AFLP标记技术在湖北钉螺遗传多样性中的应用研究

Application of amplified fragment length polymorphism in the study of genetic diversity of Oncomelania hupensis

目的探讨扩增片段长度多态性(AFLP)标记技术在湖北钉螺遗传多样性研究中的应用。方法随机抽取云南大理和湖南岳阳钉螺各1只,用异硫氰酸胍和树脂等抽取DNA,然后用64对引物对基因组DNA进行AFLP扩增,扩增产物用6%的变性聚丙烯酰胺凝胶电泳,荧光检测扩增产物。结果每对引物扩增的AFLP标记数在5～55之间,云南钉螺平均每对引物出现38.30(95%CI36.03～40.57)个标记,湖南钉螺平均每对引物扩增出39.14(95%CI36.71～41.57)个标记;每对引物扩增的多态性标记数和多态性频率分别在3～37个和28.6%～76.2%之间,分别平均为23.67(95%CI22.12～25.22)和47.36%(95%CI45.22%～49.50%);两地钉螺的遗传相似系数平均为0.69(95%CI0.67～0.70)。结论AFLP标记技术可用于湖北钉螺的分类与遗传多样性的研究。

Objective To explore the application of amplified fragment length polymorphism (AFLP) in the study of genetic diversity of Oncomelania hupensis. Methods A snail was taken at random from Dali region, Yunnan Province and Yueyang City, Hunan Province respectively, and DNA was drawn with guanidinium thiocyanate and resin et al. Genomic DNA was amplified selectively by the AFLP technique with 64 pairs of primers, and the polymorphism of PCR products was examined by SDS-PAGE. Results The number of AFLP markers was between 5 and 55 for a pair of primers. The average number of a pair of primers was 38.30 (95% CI 36.03-40.57) for the snail from Yunnan Province, and 39.14 (95% CI 36.71-41.57) for the one from Hunan Province. For a pair of primers, the number of amplified polymorphic markers was between 3 and 37, and the polymorphic rate was between 28.6% and 76.2%, the average, 23.67 (95% CI 22.12-25.22) and 47.36%(95 %CI 45.22%-49.50%), respectively. The average similarity between the two populations was 0.69 (95% CI 0.67-0.70). Conclusion The AFLP technique is a new path for classifying Oncomelania hupensis and studying the genetic diversity of it.[