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Anopheles sinensis Wiedemann sensu stricto (s.s.) is a dominant mosquito and considered a secondary malaria vector in the Republic of Korea (ROK). Despite the potential significance for malaria control, population genetics studies have been conducted using only mitochondrial DNA (mtDNA), and studies of the genetics of hybridization have never been attempted. In this study, 346 specimens from 23 localities were subject to experiments. Among them, 305 An. sinensis s.s. specimens from 20 localities were used for mtDNA analysis, and 346 specimens comprising 341 An. sinensis s.s. from 22 localities and five Anopheles kleini Rueda from one locality were examined in the microsatellite study. Neighbor-joining analysis of pairwise FST and RST based on microsatellite results showed that the populations are divided into two groups, as did the mtDNA results. However, the Bayesian analysis and factorial correspondence analysis plots showed three distinct clusters. Among the mtDNA and microsatellite results, only microsatellites represented small but positive and significant isolation-by-distance patterns. Both molecular markers show the Taebaek and Sobaek Mountain ranges as barriers between the northern and southern parts of the ROK. The newly recognized third group suggests possible introgressive hybridization of An. sinensis s.s. with closely related species. The slightly different composition of populations in each group based on different markers is probably because of different population dynamics in each group. These results imply that there is restricted gene flow of epidemiologically important malaria-related genes between the northern and southern parts of the ROK., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Background: In the Republic of Korea (ROK), six sibling species of the Anopheles sinensis complex are considered the vector species of malaria, but data on their susceptibilities to malaria and vector capacities have been controversial. The intensive use of insecticides has contributed to the rapid development and spread of insecticide resistance in the An. sinensis complex. Knockdown resistance (kdr) to pyrethroids and DDT in the An. sinensis complex is associated with a mutation in codon 1014 of the voltage-gated sodium channel (VGSC) gene. Because the degree of insecticide resistance varies among mosquito species and populations, the detection of kdr mutations among the six sibling species of the An. sinensis complex is a prerequisite for establishing effective long-term vector control strategies in the ROK METHODS: In order to investigate species-specific kdr mutations, An. sinensis complex specimens have been collected from 22 sites in the ROK. Because of the difficulties with species identifications that are based only on morphological characteristics, molecular identification methods have been conducted on every specimen. Part of the IIS6 domain of the VGSC was polymerase chain reaction-amplified and directly sequenced., Results: The molecular analyses revealed that mutations existed at codon 1014 only in An. sinensis sensu stricto and no mutations were found in the other five Anopheles species. In An. sinensis s.s., one wild type (TTG L1014) and three mutant types (TTT L1014F, TTC L1014F, and TGT L1014C) of kdr alleles were detected. The TTC L1014F mutation was observed for the first time in this species., Conclusions: The fact that the highly polymorphic kdr gene is only observed in An. sinensis s.s., out of the six Anopheles species and their geographical distribution suggest the need for future studies of insecticide resistance monitoring and investigations of species-specific resistance mechanisms in order to build successful malaria vector control programmes in the ROK.