缢蛏(Sinonovacula constricta)微卫星标记的分离及近缘物种通用性

ISOLATION OF MICROSATELLITE LOCI FROM RAZOR CLAM SINONOVACULA CONSTRICTA AND TRANSFERABILITY TO RELATED SPECIES

采用磁珠富集和PCR筛选相结合的方法,得到12对缢蛏(Sinonovacula constricta)的多态性微卫星引物。每个位点的观测等位基因数为2—15个,有效等位基因数为1.0339—8.8063个;观测杂合度(Ho)为0.0333—1.0000,平均观测杂合度为0.7525;期望杂合度(He)为0.0333—0.9020,平均期望杂合度为0.6866;多态信息含量(PIC)为0.0320—0.8680。所有位点中,有8个位点属于高度多态位点(PIC>0.5),SC1-5和SC4-6属于低度多态位点(PIC<0.25);经Bonferroni校正后,无显著偏离哈迪-温伯格平衡的位点;连锁不平衡检验结果表明,位点间不存在连锁不平衡现象。此外,分析了这些引物在近缘种长竹蛏(Solen strictus)、大竹蛏(Solen grandis)和小刀蛏(Cultellus attenuatus)的通用性情况,结果显示:长竹蛏中,位点SC1-4、SC2-8、SC3-1、SC3-14表现出了高度多态(PIC>0.5);在大竹蛏中,位点SC3-4、SC3-7、SC4-6表现出了高度多态性(PIC>0.5),SC2-9为低态位点(PIC<0.25);在小刀蛏中,位点SC1-7、SC2-9、SC3-4、SC3-14表现出了高度多态性(PIC>0.5),SC4-6为低态位点(PIC<0.25)。

Using magnetic-bead enrichment and PCR screening method, we obtained 12 pairs of microsatellite primers from razor clam Sinonovacula constricta. The allele number and the effective allele number of these markers ranged from 2 to 15 and from 1.0339 to 0.8063; the observed and expected heterozygosity values ranged from 0.0333 to 1.0000(average 0.7525) and 0.0333 to 0.9020(average 0.6866), respectively. The polymorphic information content(PIC) of these loci changed from 0.0320 to 0.8680. Among them, eight were highly polymorphic loci, and SC1-5 and SC4-6 were slightly polymorphic loci. No loci was significantly deviated from Hardy-Weinberg equilibrium after Bonferroni correction, and no significant linkage disequilibria was detected between the comparisons of these loci. In addition, the transferability of all polymorphic short sequence repeats(SSRs) was assessed in three closely-related species. SC1-4, SC2-8, SC3-1, and SC3-14 were highly polymorphic loci in Solen strictus, SC3-4, SC3-7, SC4-6 were highly polymorphic loci in Solen grandis, and SC1-7, SC2-9, SC3-4, SC3-14 highly polymorphic in Cultellus attenuatus.