Morphological and molecular biological identification of Culicoides kelinensis
Objective To systematically identify Culicoides kelinensis collected in Gongshan county of Yunnan province by using morphological and molecular biological methods.Methods In August 2017,the midges were collected by using the trap lamp in Gongshan county.Culicoides kelinensis was selected by morphological identification,DNA was extracted by non-destructive tissue digestion,COI gene and ITS-1 gene were amplified for sequence analysis.Culicoides kelinensis tablet was made for identification based on morphology.Results A total of 5 females and 1 male Culicoides kelinensis were collected from Gongshan county in Yunnan province.Diagnosis:the female Culicoides kelinensis was eyes continguous,bare,AR 1.04,PR 4.50 and cell R2 with pale,the wing surface had 3 transverse bands of light color formed by pale spots,spermathecae 2,unequal.The male hypopygium Culicoides kelinensis with 1 pair lateral process of small and pointed,the adeagus with broad in the middle,a slender rod at the end,parameresis separatedand end with small branches.The phylogenetic analysis based on COI gene and ITS-1 gene sequences showed that Culicoides kelinensis was located in an independent evolutionary branch.The nucleotide homology of COI gene was 100.0％,and the nucleotide homology of ITS-1 gene was 99.2％-100.0％between them.Conclusion The results of morphological identification of Culicoides kelinensis were the same as those of types specimens.The sequence information of COI gene and ITS-1 gene of female and male Culicoides kelinensis were obtained,which will provide the basis for identification of Culicoides kelinensis in the future.
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