摘要
本研究利用通用引物的多重PCR方法开展小圆胸小蠹Euwallacea fornicatus的分子鉴定,以期探究多重PCR在昆虫分子鉴定中的可行性,并为开展小圆胸小蠹的有效、准确鉴定及综合防治等提供重要依据。使用多重PCR方法扩增了小圆胸小蠹的COI、16S和28S的3个分子片段,并将获得的目的序列在GenBank中进行BLAST比对;利用MEGA 7计算方胸小蠹属不同种间的遗传距离,并基于邻接法和最大似然法分别构建单基因系统发育树。结果表明:多重PCR可以用于小圆胸小蠹分子序列的获取;基于COI和16S的遗传距离分析表明了小圆胸小蠹的种内遗传距离均小于2%;基于单个基因构建的系统发育树均显示本研究扩增的小圆胸小蠹COI和16S序列与GenBank中获取的小圆胸小蠹COI和16S序列聚为一支。多重PCR可以应用于小圆胸小蠹的分子鉴定,该方法不仅可以提高物种鉴定的准确率,还可以减少PCR过程中的时间和DNA消耗。
In this study,the multiplex PCR with universal primers was used to conduct molecular identification of Euwallacea fornicatus,so as to explore the feasibility of multiplex PCR in the molecular identification of insect and provided an important basis for the effective and accurate identification and integrated control of E.fornicatus.Three molecular fragments of COI,16S and 28S were amplified by multiplex PCR,and the target sequences were blasted in GenBank.The genetic distance of Euwallacea was calculated using MEGA 7.Phylogenetic analyses were performed based single-locus using neighbor-joining and maximum likelihood approaches.Our results indicated that the multiplex PCR method could be used to obtain molecular sequences of E.fornicatus.The genetic distance analysis based upon single-locus showed that the intra-specific genetic distance within E.fornicatus were all less than 2%。Phylogenetic trees constructed based on a single gene showed that COI and 16S sequences of E.fornicatus amplified in this study were clustered together with those COI and 16S sequences of E.fornicatus obtained from GenBank.Multiplex PCR could be applied to the molecular identification of E.fornicatus,which could not only improve the accuracy of species identification,but also reduced the time and DNA consumption in the PCR process.
作者
黄卫东
梁建锋
彭锋
桑文
陈晓胜
王兴民
HUANG Wei-Dong;LIANG Jian-Feng;PENG Feng;SANG Wen;CHEN Xiao-Sheng;WANG Xing-Min(Key Laboratory of Bio-Pesticide Innovation and Application,Guangdong Province,Engineering Research Center of Biological Control,Ministry of Education,Guangzhou 510640,China;College of Forestry and Landscape Aechitecture,South China Agricultural University,Guangzhou 510640,China)
出处
《环境昆虫学报》
CSCD
北大核心
2022年第4期994-1001,共8页
Journal of Environmental Entomology
基金
广州市科技计划重点项目(201804020070)
广东省潮州市农业农村局项目(PZH019D021)。
