Species identification of Anopheles mosquitoes from the Tumen river basin in Yanbian area of Jilin province based on morphological and molecular characteristics
LI Ji-xu1, SONG Zhen-hai2, PIAO Guang-ming3
1 Yanbian Center for Disease Control and Prevention, Yanbian 133001, Jilin Province, China; 2 Tumen Center for Disease Control and Prevention; 3 Longjing Center for Disease Control and Prevention
Abstract:Objective To understand the species of Anopheles mosquitoes in the Tumen river basin in Yanbian area of Jilin province, and master the morphological and gene characteristics of common Anopheles mosquitoes. Methods The three border towns were chosen in the Tumen river basin in Yanbian area, Yueqing, Qushui, Kaishan in 2012-2013. From May to October every year, the black light trap was used to collect the female adult of Anopheles monthly, from which the genome DNA was extracted. The PCR gene identification technique was used to identify the Anopheles species according to the characterization of its ribosomal nucleic acid internal transcribed spacer 2 segment (rDNA ITS2) gene. The genome DNA amplification products of Anopheles were sequenced, and homology analysis with gene sequence of Anopheles in GenBank of NCBI was carried out to further determine the genetic characteristics of Anopheles mosquitoes in Tumen river basin of Yanbian area. Finally, according to the molecular identification results, reverse analyses was conducted to provide the confirmation for morphological classification. Results Five Anopheles species including An. hycranus group An. sineroides, An. kleini, An. lesteri, An. pullus, and An. belenrae were found in the study area, and the homology between their genomic DNA amplification sequences and that of Anopheles published in GenBank reached 97%-100%. The subcostal, preapical, apical fringe and fringe had obvious species specificity in morphology among An. kleini, An. lesteri, and An. pullus. ConclusionAnopheles sinensis and An. koreicus were not found in Tumen river basin of Yanbian area, An. belenrae was found for the first time in this area and became the new recorded species with An. kleini. The initial recorded species of An. anthropophagus was revised to An. lesteri. The An. yatsushiroensis was revised to An. pullus. According to the white as spots of wings as species specificity of female adults Anopheles mosquitoes, we can identify the morphology of new and related species rapidly and conveniently, such as An. kleini, An. lesteri, and An. pullus.
李基旭, 宋振海, 朴光明. 吉林省延边地区图们江流域按蚊形态学及基因特征分析[J]. 中国媒介生物学及控制杂志, 2018, 29(1): 42-46.
LI Ji-xu, SONG Zhen-hai, PIAO Guang-ming. Species identification of Anopheles mosquitoes from the Tumen river basin in Yanbian area of Jilin province based on morphological and molecular characteristics. Chines Journal of Vector Biology and Control, 2018, 29(1): 42-46.
[1] 马雅军,徐建农. 中国按蚊的分类研究进展[J]. 中国媒介生物学及控制杂志,2015,26(5):433-438. [2] 陆宝麟,许锦江,俞渊,等. 中国动物志. 昆虫纲. 第9卷. 双翅目:蚊科(下卷)[M]. 北京:科学出版社,1997:2-113. [3] 瞿逢伊,朱淮民. 我国按蚊分类进展和若干蚊种学名的订正[J]. 中国寄生虫学与寄生虫病杂志,2008,26(3):210-216. [4] 王钊. 疟疾学(下)[M]. 青岛:青岛海洋大学出版社,1996:34-44. [5] 郭传坤. 我国嗜人按蚊研究进展[J]. 中国热带医学,2005,5(2):396-398,384. [6] 方圆,施文琦,张仪. 赫坎按蚊种团(双翅目:蚊科)分类研究进展[J]. 中国寄生虫学与寄生虫病杂志,2016,34(6):565-570. [7] Collins FH,Paskewitz SM. A review of the use of ribosomal DNA (rDNA) to differentiate among cryptic Anopheles species[J]. Insect Mol Biol,1996,5(1):1-9. [8] 吴静,马雅军,马颖. 基于mtDNA和rDNA基因序列的中国按蚊属塞蚊亚属种类的系统发育研究(双翅目:蚊科)[J]. 昆虫学报,2010,53(9):1030-1038. [9] 林琳,杨曼尼,南春燕,等. 中国部分地区赫坎按蚊种团的分子鉴别研究[J]. 中国媒介生物学及控制杂志,2013,24(2):92-97. [10] 陆宝麟. 中国动物志. 昆虫纲. 第8卷. 双翅目:蚊科(上、下卷)[M]. 北京:科学出版社,1997:60-566,2-113. [11] Lee KW. A revision of the illustrated taxonomic keys to genera and species of female mosquitoes of Korea (Diptera:Culicidae)[C]//5th Medical Detachment,168 Medical Battalion,18th Medical Command,U.S. Army,1998:40. http://wrbu.si.edu. [12] 李石柱,马雅军. 分子鉴别技术在蚊虫分类中的应用[J]. 国外医学寄生虫病分册,2004,31(5):198-201. [13] Rueda LM. Two new species of Anopheles (Anopheles) Hyrcanus Group (Diptera:Culicidae) from the Republic of South Korea[J]. Zootaxa,2005,941(1):1-26. [14] 马颖,马雅军,孟祥梅,等. 我国辽宁省赫坎按蚊种团的分子鉴别研究[J]. 寄生虫与医学昆虫学报,2009,16(4):224-227. [15] Wang Y, Yu WQ, Shi H, et al. Historical survey of the kdr mutations in the populations of Anopheles sinensis in China in 1996-2014[J]. Malar J,2015,14:120. [16] Wilkerson RC, Cong L,Rueda LM,et al. Molecular confirmation of Anopheles (Anopheles) lesteri from the Republic of South Korea and its genetic identity with An. (Ano.) anthropophagus from China (Diptera:Culicidae)[J]. Zootaxa,2003,378(1):1-14. [17] Ma YJ, Xu JN. The Hyrcanus Group of Anopheles (Anopheles) in China (Diptera:Culicidae):species discrimination and phylogenetic relationships inferred by ribosomal DNA internal transcribed spacer 2 sequences[J]. J Med Entomol,2005,42(4):610-619. [18] Shin EH, Hong HK. A new synonym of Anopheles (Anopheles) pullus Yamada, 1937-A.(A.) yatsushiroensis Miyazaki,1951[J]. Kor J Entomol,2001,31(1):1-5. [19] Shin EH, Lee WG, Chang KS, et al. Black light trap collections in Wangging county and Yanji city,Jilin province,China, August 2006-2007[J]. Entomol Res,2011,41(1):36-38.