论文部分内容阅读
目的:建立应用DNA条形码技术鉴定蛆症异蚤蝇的方法。方法:分别提取在口岸检疫中截获的双翅目幼虫和成虫的基因组DNA,利用节肢动物线粒体细胞色素氧化酶I(COI)的通用引物进行DNA扩增。PCR产物直接测序,测序引物同扩增引物,得到清晰可读的665 bp COI的DNA片段碱基序列。与GenBank中相近的序列进行比较,并用ClustalW2在线建立NJ系统发育树。结果:成虫和幼虫COI的DNA序列100%相同,与GenBank中公布的长度为559 bp的蛆症异蚤蝇的COI相应片段相似性高达98.9%,与同属其他种类的可比片段的DNA序列相似性则低于90%。结论:根据结果可以判断所截获的双翅目昆虫为蛆症异蚤蝇。本文的研究再一次证明了DNA条形码技术可以直接应用于昆虫种类的鉴定。
Objective: To establish a method of DNA maggot identification in different maggot disease fleas. Methods: The genomic DNA of Diptera larvae and adults intercepted in port quarantine were extracted separately. DNA amplification was performed by using common primers of arthropod mitochondrial cytochrome oxidase I (COI). The PCR product was sequenced directly, and the sequencing primer and the amplification primer were used to obtain a clear and readable DNA fragment base sequence of 665 bp COI. Similar sequences to GenBank were compared and NJ phylogenetic tree was established online with ClustalW2. Results: The DNA sequences of COIs of adult and larvae were 100% identical with the corresponding COI fragments of maggots with the length of 559 bp in GenBank, reaching 98.9%. Compared with the DNA sequence similarity of other species Then less than 90%. Conclusion: Based on the results, it can be concluded that the captured Diptera is maggot-like flea. This study once again proves that DNA barcoding technology can be directly applied to the identification of insect species.