不同线粒体基因作为DNA条形码鉴定按蚊的比较分析  

Comparative analysis of different mitochondrial genes as DNA barcodes for identifying Anopheles mosquitoes

作　　者：康泽辉 张乐宁 王彦力 张晓 KANG Zehui;ZHANG Lening;WANG Yanli;ZHANG Xiao(College of Plant Health and Medicine,Qingdao Agricultural University,Qingdao 266109,China;Lingcheng Bureau of Agriculture and Rural Affairs,Dezhou 253052,China)

机构地区：[1]青岛农业大学植物医学学院,山东青岛266109 [2]陵城区农业农村局,山东德州253052

出　　处：《实验技术与管理》2023年第12期37-43,共7页Experimental Technology and Management

基　　金：青岛农业大学实验技术研究课题“生物前沿技术与成果融入普通昆虫学实验实践教学的探索与应用”(SYJS202325);青岛农业大学高层次人才科研基金(663-1118015,663-1119008)。

摘　　要：蚊虫是最为重要的医学昆虫之一,准确鉴定其种类是防控的重要基础,但基于形态学鉴定困难较多。线粒体基因组序列获取便捷、已公布数据全面,作为DNA条形码开展分子鉴定具有突出优势。为进一步评估不同线粒体基因序列作为DNA条形码在蚊科昆虫鉴定中的作用,本研究以按蚊属为例,选取在GenBank数据库中线粒体全基因组序列公布最多的5种,每种随机下载5条序列,通过进化速率、遗传距离、条形码间隙和系统发育分析,比较了线粒体全部13个蛋白质编码基因分别作为DNA条形码鉴定种类的可靠性。结果表明按蚊线粒体的13个蛋白质编码基因中,ND5基因进化速率最快,而COI基因最为保守。除COIII基因外,其余基因均可作为DNA条形码开展分子鉴定,但其中ND4L基因的条形码间隙不显著,可能存在鉴定不准确的情况。剩余线粒体基因中,ATP8、COI、COII、CytB和ND5基因间隙显著且在种内和种间均具有较强的稳定性,优先推荐选用为DNA条形码,ATP6、ND1、ND2、ND3、ND4和ND6基因则存在种内或(和)种间不保守的情况,可以作为备选序列。研究结果为科学选用DNA条形码序列进行蚊科昆虫的鉴定提供了参考依据。Mosquitoes are one of the most important medical insects,and accurate identification of species is an important foundation for prevention and control.However,there are many difficulties in morphological identification.The convenience of obtaining mitochondrial(mt)genome sequences and comprehensive published data make it a prominent advantage for molecular identification as a DNA barcode.In order to further evaluate the role of different mt gene sequences as DNA barcodes in the identification of mosquitoes,we took Anopheles as an example in this study,and selected 5 species with the most published mt genome sequences in the GenBank database,and randomly downloaded 5 sequences for each species.Through evolutionary rate,genetic distance,barcode gap and phylogenetic analysis,the reliability of all 13 mt protein coding genes as DNA barcodes for species identification is compared.The results indicate that among the 13 protein coding genes in the mt genome of Anopheles mosquitoes,the ND5 gene has the fastest evolutionary rate,while the COI gene is the most conservedgene.Except for the COIII gene,all other genes can be used as DNA barcodes for molecular identification,but the barcode gap of the ND4L gene is not significant,which may lead to inaccurate identification.Among the remaining mt genes,there are significant barcode gaps between ATP8,COI,COII,CytB,and ND5 genes,and these genes have strong stability both within and between species which are preferred as DNA barcodes.The ATP6,ND1,ND2,ND3,ND4,and ND6 genes exhibit intraspecific or(and)interspecific non-conservation and can be used as candidate DNA barcode sequences.This research provides a reference basis for the scientific selection of DNA barcode sequences for the identification of mosquitos.

关 键 词：按蚊属 分子鉴定 遗传距离 条形码间隙 系统发育 

分 类 号：R384.1[医药卫生—医学寄生虫学]