Abstract: 【Objective】 The experiment was aimed to investigate and identify the species of Culex pipiens complex in Kunming, Yunnan, and provide data for the prevention and control of mosquito borne infectious diseases. 【Method】 Morphological and molecular biological identification were conducted to the male Culex pipiens complex collected from Qinglong mountain next to the Jindian and Huangtupo，Kunming，in June 2022 and May 2023.The morphological characteristics of mosquitoes were observed under stereomicroscope，and the male mosquito phallosomes was cut off for observation, and the mosquito species were identified according to the morphological characteristics.The mitochondrial cytochrome C oxidase subunit Ⅰ(COⅠ) gene and acetylcholinesterase gene 2 (AChE2) of mosquitoes were amplified and their sequences were analyzed. 【Result】 Morphology showed that the middle end of the phallosomes of mosquitoes collected from Qinglong mountain next to the Jingdian was pointed，and the extension part of the inner leaf was broad leaf.The middle end of the phallosomes of mosquitoes collected from Huangtupo of Kunming is flush，and the inner leaf of the stem was blade like.The results of gene sequence similarity analysis showed that there was no significant difference in the COⅠ gene of the collected mosquitoes.The AChE2 gene distinguished mosquitoes into two types.Mosquitoes collected from Qinglong mountain next to the Jingdian had high similarity with Culex pipiens quinquefasciatus，with the highest similarity ranging from 98.1% to 100%.Mosquitoes collected from the Huangtuppo had high similarity with Culex pipiens pipiens，with the highest similarity of 99.7%.The results of COⅠ gene phylogenetic analysis showed that the six mosquitoes were in the same evolutionary branch as Culex quinquefasciatus, Culex pipiens, Culex pipiens molestus, and Culex pipiens pallens, without significant genetic evolutionary differences.The phylogenetic analysis of AChE2 gene showed that mosquitoes collected from Qinglong Mountain next to the Jingdian were clustered with Culex quinquefasciatus, while mosquitoes collected from Huangtupo were clustered with Culex pipiens, located in different evolutionary branches. 【Conclusion】 Two kinds of the Culex pipiens complex were collected.The mosquitoes collected from Qinglong mountain next to the Jingdian were Culex pipiens quinquefasciatus，while the mosquitoes collected from the Huangtupo were Culex pipiens pipiens.Research had shown the discovery of Culex pipiens pipiens in Southern regions such as Kunming in China, and the monitoring of West Nile virus and related viruses should be strengthened.

Key words: Kunming; Culex pipiens complex; Culex quinquefasciatus; Culex pipiens pipiens; morphological identification; biological identification