中缅边境地区部分传疟按蚊蚊种的形态学及分子鉴定

Species identification of anopheles mosquitoes from the bordering area of China and Myanmar based on morphological and molecular characters

目的通过形态学特征和ITS2序列扩增分析对中缅边境地区部分按蚊进行蚊种鉴定,分析其传疟潜力。方法从中缅边境地区采集部分按蚊标本,首先根据其形态学特征进行蚊种鉴定;然后提取基因组DNA,进行ITS2区段PCR扩增和测序;通过NCBI Blast分析所测序列与Gen Bank中不同按蚊的序列同源性;最后利用MEGA5软件将测序结果与Gen Bank中其他按蚊的ITS2序列进行序列比对,并构建系统进化树。综合以上形态学特征和分子鉴定结果,确定按蚊种类,分析其传疟潜力。结果经形态学鉴定为微小按蚊、带足按蚊及可赫按蚊的ITS2序列分别为506、575 bp和517 bp。BLAST结果提示,根据ITS2序列进行的按蚊蚊种分子鉴定结果均与之前的形态学鉴定一致。通过与Gen Bank中其他按蚊的ITS2序列进行比对,系统进化树显示捕获的微小按蚊、带足按蚊及可赫按蚊分别形成单独的支系。结论本研究综合利用传统形态学鉴定和分子生物学技术明确了中缅边境地区部分按蚊蚊种为微小按蚊、带足按蚊及可赫按蚊。

Objective To identify the species of anopheles mosquitoes from the China-Myanmar bordering area based on morphological characters and sequence analysis of the second internal transcription spacer（ITS2） of ribosomal DNA.Methods The anopheles mosquitoes were collected from the bordering area of China and Myanmar.Firstly,the mosquito species were identified according to the morphological characters.Then,the genomic DNA was extracted from the anopheles mosquitoes.The ITS2 sequences of the anopheles mosquitoes were obtained by PCR amplification and sequencing.And the ITS2 sequences were analyzed by online BLAST method.Finally,the ITS2 sequences were aligned with their homologs of other anopheles obtained from Gen Bank,and the phylogenetic tree was built using MEGA5 software.Results The ITS2 sequence lengths of Anopheles minimus,Anopheles peditaeniatus and Anopheles kochi identified morphologically were 506 bp,575 bp and 517 bp,respectively.The results of BLAST showed that the molecular identification results of the anopheles species according to the ITS2 sequences were the same as the morphological identification results.The phylogenetic tree showed that the collected Anopheles minimus,Anopheles peditaeniatus and Anopheles kochi formed the separate branches.Conclusion In the present study,some anopheles species in the bordering area of China and Myanmar are identified as Anopheles minimus,Anopheles peditaeniatus and Anopheles kochi by both morphological and molecular biological methods.