应用形态和分子特征对我国海南省按蚊的鉴定研究

中国媒介生物学及控制杂志 ›› 2011, Vol. 22 ›› Issue (3): 205-208,217.

应用形态和分子特征对我国海南省按蚊的鉴定研究

朱钊民¹, 林琳², 曾林海³, 赵伟³, 马雅军²

1 第二军医大学研究生管理大队临床1队，上海 200433;
2 第二军医大学病原生物学教研室，上海 200433;
3 海南省疾病预防控制中心，海口 570203

收稿日期:2010-12-30 出版日期:2011-06-20 发布日期:2011-06-20
通讯作者: 马雅军，Email:yajunm@yahoo.com.cn; 赵伟，Email:hainahbm@163.com
作者简介:朱钊民（1988-），男，现为第二军医大学8年制临床本科2007级学生
基金资助:
国家自然科学基金委员会-云南省人民政府联合基金（U0932604）；国家科技重大专项课题（2008ZX10004-010）

Morphology and moleculer identification of anopheline mosquitoes in Hainan province

ZHU Zhao-min¹, LIN Lin², ZENG Lin-hai³, ZHAO Wei³, MA Ya-jun²

1 Clinical Company 1, Graduate Administration Division, Second Military Medical University, Shanghai 200433, China;
2 Department of Pathogen Biology, Second Military Medical University, Shanghai 200433, China;
3 Hainan Center for Disease Control and Prevention, Haikou 570203, Hainan Province, China

Received:2010-12-30 Online:2011-06-20 Published:2011-06-20
Supported by:
Supported by the National Natural Science Foundation of China-Yunnan Joint Fund （No. U0932604） and Major National Science and Technology Projects of China （No. 2008ZX10004-010）

摘要/Abstract

摘要：

目的　利用形态特征与分子特征相结合对我国海南省按蚊进行种类鉴定。方法　研究样本为海南省4个不同地理环境所采集的按蚊成虫和幼虫，依据形态特征鉴定成虫后，测定和分析部分成虫和幼虫的rDNA-ITS2和28S-D3序列以进行分子鉴定。结果　对采集的336只按蚊成虫进行形态鉴定，结果显示总共分为9种，其中迷走按蚊所占比例最高，为81.55%。测序获得成虫和幼虫的ITS2序列37条，D3序列41条，Blast对比的结果显示，形态鉴定的成虫中，1只乌头按蚊错定为微小按蚊；分子鉴定的幼虫包括中华按蚊、迷走按蚊、腹簇按蚊和未定名种（与圣代克按蚊和浅色按蚊同源性高）；序列分析发现在ITS2序列中迷走按蚊和微小按蚊分别存在1处（G/A）和2处（A/T）单碱基双峰，D3序列中迷走按蚊和腹簇按蚊存在3处（G/A）和1处（G/A）单碱基双峰。结论　应用形态特征结合分子特征鉴定按蚊的可靠性和实用性强；分子特征提示迷走按蚊、微小按蚊和腹簇按蚊等处于隐种的分化中。

关键词: 按蚊, 形态, rDNA, 海南省

Abstract:

Objective To identify anopheline mosquitoes in Hainan province based on morphological and molecular characteristics. Methods　Anopheline adults and larval specimens were collected from four different geographic locations in Hainan province. The adults were morphologically identified, and the rDNA-ITS2 and 28S-D3 fragments of selected adults and larvae were amplified and sequenced for molecular identification. Results　The morphological identification of 336 anopheline adults revealed nine species, predominantly Anopheles vagus, which comprised 81.55%. Thirty-seven ITS2 and 41 D3 sequences were amplified. Blast results were consistent with morphological identification; excluding one An. aconitus identified as An. minimus. Molecular identification verified An. sinensis, An. vagus, An. kochi and other anopheline species (highly homogeneous with An. sundaicus and An. subpictus). Double peaks of one (G/A) and two (A/T) bases were found within the ITS2 sequences of An. vagus and An. minimus and three (G/A) and one (G/A) bases were observed within the D3 sequences of An. vagus and An. kochi. Conclusion　The morphological and molecular identification of anopheline mosquitoes is reliable and feasible but suggests ongoing cryptic speciation in An. vagus, An. minimus and An. kochi.

Key words: Anopheline mosquitoes, Morphology, rDNA, Hainan province

中图分类号:

R384.1

朱钊民, 林琳, 曾林海, 赵伟, 马雅军. 应用形态和分子特征对我国海南省按蚊的鉴定研究[J]. 中国媒介生物学及控制杂志, 2011, 22(3): 205-208,217.

ZHU Zhao-min, LIN Lin, ZENG Lin-hai, ZHAO Wei, MA Ya-jun. Morphology and moleculer identification of anopheline mosquitoes in Hainan province[J]. Chines Journal of Vector Biology and Control, 2011, 22(3): 205-208,217.

