三种摇蚊幼虫RAPD扩增条件的优化及在昆虫系统发育分析中的应用

Random amplified polymorphic DNA analysis of three chironomid species

为了研究摇蚊科昆虫种群遗传的多样性,以促进对其资源的合理保护,以萨摩亚摇蚊Chironomus samoensisEdwards基因组DNA为模板,对摇蚊幼虫的RAPD扩增条件进行优化,建立了摇蚊幼虫RAPD扩增反应的最佳体系:按照利用优化的RAPD扩增条件进行研究,实验有着良好的重现性。用16个随机引物对3种摇蚊幼虫类群各10个个体进行RAPD扩增,其中萨摩亚摇蚊共扩增出78个条带,多态座位率为41.03%,Shannon遗传多样性指数为0.2570,群体内相似度为0.8730;红裸须摇蚊Propsilocerus akamusi(Tokunaga)共75个条带,多态座位率为44.0%,Shannon遗传多样性指数为0.2472,群体内相似度为0.8731;刺铗长足摇蚊Tanypus punctipennis(Fabricius)共67个条带,多态座位率为41.79%,Shannon遗传多样性指数为0.1943,群体内相似度为0.9066。聚类分析结果表明,刺铗长足摇蚊与红裸须摇蚊的亲缘关系较近。

In order to reveal the genetic diversity of chironomid population, and the protect of its resource, the genomic DNA of Chironomus samoensis Edwards was used as template to optimize RAPD amplified reaction conditions for chironomid larvae. A high reproducibility was obtained with the optimized experiment conditions. The genetic diversity of three chironomid larvae populations （C, samoensis, Propsilocerus akamusi （Tokuraga） and Tanypus punctipennis Fabricius） were analyzed. Sixteen random primers were used to amplifying 10 individuals DNA of each populations. Total 78 bands were amplified, and the percent of polymorphic loci was 41.03 in C. samoensis, 75 bands and 44.00 percent of polymorphic loci in, P. akamusi, and 67 bands and 41.79 percent of polymorphic loci in T. punctipennis. The similarity of their RAPD bands accounted for 0.8730, 0.8731 and 0.9066, respectively. The relationship between T. punctipennis and P. akamusi is relative close by UPGM, A method.