FIASCO法筛选铜锈环棱螺微卫星标记
摘要： 2011年9-10月，从武昌南湖和江苏太湖分别采集了50个鲜活的铜锈环棱螺(Bellamya aeruginosa)和10～20个不等的铜锈环棱螺(Bellamya aeruginosa)，梨形环棱螺(Bellamya purificata)，方形环棱螺(Bellamya quadrata)，角形环棱螺(Bellamya angularia)和坚环棱螺(Bellamya lapillorm)的鲜活个体。从足肌中提取基因组DNA，以其中铜锈环棱螺的基因组DNA为模板，采用Fast isolation by AFLP sequences containing repeats (FIASCO)法构建铜锈环棱螺微卫星文库。结果表明，筛选出的15个微卫星标记具有很高的多态性，其中多态信息含量(PIC)值在0.3174～0.8886之间变化，平均值0.7074；每个位点有5～12个等位基因，平均值8.4；期望杂合度(HE)和观测杂合度(HO)分别在0.6674～0.9073和0.0714～0.9130之间变化，平均值分别是0.7985和0.6779。6个微卫星位点显著偏离哈代温伯格平衡(Hardy-Weinberg equilibrium)，并有8对位点间存在连锁不平衡(linkage disequilibrium)。这15个微卫星标记在环棱螺种间扩增也表现出较高的多态性，可为我国环棱螺遗传多样性研究提供依据。
关键词： 铜锈环棱螺；微卫星标记；多态性；杂合体缺失
（ 1、College of Fisheries, Huazhong Agricultural University, WuHan 430070； 2、College of Fisheries, Huazhong Agricultural University； ）
Abstract： September-October 2011, we collected 50 samples of fresh individual Bellamya aeruginosa in Nanhu, and 10-20 samples of Bellamya aeruginosa, Bellamya purificata, Bellamya quadrata, Bellamya angularia, Bellamya lapillorm in Taihu, Yixing city. We extracted genomic DNA from foot muscle in fresh individuals. Taking one of Bellamya aeruginosa genomic DNA as a template to build the Bellamya aeruginosa microsatellite library according to the fast isolation by AFLP sequences containing repeats (FIASCO) method. And then we screened highly polymorphic microsatellite markers, which can provide important information for the study of genetic diversity, genetic structure and classification of Bellamya. The results showed that 15 microsatellites loci were screened from the freshwater snail Bellamya aeruginosa which had highly polymorphic. The average polymorphic information content (PIC) was 0.7074 ranged from 0.3174 to 0.8886. Allele number per locus ranged from 5 to 12 and the average was 8.4. The observed heterozygosity (HO) and expected heterozygosity (HE) varied from 0.6674 to 0.9073 and 0.0714 to 0.9130 with an average of 0.6779 and 0.7985 respectively. There were six microsatellites loci significantly deviation from Hardy-Weinberg equilibrium (HWE) after Bonferroni correction (P < 0.0034) and there were eight loci existent linkage disequilibrium. In addition, 15 microsatellite markers also showed a moderate or high polymorphism in Bellamya population amplification. These fifteen polymorphic microsatellites loci would be valuable for species identification and population genetics analysis of Bellamya.
Keywords： Bellamya aeruginosa; microsatellites loci; polymorphism; Heterozygote deficiency
