-nucleotide bases for primer synthesis and polymorphism detection. Finally, thirty-one highly polymorphic markers were developed. The population genetic analysis resulted a high polymorphism with a range of 4 to 29 alleles (mean 13.48). Observed and expected heterozygosities varied from 0.281 to 0.875 (mean 0.650) and 0.557 to 0.958 (mean 0.838), respectively. The average of polymorphism information content was 0.803 (ranged from 0.456 to 0.939) indicating that the 31 microsatellite markers were highly polymorphic. The results of Hardy-Weinberg equilibrium test showed that 17 microsatellite markers conformed to HWE, and no chain imbalance was detected. Cross-amplifications discovered a total of 27, 20, 20, 18, and 9 out of 31 microsatellite markers were successfully across in Decapterus macarellus, Decapterus macrosoma, Decapterus kurroides, Trachurus japonicus and Selaroides leptolepis, respectively. These long repeat microsatellites can provide a powerful genetic tool for assessment of scientific management and conservation of the blue scad fishery resources. In addition, it provides a new tool in studying phylogenetic relationship among the Decapterus and Carangidae species.
Key words:  marine biology  Decapterus maruadsi  microsatellite loci  SLAF-seq  cross-amplification

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