参考文献

[1] 蔡贤铮.海南岛按蚊名录
[J].海南医学,1992,8(4):8-10.

[2] ChenB,ButlinRK,PedroPM,etal.Molecularvariation,systematicsanddistributionoftheAnophelesfluviatiliscomplexinsouthernAsia
[J].MedVetEntomol,2006,20(1):33-43.

[3] ChenB,HarbachRE,ButlinRK.MolecularandmorphologicalstudiesontheAnophelesminimusgroupofmosquitoesinsouthernChina:taxonomicreview,distributionandmalariavectorstatus
[J].MedVetEntomol,2002,16(3):253-265.

[4] 黄捷敏,王善青,林世千.海南省2000－2008年疟疾防治、监测及疫情分析
[J].中国热带医学,2010,10(2):146-148.

[5] 吴静,马雅军,张迎春.蚊科系统发育关系重建研究进展
[J].国际医学寄生虫病杂志,2008,35(3):153-156.

[6] 陆宝麟.中国动物志.昆虫纲.双翅目.蚊科
[M].第9卷(下卷).北京:科学出版社,1997:12-15.

[7] CollinsFH,MendezMA,RasmussenMO,etal.AribosomalRNAgeneprobedifferentiatesmemberspeciesoftheAnophelesgambiaecomplex
[J].AmJTropMedHyg,1987,37(1):37-41.

[8] 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鉴定研究
[J].寄生虫与医学昆虫学报,2009,16(4):224-227.

[9] 马雅军,瞿逢伊,曹毓存,等.我国雷氏按蚊和嗜人按蚊的分子鉴别鉴定和分类地位的探讨
[J].中国寄生虫学与寄生虫病杂志,2000,18(6):325-328.

[10] MaYJ,QuFY,XuJN,etal.SequencedifferencesofrDNA-ITS2andspecies-diagnosticPCRassayofAnophelessinensisandAnophelesanthropophagus
[J].MedCollPLA,1998,13(2):123-128.

[11] SharpeRG,HarbachRE,ButlinRK.MolecularvariationandphylogenyofmembersoftheMinimusgroupsofAnophelessubgenusCellia(Diptera:Culicidae)
[J].SystEntomol,2000,25(2):263-272.

[12] Alam MT, Das MK, Ansari MA, et al. Molecular identification of Anopheles (Cellia) sundaicus from the Andaman and Nicobar islands of India
[J]. Acta Trop,2006,97(1):10-18.

[13] Joshi D, Choochote W, Min GS. Short report: Natural hybrid between Anopheles kleini and Anopheles sinensis
[J]. Am J Trop Med Hyg,2009,81(6):1020-1022.

[14] 郑彬,汤林华,马雅军,等. 云南微小按蚊A和C栖性的比较
[J].中国寄生虫学与寄生虫病杂志,2005,23(3):103-107.

[15] Harbach RE, Parkin E, Chen B, et al. Anopheles (Cellia) minimus Theobald (Diptera:Culicidae):neotype designation, characterization, and systematics
[J]. Proc Entomol Soc Wash,2006,108(1):198-209.

[16] Harbach RE,Garros C,Manh ND,et al. Formal taxonomy of species C of the Anopheles minimus sibling species complex (Diptera: Culicidae)
[J]. Zootaxa,2007,1654:41-54.

[17] Manguin S, Garros C, Dusfour I, et al. Bionomics taxonomy and distribution of the major malaria vector taxa of Anopheles subgenus Cellia in Southeast Asia: an updated review
[J]. Inf Genet Evol, 2008,8(4):489-503.

[18] 周水森,汤林华,顾政诚,等. 不同地区微小按蚊rDNA-ITS2序列差异
[J]. 中国寄生虫学与寄生虫病杂志,2002,20(1):29-31.

[19] 周水森,汤林华,夏明仪,等. 核糖体28S-D3等位基因特异扩增鉴定微小按蚊亲缘种A和C
[J]. 中国媒介生物学及控制杂志, 2003,14(6):408-410.

[20] 郑彬,汤林华,王学忠,等. 云南省微小按蚊A、C 种群密度高峰及嗜血习性的观察
[J]. 国际医学寄生虫病杂志,2007,7(6):171-173.

[21] Yu Y. Studies on the two forms of Anopheles (Cellia) minimus Theobald,1901 in China (Diptera:Culicidae)
[J]. Mosq Syst,1987,19(2):143-145.

[22] Chen B, Pedro PM, Harbach RE, et al. Mitochondrial DNA variation in the malaria vector Anopheles minimus across China, Thailand and Vietnam:evolutionary hypothesis,population structure and population history
[J]. Heredity,2010,106(2):241-252.

[23] Harbach RE. TheclassificationofgenusAnopheles (Diptera: Culicidae): a working hypothesis of phylogenetics relationships
[J]. Bull Entomol Res,2004,94(6):537-553.