基于高通量测序的鹿角杯形珊瑚微卫星标记的开发

Development of microsatellite in Pocillopora damicornis using high-throughput sequencing

为了更深入开展珊瑚遗传结构及其多样性研究,本研究采用高通量测序对鹿角杯形珊瑚进行微卫星标记的开发。首先用人工白化的方法获得污染较低的鹿角杯形珊瑚,然后使用该样品DNA,基于高通量测序平台对鹿角杯形珊瑚简化基因组进行了测序,最终设计得到鹿角杯形珊瑚微卫星引物2786对。利用海南三亚鹿角杯形珊瑚对其中6对多态性较高的微卫星标记进行了评价。每个微卫星位点的有效等位基因数(Ne)为2.198~5.000;观测杂合度(Ho)为0.531~0.875;期望杂合度(He)为0.512~0.816;Shannon多样性指数为0.856~1.829;多态信息含量(PIC)为0.441~0.777,其中有4对微卫星标记呈现为高度多态性(PIC>0.5),2对微卫星标记为中度多态性。经过Bonferroni校正后,只有微卫星标记LB11偏离哈迪—温伯格平衡;连锁不平衡检验结果表明,位点之间不存在连锁不平衡现象。

To get a further study in coral genetic structure and diversity,this study used high-throughput sequencing to the development of Pocillopora damicornis microsatellite.Artificial bleaching was conducted to get the less contaminated coral genomic DNA.Then,we obtained 2786 microsatellite loci after next generation sequencing.This study examined 6 polymorphic markers in Sanya P.damicornis population.Results revealed that effective allele number of these markers ranged from 2.198 to 5.000 per locus,the observed heterozygosity value is from 0.531 to 0.875,the expected heterozygosity value ranged from 0.512 to 0.816.The Shannon index ranged from 0.856 to 1.829 and the polymorphic information content(PIC)of these loci changed from 0.441 to 0.777.There are 4 high polymorphic loci and 2 moderate polymorphic locus.Only LB11 loci was deviated from Hardy-Weinberg equilibrium after Bonferroni correction but no significant linkage disequilibria was detected between the comparisons of these loci.