Genetic Diversity Analysis Of Culex Pipiens From Different Regions In Jiangsu Province

Objective AFLP (Amplified Fragment Length Polymorphism) technique was used to analysis Genetic diversity and genetic variation of Culex pipiens in different regions in jiangsu province, the similarity index and genetic distance were calculate among samples and UPGMA phylogenetic tree was constructed.Method Culex pipiens of eight regions of Jiangsu Province were Collected, respectively from Changshu, Zhangjiagang, Yancheng, Wuxi, Xuzhou, Nanjing, Nantong, Kunshan, the breeding of Pathogen Biology Laboratory of Nanjing Medical University of typical Culex pipiens also was collected, identified them by the extraction of nucleic acid, gene amplification, sequencing and cloning plasmid vector, Multiple PCR and gel electrophoresis.then start the analysis of AFLP, namely by enzyme digestion and connection, pre-amplification, selective amplification, and then detection the amplication effect by capillary electrophoresis; band will appear on the electropherogram denoted with1, no band is denoted with0, establish0,1matrix, the result were input into NTSYS2.10software and computing Nei-Li similarity coefficient.genetic distance (GD) using the formula:GD=1-2Nab/(Na+Nb), using the UPGMA (unweighted pair-group method with arithmetic means) cluster analysis.Result10pairs of primer combinations were screened from the80pairs of primer combinations. A total of262loci were amplified from10pairs of primer in nine samples, ranged located100-1500bp,185of which were polymorphic loci, polymorphism rate (PP)was up to70.6%, and14-30polymorphic loci were detected per pair of primers combinations, the average polymorphic loci was18.5. E2M7primer combinations amplified polymorphism was maximum,ratio of polymorphism was87.5%, E4M7primer combinations amplified polymorphism was minimum, ratio of polymorphism was48.3%, among10pairs of primers Kunshan amplified bands was maximum (176), Nantong region amplified band was minimum (48); use NTSYS2.10software calculations show that the genetic similarity index between zhangjiagang and Kunshan were the highest (0.8478), the genetic distance were the lowest(0.1522); Wuxi and Xuzhou genetic similarity index were the lowest (0.2139), the genetic distance were the highest (0.7861). Nei genetic distances based on UPGMA system constructed phylogenetic tree showed that the similarity coefficient value of0.64for the domain, nine regions clustered into three groups, which Nanjing Medical University, Yancheng, Xuzhou,, clustered into one group; Changshu, Zhangjiagang, Kunshan clustered into one group;Wuxi, Nantong, Nanjing clustered into one group.Conclusion some genetic diversity and genetic differences exist among Culex pipiens from different regions in Jiangsu Province and show AFLP can be used to investigate effectively genetic diversity of Culex pipiens.