摘要
传统分类学鉴定需要有完整无残缺的成虫标本,对于非昆虫分类学研究人员应用传统分类学方法鉴定蚊虫较为困难;本研究旨在发掘更多适用于蚊类鉴定的DNA条形码,实现口岸一线蚊类的快速准确鉴定。本文通过提取14个蚊虫样品的基因组DNA,使用通用引物PCR扩增ITS1、ITS2、Cytb、COI基因及序列,PCR产物双脱氧法测序,结合NCBI中相关数据分析部分ITS2、Cytb、COI序列的相似性和遗传关系。结果显示:获得了14个蚊虫样品的部分ITS1、ITS2、Cytb、COI序列的非全长碱基序列共计39条;依据种内遗传距离和相似度将14个未知蚊虫分子分类鉴定为6种蚊虫:凶小库蚊、致倦库蚊、尖音库蚊、赫坎按蚊、白纹伊蚊、里海伊蚊;碱基使用率统计表明所测样品的Cytb、COI基因符合昆虫线粒体DNA碱基组成;构建基因ITS2、Cytb、COI系统进化树,符合蚊虫系统发育遗传进化理论。本研究对14个未知蚊虫样品的进行了分子鉴定,为蚊类的快速准确识别鉴定提供基础数据。
Traditional taxonomical identification requires complete and uncrippled adult specimens, which makes difficult for non-insect taxonomical researchers to identify mosquitoes with traditional taxonomical methods, can not achieve rapid and accurate identification. The purpose of present study is to discover more DNA barcodes suitable for mosquito identification, so as to achieve rapid and accurate identification of mosquitoes at the port. Traditional taxonomic methods were used to determine the general group of unknown mosquitoes. The genomic DNA of 14 mosquito samples were extracted by molecular biology, and the ITS1, ITS2, Cytb and COI gene sequences were amplified via PCR using universal primers, and the PCR products were sequenced with the method of double deoxidization, the genetic relationship and similarity of some ITS2, Cytb and COI were analyzed together with related NCBI data. The results showed that a total of 39 non-full-length base sequences of ITS1, ITS2, Cytb and COI genes of 14 mosquito samples were obtained, of which 21 were new sequences. 14 unknown mosquito molecules were classified and identified as 6 mosquito species according to their intra-species genetic distance and similarity: Culex modestus, Cx. pipiens quinquefasciatus, Cx. pipiens pallens, Anopheles hyrcanus, Aedes albopictus and Ae. caspius. The statistics of base utilization showed that the Cytb and COI genes of the tested samples was in accordance with the composition of insect mitochondrial DNA base. The phylogenetic tree of genes ITS2, Cytb and COI were constructed, which was in accordance with the theory of mosquito phylogenetic evolution. The data of genes ITS1, ITS2 and Cytb studied in present paper need to be further enriched.
作者
陈光辉
王孟文
李焱
刘东
廖凌
CHEN Guang-Hui;WANG Meng-Wen;LI Yan;LIU Dong;LIAO Ling(College of life Science and Technology,Xinjiang University,Urumqi 830046,China;Kashi Customs Technology Center,Kashi 844000,China;College of Life and Geographic Sciences,Kashi University,Kashi 844000,China)
出处
《寄生虫与医学昆虫学报》
CAS
2020年第1期26-36,共11页
Acta Parasitologica et Medica Entomologica Sinica
基金
新疆南部地区媒介和人群中西尼罗病毒感染状况调查及风险研究(项目编号:2016IK265)。
关键词
蚊类
DNA条形码
内转录间隔子
细胞色素氧化酶b
线粒体细胞色素c氧化酶亚基Ⅰ
Mosquitoes
DNA barcoding
Rapid identification
Internal transcribed spacer1/2(ITS1,ITS2)
Cytochrome b(Cytb)
Mitochondrion cytochrome c oxidase subunitⅠ(COI)
