Abstract：We analysed the phylogenetic relationship and establish molecular identification of Culicoides species by sequencing the nuclear ribosomal DNA internal transcribed spacer 1 (ITS1) sequences. The ITS1 sequences of six closely related species of the genus Culicoides, C. arakawai, C. holcus, C. jacobsoni, C. indianus, C. huffi and C. humeralis, were obtained by DNA sequencing. The calculation of genetic distance was based on Kimura 2-parameter formula. The base composition of ITS1 sequences and construction of phylogenetic trees which took the Allohelea annulate as outgroup were analyzed by MEGA 6.06v. Approximately 352 bp of ITS1 sequences were sequenced for those 6 species and the content of T, C, A, G of the sequences were 36.8%, 13.0%, 30.0%, 20.1%, respectively. Statistical analysis of genetic distances showed that intraspecific genetic distances was different from that of interspecies significantly (t=32.430,P<0.05). The results of phylogenetic relationship were consistent with the morphological identification which indicated that the topologies of Maximum Likelihood (ML) tree and Neighbor-Joining (NJ) tree were quite similar; distinguished between different species of Culicoides clearly and no cross between different species; the same species gathered closely with each other. This study confirmed that rDNA-ITS1 sequences can be used for molecular identification and phylogenetic analysis in the genus Culicoides.
岑常活, 韩晓静, 常琼琼, 段琛, 侯晓晖. 库蠓核糖体DNA内转录间隔区1序列的测定与分析[J]. 中国人兽共患病学报, 2018, 34(12): 1087-1094.
CEN Chang-huo, HAN Xiao-jing, CHANG Qiong-qiong, DUAN Chen, HOU Xiao-hui. Sequence and analysis of rDNA-ITS1 of the genus Culicoides (diptera: ceratopogonidae). Chinese Journal of Zoonoses, 2018, 34(12): 1087-1094.
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